Modern perspectives of heavy metals alleviation from oil contaminated soil: A review.

Heavy metal poisoning of soil from oil spills causes serious environmental problems worldwide. Various causes and effects of heavy metal pollution in the soil environment are discussed in this article. In addition, this study explores new approaches to cleaning up soil that has been contaminated with heavy metals as a result of oil spills. Furthermore, it provides a thorough analysis of recent developments in remediation methods, such as novel nano-based approaches, chemical amendments, bioremediation, and phytoremediation. The objective of this review is to provide a comprehensive overview of the removal of heavy metals from oil-contaminated soils. This review emphasizes on the integration of various approaches and the development of hybrid approaches that combine various remediation techniques in a synergistic way to improve sustainability and efficacy. The study places a strong emphasis on each remediation strategy that can be applied in the real-world circumstances while critically evaluating its effectiveness, drawbacks, and environmental repercussions. Additionally, it discusses the processes that reduce heavy metal toxicity and improve soil health, taking into account elements like interactions between plants and microbes, bioavailability, and pollutant uptake pathways. Furthermore, the current study suggests that more research and development is needed in this area, particularly to overcome current barriers, improve our understanding of underlying mechanisms, and investigate cutting-edge ideas that have the potential to completely transform the heavy metal clean up industry.

Flexible Piezoresistive Sensors Based on PPy Granule-Anchored Multilayer Fibrous Membranes with a Wide Operating Range and High Sensitivity.

The employment of flexible piezoresistive sensors has sparked growing interest within the realm of wearable electronic devices, specifically in the fields of health detection and e-skin. Nevertheless, the advancement of piezoresistive sensors has been impeded by their limited sensitivity and restricted operating ranges. Consequently, it is imperative to fabricate sensors with heightened sensitivity and expanded operating ranges through the utilization of the appropriate methodologies. In this paper, piezoresistive sensors were fabricated utilizing electrospun polyvinylidene fluoride/polyacrylonitrile/polyethylene-polypropylene glycol multilayer fibrous membranes anchored with polypyrrole granules as the sensing layer, while electrospun thermoplastic polyurethane (TPU) fibers were employed as the flexible substrate. The sensitivity of the sensor is investigated by varying the fiber diameter of the sensing layer. The experimental findings reveal that a concentration of 14 wt % in the spinning solution exhibits high sensitivity (996.7 kPa-1) within a wide working range (0-10 kPa). This is attributed to the favorable diameter of the fibers prepared at this concentration, which facilitates the uniform in situ growth of pyrrole. The highly deformable TPU flexible fibers and multilayer sensing layer structure enable different linear responses across a broad pressure range (0-1 MPa). Furthermore, the sensor demonstrates good cyclic stability and can detect human movements under different pressures. These results suggest that the piezoresistive sensor with a wide operating range and high sensitivity has significant potential for future health monitoring and artificial intelligence applications.

Combined effect of endophytic Bacillus mycoides and rock phosphate on the amelioration of heavy metal stress in wheat plants.

