DIRECT SCIENCES: SOLVING PROBLEMS THROUGH SCIENTIFIC NETWORKING

Commercialization and Indiscriminate growth of industrial sector has led down great impact not only on our living standard but also on environment. Indiscriminate release of heavy metals as industrial pollutant has been tremendously increased. The bio-availability of toxic heavy metals ions directs biomagnification that resulted in severe health hazards. Microorganism can offers attractive, eco-friendly and possible solution to this concern. Bacteria combat metal toxicity through adapted resistance mechanisms that can be exploited to minimize the alarming effects of heavy metals. Bioremediation through resistant bacteria serve as great temptation in modern research due to its promising results. This technology still needs more exploration in order to yield maximal results. Further research on different parameters of research design for bioremediation of heavy metals can led to innovation and even more sustainable model of pollution control. This review comprehensively highlights the sources, importance, consequences of heavy metals pollution. This review also described the underlying mechanisms of metal resistance by bacterial species to combat their devastating effects in terms of environmental pollution.

Heavy metals, Pollution, Resistance mechanism, health hazards, Bioremediation, Regulatory standards

Light is fundamental precursor for plant growth and development. LED provides a uniform source of light for closed plant farming practices. The present study was carried out to investigate the effect of various wavelength light-emitting diodes (LEDs) on photosynthetic parameters of Brassica juncea. Under red light treatment, number of leaves, fresh and dry mass of roots and shoots was increased. While under blue light, stem length was longer than other lights. In combination of both lights, root length, area of leaves was greater as compared to others. Moisture content was more in plant growing under red light. The contents of chlorophyll a were higher under blue LEDs light while chlorophyll b concentration was higher under red. However, the plants grown under blue and red light exhibited closed stomata and fewer in number. The concentration of total soluble protein and soluble sugar were found highest under blue LEDs. From these results, we can conclude that red light is most significant for biomass production and chlorophyll b contents while blue light is preferable for soluble proteins and sugar concentration and chlorophyll a.

LEDs, Growth, Chlorophyll, Stomata, Brassica

Hypertension, an outstanding danger factor for different cardiovascular, fringe vascular and renal occasions is a significant general wellbeing challenge. Reninangiotensin framework being the most essential pathogenic system of hypertension is interceded by a key segment, the angiotensin converting enzyme. The aim of current study was to know the relationship of hypertensive patients with angiotensin converting enzyme gene polymorphism. Total 65 clinically analyzed hypertensive patients with no related sickness condition and five age and sex coordinated clearly sound controls were included in the current study. Polymerase chain reaction was used for the amplification of intron number 16 of Angiotensin converting enzy megene harboring the 287 bp Alu repeat sequence. Homozygous Deletion/Deletion, Insertion/Insertion and heterozygous Insertion/Deletion were the three possible genotypes which were analyzed. After the amplification of products, 2% agarose gel was used for the separation of products. Statistical Package for the Social Sciences 20 (SPSS 20) was used for the statistical analysis; the P value was found non-significant and to avoid any false positive or false negative significance we cannot conclude any result.

Insertion; deletion; renin angiotensin system; total peripheral resistance; blood pressure

Hyphaene thebaica and Medemia argun are very important desert plants not only as sources of nutrients for human and animal nutrition, but also as potential sources of antioxidant and antimicrobial secondary metabolites agents such as polyphenolics including protoanthocyanidins, flavonoids and phenolics in addition to other classes of secondary metabolites such as alkaloids and saponins. The recently discovering biological activity of H. thebaica has increased its importance. Having very narrow ecological amplitude, M. argun has received very little attention from researchers. The description of M. argun as endangered species may also contributed to decreasing the number of studies on Medemia. The objective of this paper is to review the phytochemistry and biological activity of both H. thebaica and M. argun. The biological activity of both plants will be discussed and evaluated in terms of the content of secondary metabolites in their extracts. Till now M. argun has not been biologically exploited yet although it has diverse arrays from active secondary metabolites in different parts of the plant.

Hyphaene thebaica; Medemia argun; Secondary metabolites; Polyphenolics; Antioxidants; Anticancer activity; Antimicrobial activity

Mango is a major fruit crop of Sindh. In recent past death in mango has caused massive losses in its production. In order to find the preventive measures for this disease efficacy of plant extracts was evaluated in this experiment. Botanical extracts of five plants i.e. neem (Azadirachta indica), ak (Calotropis procera), dhatura ( Nicotiana tabacum) and onion (Allium cepa) were used. These extracts were tested in vitro in four concentrations 2ml, 5ml, 10ml, 20ml per liter of nutrient media in three replicates using completely randomized design. The results revealed that plant extracts considerably inhibited mycellial growth of test fungus as compared to control. Neem extract was the most effective and it produced 84.29% inhibition followed by tobacco with 74.84% reduction in fungal growth. Minimum reduction (53.428%) of Ceratocystis fimbriata was observed in dhatura extracts. The use of these extracts especially neem can be fruitful in eco management of sudden death in mango

Sudden death, Mango, Plant extracts, Management

The effect of allelochemicals from Casuarina cunninghamiana branchlets was assessed on the growth and metabolomic content of three recipient plants: (cucumber), Lycopersicon esculentum Mill.(tomato) and Brassica nigra L. (black mustard). The aqueous extract delayed the germination rates of the three species with pronounced effect on B nigra and L. esculentum.The aqueous extract significantly reduced the seeds germination percentage of B. nigra. The shoot and root lengths were reduced significantly under these concentrations and the germination index (GI) registered lower value due to the effect of the extract on root length. Under these concentrations (1-4 %) the seeds germination percentage of L. esculentum was not significantly affected but the root length was dramatically decreased. No significant change either in seeds germination percentage or in the root length up to the concentration of 4 % was recognized in .Application of allelochemicals from germination and growth criteria but also affected the biochemical and metabolomic content in C. sativus. Application of allelochemicals from C. cunninghamiana branchlets not only affected the germination and growth criteria but also affected the biochemical and metabolomic content in the recipient crop species. A significant increase in carbohydrates, proteins and was also observed in L. esculentum and proline content was observed in both plants. Stress on germinated crop seeds resulted in a significant decrease in photosynthetic pigmen cunninghamiana branchets aqueous extract in both plants but the content was increased significantly under the highest concentration in tomato. Sensitivity of the weed allelochemicals from the donner plant may open the door to exploit overcome the growth of weeds species associated to crop plants through using this plant as natural herbicides

