Advanced methods of algal pigments extraction: A review.

Algae are exclusively aquatic photosynthetic organisms that are microscopic or macroscopic, unicellular or multicellular and distributed across the globe. They are a potential source of food, feed, medicine and natural pigments. A variety of natural pigments are available from algae including chlorophyll a, b, c d, phycobiliproteins, carotenes and xanthophylls. The xanthophylls include acyloxyfucoxanthin, alloxanthin, astaxanthin, crocoxanthin, diadinoxanthin, diatoxanthin, fucoxanthin, loroxanthin, monadoxanthin, neoxanthin, nostoxanthin, perdinin, Prasinoxanthin, siphonaxanthin, vaucheriaxanthin, violaxanthin, lutein, zeaxanthin, ß-cryptoxanthin, while carotenes include echinenone, α-carotene, ß-carotene, γ-carotene, lycopene, phytoene, phytofluene. These pigments have applications as pharmaceuticals and nutraceuticals and in the food industry for beverages and animal feed production. The conventional methods for the extraction of pigments are solid-liquid extraction, liquid-liquid extraction and soxhlet extraction. All these methods are less efficient, time-consuming and have higher solvent consumption. For a standardized extraction of natural pigments from algal biomass advanced procedures are in practice which includes Supercritical fluid extraction, Pressurized liquid extraction, Microwave-assisted extraction, Pulsed electric field, Moderate electric field, Ultrahigh pressure extraction, Ultrasound-assisted extraction, Subcritical dimethyl ether extraction, Enzyme assisted extraction and Natural deep eutectic solvents. In the present review, these methods for pigment extraction from algae are discussed in detail.

Bioremediation of heavy metals polluted environment and decolourization of black liquor using microbial biofilms.

BACKGROUND: With increased urbanization and industrialization, modern life has led to an anthropogenic impact on the biosphere. Heavy metals pollution and pollutants from black liquor (BL) have caused severe effects on environment and living organisms. Bacterial biofilm has potential to remediate heavy metals and remove BL from the environment. Hence, this study was planned to investigate the potential of microbial biofilms for the bioremediation of heavy metals and BL polluted environments. METHODS AND RESULTS: Eleven biofilm forming bacterial strains (SB1, SB2, SC1, AF1, 5A, BC-1, BC-2, BC-3, BC-4, BC-5 and BC-6) were isolated and identified upto species level via 16S rRNA gene sequencing. Biofilm strains belonging to Bacillus and Lysinibacillus sphaericus were used to remediate heavy metals (Pb, Ni, Mn, Zn, Cu, and Co). Atomic absorption spectroscopy showed significantly high (P ≤ 0.05) bioremediation potential by L. sphaericus biofilm (1462.0 ± 0.67 µgmL-1) against zinc (Zn). Similarly, Pseudomonas putida biofilm significantly (P ≤ 0.05) decolourized (65.1%) BL. Fourier transform infrared (FTIR) analysis of treated heavy metals showed the shifting of major peaks (1637 & 1629-1647, 1633 & 1635-1643, and 1638-1633 cm-1) corresponding to specific amide groups due to C = O stretching. CONCLUSION: The study suggested that biofilm of the microbial flora from tanneries and pulp paper effluents possesses a strong potential for heavy metals bioremediation and BL decolourization. To our knowledge, this is the first report showing promising biofilm remediation potential of bacterial flora of tanneries and pulp-paper effluent from Kasur and Sheikhupura, Punjab, Pakistan, against heavy metals and BL.

Evaluation of Safety of Stewart's Wood Fern (Dryopteris stewartii) and Its Anti-Hyperglycemic Potential in Alloxan-Induced Diabetic Mice.

