Search results for: wild bees

Authors: Eldah Ephraim Buba, Sanusi Abubakar Sadiq

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Habitat can be degraded by tourism leisure activities for example wildlife viewing can bring abrupt stress for animals and alter their natural behaviors when tourist come too close and wildlife watching have degradation effects on the habitats as they often are accompanied by the noise and commotion created by tourist as they chase wild animals. It is observed that Jos Wild Life Park is usually congested during on-peak periods which causes littering and contamination of the environment by tourist which may lead to changes in the soil nutrient. The issue of unauthorized feeding of animals by a tourist in which the food might be dangerous and harmful to their health and making them be so aggressive is also observed. The aim of the study is to investigate the environmental impact of tourists’ activities in Jos Wild Life Park, Nigeria. The study used survey questionnaires to both tourists and the staff of the wildlife park. One hundred questionnaires were self-administered to randomly selected tourists as the visit the park and some staff. The average mean score of the response was used to show agreement or disagreement. Major findings show the negative impact of tourist’s activities to the environment as air pollution, overcrowding, and congestion, solid littering of the environment, distress to animals and alteration of the ecosystem. Furthermore, the study found the positive impact of tourists activities on the environment to be income generation through tourists activities and infrastructural development. It is recommended that the impact of tourism should be minimized through admitting the right carrying capacity and impact assessment.

Keywords: environmental, impact, investigation, tourists, activities

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Authors: T. N. M. El Sebai, A. A. Khattab, Wafaa M. Abd-El Rahim, H. Moawad

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Fungal mutant strains have produced cellulase and xylanase enzymes, and have induced high hydrolysis with enhanced of rice straw. The mutants were obtained by exposing Penicillium strain to UV-light treatments. Screening and selection after treatment with UV-light were carried out using cellulolytic and xylanolytic clear zones method to select the hypercellulolytic and hyperxylanolytic mutants. These mutants were evaluated for their cellulase and xylanase enzyme production as well as their abilities for biodegradation of rice straw. The mutant 12 UV/1 produced 306.21% and 209.91% cellulase and xylanase, respectively, as compared with the original wild type strain. This mutant showed high capacity of rice straw degradation. The effectiveness of tested mutant strain and that of wild strain was compared in relation to enhancing the composting process of rice straw and animal manures mixture. The results obtained showed that the compost product of inoculated mixture with mutant strain (12 UV/1) was the best compared to the wild strain and un-inoculated mixture. Analysis of the composted materials showed that the characteristics of the produced compost were close to those of the high quality standard compost. The results obtained in the present work suggest that the combination between rice straw and animal manure could be used for enhancing the composting process of rice straw and particularly when applied with fungal decomposer accelerating the composting process.

Keywords: rice straw, composting, UV mutants, Penicillium

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Authors: Ahmed Bahieldin, Ahmed Atef, Jamal S. M. Sabir, Nour O. Gadalla, Sherif Edris, Ahmed M. Alzohairy, Nezar A. Radhwan, Mohammed N. Baeshen, Ahmed M. Ramadan, Hala F. Eissa, Sabah M. Hassan, Nabih A. Baeshen, Osama Abuzinadah, Magdy A. Al-Kordy, Fotouh M. El-Domyati, Robert K. Jansen

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Wild salt-tolerant barley (Hordeum spontaneum) is the ancestor of cultivated barley (Hordeum vulgare or H. vulgare). Although the cultivated barley genome is well studied, little is known about genome structure and function of its wild ancestor. In the present study, RNA-Seq analysis was performed on young leaves of wild barley treated with salt (500 mM NaCl) at four different time intervals. Transcriptome sequencing yielded 103 to 115 million reads for all replicates of each treatment, corresponding to over 10 billion nucleotides per sample. Of the total reads, between 74.8 and 80.3% could be mapped and 77.4 to 81.7% of the transcripts were found in the H. vulgare unigene database (unigene-mapped). The unmapped wild barley reads for all treatments and replicates were assembled de novo and the resulting contigs were used as a new reference genome. This resultedin94.3 to 95.3%oftheunmapped reads mapping to the new reference. The number of differentially expressed transcripts was 9277, 3861 of which were uni gene-mapped. The annotated unigene- and de novo-mapped transcripts (5100) were utilized to generate expression clusters across time of salt stress treatment. Two-dimensional hierarchical clustering classified differential expression profiles into nine expression clusters, four of which were selected for further analysis. Differentially expressed transcripts were assigned to the main functional categories. The most important groups were ‘response to external stimulus’ and ‘electron-carrier activity’. Highly expressed transcripts are involved in several biological processes, including electron transport and exchanger mechanisms, flavonoid biosynthesis, reactive oxygen species (ROS) scavenging, ethylene production, signaling network and protein refolding. The comparisons demonstrated that mRNA-Seq is an efficient method for the analysis of differentially expressed genes and biological processes under salt stress.