Allelopathy,Casuarina cunninghamiana,Cucumis sativus,Germination percentage,Lycopersicon esculentum, Root length, Shoot length, PCA

The present work aimed to study the floristic composition and species diversity of weed assemblages in citrus orchards in Aljouf region, Kingdom of Saudi Arabia. Floristic lists were prepared and species coverage values were determined in 20 stands, using ten quadrats (1 � 1 m) per stand. In total, 33 plant species belonging to 15 families and 30 genera were identified. The largest families were Asteraceae and Poaceae (5 species for each), followed by Fabaceae (4 species), Chenopodiaceae and Polygonaceae (3 species for each). Therophytes represented the dominant life form (66.7% of the total flora), followed by geophytes (12.1%), chamaephytes, hemicryptophytes (9.1% for each) and phanerophytes (3%). The application of TWINSPAN (Two Way Indicator Species Analysis) classified the weed vegetation into four groups dominated by Plantago lagopus, Cynodon dactylon, Imperata cylindrica and Convolvulus arvensis. The species richness for the vegetation group dominated by Plantago lagopus was significantly higher (15.8 species/stand) than for the other vegetation groups. Shannon index and evenness did not show significant differences among the different vegetation groups. The species richness of citrus orchards was relatively high which could be related to the environmental heterogeneity due to different light conditions in citrus orchards.

Agricultural systems, Species diversity, Life form, Plant communities

Castor oil plant (Ricinus communis L.) having imperative medicinal and industrial significances. In the present study, we observed the biochemical and molecular characterization of castor bean (Ricinus communis L.) collected from different climatic zones of Pakistan (Lahore, Peshawar, Rawalpindi, Dera Ismail Khan, Swat and Kohat). The protein banding pattern of all 6 accessions was found same and no specific variation was noticed among the proteins of high molecular weight. However, small amount of variation was found in the number and intensity of low molecular weight. RAPD analysis of four primers (OPA-01, OPA-03, OPA- 09 and OPA-10) bares no variation in the genomic DNA banding patterns. Variations were found in the photosynthetic pigments attributes of the studied genotypes with high Chlorophyll a, b and total Carotenoids contents in plants of Kohat zone while lower level was found in plants of Swat zones. The present study revealed no variation among the genotypes by SDS-PAGE and RAPD. However it is useful to distinguish different form of castor with special reference to photosynthetic pigments grown in diverse climatic zone.

Castor oil plant, SDS-PAGE, RAPD, Photosynthetic Pigments, Pakistan

Medicinal plants have an appreciable value in the development of modern therapeutics. In the present study ethanolic extracts from the leaves of nine selected medicinal plants, including Calendula arvensis, Dodonaea viscosa, Olea europaea, Ficus carica, Otostegia limbata, Withania somnifera, Eucalyptus camaldulensis, Mentha longifolia and Cannabis sativa were assessed against Escherichia coli, Klebsiella species, Pseudomonas aeruginosa, Salmonella species and Staphylococcus aureus using agar well diffusion method. All the selected bacterial pathogens are the major causative agent of human infectious diseases. Plant extracts used in the present study have shown potential activity against E. coli, P. aeruginosa, Salmonella species and S. aureus while inhibitory activity was exhibited against Klebsiella species. In comparison to ampicillin most of the plants showed increased activity against P. aeruginosa. Among the nine plant extracts, the Ficus carica showed highest activity against all the pathogens while exhibited increased activity (29 � 1 mm) against P. aeruginosa as compared to ampicillin (6 � 1 mm). The medicinal potential of selected plants was for the first time tested against a set of infectious disease pathogens and have shown remarkable activity. Further phytochemical analysis will be helpful for the future elucidation of novel therapeutic agents from these plants.

Nine ethnomedicinal plants, Gram negative /Gram positive bacteria, ethanolic extracts, antibacterial activity

American Foulbrood (AFB) disease caused by the bacteria Paenibacillus larvae, is a serious disease of honeybee throughout the world, that bring down a considerable economic loss to the beekeepers. All work was undertaken in the commercial Apiaries of Kohat and Peshawar regions that showed visual signs of AFB infection. In the investigated 32 Apiaries (containing 2100 colonies), 26 colonies were identified positive for AFB. This work was valuable for the over production of honey bees to kill the AFB bacteria which were identified in the current study.

AFB, Honey Bee, Beekeepers, Kohat, Peshawar

Malaria has been one of the leading public health concerns worldwide particularly in developing countries. This study was aimed to determine malarial prevalence in local population of district Kohat in Khyber-Pakhthunkhwa province of Pakistan, within the study tenure of three months (April to June). Total 560 blood samples from suspected individuals were collected. These samples were then subjected to microscopic detection of Plasmodium by developing thin and thick blood smears followed by giemsa staining. Among all collected samples, only 137 (24.46%) samples were positive for malaria. Further investigation showed that out of positive samples, 129 persons (94.16%) were infected with Plasmodium vivax while remaining 8 persons (5.84%) were infected with Plasmodium falciparum. Highest number of infected-ratio (35.77%) belonged to the age group “11-20 years”, while least prevalence (2.19%) was found in age group “51 years and above”. Malarial infection ratio was higher in males 54.74% compared to females 45.26%. It has been concluded from the study that the malarial prevalence is high in Kohat, which may be due to the availability of favorable breeding places for malarial parasites and lack of effective preventive measures in study area.

