Toxicity and bioassimilation of lead and nickel in farm ruminants fed on diversified forage crops grown on contaminated soil.

The cultivation of forage crops on wastewater-irrigated soils, while common in many developing countries, poses significant risks due to heavy metal pollution, particularly Lead (Pb) and Nickel (Ni). This practice, aimed at addressing water scarcity challenges and providing affordable irrigation, was investigated for its ecological and human health implications across three diverse sites (site A, site B, and site C). Our study unveiled increases in Pb concentrations in contaminated soil, cultivated with Sesbania bispinosa showing the highest Pb accumulation. The Ni concentrations ranged from 5.34 to 10.43 across all forage crop samples, with S. fruticosa from site C displaying the highest Ni concentration and S. bicolor from site A exhibiting the lowest. Trace element concentrations in the specimens were determined using an atomic absorption spectrophotometer. The Pb levels in the blood, hair, and feces of farm ruminants (cows, buffaloes, and sheep) varied across the sites, with buffaloes consistently displaying the highest Pb levels. Insights into daily Pb intake by ruminant's highlighted variations influenced by plant species, animal types, and sites, with site C, the cows exhibiting the highest Health Risk Index (HRI) associated with lead exposure from consuming forage crops. Soil and forage samples showed Pb concentrations ranging from 8.003 to 12.29 mg/kg and 6.69-10.52 mg/kg, respectively, emphasizing the severe health risks associated with continuous sewage usage. Variations in Ni concentrations across animal blood, hair, and feces samples underscored the importance of monitoring Ni exposure in livestock, with sheep at site B consistently showing the highest Ni levels. These findings highlight the necessity of vigilance in monitoring trace element (Pb and Ni) exposure in forage crops and livestock, to mitigate potential health risks associated with their consumption, with variations dependent on species, site, and trace element concentrations.

Health risk implications of iron in wastewater soil-food crops grown in the vicinity of peri urban areas of the District Sargodha.

Irrigation using sewage water can be beneficial, as it can increase the productivity of crops but has negative consequences on crops, soil contamination, and human health. It contains a variety of toxins, such as chemicals and heavy metals, which damage the soil and crops. In this regard, the aim of the research was to assess the potential health hazards of iron (Fe) metal in food crops (leafy and root crops) treated with wastewater (T_1), canal water (T_2), and tube well water (T_3). Water, soil, and edible components of food crops were collected at random from three distinct locations. Fe concentration in samples was estimated using atomic absorption spectrophotometer, following wet digestion method. The Fe concentrations, ranged from 0.408 to 1.03 mg/l in water, 31.55 to 187.47 mgkg-1 in soil and 4.09 to 32.583 mgkg-1 in crop samples; which were within permissible limits of the World Health Organization (WHO). There was a positive correlation between soils and crops. The bioconcentration factor, enrichment factor (EF), daily intake of metals (DIM), health risk index (HRI), and target hazard quotient (THQ) all values were <1, except for a pollution load index >1, which indicated soil contamination, but there was no Fe toxicity in crops, no health risk, and no-carcinogenic risk for these food crops in humans. To prevent the excessive accumulation of Fe metal in the food chain, regular monitoring is needed.

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.

Evaluation of nickel toxicity in wheat as function of diverse type of fertilizers: implications for public health.

Many studies have described the physiological, biochemical, and molecular responses to heavy metal toxicity and deficiencies individually in plants. The present study assess nickel (Ni) concentration in amended soil, plant vegetative parts, and wheat grains, grown under diverse types of fertilizers in Sargodha, Punjab, Pakistan. Different varieties of wheat were grown in pot and fields. Different treatments (municipal solid waste, poultry waste, press mud, farm yard manure) of fertilizers were applied in order to study the metal level increased in different parts (root, shoot, grain) of wheat due to fertilization. Results indicated that metal level was found highest in roots followed by shoot and grain. The highest level of nickel in root was present in V1 (2.35 mg/kg) due to T2 (2.60 mg/kg) treatment. Higher nickel levels in wheat shoot and grains were observed in V5 (2.36 mg/kg) and V8 (2.29 mg/kg), respectively, due to applied treatment T2 (2.57 mg/kg). This study concluded that treatment T9 was proven safe in view of the observed Ni concentration, while treatment T2 (municipal solid waste) resulted in higher accumulation of nickel in wheat grains which showed that municipal solid waste should be treated before their application in agriculture fields to secure the public health. This study recommended that although application of fertilizers increased the plant growth and nutritional value, it also enhanced metal accumulation in the wheat grains which could be harmful for consumers especially human being. Government should take actions to prevent metal toxicity in human food chain.