Keywords: electron transport, flavonoid biosynthesis, reactive oxygen species, rnaseq

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Authors: C. V. Nnamani, O. L. Adedeji

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The contemporary global economic meltdown has devastating effect on the Nigerian’s economy and its frantic search for alternative source of national revenue aside from oil and gas has become imperative for economic emancipation for Nigerians. Apicultural entrepreneurship could provide a source of livelihood if the basic knowledge of those plant genetic resources needed by bees is made available. A palynological evaluation of those palynotaxa which honey bees forage for pollen and nectar was carried out after standard acetolysis method. Results showed that the honey samples were highly diversified and rich in honey plants. A total of 9544.3 honey pollen, consisting of 39 honey plants belonging to 21 plant families and distributed within 38 genera were identified excluding 238 unidentified pollen grains. Data from the analysis equally revealed that Elaeis guineensis Jacq, Anacardium occidentale L, Diospyros mespiliformis Hochist xe ADC, Alchornea cordifolia Muell, Arg, Daniella oliveri (Rolfe) Hutch & Dalz, Irvingia wombolu Okafor ex Baill, Treculia africana Decne, Nauclea latifolia Smith and Crossopteryx febrifuga Afzil ex Benth were the predominant honey plants. It provided a guide to the optimal utilization of floral resources by honeybees in these regions, showing the opportunity and amazing potentials for apiculture entrepreneurship of these palytaxa. Most of these plants are rare, threatened and endangered. It calls for urgent conservation techniques and step by all players. Critical awareness creation to ensure farmers knowledge of these palynotaxa to ensure proper understanding and attendance boost from them as economic empowerment is needed.

Keywords: palynotaxa, acetolysis, enterprise, livelihood, Nigeria

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Authors: Perry Ayi Mankattah

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Introduction: Africa, which should lead the world in honey production, is importing three times the honey it produces even though it has a healthy, industrious and large population of bees. This is due to the mechanism of honey harvesting that destroys the combs and thereby reducing honey production and rate of harvesting. For Africa to take its place in the world of honey production, Africa should adopt a method that enables a higher rate of honey harvesting. The Advance Top Bar is, therefore, a simplified framework that provides that answer. It can be made of wood, plastic and metal that can be fabricated by tin/metal smiths, wielders and carpenters at the village level without any very sophisticated machines. Material and Methods: ATB is a top bar-like hollow framework of dimension 3.2*48 cm that can be made of wood, plastic and metal. It is made up of three parts of a constant hollow top bar, a variable grooved bottom bar with both bars being joined through synchronized holes (that align both the top and bottom bars ) by either metal or plastic rods of length 22cm and diameter of 5 mm with rounded balls at both ends It could be used with foundation combs or without and also other accessories to have about ten (10) function which includes commercial propolis harvesting queen rearing etc. The variable bottom bar length depends on the width of the hive, as most African beehives are somehow not standardized. Results: Foundation combs are placed within the Advance Top Bar for the bees to form their combs over its mesh to prevent comb breakage during honey harvesting. Similarly, honeycombs on top bars will produce natural foundation combs when also placed in the Advance top bar system just as they are re-used in the Langstroth Frames. Discussions and Conclusions: Any modification that will promote non-comb destruction during honey harvesting in Top bars shall cause Africa to increase honey production by over 100% as beekeepers adopt the mechanism. Honey-laden combs from the current normal top bars could be placed in the Advance Top Bar to harvest without comb destruction; hence the same system could be used as a transition to the adoption of the Advance Top Bar with less cost.