Malaria, Palsmodium falciparum, Plasmodium vivax, Kohat District, Epidemiology

To study the Antibiotic resistance determination of S. aureus isolates and screening for MRSA. This was in-vitro study on MRSA isolates received from clinical samples in the department of microbiology during three months (January 2015-March 2015). Staphylococci were identified by catalase, coagulase and DNAse tests. The samples were cultured on blood agar and MacConkey-s agar plates. Antibiotic susceptibility tests were carried out by disc diffusion method. The disc used was Amikacin (30μg), Doxycycline (30μg), Co-trim0oxazole (25μg), Gentamicin (10μg), Penicilin (10μg), Erythromycin (15μg), Clindamycin (2μg), Vancomycin (30μg), Linezolid (30μg), Cefoxitin (30μg), Fusidic acid (10 g), Chloramphenicol (30μg) and Ciprofloxacin (5μg). Our results indicate that out of 112 positive isolates of S. aureus, 34 (30.35%) were found to be MRSA. These isolates were also resistant to different commonly prescribed other anti-staphylococcal antibiotics. A total of 34 (30.35%) MRSA was isolated from various clinical samples. The susceptibility of MRSA to antibiotics is changing. Upon the isolation of MRSA isolates many of them showed resistance against different antibiotics.

Methicillin Resistant Staphylococcus aureus, Antimicrobial susceptibility pattern, Pus swab, Rawalpindi, Pakistan

Emissions from vehicles possess various pollutants which cause worst form of environmental pollution. Various pollutants of vehicles altered parameters of exposed vegetation. Increase in vehicles pollution may also increase effect of vehicles-emitted pollutants on vegetation. It affects mainly the photosynthetic pigments, respiratory activities, enzymatic activities, uptake of water etc. Present review highlights the impacts of road side pollutants like sulphur dioxide, carbon mono oxide, nitrous oxide, lead compounds and some heavy metals e.g. cadmium, copper, chromium and zinc on aerial parts of plants. Cited literature reveals that the commonly affected plants by road-side traffic pollution are Azadirachta indica , Dalbergia sissoo, Cassia surattensis, Leucaenaleu cocephala and Parkinsonia aculeates, Alstonias cholaris, Cassia siamea, Allium sativum etc. in Pakistan. It is concluded that vehicles emission had significant effect on the various parameters of the plants.

Road side pollution, Vehicles, Environment, Vegetation

Malaria has been one of the leading public health concerns worldwide particularly in developing countries. This study was aimed to determine malarial prevalence in local population of district Kohat in Khyber-Pakhthunkhwa province of Pakistan, within the study tenure of three months (April to June). Total 560 blood samples from suspected individuals were collected. These samples were then subjected to microscopic detection of Plasmodium by developing thin and thick blood smears followed by giemsa staining. Among all collected samples, only 137 (24.46%) samples were positive for malaria. Further investigation showed that out of positive samples, 129 persons (94.16%) were infected with Plasmodium vivax while remaining 8 persons (5.84%) were infected with Plasmodium falciparum. Highest number of infected-ratio (35.77%) belonged to the age group “11-20 years”, while least prevalence (2.19%) was found in age group “51 years and above”. Malarial infection ratio was higher in males 54.74% compared to females 45.26%. It has been concluded from the study that the malarial prevalence is high in Kohat, which may be due to the availability of favorable breeding places for malarial parasites and lack of effective preventive measures in study area.

Malaria, Palsmodium falciparum, Plasmodium vivax, Kohat District, Epidemiology

To study the Antibiotic resistance determination of S. aureus isolates and screening for MRSA. This was in-vitro study on MRSA isolates received from clinical samples in the department of microbiology during three months (January 2015-March 2015). Staphylococci were identified by catalase, coagulase and DNAse tests. The samples were cultured on blood agar and MacConkey-s agar plates. Antibiotic susceptibility tests were carried out by disc diffusion method. The disc used was Amikacin (30μg), Doxycycline (30μg), Co-trim0oxazole (25μg), Gentamicin (10μg), Penicilin (10μg), Erythromycin (15μg), Clindamycin (2μg), Vancomycin (30μg), Linezolid (30μg), Cefoxitin (30μg), Fusidic acid (10 g), Chloramphenicol (30μg) and Ciprofloxacin (5μg). Our results indicate that out of 112 positive isolates of S. aureus, 34 (30.35%) were found to be MRSA. These isolates were also resistant to different commonly prescribed other anti-staphylococcal antibiotics. A total of 34 (30.35%) MRSA was isolated from various clinical samples. The susceptibility of MRSA to antibiotics is changing. Upon the isolation of MRSA isolates many of them showed resistance against different antibiotics.

Methicillin Resistant Staphylococcus aureus, Antimicrobial susceptibility pattern, Pus swab, Rawalpindi, Pakistan

Emissions from vehicles possess various pollutants which cause worst form of environmental pollution. Various pollutants of vehicles altered parameters of exposed vegetation. Increase in vehicles pollution may also increase effect of vehicles-emitted pollutants on vegetation. It affects mainly the photosynthetic pigments, respiratory activities, enzymatic activities, uptake of water etc. Present review highlights the impacts of road side pollutants like sulphur dioxide, carbon mono oxide, nitrous oxide, lead compounds and some heavy metals e.g. cadmium, copper, chromium and zinc on aerial parts of plants. Cited literature reveals that the commonly affected plants by road-side traffic pollution are Azadirachta indica , Dalbergia sissoo, Cassia surattensis, Leucaenaleu cocephala and Parkinsonia aculeates, Alstonias cholaris, Cassia siamea, Allium sativum etc. in Pakistan. It is concluded that vehicles emission had significant effect on the various parameters of the plants.

Road side pollution, Vehicles, Environment, Vegetation

Main cause of crops yield reduction in the world is the presence of weeds. Plant-derived smoke solution is well known for its promotry effect on plant growth. In the present study smoke solution derived from six different weeds Asphodelus tenuifolius (Cav.), Avena sativa (L.), Galium tricornutum (Dandy). Parthenium hysterophurs (L.), Phalaris minor (Retz.) and Scandix pecten-veneris (L.) were applied to find their effect on wheat seed germination, seedling vigor and seedling mass. It was observed that all the weeds-derived smoke solution significantly increased seed germination at 12, 24 and 36h at higher dilutions (1:1000, 1:3000, 1:5000 and 1:10000). Seedling length of wheat was also increased by Asphodelus, Avena, Galium, Parthenium and Phalaris at 1:1000, 1:3000 and 1:10000 dilutions. Similar effects were observed on seedling mass of wheat and increased at different dilutions by all weeds smoke solutions. Concentrated smoke solutions significantly inhibited plant growth of wheat. It might be concluded that effect of smoke solution is concentration dependent and it increases with the increase of smoke dilutions. It is also concluded that smoke solution application is environment friendly.