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.

Appraisal of iron accumulation in soil, forages, and blood plasma of sheep and goats: a case study in different districts of Punjab, Pakistan.

Minerals are essential for ruminants affecting significantly the production of grazing livestock. Iron level in forages, soil, and blood plasma of the small ruminants (goat and sheep) was investigated in three districts of Punjab. Atomic absorption spectrophotometer was used to determine the concentration of iron in collected samples. The results revealed that the mean Fe concentrations in soil of districts Sargodha, Mianwali, and Bhakhar were significantly varied and ranged from 21.85 to 23.78, 28.45 to 31.2, and 18.079 to 24.33 mg/kg, respectively. The Fe level in soil of Mianwali significantly varied and was higher than Sargodha and Bhakkar. The mean Fe concentration in forages which were used for feeding purpose were significantly varied and found between 10.95-14.49, 23.63-25.65, and 6.616-9.45 mg/kg for Sargodha, Mianwali, and Bhakhar, respectively. The mean Fe concentrations in blood plasma of goat which consumed the contaminated forages were 8.5026-11.763 mg/L in district Sargodha, 19.77-20.19 mg/L in Mianwali, and 5.508-5.858mg/L in Bhakkar. In blood plasma of sheep, the residual levels of Fe in districts Sargodha, Mianwali, and Bhakhar were ranged from 9.987 to 12.455, 15.8 to 19.785, and 3.425 to 6.383 mg/L, respectively. This study provides the data of metals effected by different sites and also their mobility from low to higher trophic level which enables us to study the iron toxicity in different trophic levels, and we recommend different safe limits and treatment in case of low and high metal profile.

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.

Bioaccumulation of cadmium in different genotypes of wheat crops irrigated with different sources of water in agricultural regions.

The study was carried out to evaluate the health risks associated with accumulation of cadmium in the different genotypes of wheat, grown in agricultural regions of Punjab, Pakistan. Five genotypes irrigated with three varied water sources were selected randomly from each region. Among all sources of water, types of soil, and grain samples, the cadmium (Cd) quantities were found (2.24-2.82 mg/L, 1.75-4.16 mg/kg, 0.86-1.90, respectively), exceeding the maximum permissible limits (0.01 mg/L, 1.1 mg/kg, 0.2 mg/kg, respectively) described by FAO/WHO. The pollution load index (PLI) exhibited by all of the samples was higher than 1.00, the permissible limit; however, other factors including bioaccumulation, translocation, bio-concentration, daily intake, and enrichment values of Cd were less than 1.00. Moreover, the health risk index for cadmium in all types of wheat grain samples was less than 1.00. The study concluded that the continuous use of wastewater resources may lead to the accumulation of cadmium in the vital body organs that may cause severe health hazards.

Level and speciation of nickel in some forages in relation to spatial and temporal fluctuations.