Diabetes has become a critical challenge to the global health concerns. Cytotoxicity and development of resistance against available drugs for management of diabetes have shifted the focus of global scientific researchers from synthetic to herbal medications. Therefore, the current study was conducted to investigate the possible anti-hyperglycemic potential of Dryopteris stewartii using Swiss albino mice. To evaluate any possible toxic effect of the plant, acute oral toxicity test was performed while the anti-diabetic effects of aqueous and ethanol extracts at 500 mg/kg, positive, negative and normal control were assessed simultaneously. The anti-diabetic study revealed that aqueous extract has higher anti-diabetic potential than ethanol extract while lowered blood glucose level at second week reaching 150 mg/dL, exerting stronger anti-diabetic effects, compared to ethanol extract (190 mg/dL). Oral glucose tolerance findings revealed that aqueous extract decreased blood glucose level by -0.41-fold, compared to ethanol extract showing a decrease by only -0.29-folds. The histopathological evaluation of liver and pancreas of all groups revealed normal cell architecture with no morphological abnormalities. These results suggested the possible use of D. stewartii as anti-diabetic herbal drug in near future. However, these recommendations are conditioned by deep mechanistic studies.

Antioxidant potential and chemical characterization of bioactive compounds from a medicinal plant Colebrokea oppositifolia Sm.

Colebrookea oppositifolia is a highly used medicinal plant and an enriched source of essential oils. Therefore, the present study was designed with the aim to extract the chemical constituents and to evaluate its antioxidant potential. Fresh plant parts were subjected to the extraction of volatile chemical constituents by maceration using n-hexane as the menstruum. The resulting n-hexane fractions were purified and then subjected to GC-MS and FTIR analysis. In-vitro antioxidant abilities were evaluated by, DPPH, total phenolic content (TPC), total flavonoid content (TFC) method against the standard solutions of (Gallic acid, Quercetin) as a positive control. The GC-MS analysis of leaves, stem and inflorescence showed a total of 100, 98 and 48 components out of which 47, 16 and 17 peaks were identified representing the 67.64 %, 73.16 % and 61.93 % of the total oily fractions, respectively. The FTIR spectrum indicated the presence of various functional groups. In-vitro antioxidant results exhibited that leaves showed the highest antioxidant potential by DPPH (3.365 ± 0.002), and the highest total phenolic content by FC method (203.00 ± 0.091). Foliar micromorphological features were found significant in the authentication of C. oppositifolia. Further pharmacognostic studies of this plant are recommended to evaluate its therapeutic potential.

Acute and sub-acute toxicity study of a Pakistani polyherbal formulation.

BACKGROUND: Herbology is the prevailing system among the nationally-accepted alternative or complementary systems of medicine. The system is due to its general and patient-oriented methodology, is widely used in the general population exposing them to the risk of the side effects of the herbal medicines. METHOD: The aim of study was to assess the acute and sub-acute toxicity of the polyherbal formulation Hab-e-Kabad Noshadri tablets. In the acute arm of the study, a single dose of 2000 mg/kg was administered to Swiss Albino mice which were observed for physical symptoms and behavioral changes for 72 h. In sub-acute toxicity study repeated doses of the polyherbal preparation was administered to Wistar rats of both genders, separately. The animals received three doses of polyherbal product (50 mg/kg/day, 100 mg/kg/day and 200 mg/kg/day) for a period of 28 days. On 28th day of experiment, blood sampling of animals was done for hematological and biochemical analysis i.e. liver and renal function parameters, lipid profile and then sacrificed for histopathological examination of liver and kidney. RESULT: There was no morbidity and mortality noticed with single dose administration in acute toxicity study in mice. In sub-acute toxicity study, morphological changes with some damage in liver and kidney tissues of male and female animals were recorded at dose of 100 mg/kg/day and 200 mg/kg/day. CONCLUSIONS: It was found that prolonged use at higher dose i.e. 200 mg/kg/day of this polyherbal formulation should be avoided and practitioners should cautiously prescribe this formulation in patients with hepatic and renal impairment.

Building an environmentally accountable medical curriculum through international collaboration.