BACKGROUND: Zinc (Zn) and nickel (Ni) are nutrients that are crucial for plant growth; however, when they are present at higher concentrations, they can cause toxicity in plants. The present study aimed to isolate plant growth promoting endophytic bacteria from Viburnum grandiflorum and assess its plant and defense promoting potential alone and in combination with RP in zinc (Zn) and nickel (Ni) toxic soil. The isolated endophytic bacteria were identified using 16s rRNA gene sequencing. For the experiment, twelve different treatments were applied using Zn, Ni, isolated endophytic Bacillus mycoides (Accession # MW979613), and rock phosphate (RP). The Ni, Zn and RP were used at the rate of (100 mg/kg) and (0.2 g/kg) respectively. A pot experiment with three replicates of each treatment was conducted using a complete randomized design (CRD). RESULTS: The results indicated that Ni (T5 = seed + 100 mg/kg Ni and T9 = seed + 100 mg/kg Zn) and Zn concentrations inhibited plant growth, but the intensity of growth inhibition was higher in Ni-contaminated soil. Bacillus mycoides and RP at 100 mg/Kg Zn (T12 = inoculated seed + 100 mg/kg Zn + RP0.2 g/kg.) increased the shoot length, leaf width, protein and sugar content by 57%, 13%, 20% and 34%, respectively, compared to the control. The antioxidant enzymes superoxide dismutases (SOD), peroxidase (POD) were decreased in contaminated soil. Furthermore, Ni and Zn accumulation was inhibited in T11 (seed + 100 mg/kg Zn + RP0.2 g/Kg) and T12 (inoculated seed + 100 mg/kg Zn + RP0.2 g/Kg) by 62 and 63% respectively. The Cu, Ca, and K, contents increased by 128, 219 and 85, Mn, Na, and K by 326, 449, and 84% in (T3 = inoculated seed) and (T4 = inoculated seed + RP 0.2 g/Kg) respectively. CONCLUSIONS: Ni was more toxic to plants than Zn, but endophytic bacteria isolated from Viburnum grandiflorum, helped wheat (Triticum aestivum) plants and reduced the toxic effects of Ni and Zn. The effect of Bacillus mycoides was more prominent in combination with RP which promoted and suppressed heavy-metal toxicity. The reported combination of Bacillus mycoides and RP may be useful for improving plant growth and overcoming metal stress.

Heavy metals mitigation and growth promoting effect of endophytic Agrococcus terreus (MW 979614) in maize plants under zinc and nickel contaminated soil.

Introduction: Heavy metals such as iron, copper, manganese, cobalt, silver, zinc, nickel, and arsenic have accumulated in soils for a long time due to the dumping of industrial waste and sewage. Various techniques have been adapted to overcome metal toxicity in agricultural land but utilizing a biological application using potential microorganisms in heavy metals contaminated soil may be a successful approach to decontaminate heavy metals soil. Therefore, the current study aimed to isolate endophytic bacteria from a medicinal plant (Viburnum grandiflorum) and to investigate the growth-promoting and heavy metal detoxification potential of the isolated endophytic bacteria Agrococus tereus (GenBank accession number MW 979614) under nickel and zinc contamination. Methods: Zinc sulfate and nickel sulfate solutions were prepared at the rate of 100 mg/kg and 50 mg/kg in sterilized distilled water. The experiment was conducted using a completely random design (CRD) with three replicates for each treatment. Results and Discussion: Inoculation of seeds with A. tereus significantly increased the plant growth, nutrient uptake, and defense system. Treatment T4 (inoculated seeds), T5 (inoculated seeds + Zn100 mg/kg), and T6 (inoculated seeds + Ni 100 mg/kg) were effective, but T5 (inoculated seeds + Zn100 mg/kg) was the most pronounced and increased shoot length, root length, leaf width, plant height, fresh weight, moisture content, and proline by 49%, 38%, 89%, 31%, 113%, and 146%, respectively. Moreover the antioxidant enzymes peroxidase and super oxidase dismutase were accelerated by 211 and 68% in contaminated soil when plants were inoculated by A. tereus respectively. Similarly the inoculation of A. tereus also enhanced maize plants' absorption of Cu, Mn, Ni, Na, Cr, Fe, Ca, Mg, and K significantly. Results of the findings concluded that 100 mg/kg of Zn and Ni were toxic to maize growth, but seed inoculation with A. tereus helped the plants significantly in reducing zinc and nickel stress. The A. tereus strain may be employed as a potential strain for the detoxification of heavy metals.

Employee green behavior: a study on the impact of corporate social responsibility (CSR) on employee green behavior, green culture: the moderating role of green innovation.