Keywords: honey, harvest, increase, production

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Authors: Yahaya Tajudeen, Joy Okpuzor, Tolu Ajayi

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The cell-rebuilding efficacy of four food plants eating as vegetables and spices in Nigeria was assessed in the lungs of wild rats (Rattus rattus) living in a polluted environment. The plants are roselle (Hibiscus sabdarrifa), moringa (Moringa oleifera), ginger (Zingiber officinale) and ugwu (Telfairia occidentalis). Sixty rats were caught from the vicinity of a cement factory in Sagamu, Southwestern-Nigeria and grouped into 6. The control group was administered distilled water, while the test groups were given ethanolic extracts of roselle, moringa, ginger, ugwu and the mixture of the extracts for 180 days. The histopathology of the rats was conducted before and at the end of 180 days extracts administration. Before administering the extracts, the lungs of the rats showed vascular congestion, severe fibrosis and congested alveolus; all which were also observed in the lungs of control rats at the end of the treatment. However, the lungs of rats that were treated with the extracts of the plants showed moderate, mild or no histological damage compared to the control rats. The extract of the mixture of the plants performed best, followed by ginger, ugwu and roselle, respectively. These findings suggest that the food plants contain phytonutrients and phytochemicals, which repaired damaged cells and tissues in the exposed rats. Consequently, the plants could play a role in ameliorating health effects of environmental pollution.

Keywords: food plants, wild rats, lung, histopathology, fibrosis, cell-rebuilding

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Authors: Dedy Pratama, Akhmadu Muradi, Hilman Ibrahim, Patrianef Darwis, Alexander Jayadi Utama, Raden Suhartono, D. Suryandari, Luluk Yunaini, Tom Ch Adriani

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Objective: Diabetic Foot Ulcer (DFU) is one of the complications of Type 2 Diabetes Mellitus (T2DM) that can lead to disability and death. Inadequate vascularization condition will affect healing process of DFU. Therefore, we investigated the expression of polymorphism TGF- β1 in the relation of the occurrence of DFU in T2DM. Methods: We designed a case-control study to investigate the polymorphism TGF- β1 gene 1800469 C > T and 1982073 C > T in T2DM in Cipto Mangunkusumo National Hospital (RSCM) Jakarta from June to December 2016. We used PCR techniques and compared the results in a group of T2DM patients with DFU as the case study and without DFU as the control group. Results: There were 203 patients, 102 patients with DFU and 101 patients control without DFU. 49,8% is male and 50,2% female with mean age about 56 years. Distribution of wild-type genotype TGF-B1 1800469 C > T wild type CC was found in 44,8%, the number of mutant heterozygote CT was 10,8% and mutant homozygote is 11,3%. Distribution of TGF-B1 1982073 C>T wild type CC was 32,5%, mutant heterozygote is 38,9% and mutant homozygote 25,1%. Conclusion: Distribution of alleles from TGF-B1 1800469 C > T is C 75% and T 25% and from TGF-B1 1982073 C > T is C53,8% and T 46,2%. In the other word polymorphism TGF- β1 plays a role in the occurrence and healing process of the DFU in T2DM patients.

Keywords: diabetic foot ulcers, diabetes mellitus, polymorphism, TGF-β1

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Authors: Rachel Thomas Tharmabalan

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The growing unease over the matter of food security in the world is the result of a maturing realization that the genetic base of most human caloric intake from plants is dangerously narrow. Malaysia’s tropical rainforests have the potential to contribute to diet diversification and provide a source of nutrient-rich food as the Orang Asli communities in Malaysia have relied almost entirely on the jungle for food, fodder, medicine and fuel antithetical to what is happening today. This segregation of the Orang Asli from traditional lands and resources leads to severe loss of knowledge of biodiversity. In order to preserve these wild edibles, four different types of vegetables that are frequently consumed by the Orang Asli which consists of Rebu, Meranti, Saya and Pama were selected. These vegetables were then analysed to determine its proximate and mineral content to help ascertain claims and reaffirm the impact it can play in ensuring food and nutrition security, in addition to combating chronic diseases. From the results obtained, the Meranti had the highest crude fiber, iron and calcium content. Other minerals such as potassium, magnesium and copper were also found in varying content. These wild edibles could also contribute to education and bring awareness to younger generations as well as urban populations to start consuming more of these in their daily life as it could prevent various chronic diseases in Malaysia.