BACKGROUND: Global environmental change is associated with significant health threats. The medical profession can address this challenge through advocacy, health system adaptation and workforce preparedness. Stewardship of health systems with attention to their environmental impacts can contribute to mitigation of and adaptation to negative health impacts of environmental change. Medical schools have an integral role in training doctors who understand the interdependence of ecosystems and human health. Yet integrating environmental perspectives into busy medical curricula is not a simple task. CONTENT: At the 2016 Association for Medical Education in Europe conference, medical educators, students and clinicians from six continents discussed these challenges in a participatory workshop. Here we reflect on emerging themes from the workshop and how to plan for curricular change. Firstly, we outline recent developments in environmental health and associated medical education. Secondly, we reflect on our process and outcomes during this innovative approach to international collaboration. Thirdly, we present learning objectives which cover core content for environmentally accountable medical curricula, developed through a reflective process of international collaboration integrating current literature and the workshop outcomes. CONCLUSIONS: International collaboration can bring together diverse perspectives and provide critical insights for the inclusion of environmental health into basic education for medical practitioners.

In Vivo Study of Moringa oleifera Seed Extracts as Potential Sources of Neuroprotection against Rotenone-Induced Neurotoxicity.

Parkinson's disease (PD) is a leading neurodegenerative disorder affecting 1-3 percent of the elderly population. Oxidative stress is the primary factor for the neurodegeneration of Substantia Nigra (SN). The current study aims to assess the seed extracts of Moringa oleifera (MO) on rotenone-mediated motor function impairments in a PD mouse model. For this purpose, two different seed extracts of MO were prepared, including aqueous MO (AqMO) and ethanolic MO (EthMO). Male Swiss albino mice were grouped into five groups. Mice received 2.5 mg/kg rotenone for 21 consecutive days, and control mice received the vehicle. Extract-treated mice received 200 mg/kg AqMO and EthMO separately, orally and daily for 28 days. Sinemet-treated mice received 20 mg/kg, oral dose, as a positive group. The motor function performance was evaluated using standard neurobehavioral tests. The antioxidant potentials of MO seed extracts were estimated by lipid peroxidation (LPO), reduced glutathione (GSH), glutathione-s-transferase (GST) and catalase (CAT) activities in mice brain homogenates. The PD mice brain SN sections were investigated for neurodegeneration. MO seed extract-treated mice showed a significant reduction in motor dysfunction compared to rotenone-treated mice as assessed through the open field, beam walk, pole climb-down, tail suspension, stride length and stepping tests. Increased antioxidant capacities of the PD mice brains of MO extract-administered groups were observed compared to the control. A histological study showed reduced signs of neurodegeneration, vacuolation around multipolar cells and cytoplasmic shrinkage in MO extract-treated mice SN brain sections. Collectively, MO seed extracts protected the animals from locomotor deficits induced by rotenone, possibly through antioxidant means, and seem to have potential applications in neurodegenerative diseases.

Comparative Microstructural Evaluation of Wood in Three Dominant Ziziphus Species of Desert Ecosystem (Cholistan), Pakistan.

The present microstructural evaluation was carried out on the woods of three ethnobotanically important local fruit trees, namely, Ziziphus mauritiana Lam., Z. spina-christi (L.) Willd., and Z. nummularia (Burm.f.) Wight and Arn., of family Rhamnaceae from Cholistan Desert of Pakistan. Wood samples were sectioned with sliding sledge microtome to make permanent slides for observing different anatomical parameters under the light microscope. All selected species were observed to have diffuse-porous wood with indistinct growth rings. The vessels were rounded in outline in all the species studied and found mostly solitary or in radial multiples of 2 in Ziziphus mauritiana and Z. nummularia, while in radial multiples of 2 to 5 in Z. spina-christi. The intervessel pits were scalariform to opposite. The rays were uniseriate in Ziziphus mauritiana, while mostly were biseriate in Ziziphus spina-christi. Simple perforation plates and diffuse, confluent, and vasicentric types of axial parenchyma were present in all the selected species. The fibers were thin-walled and nonseptate. One-way ANOVA followed by the Tukey test was conducted to observe different anatomical variations within selected species. Principal component analysis revealed correlations among studied anatomical parameters. The number of rays per mm was comparatively larger in Ziziphus nummularia, showing its greater susceptibility to wood-deteriorating agents than in other selected species. The Runkel ratio indicated the selected species suitable for making paper.