This study adopts the natural resource-based view (NRBV) of the firm to investigate the relationships between corporate social responsibility (CSR), employees' green behavior (EGB), green culture, environmental performance (EP), and green innovation (GI) in China's automobile manufacturing industry. A survey of 465 employees was conducted. The data were analyzed using partial least squares structural equation modeling. The results showed that CSR positively influences EGB and GC, which in turn improves EP. CSR also directly impacts EP. Furthermore, GI enhances the positive effect of CSR on EP through the mediators EGB and GC. The findings contribute to the literature by elucidating the mechanisms linking CSR and EP. They provide an integrated framework incorporating direct, mediated, and moderated effects. For practice, the study highlights the importance of CSR, EGB, and GC for manufacturers aiming to improve environmental sustainability. It suggests managers should promote green values, foster eco-innovation, and encourage employees' discretionary green actions. The moderating effect of GI implies embedding sustainability in corporate culture and operations to magnify the gains from CSR. Overall, the research offers insights into translating green strategic intent into performance outcomes through cultural and behavioral transformation.

The Re-/Up-Cycling of Wood Waste in Wood-Polymer Composites (WPCs) for Common Applications.

Wood-polymer composites (WPCs) are a class of materials intensively studied and promoted in the context of sustainable development, mainly when aspects related to the increasing awareness of environmental issues and waste management are considered. Feasible opportunities for producing WPCs with value-added properties intended for common applications emerge when polymers, either synthetic or from renewable resources, raw or waste, are employed in re-/up-cycling approaches. In this context, some examples of easily achievable WPCs are presented herein, namely, formulations based on different wood waste and polymer matrices (synthetic: polypropylene and malleated polypropylene as a compatibilizer; natural: plasticized starch). Their level of performance was assessed through different characterization methods (FTIR, WAXD, TGA, DSC, mechanical test, etc.). The benefits and limitations of this approach are also discussed.

Fecal microbiota transplantation attenuates Escherichia coli infected outgrowth by modulating the intestinal microbiome.

BACKGROUND: Given the crucial role of gut microbiota in animal and human health, studies on modulating the intestinal microbiome for therapeutic purposes have grasped a significant attention, of which the role of fecal microbiota transplantation (FMT) has been emphasized. METHODS: In the current study, we evaluated the effect of FMT on gut functions in Escherichia coli (E. coli) infection by using mice model. Moreover, we also investigated the subsequently dependent variables of infection, i.e., body weight, mortality, intestinal histopathology, and the expression changes in tight junction proteins (TJPs). RESULTS: The FMT effectively decreased weight loss and mortality to a certain extent with the restoration of intestinal villi that resulted in high histological scores for jejunum tissue damage (p < 0.05). The effect of FMT on alleviating the reduction of intestinal TJPs was also proved by immunohistochemistry analysis and mRNA expression levels. Moreover, the abundance of health-threatening bacteria, belonging to phylum Proteobacteria, family Enterobacteriaceae and Tannerellaceae, genus Escherichia-Shigella, Sphingomonas, Collinsella, etc., were significantly increased, whereas beneficial bacteria, belonging to phylum Firmicutes, family Lactobacillaceae, genus Lactobacillus were decreased in the gut of infected mice. Furthermore, we sought to investigate the association of clinical symptoms with FMT treatment with modulation in gut microbiota. According to beta diversity, the microbial community of gut microbiota results reflected the similarities between non-infected and FMT groups. The improvement of the intestinal microbiota in FMT group was characterized by the significant high level of beneficial microorganisms with the synergistic decrease of Escherichia-Shigella, Acinetobacter, and other taxa. CONCLUSION: The findings suggest a beneficial host-microbiome correlation following fecal microbiota transplanatation for controlling gut infections and pathogens-associated diseases.

The Synergistic Effect of Polystyrene/Modified Boron Nitride Composites for Enhanced Mechanical, Thermal and Conductive Properties.

Thermal conductivity (TC) and thermal stability are the basic requirements and highly desirable properties in thermal management, heat storage and heat transfer applications. This work is regarding the fabrication of polystyrene/boron nitride composites and melt extruded to produce good thermal stability, increased thermal conductivity and enhanced mechanical properties. Our strategy is potentially applicable to produce thermally conductive composites of low cost over large scale. Boron nitride powder is bath sonicated in 10% NH3 solution to avoid its agglomeration and tendency toward entanglement in a polymer matrix. An approximately 67.43% increase in thermal conductivity and 69.37% increase in tensile strength as well as 56 multiple increases in thermal stability of the optimum samples were achieved. The developed polymeric composites are potentially applicable in the electronic industry, especially in electronic devices used for 5G, heat sink and several other aviation applications.