Smoke solutions, Plant growth, Seed germination, Seedling vigor, Weeds

Soymilk was prepared by soaking soybeans fortnight with sodium bicarbonate (baking soda), drained, and then blanched to inactivate lipoxygenase enzyme. Beans were passed through blender and extract was squeezed through muslin cloth, adjusted TSS of the remaining milk to 12% with distilled water, and homogenized. The milk was sweetened with 5% cane sugar and divided to four equal lots. Three lots were flavoured separately with most acceptable quantity (10%) of mango, banana and guava fruit pulp and one lot was kept as control. All the samples were pasteurized at 161F for 16 minutes, cooled packed in 250ml sterilized glass bottles and stored in refrigerator (40-45F). All the samples were analyzed for pH, acidity, TSS, protein and fat and organoleptically evaluated at an interval of one week till coagulation. It was observed that pH decreased while acidity and TSS significantly (P<0.05) increased in all samples. There was no effect of storage on protein content of all samples. The fat content remained almost the same. The score for color and flavour significantly (P<0.05) decreased during storage. All the samples coagulated after four week storage.

Soymilk, Fruit flavoured soymilk, flavoured milk, Physicochemical analyses

Frequent environmental contaminants within microbiology laboratory create not only diagnostic quandaries but also poses major risk for health care workers and patients. Objective of our study was to isolate and identify the common laboratory contaminant bacteria with an ultimate goal to reduce false positive culture reports as well as Laboratory acquired infections. The study was conducted in laboratory of Microbiology Department Kohat University of Science and Technology, Kohat from March 2014 to June 2014. Total 5 samples were collected from different areas of laboratory including table, floor, clothing, air, and incubator. Out of 5 collected samples, total 22 bacterial contaminants were isolated and identified by various biochemical tests. Out of 22 bacterial contaminants, 8 (36.36%) was Staphylococcus epidermis which was the most common contaminants. 7 (31.81%) was Bacillus subtilis which was the second most common contaminants. 4 (18.18%) was Staphylococcus aerous while 3 (13.63%) was Deptheriods which showed minimum bacterial contaminants. Precaution should be taken to get rid of these organisms from laboratory by means of proper laboratory disinfection and sterilization as well as personal hygiene of laboratory workers. From our study we conclude that laboratory can be a potential source of contamination to give false positive results.

Laboratory contaminant, Staphylococcus epidermises, Bacillus subtilis

High degree of sequence variations are present throughout in the coding regions of Hepatitis C virus (HCV) genome. However, a high degree of sequence conservation within the 5-untranslated region (UTR) has made this region a target of choice for most of detection assays based on nucleic acid amplification. The current study was designed to determine the HCV genotypes in samples of HCV chronically infected patients in Pakistan. Specific primers targeting 5-UTR region were designed, which were then used for both amplification and sequencing of all isolates. All HCV isolates were sequenced and genotyped based upon phylogenetically informative regions within the 5-UTR of HCV genome. Out of 15 samples, 9 (60 %) samples 5-UTR of HCV genome was successfully sequenced. However, only 6 (66.7 %) samples were assigned genotypes based upon sequence comparison with reference sequences of database. Results of this study revealed that five isolates that were assigned genotype 3a, showed 97 to 99 % sequence conservation to isolates of genotype 3a previously reported from Pakistan as well as from rest of world, grouped together and coincided with their positions in the phylogenetic tree. Moreover, one isolate that was assigned genotype 4a, showed 97 % sequence conservation to isolates of genotype 4a previously reported from rest of world, coincided with its position in the phylogenetic tree. Our findings suggest that direct sequencing analysis of the 5-UTR of HCV genome is a sensitive and efficient approach of HCV genotyping; it may be adopted as a routine diagnostic procedure for HCV genotyping in clinical settings.

HCV genotypes, 5-Untranslated region, Sequencing, Phylogenetic analysis, Pakistan

Rice (Oryza sativa) is one of the principal foods for a large part of the worlds human population, but its productivity is reduced by the toxic effects of heavy metals such as lead. In the present study seeds of the rice cultivars KSK-133, NIAB-IR-9, Basmati-385 (B-385) and Shaheen Basmati (SB) were treated with different concentrations of lead chloride (PbCl2) (0, 250, 500, 1000 and 2000 ppm) to evaluate the effect on germination, seedling growth and some biochemical attributes. Lead had no effect on germination percentage, except cv. SB at 2000 ppm. Germination rate, root and shoot length and dry weight decreased with increasing lead concentrations. A sand culture experiment was conducted to measure the ion contents (Na, K, Ca), photosynthetic pigments, total protein and nitrogen contents. Chlorophyll a, chlorophyll b and carotenoids decreased with the increasing concentration of lead. Three varieties (NIAB-IR-9, KSK-133 and SB) had a decreasing trend in potassium and calcium ions and increasing in lead ion concentration. There was an increase in sodium ion concentration with the increase in lead concentration but B-385 showed contrary results. Results confirm that lead has an inhibitory effect on plant growth and development.

Oryza sativa L., Lead stress, Germination, Plant growth

The study was carried out during the chickpea growing season to study the effect of seed size on yield and yield components of chickpea. Ten chickpea genotypes NKC-5-S13, NKC-5-S24, NKC-5-S17, NDC-122, NDC-4-20-6, NKC-5-S16, KARAK-2, NDC-4-20-1, NIFA-88 and NIFA-95 were used in this study. Randomized complete design (RCBD) was used with three replications. Data were recorded on days to 50% flowering, plant height, primary branches plant-1, secondary branches plant-1, pods plant-1, seeds pod-1, number of seeds plant-1, 100 seed weight, biological yield plant-1, harvest index, leaf area, leaflet leaf-1, seed yield plant-1, seed yield plot-1. Significant differences were observed for all traits. Ranges of different parameters were recorded as 138-149.3 for days 50% flowering, 53.3-67.8cm for plant height, 1.6-4.5 for primary branches plant-1, 4.1-9.7 secondary branches plant-1, 25-69.2 for pods plant-1, 1-1.7 for number of seeds pod-1, 34.8-69.5 for number of seed plant-1, 18.4-31.1g for 100-seed weight, 9.9-40.5g for biological yield plant-1, 29-51.1g for harvest index, 290.1-716g seed yield plot-1, 4.5-20.5g for seed yield plant-1, 3.8-8cm for leaf area and 12-14.9 for number of leaflets leaf-1.The effect of seed size on yield and yield components was significant for most of the traits.