Medicinal Perspective of Antibacterial Bioactive Agents in Earthworms (Clitellata, Annelida): A Comprehensive Review.

Humoral practice is a fundamental natural biological phenomenon in earthworm defensive system which protects them from infectious bacteria and irritating agents by different mechanisms. The defensive system of earthworms is highly complicated because they lack antibodies in their blood circulatory system but their body extracts and coelomic fluid comprise of different bioactive agents (i.e. peptides and proteins) that defend these worms. There are various groups of bioactive agents such as proteases (name depends on proteins/peptide function or formal earthworm species name), metabolites (total 59 metabolites found in Eisenia fetida), metal binding proteins (2 proteins such as Ca2+ binding calmodulin and metallothionein), active proteins (include lysozyme, lysenin and eiseniapore etc.), antimicrobial peptides (antibacterial vermi-peptides family (AVPF), antimicrobial peptide I (PP-I), coelomic cytolytic factor (CCF, CCF-I and CCF like protein), fetidin, lysenin, lumbricin (lumbricin I, lumbricin PG, and lumbricusin), organic acids (fatty acids, succinic acids, and lauric acid) and other organic compounds (such as purine and vitamin D). The presence of above mentioned molecules confer therapeutic potential that affect energy intake and involve in decreasing oxidative stress, metabolic disturbances and pro-inflammatory conditions. The future perspectives of earthworm bioactive compounds are concerned with the development of provisional standards, purification and classification for utilizations in pharma industry.

Light and scanning electron microscopy-based foliar morpho-anatomical comparison of selected family Rosaceae members distributed in District Lahore, Punjab, Pakistan.

In the present study morpho-anatomical characterization of selected Rosaceae members distributed in District Lahore was performed. Light and scanning electron microscopy was used for systematic characterization of the selected 19 species. Distinguished morpho-anatomical features such as size and shape of epidermal cells, size and type of stomatal cells, size and shape of trichomes, oil droplets, and silica bodies were contrasted. Results reported remarkable variations which could be taxonomically useful in identification of these members. Polygonal epidermal cells were observed in Eriyobotraya japonica, Potentilla bifurca, Potentilla supina, and Prunus amygdalus. However, Prunus cerasus possessed irregular-shaped epidermal cells that can be distinguished from hexagonal epidermal cells of Prunus persica. Similarly, stomatal type varied among some members. Paracytic or perisocytic stomata were observed in E. japonica whereas P. bifurca observed paracytic and anisocytic stomata. Lengths of guard cells were also of variable sizes. The average length of guard cells ranged from 53 (52-54) µm to 74 (73-75) µm in abaxial view. Potentilla supina had biggest, while Rosmarinus officinalis had tiny guard cells. Trichomes were tubular, stellate, cylindrical, ribbon-like, glandular, and nonglandular. Silica bodies in the present investigation were bilobed, rounded, and oval-shaped. It is inferred that diverse anatomical features proved to be valuable taxonomic tools that could be fruitfully helpful in identification of plants at specific as well as generic level.

Application of green silver nanoparticles synthesized from the red seaweeds Halymenia porphyriformis and Solieria robusta against oral pathogenic bacteria by using microscopic technique.

Aqueous extracts of two red seaweeds Halymenia porphyriformis and Solieria robusta were used to synthesize green silver nanoparticles. These biogenic nanoparticles were tested against four strains of oral pathogenic bacteria which cause tooth decay or cavities. Staphylococcus aureus (MT416445), Streptococcus viridans (MT416448), Lactobacillus acidophilus (MT416447) and Lactobacillus brevis (MT416446) were used. Characterization of AgNPs was done by UV-Visible spectroscopy, SEM, XRD and FTIR. XRD analysis revealed the crystalline nature of the particles. The size analysis by XRD of the green AgNPs by H. porphyriformis indicated it had smaller particles, 15.23 nm, when compared to AgNPs by S. robusta (17 nm). Both green synthesized silver nanoparticles showed moderate antibacterial activity against all strains of bacteria, except L. acidophilus. Both particles showed their maximum zone of inhibition against L. acidophilus at a lower concentration of 50 and 100 µg. However, it was concluded that silver nanoparticles of H. porphyriformis are more effective than that of S. robusta due to their smaller size.