Keywords: food and nutrition security, Orang Asli, underutilized plants, wild edible food systems

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Authors: Murad Aydin Şanda, Hasan Hüseyi̇n Doğan

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Twenty-two element contents were analyzed in five wild Agaricus and Agrocybe taxa [Agaricus bresadolanus Bohus, Agaricus essettei Bon, Agaricus xanthoderma Genev. Agrocybe paludosa (J.E. Lange) Kühner & Romagn. Ex Bon and Agrocybe praecox (Pers.) Fayod] from Marmara region of Turkey by ICP-AES equipment. The element uptake levels were observed at different amounts in each Agaricus and Agrocybe species. The highest Pb and P concentrations were determined as 16.74 and 1.501 mg.kg-1 in A. essettei and A.bresadolanus respectively. Ag, P, and Hg concentrations were determined as 30685, 1,501, and 5978 mg.kg-1 in A. bresadolanus respectively. A. essettei has highest Ni, Cu, and Mn concentrations as 37.1, 43.63 and 1476 mg.kg-1 respectively, whereas A. praecox has highest Mo, Ni and P as 0.54, 10.20 and 27.9 mg.kg-1 respectively. A. paludosa has highest Zn, Cd, and Ba concentration as 336.8, 2.26 and 571.5 mg.kg-1 respectively. The highest K concentration was found in A. xanthoderma with 5.31 mg.kg-1. According to Who and FAO critters, identified metals in Agaricus and Agrocybe genera are not harmful to People if they would be consumed.

Keywords: agaricus, element, macrofungi, Turkey

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Authors: Sumira Jan, Javid Iqbal Mir, Desh Beer Singh, Anil Sharma, Shafia Zaffar Faktoo

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Background and Aims: Climatic and edaphic factors immensely influence crop quality and proper development. Regardless of economic potential, Himalayan Oregano has not subjected to phytonutrient and chemometric evaluation and its relationship with environmental conditions are scarce. The central objective of this research was to investigate microclimatic variation among wild and cultivated populations located in a microclimatic gradient in north-western Himalaya, Kashmir and analyse if such disparity was related with diverse climatic and edaphic conditions. Methods: Micrometeorological, Atomic absorption spectroscopy for micro elemental analysis was carried for soil. HPLC was carried out to estimate variation in phytonutrients and phytochemicals. Results: Geographic variation in phytonutrient was observed among cultivated and wild populations and among populations diverse within regions. Cultivated populations exhibited comparatively lesser phytonutrient value than wild populations. Moreover, our results observed higher vegetative growth of O. vulgare L. with higher pH (6-7), elevated organic carbon (2.42%), high nitrogen (97.41Kg/ha) and manganese (10-12ppm) and zinc contents (0.39-0.50) produce higher phytonutrients. HPLC data of phytonutrients like quercetin, betacarotene, ascorbic acid, arbutin and catechin revealed direct relationship with UV-B flux (r2=0.82), potassium (r2=0.97) displaying parallel relationship with phytonutrient value. Conclusions: Catechin was found as predominant phytonutrient among all populations with maximum accumulation of 163.8 ppm while as quercetin exhibited lesser value. Maximum arbutin (53.42ppm) and quercetin (2.87ppm) accumulated in plants thriving under intense and high UV-B flux. Minimum variation was demonstrated by beta carotene and ascorbic acid.

Keywords: phytonutrient, ascorbic acid, beta carotene, quercetin, catechin

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Authors: Thilivhali Emmanuel Tshikalange, Darky Cheron Modishane, Frederick Tawi Tabit

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The fruit pulp extracts of twelve selected ethnobotanical wild edible fruits from Mutale local municipality in Venda (Limpopo Province, South Africa) were investigated for their antimicrobial, antioxidant and cytotoxicity activities. Methanol extracts were prepared and tested against six micro-organisms (Salmonella typhi, Streptococcus pyogenes, Bacillus cereus, Klebsiella pneumoniae, Prevotella intermedia and Candida albicans). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined using the micro-dilution method, while for antioxidant activity the 2,2-diphenyl-1-picrylhydrazyl method was used. Of the 12 extracts tested, Adonsonia digitata, Berchemia discolor, Manilkara mochisia, Xanthocercis zambesiaca, Landolphia kirkii and Garcinia livingstonei showed antimicrobial activity, with MIC values ranging from 12.5 to 0.4 mg/ml. Gram negative bacteria were more resistant to the extracts in comparison to Gram positive bacteria. Antioxidant activity was only detected in Adonsonia digitata extract and the IC50 (substrate concentration to produce 50% reduction) was found to be 16.18µg/ml. The cytotoxicity of the extracts that showed antimicrobial and antioxidant activities was also determined. All plant extracts tested were non-toxic against human kidney cells (HEK293), with IC50 values of >400 µg/ml. The results presented in this study provide support to some traditional uses of wild edible fruits.