Cicer arietinum, Seed size, Yield components

Efforts were made to develop proper protocol for establishment of aseptic conditions as well as efficient initiation and multiplication of in vitro cultures of olive cultivar “Uslu”. Our work inquires that the main factors such as media composition; growth regulators, fungicide (mancozeb) and bacteriocide (Clorox) have prominent effects on the micropropagation of the selected olive cultivar. The effectiveness of different treatments, in terms of concentration and time, was investigated for optimization of aseptic conditions.For controlling fungal contamination; the best results were achieved when 5gm mancozeb was used for 75 minutes, while 50% Clorox treatment for 5 minutes was found to be the best for controlling bacterial contamination. After sterilization, medium for direct shoot regeneration and proliferation were optimized. The explants were cultured on olive media containing different concentrations and combinations of auxins and cytokinins. Among the treatments used, the highest sprouting rates were observed in the media containing 6-Benzyleaminopurine3.5 mg/L and 1-Naphthaleneacetic acid 1.5 mg/L. Our findings accentuate on the fact that the applicability of the optimized methods should be evaluated for olive cultivars of commercial value.

Browning, Callus, Contamination, Olive media, Phytohormone, Proliferation, Sprouting

Berberis lyceum is an economically and medicinally important genus distributed abundantly in the mountainous ranges of Pakistan. The color of hairs on the leaves of B. lyceum genotypes was pale to brown while the branch color was brown in all B. lyceum genotypes. Similarly, flower color was pale yellow while the plant height was ranged from 2.0 to 5.0 meters. Average length of lamina was 2.4 cm, average numbers of thorns per branch were 50.0 and average number of berries per branch was 18. Leaves tend to have comparatively high phenolic compounds that can interfere with DNA isolation. DNA-based markers are becoming popular for the identification of plants. Molecular genetic diversity studies of Berberidaceae family are almost lacking, which revealed the neglected status and the extent of the gap in the knowledge of this family. Mercaptoethanol being a reducing agent directly or indirectly causes damage to DNA. It is one of the limiting factors found in our study that can inhibit the oxidation process and at its low concentration brown viscous DNA pellet was extracted from fresh leaves while at high concentration intact whitish DNA pellet of good quality was observed that found suitable for PCR amplification. The DNA quantity and quality from dry leaves were quite low that was improved with the increase in the concentration of β 2-mercaptoethanol. It is concluded that a combination of 1% PVP and 0.01 to 0.06 % β mercaptoethanol was found satisfactory for extracting DNA from fresh and dry leaves of B. lyceum genotypes. Further, fresh and dry leaves were not able to exhibit any significant effects.

Berberis lyceum , Barberry, Morphological variations, DNA isolation, RAPD

Microorganisms are important source of proteases which are employed in detergent, food, pharmaceutical, leather and photographic industries. Among microorganisms, bacteria have been widely explored globally for their protease production that leads to discovery of several novel proteases. The present research work was carried out to explore the indigenous protease producing soil bacterial flora of Karak, Khyber Pakhtunkhwa, Pakistan. A total of 60 bacteria were isolated from 16 soil samples collected from different sites in district Karak. The isolates were screened for protease production by using skimmed milk agar. Among all the isolates, 18 bacterial isolates were found protease producers with various degree of protease activity. These isolates were further identified through their morphological, microscopic and biochemical examination. It was concluded that soil of Karak is rich in protease producing bacteria that exhibit diversified proteolytic activities and biochemical features. Further optimization of protease activity and their structure elucidation may be performed which will be helpful to identify novel protease.

Soil sample, Protease producing bacteria, Karak, Skim milk agar

Nanotechnology from the very beginning has been implicated to be useful in various fields of science, owing to its interdisciplinary dimensions. Various studies around the globe have resulted in a plethora of information regarding the diagnostic and therapeutic potentials of nanotechnology. Different kinds of nano-structures have been produced with unique properties due to their peculiar sizes with propositions in medicine and healthcare. Such nano-scale structures with their prospective geometries are very important in relation to their functionalities. Moreover, modifications on surface of nanoparticles have been impregnated to bring changes that can provide desired effects. Targeted drug therapy by nanoparticles is a subject of recent interest, as it focuses on removing the side effects of certain conventional therapies. Various viral infections have been explored by targeting effects of nanotechnology. Conventional methods, targeting viral infections have certain drawbacks, which can be tackled by nanotechnological approaches. This review focused on the current status of nanoparticles in specific systems that work against viruses, using the conceptual translation of the nano-scale moieties. Different targets for various virology related entities have been exploited and resulted in promising aspects of targeting agents and nano-therapeutics.

Nanotechnology, Viruses, Nanoparticles, Nano-Carriers, Targeted Drug Delivery

Abiotic stress is a major problem around the world causing huge losses to cultivated crops. To overcome this problem, a study was carried out to examine the effect of Sodium chloride (NaCl) stress on germination of seeds, plantlet development (root, shoot length and fresh, dry weight), cell damage (cell membrane stability) and SDS-PAGE analysis of two rice cultivars i.e. Basmati-385 and Shaheen Basmati. It was observed that salt stress had negative effects on seed germination and germination rate. In both rice varieties, gradual reduction in root and shoot length was observed from control to 100 mM Sodium chloride stress while a considerable decrease in shoot and root length was examined at 150 mM. Root and shoot dry weight showed progressive decrease with increasing salt stress. Cell injury also showed progressive increase from control to 150 mM salt concentration. In SDS-PAGE analysis, no variation was found in proteins banding pattern of rice leaves proteins under salt stress and control.