Composition, impacts, and removal of liquid petroleum waste through bioremediation as an alternative clean-up technology: A review.

Exposure to hazardous wastes, especially petroleum wastes hydrocarbon (PWHCs), can damage human health and biological diversity. A huge amount of petroleum waste along with persistent organic pollutants is being generated during exploration and processing of crude oil. The dumping of petroleum waste hydrocarbons in an open pit contaminates the soil which can cause severe threats to human health and agro-geo-environmental ecosystem. The current study aimed to evaluate the mode of occurrence, composition, environmental, and health impacts of petroleum waste by using recent literature. The extracted results show that oil emulsion contains 48% oil, suspension 23%, settled emulsion 42%, and sludge emulsion 36%. The study discusses the possible biological techniques for rehabilitation of petroleum waste-contaminated areas. Several physical and chemical techniques are available for remediation of petroleum waste, but they are either costly or environmentally not feasible. Whereas, biological remediation namely, Bioremediation (Biostimulation and Bioaugmentation), Phytoremediation (Phytodegradation, Rhizoremediation, Phytovolatilization, and Rhizo-filtration) is a cheap and environmentally friendly way to remove petroleum waste hydrocarbons from contaminated soil and water. Some important enzymes (i.e., peroxidase, nitrilase, nitroreductase, phosphatase) and plant species i.e., Acacia and Chloris species are prominent methods to remediate the PWHCs. The knowledge assembled in this review is expected to create new doors for researchers to develop more efficient techniques to control the harmful impacts of PWHCs on the environment and health.

Catalytic potential of endophytes facilitates synthesis of biometallic zinc oxide nanoparticles for agricultural application.

Recent advances fascinated the use of biological resources in combination with metals to obtain high quality biometals and apply its advantages in different fields. Endophytic mediated Zinc oxide nanoparticles (ZnO-NPs) is an economical and ecofriendly way for farmers to avoid Zn deficiency in rice crop and obtain high yield. Here we synthesized ZnO-NPs utilizing endophytic bacterial strain of Enterobacter hormaechei (E. hormaechei). The physiochemical properties of the prepared NPs were determined through UV-Vis spectroscopy, XRD, FT-IR, SEM and TEM. The prepared NPs revealed surface plasmon resonance (SPR) at 320 nm (nm) and crystalline structure with 21 nm average crystalline size. FT-IR spectra showed the presence of carboxylic, alcohol and amine functional groups, which confirm the biometallic assembling of the ZnO and endophytic bacterial functional groups. SEM showed pyramidal symmetry whereas TEM revealed poly dispersed spherical shape with particle size distribution 18-48 nm. Our results showed that prepared NPs possess significant antifungal, antibacterial and antioxidant potential at 25, 50 and 100 µg/mL concentrations. Moreover, Cytotoxic and hemolytic assay showed significant results (less % viability and hemolysis activity) at 50 and 100 µg/mL (ZnO-NP's) concentrations as compared to control. The prepared ZnO-NPs were used as bio fertilizer in various concentrations as a foliar spray, which showed significant enhancement of the rice plant growth, along with chlorophyll, proteins and carotenoid contents. These results recommend that endophytic mediated ZnO-NPs are biocompatible and possess significant potential for agricultural applications.

Advances in Biochar and PGPR engineering system for hydrocarbon degradation: A promising strategy for environmental remediation.