The present experimental work was conducted at different sites of district Bhakkar, a semiarid region of Pakistan, to assess whether the goats are suffering nickel deficiency or toxicity and what are the possible seasonal effects on the availability and translocation of nickel in food chain. A total of 27 forage and 320 goats according to four physiological stages [does (she goat), bucks (he goat), wether (castrated), juvenile (6 month)] were recruited for this study. To fulfill this objective, soil, forage, blood plasma, urine, and feces samples were collected in 4 seasons of the year at 2 sites and were analyzed by atomic absorption spectrophotometer for nickel concentration. Different indices BCF, EF, and PLI were also studied to check the metal transfer. The results showed that sites had significant (P < 0.05) effect on nickel concentration in soil, forage, and goats. On the other hand, season and site x season had nonsignificant (P > 0.05) effects on nickel level in soil and goats. The soil (0.68-0.71 mg kg-1), forage (3.41-3.70 mg/kg), and blood (0.21-0.28 mg/l) level was lower than the permissible limits, while feces (0.57-1.34 mg/kg) and urine (0.35-1.32 mg/l) had enough concentration of nickel. Sources showed significant (P < 0.05) effects on Ni level in all stages of goats. All stages of goats except Wether (castrated) showed low level of nickel in blood. Most fluctuations in nickel concentration were observed in (S1) summer (low) and spring (S4) (high) season as a whole, while overall site 2 had high level of nickel than site 1. Thus, nickel showed deficiency in soil, forage, as well as in all stages of goats except wether goats. Nickel containing mineral mixtures are essential for does (she goat), bucks (he goat), and juveniles (6 months old), so application of Ni containing fertilizers to the soil and forage of that region and supplementation of Ni mineral mixture for grazing ruminants should be done.

Health risk assessment through determining bioaccumulation of iron in forages grown in soil irrigated with city effluent.

The irrigation with sewage water can be useful if it has no negative effects on food crop yield, soil pollution, and health of humans. However, it includes various types of contaminants like heavy metals that pollute the soil and crops. In this regard, the aim of this study was to evaluate the possible health risks of heavy metals in forages. Forages both of summer and winter were grown with different water treatments (sewage water and tap water) in Department of Botany, University of Sargodha. The concentration of iron (Fe) in water, soil, and plant samples was determined. The Fe values in tap and sewage water were observed as 0.090 and 0.115 mg/L, respectively. The highest mean concentration of Fe was 9.608 mg/kg in the soil where Trifolium alexandrinum is grown, and the lowest mean concentration was 0.154 mg/kg which occurred in the soil where Trifolium resupinatum is grown in winter. The maximum mean concentration of Fe in the root samples of plants was observed as 2.483 mg/kg in Pennisetum typhoideum, and the minimum mean concentration occurred as 0.390 mg/kg in Zea mays grown in summer. The maximum bioconcentration factor value of Fe was observed for T. resupinatum (5.259) grown in winter. The maximum pollution load index value of Fe was observed for T. alexandrinum (0.1688). The maximum value of daily intake of metals was observed as 0.0731 in Medicago sativa, and the maximum health risk index value was determined as 0.1091 in P. typhoideum.

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.

Expression of four S. pneumoniae type 2 polysaccharide biosynthetic enzymes utilising the endogenous Kex2 protease activity in tobacco.

In order to express multisubunit proteins, or to manipulate metabolic pathways in plants it is essential to be able to efficiently express multiple proteins within the same plant cell. To increase the efficiency of multi-protein expression, we demonstrate the use of the Golgi localized Kex2 protease activity in tobacco to process a large polyprotein precursor consisting of four individual protein domains into its individual protein constituents. Four genes encoding enzymes involved in the biosynthesis of S. pneumoniae type 2 polysaccharide were assembled into a single expression cassette as a large polyprotein driven by a single cauliflower mosaic virus (CaMV) 35S promoter. Each of the individual protein domains were separated by three sequential Kex2 protease digestion sites. At the N-terminus a Pr1b signal peptide was incorporated for efficient targeting of the polyprotein to the apoplast. Each individual protein domain was tagged with its own immuno-tag. The construct was used for the transformation of Nicotiana tabacum and stable lines were selected. All four processed proteins could be immunologically detected in protein extracts using Western blotting indicating correct expression and Kex2 processing. Utilisation of the Kex2 protease system represents an efficient way of expressing multiple proteins in the same plant. This method simplifies the transformation procedures, and presents a method for expression of multiple proteins within the same plant.

Characterization of water treatment sludge and its reuse as coagulant.