Salinity, Cell membrane stability, Rice, SDS-PAGE

Ten olive (Olea europaea L.) genotypes were collected from Agriculture Research Institute Tarnab at Peshawar and Sang Bhatti at Sawabi, KPK, Pakistan to evaluate their genetic diversity. Genomic DNA was isolated from fresh leaves, using a CTAB extraction method with few modifications. Three RAPD primers OPC-01, OPC-09 and OPE-07 obtained from Biron Technologies were tested by RAPD-PCR. Totally 27 bands were amplified and these amplified fragments ranged in size from 100 to 400 bp. Comparatively, more bands were observed in OPC-09 primer, with 12 bands and least number of bands were observed in OPC-01 primer with 7 bands from the electrophoresis results of the olive genotypes. Among three RAPD primers, only OPC-09 was polymorphic, showing (42%) polymorphism.

Olive, Genetic Diversity, Germplasm, RAPD Markers

Mastitis is the inflammation of mammary glands of the dairy animals and tends to be a serious problem in the world while brucellosis is an important zoonotic disease caused by bacteria of the genus Brucella, found globally and is transmitted directly or indirectly through contact with infected secretions, milk and body fluids. Milk when contaminated by brucellae presents a high threat to the new born calves and also to the human as it can spread through ingestion and cause undulant fever or abortion. The current study was conducted to determine the prevalence of mastitis, and its correlation with brucellosis in bovine. A total of 196 milk samples were collected randomly and the overall incidence of mastitis was 25.5% by Surf Field Mastitis Test (SFMT) and brucellosis 6.1% by a routine Milk Ring Test (MRT). The results showed significant co-relationship between brucellosis and mastitis positive samples in accordance to the point prevalence analysis.

Mastitis, Brucellosis, Bovine

Morphological characterization is the first step in description and classification of the germplasm. But biochemical markers like isozymes, resistance to chemicals and seed storage proteins are also being used for genotyping studies in plant breeding. Now days, molecular markers are the markers of attention for advanced research. These can be classified into two categories including non-Polymerase Chain Reaction (PCR) based techniques and PCR-based techniques. Several new array chip techniques that utilize DNA hybridization coupled with labeled nucleotides and Next-Generation sequencing techniques have also been developed. The techniques that are promising for plant genotyping are the Restriction fragment length polymorphism (RFLP), Amplified Fragment Length Polymorphism (AFLP), Random Amplified Polymorphic DNA (RAPD), Microsatellites or Simple Sequence Repeat (SSR), and Single Nucleotide Polymorphism (SNP). The choice of a molecular marker technique depends upon the reliability, ease of analysis and level of polymorphisms. The merger of genetics and genomics has unraveled the new insights by defining various metabolic, regulatory and developmental pathways, but rigorous investigations still need to be completed.

Markers, genotyping, plants

HCV is highly prevalent in Khyber Pakhtunkhwa (KPK) Pakistan. To check the Serological prevalence of Hepatitis C infection in IDPs of Waziristan residing at Bannu Camp, this Research study was conducted in the Blood Bank main laboratory of Hayatabad Medical Complex Peshawar Khyber Pakhtunkhwa. Samples were collected from the IDPs Patients in the month of July and brought it in closed icebox container to the laboratory of Hayatabad Medical Complex, Peshawar. HCV infection was diagnosed on the basis of RDT Rapid Diagnostic Test and Serologic test by using Enzyme linked Immuno Sorbent Assay (ELISA). Out of 500 patients high prevalence was observed in age group of 40-60 years which was 82.17% while in age group 10-20 years the prevalence was 61.11%. The lowest prevalence was observed in the age group of 20-40, which was 55%. The data was significant when analyzed by chi square test with P<0.05.In gender wise, high prevalence was observed in females which were 72.69% while in children it was 50%. The lowest prevalence was observed in the males (66.66%). It was concluded from the present study that Hepatitis C Virus infection was more prevalent in IDPs of North Waziristan. This study will be helpful to the health care policy makers to design strategies for controlling and eradication of Hepatitis C infection.

Hepatitis C, Serological, Anti-HCV, Patients

Metabolomics is well recognized technique for the analysis of biological system. Recently NMR is becoming an important tool of interest for researchers in the area of bimolecular analysis, especially for drug discovery. This technique is a significant step forward for identifying biomarkers, ligand interaction to the target, pathway recognition and metabolomic mapping. NMR based metabolomic is not only being used for the identification and structure elucidation of synthetic or natural compounds but also is considered as an imperative tool for biomolecular drug discovery. Significant advancements in methodological developments, software (including bioinformatics tools), and hardware (including instrumentation) is exploring new areas for industrial drug discovery. In this review, we discussed the importance of NMR technique in bio-molecular sciences.

Bimolecular analysis, NMR, Metabolomics

Genomics and its related studies boosted explorations when applied in various dimensions of biology. The most common concept employed is to mix natural abilities of various living organisms or distant biological sources in the form of genes targeted for their products. With the advent of 21st century, this field gained a pace due to the attention by various scientific communities worldwide. Though, many hurdles still exist on its way but synthetic biology has led the basis for advanced outcomes by merging the potentials of genetic engineering and electronic techniques. This piece of literature reviews the research and development of synthetic biology accomplished since past in various life sciences with emphasis on pharmaceuticals, vaccines and biofuel development. The efforts of international scientific community and international organizations are also highlighted, who developed regulations and transmitted the importance to applied level. The production of biofuel, anti-microbial drugs, vaccines or other biological components with the help of genetic engineering technology was the first generation which after integration in synthetic biology has successfully transferred to a new generation. Along with the past, this paper also forecasts the future of synthetic biology in minimizing the limitations and problems faced in biological research with the help of synthetic biology.