In soil, polycyclic aromatic hydrocarbons (PAHs) have resulted in severe environmental deterioration, compromised soil characteristics, and negatively affect all life forms, including humans. Developing appropriate and effective clean-up technology is crucial in solving the contamination issues. The traditional methods to treat PHAs contaminated soil are less effective and not ecofriendly. Bioremediation, based on bioaugmentation and biostimulation approaches, is a promising strategy for remediating contaminated soil. The use of plant growth-promoting rhizobacteria (PGPR) as a bioaugmentation tool is an effective technique for treating hydrocarbon contaminated soil. Plant growth-promoting rhizobacteria (PGPR) are group of rhizospheric bacteria that colonize the roots of plants. Biochar is a carbon-rich residue, which acts as a source of nutrients, and is also a bio-stimulating candidate to enhance the activities of oil-degrading bacteria. The application of biochar as a nutrient source to bioremediate oil-contaminated soil is a promising approach for reducing PHA contamination. Biochar induces polyaromatic hydrocarbons (PAHs) immobilization and removes the contaminants by various methods such as ion exchange electrostatic attractions and volatilization. In comparison, PGPR produce multiple types of biosurfactants to enhance the adsorption of hydrocarbons and mineralize the hydrocarbons with the conversion to less toxic substances. During the last few decades, the use of PGPR and biochar in the bioremediation of hydrocarbons-contaminated soil has gained greater importance. Therefore, developing and applying a PGPR-biochar-based remediating system can help manage hazardous PAH contaminated soil. The goal of this review paper is to (i) provide an overview of the PGPR mechanism for degradation of hydrocarbons and (ii) discuss the contaminants absorbent by biochar and its characteristics (iii) critically discuss the combined effect of PGPR and biochar for degradation of hydrocarbons by decreasing their mobility and bioavailability. The present review focuses on techniques of bioaugmentation and biostimulation based on use of PGPR and biochar in remediating the oil-contaminated soil.

Comparative Analysis of Microbial Consortiums and Nanoparticles for Rehabilitating Petroleum Waste Contaminated Soils.

Nano-bioremediation application is an ecologically and environmentally friendly technique to overcome the catastrophic situation in soil because of petroleum waste contamination. We evaluated the efficiency of oil-degrading bacterial consortium and silver nanoparticles (AgNPs) with or without fertilizer to remediate soils collected from petroleum waste contaminated oil fields. Physicochemical characteristics of control soil and petroleum contaminated soils were assessed. Four oil-degrading strains, namely Bacillus pumilus (KY010576), Exiguobacteriaum aurantiacum (KY010578), Lysinibacillus fusiformis (KY010586), and Pseudomonas putida (KX580766), were selected based on their in vitrohydrocarbon-degrading efficiency. In a lab experiment, contaminated soils were treated alone and with combined amendments of the bacterial consortium, AgNPs, and fertilizers (ammonium nitrate and diammonium phosphate). We detected the degradation rate of total petroleum hydrocarbons (TPHs) of the soil samples with GC-FID at different intervals of the incubation period (0, 5, 20, 60, 240 days). The bacterial population (CFU/g) was also monitored during the entire period of incubation. The results showed that 70% more TPH was degraded with a consortium with their sole application in 20 days of incubation. There was a positive correlation between TPH degradation and the 100-fold increase in bacterial population in contaminated soils. This study revealed that bacterial consortiums alone showed the maximum increase in the degradation of TPHs at 20 days. The application of nanoparticles and fertilizer has non-significant effects on the consortium degradation potential. Moreover, fertilizer alone or in combination with AgNPs and consortium slows the rate of degradation of TPHs over a short period. Still, it subsequently accelerates the rate of degradation of TPHs, and a negligible amount remains at the end of the incubation period.

Coactive role of zinc oxide nanoparticles and plant growth promoting rhizobacteria for mitigation of synchronized effects of heat and drought stress in wheat plants.

This study intended to investigate the potential of the plant growth-promoting rhizobacteria (PGPR) and green synthesized zinc oxide nanoparticles (ZnO-NPs) (fruit extract of Papaya) against heat and drought stress in wheat. The characterization of green-synthesized ZnO-NPs was done through UV-vis spectrophotometry, Fourier-transform infrared spectrometry, X-ray diffraction and scanning electron microscopy. Individual and combination of PGPR (Pseudomonas sp.) and ZnO-NPs (10 ppm) amendments were tested in a pot experiment to upregulate wheat defence system under three stress groups (drought, heat and combined heat and drought stress). Drought and heat stress synergistically caused higher damage to wheat plants than individual heat and drought stress. This observation was confirmed with remarkable higher MDA and hydrogen peroxide (H2O2) content. Treated plants exposed to all stress groups showed an improved wheat growth and stress resistance through better biomass, photosynthetic pigments, nutrients, soluble sugars, protein and indole acetic acid content. Combination of ZnO-NPs and Pseudomonas sp. Protects the plants from all stress groups by producing higher proline, antioxidant enzymes i. e superoxide dismutase, peroxidase, catalase, ascorbate peroxidase, glutathione reductase and dehydroascorbate reductase, and abscisic acid. Moreover, higher stress alleviation by this treatment was manifested by marked reduced electrolyte leakage, MDA and H2O2. The findings of current study confirmed that the synergistic actions of PGPR and ZnO-NPs can rescue plants from both single and combined heat and drought stress.