Coagulation-flocculation process results in the generation of large volume of waste or residue, known as water treatment sludge (WTS), in the purification of surface water for potable supplies. Sustainable management of the inevitable waste requires careful attention from the plant operators and sludge managers. In this study, WTS produced with the optimum alum dose of 30 ml/L at the laboratory scale has been treated with sulphuric acid to bring forth a product known as sludge reagent product (SRP). The performance of SRP is evaluated for its efficiency in removing the colloidal suspensions from the Yamuna river water over wide pH range of 2-13. 1% sludge acidified with sulphuric acid of normality 2.5 at the rate of 0.05 ml/ml sludge has been observed as the optimum condition for preparing SRP from WTS. The percentage turbidity removal is greater at higher pH value and increases with increasing the dosage of SRP. The optimum SRP dosage of 8 ml/L in the pH range of 6-8 performed well in removing the colloidal suspension and other impurities from the Yamuna water. The quality of treated water met the prescribed standards for most of the quality parameters. Thus, SRP has the potential to substitute the conventional coagulants partially or completely in the water treatment process, depending on the quality needed at the users end.

RAPD and SSR based genetic diversity analysis of elite-2 set of synthetic hexaploid wheats.

BACKGROUND: Synthetic hexaploid wheats are artificially reconstituted hexaploid wheats that possess high genetic variation which could be utilized for the development of new improved wheat varieties. One such group of synthetic wheats is called the Elite-II set of synthetic wheats that are derived from crossing durum wheat with different Aegilops tauschii wheats. MATERIALS AND METHODS: In the current study genetic diversity was investigated among 18 Elite-II synthetic hexaploid wheat lines at DNA level. Two types of molecular markers i.e. RAPD and SSR were used for this purpose. RESULTS: Both types of markers proved useful in estimating the overall genetic diversity among these lines. Based on RAPD data range of genetic distances in these lines was from 0 to 100 percent. Seven D-genome specific SSRs were also used to get further estimation of the genetic diversity contributed by Aegilops tauschii parent. On the basis of results obtained it is inferred that the Aegilops tauschi accessions used in the production of these synthetic lines were genetically different and they contributed to the enhancement of genetic variation in the synthetic lines. These results could be helpful for future genome mapping programs. CONCLUSION: The overall extensive genitive diversity indicates that these lines are good candidates for development of improved wheat varieties by crossing with cultivated wheat varieties.

A study on the transfer of iron in soil-plant-animal continuum under semi-arid environmental conditions in Sargodha, Pakistan.

The present investigation on the iron (Fe) transfer from soil to plant and in turn to animal (cows), as a function of sampling periods was conducted at the Livestock Experimental Station Sargodha, Pakistan which falls under semi-arid conditions. Although the iron transfer from soil to forage increased consistently, the forage Fe content decreased progressively with increase in sampling period. Highest Fe transfer from forage to cow blood plasma was observed during October and lowest during January. The transfer of Fe from forage to animal milk was maximum during the months of October and January and minimum during December. The transfer of Fe to plasma and milk was found to be dependent variably on the growth stage of forage in this investigation. Based on the findings of the present study, it is evident that mineral supplementation with higher Fe availability is urgently warranted to the animals particularly during the months of December and January to enhance plasma Fe in the cows being reared at that livestock farm during the entire grazing period. Thus, obligatory supplementation of Fe to the ruminants is highly recommended. Since the processes involved in iron management system in humans, animals, and plants are basically similar, appropriate elemental management must be provided to the living organisms, otherwise deficient or excessive levels of iron may deteriorate the developing cells of the organisms.

A study on the transfer of cadmium from soil to pasture under semi-arid conditions in Sargodha, Pakistan.

The study was conducted on the cadmium (Cd) transfer from soil to pasture at Khizerabad Livestock Farm, District Sargodha (falling under semi-arid conditions), Pakistan. The concentrations of Cd in the soil and forage ranged from 2.80 to 6.74 mg/kg and 1.14 to 4.20 mg/kg, respectively, in different sampling periods. The higher values of Cd in pasture suggested the possible risk of entering Cd into higher food chain as these concentrations of Cd can potentially be transported from soil to different animals rearing on the farm pastures, and they should be taken into account in risk assessment of chemical toxicity. Providing region-specific mineral mixture having highly bioavailable forms of other trace elements to the ruminants like Zn, Fe, and Mn, which are known to antagonize Cd, would help in overcoming the Cd toxicity. There is an urgent need of permanent monitoring of cadmium content in the feed used in animal nutrition at livestock farm.