Synthetic Biology, Drugs, Vaccines, Biofuels

The Hepatitis C virus (HCV) is an enveloped, positive strand RNA virus and has been classified as a separate genus Hepacivirus of the family Flaviviridae. The HCV genome is a linear molecule with a length of approximately 9.6 kb that is flanked by 5` and 3’ untranslated regions (UTRs) and encodes to a nearly 3,010 amino acids polyprotein precursor. It is cleaved by combination of viral and host proteases into structural (C, P7, El, and E2) and non-structural proteins (NS2, NS3, NS4A, NS4B, NS5A and NS5B), essential for viral replication and viron formation in post-translational process. The 5` and 3’ UTRs are highly conserved regions in HCV genome, crucial for molecular processes such as replication and translation. In this review, we summarize the current knowledge about HCV molecular and structural HCV biology. This knowledge may help to improve treatment strategies and development of vaccine against HCV.

HCV, replication, post-translational process, nonstructural proteins, Untranslated regions

This survey was implemented to isolate, identify and analyze the susceptibility of HA-MRSA against commonly applied antibiotics as well as to detect its prevalence. Total 1394 samples of various natures were collected from different wards of Government Lady Reading Hospital Peshawar (Khyber Pakhtunkhwa) Pakistan. Staphylococcus aureus was isolated and identified by using selective media and biochemical tests including catalase, and coagulase tests, followed by detection of MRSA against methicillin. Further, susceptibility of MRSA was investigated against 14 antibiotics using standard disc-diffusion protocol. Demographic analyses revealed that out of 1394 samples, pus were the highest-rated (540) samples followed by urine (381) and blood (354) while the lowest number were semen samples (7). From total number of samples, highest incidences of Staphylococcus aureus were detected in pus (71.48%), blood (68.08%) and urine (54.35%) while lowest count was 42.86% in semen samples. Among Staphylococcus aureus -positive samples, MRSA rated 100%, 66.67%, 62.95%, 44.44%, 29.68%, 21.16%, 20.93% and 0.00% in semen, sputum, pus, throat-swab, urine, blood HVS and stools samples, respectively. All MRSA was found to possess average resistance of 64.08% (while 35.92% susceptibility) against all antibiotics collectively. Individually, highly resistant MRSA were recorded 98.71% against gentamicin followed by norfloxacin (82.14%) and ciprofloxacin (79.34%) while mostly susceptible to vancomycin (92.09%), chloramphenicol (67.09%) and sparfloxacin (57.40%). Overall, most adult female were found infected than male and children and vancomycin showed highest bactericidal property.

Nosocomial Infection, Staphylococcus aureus, MRSA, HA-MRSA, Antibiotic Sensitivity Pattern

A study was conducted to investigate the microbial quality of food (drinking water and processed meat) at popular food street (Namak-Mandi Peshawar Pakistan). For this purpose, 30 different food samples from different shops, including processed meat (Tikka Karahi (7), Meat Kari (Gosht Karahi) (7), Mutton kari (Mutton Karahi) (5), Chicken Karahi (6) and drinking water (5) were analyzed for Escherichia coli, Salmonella spp., Shigella spp., Staphylococcus aureus and fungi isolation. Total Viable Count for all samples was also determined. The drinking water samples showed the average percentage of 20, 100, 100 and 40 for S. aureus, Salmonella, Shigella, E. coli and fungi, respectively. The highest total viable count of water samples was 4.2x107CFU ml-1. The processed meat samples showed 32.95, 51.81, 64.85 and 72.76% average presence of S. aureus, Salmonella, Shigella, E.coli and fungi, respectively. The highest Total Viable Count (2.5 x 109CFU ml-1) observed in Tikka Karahi and Mutton Karahi.

Namak Mandi; Processed Meat; E. coli; S. aureu; Shigella; Salmonella

Atrazine; a member of s-triazine group, is a preferred herbicide used globally either individually or as adjunct with other herbicides. Beside its use, problem arises because of its low degradability leading to elevation of Atrazine concentration above the permissible limit resulting in the contamination of soil and water which eventually can greatly affect the living organisms. This contamination level of Atrazine in soil and water can be effectively determined by using high performance liquid chromatography; one of the most effective tool to analyze or quantify the concentration of any compound residues and its byproducts. This study was aimed to isolate the microorganisms capable to degrade Atrazine from soil, obtained from farmed areas of Gomal University D. I. Khan in December, 2011, having history of at least eight years continuous exposure to Atrazine. Isolated fungal and bacterial strains were observed for their potential to degrade Atrazine. Fungal isolates were identified on the basis of colony morphology and spore characteristics while bacteria were identified on the basis of the biochemical parameters. Out of twelve fungal strains, Aspergilus fumigates showed the highest biodegradation rate (58.24%) while the lowest biodegradation was shown by Aspergillus terreus (17.56%). Among the bacterial isolates three were Gram negative and five were Gram positive. The highest degradation potential (62.77%) was shown by Streptococcus specie while E. coli with lowest degradation rate (26.48%). Bacteria showed more average degradation potential than fungal isolates as the total average of degradation rate observed was 44.62% by bacteria and 40% by fungal isolates.

Atrazine, Atrazine Degradation, HPLC, Atrazine Degrading Microorganisms

The effect of plant derived smoke solutions on seed germination and seedling vigor is well documented. The present study was designed to find out the effect of Cymbopogon-derived smoke solution (1:500 dilution) on root of Ipomoea marguerite under CoCl2 (10, 20 and 30 ppm). Inhibitory effect of CoCl2 was observed on number of adventitious roots, numbers of lateral roots and adventitious root length as the concentration of CoCl2 (10, 20 and 30 ppm) increases. It was observed that 1:500 dilution of smoke significantly increased number of adventitious roots, numbers of lateral roots and adventitious root length. Plant derived smoke solution alleviated significantly inhibitory effects of CoCl2 at lower concentration (10 and 20 ppm). These results suggest that plant derived smoke dilution (1:500) showed positive effects and alleviated the inhibitory effect of CoCl2 on plant cuttings.

Alleviating solution, CoCl2, seedling length, smoke solution

In this study protease producing bacteria from different regions of Kohat, Khyber Pakhtunkhwa, Pakistan were isolated and screened for protease production on skimmed milk agar and casein agar plates. Out of 100 isolates, 12 were found to be protease producing on skimmed milk/casein agar plates. These isolates were subjected to several biochemical tests and gram staining and were identified to be mainly Bacillus species. Optimum enzymatic activity was found to be at 55˚C and pH 8-9.SDS-PAGE and Zymographic analysis showed that there is mainly one protease with molecular weight of around 50 kDa.