Antagonistic, Anti-oxidant, Anti-inflammatory and Anti-diabetic Probiotic Potential of Lactobacillus agilis Isolated From the Rhizosphere of the Medicinal Plants.

Potential probiotic bacteria can be used as a biotherapeutic agent and a sustainable alternative to antibiotics, as an anti-oxidative, anti-inflammatory, and anti-diabetic agent without causing any serious side effects. Mostly human-friendly Lactic acid bacteria (LAB) have been isolated from the animal-human origin to be used as biotherapeutic agents or to produce useful metabolites (nutraceutical). However, less information is known about the role of medicinal plants associated LAB as biotherapeutic agents. The isolation of 115 human-friendly Lactobacillus strains was done from the rhizosphere of the medicinal plants Ocimum tenuiflorum, Azadirachta indica, Ficus carica. The obtained bacteria were then tested for their safe status before being using it for a beneficial purpose. Out of 115 strains, 29 (25%) were negative for blood hemolytic activities. Among these 29 isolates, three isolates did not show a breakdown of gelatin and were recognized as safe. Antibiotic resistance assay showed resistance of two of them against antibiotics discs of Streptomycin (10 µg), Ciprofloxacin (20 µg), Vancomycin (30 µg), Metronidazole (10 µg), Ampicillin (5 µg), Chloramphenicol (30 µg), Kanamycin (30 µg), Erythromycin (15 µg), Penicillin (10 µg) and Tetracycline (30 µg). The bacterial isolate (T-2) was found safe that was identified as Lactobacillus agilis by sequence analysis of 16 s rRNA gene and processed in vitro as an anti-bacterial, anti-oxidant, anti-diabetic, and anti-inflammatory agent. Free radical scavenging activities and inhibition of α-amylase activities for Lactobacillus agilis were found relative to standard drug values as 68% and 73% and 51.3% and 65.3%, respectively. The in-vitro anti-inflammatory assay showed 61.6% (Lactobacillus agilis) while showed 69% (aspirin) activity for denaturation albumin protein. The results suggested that Lactobacillus agilis can be used as a potential probiotic strain as well as can be used to produce nutraceuticals.

Variation in archaeal and bacterial community profiles and their functional metabolic predictions under the influence of pure and mixed fertilizers in paddy soil.

Impact of environmental perturbations i.e., nitrogen (N), phosphorus (P), potassium (K), and rice straw (Rs) on the dynamics of soil bacterial and archaeal communities are multifactor dependent and seeks a contemporary approach to study underlying mechanisms. The current study investigates the effect of pure and mixed fertilizers on soil physicochemical properties, the microbial community structure, and their functional metabolic predictions. It involved amendments with distinct combinations of N as C(H2N)2O, P and K as KH2PO4, K as KCl, and Rs in paddy soil microcosms with concentrations common in rice fields agriculture. Soil pH, electrical conductivity (EC), total carbon (TC), total nitrogen (TN), organic matter (OM), available K (AK), and total extractable P (TEP) were evaluated. To comprehend community variation and functional predictions, 16S rRNA-based high throughput sequencing (HTS) and phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt) were employed, respectively. Our findings showed enhanced community richness and diversity in all amendments compared to control. Proteobacteria, Actinobacteria, and Firmicutes were dominant bacterial phyla. Regarding relative abundance, Chloroflexi, Bacteroidetes, and Verrucomicrobia showed positive while Actinobacteria, Acidobacteria, and Gemmatimonadetes showed negative trends compared to controls. Thaumarchaeota and Euryarchaeota were dominant archaeal phyla and exhibited increasing and decreasing trends, respectively. The PICRUSt analysis indicated functional prediction more towards amino acid, carbohydrate, energy, and lipid metabolism while less towards others. Concerning energy metabolism, most and least responsive treatments were KP and controls, respectively. These outcomes enhanced our understanding regarding soil quality, fertilizer composition and application, and functional metabolomics of archaea and bacteria.