Antimicrobial Efficacy of Biogenic Cobalt and Copper Nanoparticles against Pathogenic Isolates.

Biogenic synthesis of cobalt (Co) and copper (Cu) nanoparticles (NPs) was performed using the bacterial strains Escherichia coli and Bacillus subtilis. Prepared NPs were confirmed by a color change to maroon for CoNPs and green for CuNPs. The NPs characterization using FTIR showed the presence of functional groups, i.e., phenols, acids, protein, and aromatics present in the Co and CuNPs. UV-vis spectroscopy of E. coli and B. subtilis CuNPs showed peaks at 550 and 625 nm, respectively. For E. coli and B. subtilis CoNPs, peaks were observed at 300 nm and 350 nm, respectively. Antibacterial and antifungal activity of B. subtilis and E. coli Co and CuNPs was determined at 100 mg/mL concentration against two bacterial strains at 5, 2.5, and 1.5 mg/mL against fungal two strains F. oxysporum and T. viridi, respectively. B. subtilis CuNPs showed significantly higher inhibition zones (ZOI=25.7-29.7 mm) against E. coli and B. subtilis compared to other biogenic NPs. Likewise, B. Subtilis CuNPs showed lower MIC (4.3 ± 6.3) and MBC (5.3 mg/mL) values against both tested isolates. Antifungal activity of B. subtilis and E. coli CuNPs and CoNPs showed a concentration-dependent decrease in ZOI. Among all biogenic NPs, B. subtilis CoNPs showed the highest ZOI (25-30 mm) against F. oxysporum followed by E. coli CuNPs with maximum ZOI (20-27 mm) against T. viridi. Again, B. subtilis CoNPs and E. coli CuNPs showed lowest MIC and MFC values against both fungal isolates. In conclusion, the current study showed that biogenically synthesized B. subtilis Cu or CoNPs can be used as effective antimicrobial agents due to their potential antibacterial and antifungal potential.

Toxicological effects of the chemical and green ZnO NPs on Cyprinus carpio L. observed under light and scanning electron microscopy.

Nanoparticles in aquatic bodies cause serious harm to the aquatic organisms when accumulated in high amounts. However, green nanoparticles synthesized using plants can be less toxic as compared to chemical nanoparticles. Hence, we designed our study to investigate the toxicological effects of chemical and green zinc oxide nanoparticles (ZnO NPs) on the biological activity of juvenile Cyprinus carpio. The green ZnO NPs were synthesized from Solieria robusta, and chemical ZnO NPs were synthesized using zinc chloride solution and ammonium hydroxide. Characterization was done by using light microscopy, scanning electron microscope (SEM), Fourier transmission infrared radiation, and X-ray diffraction (XRD) techniques. The highest absorbance of nanoparticles was observed at 360 which confirmed the synthesis of ZnO. The SEM analysis showed that green nanoparticles were hexagonal while the chemical nanoparticles were spherical to cubic in shape. Definite peaks were observed in XRD of green and chemical NPs at 2θ angles 45.84° and 32.18°, respectively. Oxidative stress was determined by chemical analysis of catalase, glutathione S-transferase (GST), glutathione (GSH), and lipid peroxidation (LPO) activities. The toxicological effects of chemical ZnO NPs on the catalase, LPO, GST, and GSH activities were more than green ZnO NPs. The histopathological investigation proved that the effect of chemical nanoparticles was worse than green ZnO NPs. More tissue damage was found in chemical nanoparticles than green synthesized nanoparticles. It was concluded that chemical nanoparticles can be replaced by green nanoparticles, as green nanoparticles are eco-friendly with less toxicological effects. This replacement can limit the toxic effect of nanoparticles when they get accumulated in high amounts in water bodies.