Protease; Bacillus species; Casein and skim milk agar

Heavy metal contamination of soil decreases microbial diversity and causes bacterial communities to lose part of their degradative capabilities. The present work is mainly related to Nickel (Ni) toxicity. Ni is the twenty fourth most abundant element of the earth’s crust and has been detected in the biosphere. High concentration of Nickel frequently inhibits enzymatic activity, DNA replication, transcription and translation by binding to proteins and nucleic acids. We investigated the impact of Ni-stress on the soil bacterial populations in artificial microcosmic conditions as well as compared the artificially created stress with the natural long term heavy metal stress to bacterial populations. Microcosmic conditions were provided by using sterile test tubes for two different intervals (24 and 48 hours) by addition of 20g soil and Ni treatments (500, 1000 and 1500 ppm). Our results showed that Ni stress significantly reduce the bacterial densities as compared to control without Ni contamination. The impact of Ni increased with increase in its concentration. The isolates were picked and characterized using different biochemical tests. The bacterial strains isolated in the study may be useful for bioremediation of heavy metals. Similarly, identifying the enzymes produced in the Ni resistant strains will also be an important future perspective.

Bacteria; Bioremediation; Biochemical tests; Microcosms; Ni

Pollution caused by heavy metals affects all forms of life. Pakistan is an agricultural country. The soil of Pakistan is rated low in fertility to fulfill our economic demand. Due to the lack of awareness in farmers, fields are irrigated by agricultural land disposals, mineral, industrial wastes and use of fertilizer. Areas especially Islamabad, Rawalpindi, Faisalabad, Lahore, Sheikhpora, Kala Shah Kaku, Multan, Kallarkahar, Kalarkahar-Lilla and Lilla-Lahore, Peshawar, Mardan, Sawabi, Nowshera, Charasda, Khyber agency, Kohat, and Karak are contaminated through industrial contamination, fertilizer, automobile, mining and natural constituents of the soil. Toxic levels of heavy metal contamination pose a serious threat to the microorganisms, animal plants and human life. Plants exposed to heavy metals result injury in terms of chlorosis along with toxic effects in the form of reduced photosynthesis, browning of root tips, growth inhibition, and finally death. Soil microbial population is highly affected by the presence of heavy metals. In human and mammals, neurological damage, immune system suppression and fetal abnormalities are reported due to the toxic effects of heavy metals. Live stock affected due to toxicity of heavy metals enters into the food chain can cause serious economic and health issue.

Heavy metal; Plant; Human; Microbe; livestock

Present study reports the Agrobacterium-mediated genetic transformation of bentazon resistant gene (Cyp81A6) in cotton genotypes (Coker 312 and Coker 201). Main aims were to genetically transform Cyp81A6 in cotton genome and to perform non-PCR and PCR-based confirmation studies regarding its stable transformation. The embryogenic calli (EC) were first inoculated with the Agrobacterium tumefaciensstrain LBA4404 carrying foreign gene (Cyp81A6) and were transferred to the selection medium having kanamycin (100 mg/L) for 4 weeks. Embryogenic calli were stunted in growth in both cultivars when transferred to selective MS medium. The kanamycin resistant (Knr) calli grew well after few sub-culturings. Knr and benr assays showed greater percentage of Knrembryogenic calli. Also EC of Coker 312 showed greater resistance than Coker 201. Both genotypes profusely produced tulip shaped somatic embryos; however, their number was more in Coker 312 as compared with Coker 201. Molecular analysis of putative transgenic calli confirmed the presence of Cyp81A6 in the genomes of Coker 312 and Coker 201. Present study reveals that both cotton genotypes were efficient in integration of the gene and grew well on normal and selective MS media.

Agrobacterium; Bentazon; Cyp81A6; Embryogenic calli

Methanolic leaf extracts of Ricinuscommunis collected from different climatic zones of Pakistan were screened for antimicrobial activity against different Gram-positive and Gram-negative human important pathogenic bacteria as Escherichia coli, Staphylococcus aureus, Salmonella typhi, Klebsiellapnemoniae, Shegella, Vibreo cholera and Proteus, by using agar cup diffusion method. It was found that all Ricinuscommunis species from different origin shows high zone of inhibition against the tested pathogens. These results demonstrated that R. comminus contain potential components that may be beneficial for evolution of pharmaceutical and can be use as a medicine against different types of bacterial phathogenic flora of human body.

Ricinus Communis; Antimicrobial; Well diffusion method; Zone of Inhibition

As staple food, Brassicaceae has global recognition and acceptance for human and animal consumption. The crops in Brassicaceae are a good source of compounds effecting health and are generally used as food, moreover they are a model for plant science research. The constituents those are important nutritionally includes, phenolic compounds, vitamins, fibers, soluble sugars, minerals, fats, and carotenoids. During growth, various biotic (e.g. herbivory, fungal, bacterial and/or viral infection) and abiotic (e.g. metals, UV, temperature, drought) stresses affects the plants. The metabolomic changes can be reasonably specific, as the plant defense-associated compounds consists of a variety of constitutive and induced metabolites. In nature plants have multitrophic interactions during growth and developmental processes. The power of metabolomics analytical methods is the analysis of wide spectra of compounds resulting in a huge data set in an unbiased and comprehensive manner. These enormous metabolomic data sets can be assessed by multivariate analysis, usually stating with an unsupervised method such as principal component analysis (PCA). To understand the specificity of the interactions of the plant and its environment a large amount of data on to Brassica was obtained concerning the effects of the defense indication compounds, for example salicylic acid, jasmonic acid and furthermore of infection with pathogenic and non pathogenic fungi, as well as human pathogenic bacteria and metals. Effects were measured at various developmental stages of the plant. Finally also the consequence of storage for various periods and temperatures was evaluated. An overlap of different treatments was observed that needs to be studied in more detail. The metabolomics changes during plant growth, characterizes the changes in metabolomic fluxes through diverse pathways. The current review shows the significance of plant stress factors as a feature for variance in nutritional value of vegetables for human utilization

Brassicaceae; plant stress; metabolomics; systems biology