Molecular detection of Shiga toxin-producing Escherichia coli (STEC) O157 in sheep, goats, cows and buffaloes.

BACKGROUND: Shiga toxin-producing E. coli (STEC) are important foodborne pathogens that causing serious public health consequences worldwide. The present study aimed to estimate the prevalence ratio and to identify the zoonotic potential of E. coli O157 isolates in slaughtered adult sheep, goats, cows and buffaloes. MATERIALS AND METHODS: A total of 400 Recto-anal samples were collected from two targeted sites Rawalpindi and Islamabad. Among them, 200 samples were collected from the slaughterhouse of Rawalpindi included sheep (n = 75) and goats (n = 125). While, 200 samples were collected from the slaughterhouse of Islamabad included cows (n = 120) and buffalos (n = 80). All samples were initially processed in buffered peptone water and then amplified by conventional PCR. Samples positive for E. coli O157 were then streaked onto SMAC media plates. From each positive sample, six different Sorbitol fermented pink-colored colonies were isolated and analyzed again via conventional PCR to confirm the presence of rfbE O157 gene. Isolates positive for rfbE O157 gene were then further analyzed by multiplex PCR for the presence of STEC other virulent genes (sxt1, stx2, eae and ehlyA) simultaneously. RESULTS: Of 400 RAJ samples only 2 (0.5%) showed positive results for E. coli O157 gene, included sheep 1/75 (1.33%) and buffalo 1/80 (1.25%). However, goats (n = 125) and cows (n = 120) found negative for E. coli O157. Only 2 isolates from each positive sample of sheep (1/6) and buffalo (1/6) harbored rfbE O157 genes, while five isolates could not. The rfbE O157 isolate (01) of sheep sample did not carry any of STEC genes, while the rfbE O157 isolate (01) of buffalo sample carried sxt1, stx2, eae and ehlyA genes simultaneously. CONCLUSION: It was concluded that healthy adult sheep and buffalo are possibly essential carriers of STEC O157. However, rfbE O157 isolate of buffalo RAJ sample carried 4 STEC virulent genes, hence considered an important source of STEC infection to humans and environment which should need to devise proper control systems.

Animal and Human Brucellosis in Pakistan.

Brucellosis is a bacterial zoonotic disease that affects many animal species and can be transmitted to humans via direct contact or via contaminated food. Although brucellosis is a serious health hazard, its public health concern has been neglected in many countries. In some developing countries, such as Pakistan, where brucellosis is endemic, this disease continues to be of importance. A literature search for the past 11 years (2011-2021) provided a comprehensive insight into brucellosis in Pakistan. In this review, particular emphasis was placed on occurrence, diagnostic tests used, and prevention, treatment, and control in the context of the "One Health" approach.

Bacillus pumilus induced tolerance of Maize (Zea mays L.) against Cadmium (Cd) stress.