Honey Potential as Antibiofilm, Antiquorum Sensing and Dispersal Agent against Multispecies Bacterial Biofilm.

This study is first to test Pakistani honey bees, Apis dorsata and A. cerana honey samples as anti biofilm, anti quorum sensing (QS) and biofilm dispersal agents honey against multispecies biofilm of bacteria (obtained from obese patients). Briefly, five previously identified isolates Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Morganella morganii and Klebsiella pneumoniae (MT448672-MT448676) were selected. Antibiogram study of all five isolates was tested against three antibiotics viz., erythromycin (20 µg/mL), lincomycin (100 µg/mL) and rifampicin (100 µg/mL). In order to form multispecies biofilm, identified bacteria were grown in batch culture by mixing equal volumes (OD590nm = 0.1) of 2, 3 and 5 bacterial isolates. In total 11 groups (g1-g11) were made. Crystal violet (CV) staining method was used to evaluate the antibiofilm potential and biofilm dispersal potential of both honey samples. QS inhibition in P. aeruginosa was measured following culture supernatant method. Antibiogram study showed significant (p < 0.05) resistance by P. aeruginosa against tested antibiotics. E. coli, M. morganii and K. pneumoniae were significantly susceptible to erythromycin and S. aureus to lincomycin. Both honey samples at 2% and 5% concentrations showed significant (p < 0.05) inhibition potential of multispecies biofilm by all test groups (g1-g11). Though A. dorsata honey significantly inhibited biofilm formation at 2 and 5% against all groups but 2% concentration was highly significant against g2-g4 groups. Regarding A. cerana honey, 2% concentration was significantly effective against g1, g4-g7 and g9-g11 groups. Both honey samples significantly inhibited QS at 2 and 5%. The 5% concentration of A. dorsata honey significantly dispersed biofilm by all groups compared to 2% which showed dispersal potential only by g2 and g3 groups. Accordingly, honey samples showed significant antibiofilm, anti-QS and biofilm dispersal potentials thus can be considered as good alternative to antibiotics.

Authentication of the therapeutic Lamiaceae taxa by using pollen traits observed under scanning electron microscopy.

Herbal medicines are gaining popularity worldwide for human healthcare because of their therapeutic potential. However, adulteration and use of unauthentic raw herbals as substitutes have become a major issue for the local communities and industry for reasons of safety and efficacy. Therefore, the authentication of medicinal plants before their use in herbal medicines is a need of time. Hence, the present study was designed with an aim, to authenticate the therapeutic Lamiaceous taxa by using pollen traits observed under scanning electron microscopy. Pollen micro-morphological studies solve the problem through discrimination and correct identification of the Lamiaceae species from the adulterants. Based on pollen features, Lamiaceae were further divided into two sub-families Lamioideae (tricolpate) and Nepetoideae (hexa-colpate). The pollen grains of Lamioideae were found as small to medium-sized, tricolpate, radially/bilateral symmetrical, sub-spheroidal and oblate shape. Besides, exine patterns bireticulate, reticulate and micro-reticulate, colpus surface sculpturing as psilate, gemmate, scabrate, and verrucate have also been reported. A significant variation was found in the pattern of the reticulum, thickness, and the number of secondary lumina per primary lumen. Similarly, Nepetoideae has a hexa zono-colpate pollen but tri and tetra zono-colpate pollen have also been observed. Hence, this study contributes to the authentication and correct identification of medicinally important Lamiaceae taxa by using scanning electron microscopic techniques and can help to solve the adulteration problem. Highlights Authentication of medicinally important Lamiceous taxa was carried out through scanning electron microscopic techniques. Chemotaxonomic characterization was used for the accurate identification of the therapeutic taxa. The medicinal, palynological and phytochemical significance of Lamiaceae taxa were evaluated. A significant variation was seen in the palynological traits that help in the determination and authentication of the therapeutic Lamiaceous species. Based on the chemotaxonomic characterization, our study can help to solve the adulteration problem for the reason of safety and efficacy.

Effect of necrotrophic fungus and PGPR on the comparative histochemistry of Vigna radiata by using multiple microscopic techniques.

Rapid advances in the field of pathogen detection have opened new opportunities and better understanding for their management approaches. Aim of this study was to elucidate histopathological observations of different tissues affected by Macrophomina phaseolina and to observe the defense responses of plant growth promoting rhizobacteria (PGPR) in mungbean plants. Sections of the stem and root were prepared and stained with ferric chloride, Lugol's iodine and Wiesner's reagent and were then observed under multiple microscopic techniques. Results revealed that both pathogen and PGPR produce responses on the plant that include colonization of xylem vessels by hyphae and sclerotia, hypertrophy and hyperplasia of the cells, destruction of xylem fibers and amyloplasts in parenchymatous cells; and production of gels by the plant were observed. There was a significant increase in lignin and phenolic compounds deposition in stem and root sections of PGPR treated and non-treated mungbean plants. Whereas the soil amended with PGPR showed very less to no starch production. Moreover, production of gels and gums were also observed in both stem and root sections. Compared to light microscopy, scanning electron microscope provided greater depth of focus and resolution of the pathogen attack on plant tissues, associated bacteria. As a whole, the data demonstrated that inoculation of PGPR can be an effective strategy to stimulate plant growth and they could significantly activate disease resistance against M. phaseolina.

Impact of Interplay between Obese Gut Microbiota and Diet in Developing Obesity in Synthetic Community Mice.

This study aims at investigating the effects of cultured gut microbiota (GM) of obese human coupled high fat diet (HFD) or chow diet (CD) in development of obesity in mice. 20 stool samples were collected from obese patients and isolated bacteria were identified morphologically and biochemically. Identified isolates were mixed in equal proportions to synthesize obese GM. In vivo study was performed using obese GM combined with HFD/CD using mouse model for three months. Albino mice were treated with ampicillin from one week prior to birth until weaning of the pups at seven weeks of age and then inoculated with obese GM. Sixteen mice were divided into four groups: i.e. group 1 (G1) mice fed with CD, group 2 (G2) mice with HFD, group 3 (G3) mice with GM + HFD and group 4 (G4) mice with GM + CD. Mice from groups 3-4 were considered synthetic community (SC) mice due to transfer of synthesize human GM. 16S rRNA sequencing identified five abundant bacteria as Pseudomonas aeruginosa, Staphylococcus sp., Escherichia coli, Morganella morganii, and Klebsiella oxytoca (accession numbers: MZ150742-MZ150746). In vivo study indicated that GM combination with either HFD/CD caused significantly increased body weight in SC mice (BMI; Kg/m2) compared to HFD or CD fed mice groups. One way ANOVA revealed highly significant increase (p ≤ 0.001) in levels of total cholesterol (TC), triglycerides and low density lipoprotein (LDL) in GM coupled diet groups (G3-G4; SC mice) compared to significant increase in HFD group (G2) versus CD group (G1). Our study is first of its kind to report significant effects of using purified strains as obese GM plus diet (HFD/CD) in inducing obesity in SC mice and elevated serum liver parameters as metabolic indicators, hence providing strong evidence about significance of modified GM combination with HFD in developing obesity in SC mice.

Comparative evaluation of the toxicological effect of silver salt (AgNO3 ) and silver nanoparticles on Cyprinus carpio synthesized by chemicals and marine algae using scanning electron microscopy.