Heavy metals contaminate the soil that alters the properties of soil and negatively affect plants growth. Using microorganism and plant can remove these pollutants from soil. The present investigation was designed to evaluate the induced effect of Bacillus pumilus on maize plant in Cadmium (Cd) contaminated soil. Three different concentrations of Cd (i.e. 0.25, 0.50 and 0.75 mg kg-1) were applied in soil under which maize plants were grown. The germination percentage, shoot length, leaf length, number of leaves, root length, fresh weight and nutrient uptake by maize plant were determined. The experiment was conducted by using complete randomized design (CRD) with three replicates. The result indicated that germination percentage, Shoot length, leaf length, root length, number of leaves, and plant fresh weight were reduced by 37, 39, 39, 32 and 59% respectively at 0.75 mg kg-1 of CdSO4 concentration but when maize seeds inoculated with Bacillus pumilus significantly increased the germination percentage, shoot length, leaf length, number of leaves, plant fresh weight at different concentrations of CdSO4. Moreover, the plant protein were significantly increased by 60% in T6 (0.25 mg kg-1 of CdSO4 + inoculated seed) and Peroxidase dismutase (POD) was also significantly higher by 346% in T6 (0.25 mg kg-1 of CdSO4 + inoculated seed), however, the Superoxide dismutase (SOD) was significantly higher in T5 (0.75 mg kg-1 of CdSO4 + uninoculated seed) and was 769% higher as compared to control. The Cd contents in Bacillus pumilus inoculated maize roots and shoots were decreased. The present investigations indicated that the inoculation of maize plant with Bacillus pumilus can help maize plants to withstand Cd stress but higher concentration of Cd can harm the plant. The Bacillus pumilus has good potential to remediate Cd from soil, and also have potential to reduce the phyto availability and toxicity of Cd.

Qualitative and quantitative analysis of steatite using calibration-free laser-induced breakdown spectroscopy in conjunction with x-ray fluorescence spectroscopy and Fourier-transform infrared spectroscopy.

The qualitative and quantitative study of raw steatite was carried out using a pulsed nanosecond Nd:YAG laser set to fundamental harmonic (1064 nm) at atmospheric pressure. To cross-validate the data, x-ray diffraction (XRD), x-ray fluorescence (XRF) spectroscopy, and Fourier-transform infrared spectroscopy (FTIR) were used. The analysis of optical emission spectra in the wavelength range 200-720 nm showed the existence of silica (Si), magnesium (Mg), and calcium (Ca). The plasma temperature and electron number density were estimated using the Boltzmann plot and Stark-broadening line profile methods, with results at 4906 K and ${1.47}\times{{10}^{17}}\;{{\rm cm}^{- 3}}$, respectively. The quantitative study was carried out using the calibration-free laser-induced breakdown spectroscopy (CF-LIBS) process, assuming local thermodynamic equilibrium and an optically thin plasma, and the following results were obtained: Si = 63.37%, Mg = 35.95%, and Ca = 0.671%. The quantitative findings obtained using the CF-LIBS approach were consistent with those obtained using XRD, XRF, and FTIR, demonstrating the applied methods' potential to be effective and complementary for the qualitative and quantitative study of multiphase raw steatite.

Serological and Molecular Detection of Bovine Brucellosis at Institutional Livestock Farms in Punjab, Pakistan.

Bovine brucellosis remains a persistent infection in ruminants in Pakistan. A total of 828 (409 buffaloes and 419 cattle) sera were collected from 11 institutional-owned livestock farms in Punjab, Pakistan. The samples were tested by rose bengal plate agglutination test (RBPT) and indirect enzyme-linked immunosorbent assay (iELISA). The seroprevalence along with 95% confidence interval (CI) was determined. Univariable and multivariable analysis of the epidemiological background data was conducted and odds ratio (OR) was calculated to understand any association between the risk factors and the seroprevalence. An overall seroprevalence of 3.9% (Positive/Tested = 32/828) and 3.3% (27/828) was detected by RBPT and iELISA, respectively. The seroprevalence of 5.6% (CI 3.6-8.3) and 4.7%, (CI 2.8-7.2) and the odds ratio of 2.63 (CI 1.20-5.77) and 2.50 (CI 1.08-5.78) for testing positive by RBPT and iELISA, respectively were significantly higher (p < 0.05) in buffaloes than in cattle. Breed, sex, history of abortion and retention of fetal membranes (RFM) in the animals were not found statistically significantly associated with the infection. RBPT and iELISA based results agreed almost perfect (k = 0.877). In total, Brucella abortus-DNA (9/27) was amplified from seropositive samples by real-time polymerase chain reaction. This study identified for the first time the etiological agents of brucellosis at a molecular level at institutional-owned livestock farms in Pakistan.