The widespread use of silver nanoparticles (AgNPs) results in the unintentional release into the water body. Therefore, understanding of the potentially harmful impacts of AgNPs and Ag-salt on aquatic animals is a need of time. This study was design to analyze the oxidative stress and histopathological damages in Cyprinus carpio. The synthesis of AgNPs from Halymenia porphyraeformis and by reduction of chemical was done. Nanoparticles were characterized with UV-Visible spectroscopy, SEM, XRD, and FTIR analysis. The comparative toxicological effect of chemically synthesized silver nanoparticles (Ch-AgNPs), green silver nanoparticles (Gr-AgNPs), and Ag-salt on C. carpio was analyzed. For oxidative stress analysis, different tests Lipid peroxidation (LPO), catalase, glutathione reduction (GST), and glutathione S-transferase (GST) were performed. The highest LPO 245.168 ± 0.034 was recorded in Ch-AgNPs-treated gills and the lowest 56.4532 ± 0.02 was found in Gr-AgNPs-treated liver. Maximum GSH 56.4065 ± 0.13 was observed in Gr-AgNPs liver and minimum 40.781 ± 0.54 was recorded in Ag-salt gills. The maximum quantity of catalase 68.0162 ± 0.09 was noted in the Ag-salt-treated liver and the minimum was calculated 17.3665 ± 0.01 in the liver of Ch-AgNPs and highest values of GST 765.829 ± 0.11 were recorded in gills of Gr-AgNPs and lowest 633.08 ± 0.26 in the liver of Ch-AgNPs-treated fish. In conclusion, maximum destruction was found in the gills and liver of the fish treated with chemical and green AgNPs followed by Ag-salt as compared to control. The adverse effects of AgNPs and Ag-salt were probably related to the oxidative stress in the fish that lead to histopathological damage of its vital organs.

Morphological and palynological assessment of some taxa of genus Echinochloa through light and scanning electron microscopy.

The present study was undertaken to gather information on selected species of Echinochloa based on their morphological and palyonological examination through light and scanning electron microscopy. As the Echinochloa species are most problematic grass weeds and exhibit high interspecific and intraspecific variability so due to their great morphological diversity it is difficult to recognize them. A lot of variation was observed in morphological features of studied taxa. In the present study, E. colona had more or less spreading dense racemes whereas in E. crus-galli racemes were linear to ovate. Upper glume surface was pubescent in E. colona, hispid in E. crus-galli and spinose in E. walteri. E. colona was comprised of five-nerved glumes whereas in E. crus-galli glumes were three-nerved. Although pollen basic characters in family Poaceae remains consistent but great variation was observed in pollen of studied taxa. The pollen size was quite variable among the species of genus Echinochloa. In polar view, the size of pollen grains ranged from 27.5 (20-35) µm to 40 (30-50) µm. E. crus-galli was appeared to be the smallest in size whereas E. colona was the largest.). P/E ratio ranged from 0.84 to 1.15 µm among the species. The highest value of pollen fertility was found in E. colona as 93.33% and lowest was in E. crus-galli as 87.50%. The morphological key of studied taxa is also given to study variations. In conclusion, it was observed that microscopy of morphological and palynological assessment can play a vital role in identification of taxonomically problematic taxa.

Characterization of the genetic basis of local adaptation of wheat landraces from Iran and Pakistan using genome-wide association study.

Characterization of genomic regions underlying adaptation of landraces can reveal a quantitative genetics framework for local wheat (Triticum aestivum L.) adaptability. A collection of 512 wheat landraces from the eastern edge of the Fertile Crescent in Iran and Pakistan were genotyped using genome-wide single nucleotide polymorphism markers generated by genotyping-by-sequencing. The minor allele frequency (MAF) and the heterozygosity (H) of Pakistani wheat landraces (MAF = 0.19, H = 0.008) were slightly higher than the Iranian wheat landraces (MAF = 0.17, H = 0.005), indicating that Pakistani landraces were slightly more genetically diverse. Population structure analysis clearly separated the Pakistani landraces from Iranian landraces, which indicates two separate adaptability trajectories. The large-scale agro-climatic data of seven variables were quite dissimilar between Iran and Pakistan as revealed by the correlation coefficients. Genome-wide association study identified 91 and 58 loci using agroclimatic data, which likely underpin local adaptability of the wheat landraces from Iran and Pakistan, respectively. Selective sweep analysis identified significant hits on chromosomes 4A, 4B, 6B, 7B, 2D, and 6D, which were colocalized with the loci associated with local adaptability and with some known genes related to flowering time and grain size. This study provides insight into the genetic diversity with emphasis on the genetic architecture of loci involved in adaptation to local environments, which has breeding implications.