Keywords: antimicrobial, antioxidant, cytotoxicity, ethnobotanical, fruits

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Authors: Nasser R. El-Brollosy, Roberta Loddo

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6-Benzyl-1-(ethoxymethyl)-5-isopropyluracil (Emivirine) and its corresponding 1-benzyloxymethyl analogue (TNK-651) showed high activity against HIV-1. The present study describes synthesis of novel emivirine analogues by reaction of chloromethyl ethyl ether with uracils having 5-ethyl / isopropyl and 6-(3,5-dimethoxybenzyl) substituents. A series of new TNK-651 analogues substituted at N-1 with phenoxyethoxymethyl moiety was prepared on treatment of the corresponding uracils with bis(phenoxyethoxy) methane. The newly synthesized non-nucleosides were tested for biological activity against wild type HIV-1 IIIB as well as the resistant strains N119 (Y181C), A17 (K103N + Y181C), and the triple mutant EFVR (K103R + V179D + P225H) in MT-4 cells. Some of the tested compounds showed good activities. Among them 6-(3,5-dimethylbenzyl)-5-ethyl-1-[2-(phenoxyethyl) oxymethyl]uracil which showed inhibitory potency higher than emivirine against both wild type HIV-1 and the tested mutant strains.

Keywords: Emivirine, HIV, non-nucleoside reverse transcriptase, uracils

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Authors: Erol Bayraktarov, Asya Ilgun, Thomas Schickl, Alexandre Campo, Nicolis Stamatios

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Traditional means of harvesting honey are often stressful for honeybees. Each time honey is collected a portion of the colony can die. In consequence, the colonies’ resilience to environmental stressors will decrease and this ultimately contributes to the global problem of honeybee colony losses. As part of the project HIVEOPOLIS, we design and build a different kind of beehive, incorporating technology to reduce negative impacts of beekeeping procedures, including honey harvesting. A first step in maintaining more sustainable honey harvesting practices is to design honey storage frames that can automate the honey collection procedures. This way, beekeepers save time, money, and labor by not having to open the hive and remove frames, and the honeybees' nest stays undisturbed.This system shows promising features, e.g., high reliability which could be a key advantage compared to current honey harvesting technologies.Our original concept of fractional honey harvesting has been to encourage the removal of honey only from "safe" locations and at levels that would leave the bees enough high-nutritional-value honey. In this abstract, we describe the current state of our honey harvester, its technology and areas to improve. The honey harvester works by separating the honeycomb cells away from the comb foundation; the movement and the elastic nature of honey supports this functionality. The honey sticks to the foundation, because of the surface tension forces amplified by the geometry. In the future, by monitoring the weight and therefore the capped honey cells on our honey harvester frames, we will be able to remove honey as soon as the weight measuring system reports that the comb is ready for harvesting. Higher viscosity honey or crystalized honey cause challenges in temperate locations when a smooth flow of honey is required. We use resistive heaters to soften the propolis and wax to unglue the moving parts during extraction. These heaters can also melt the honey slightly to the needed flow state. Precise control of these heaters allows us to operate the device for several purposes. We use ‘Nitinol’ springs that are activated by heat as an actuation method. Unlike conventional stepper or servo motors, which we also evaluated throughout development, the springs and heaters take up less space and reduce the overall system complexity. Honeybee acceptance was unknown until we actually inserted a device inside a hive. We not only observed bees walking on the artificial comb but also building wax, filling gaps with propolis and storing honey. This also shows that bees don’t mind living in spaces and hives built from 3D printed materials. We do not have data yet to prove that the plastic materials do not affect the chemical composition of the honey. We succeeded in automatically extracting stored honey from the device, demonstrating a useful extraction flow and overall effective operation this way.

Keywords: honey harvesting, honeybee, hiveopolis, nitinol

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Authors: Mayank, Navneet Kaur, Narinder Singh

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The V599 mutations in the BRAF protein are extremely oncogenic, responsible for countless of malignant conditions. Along with wild type, V599E, V599D, and V599R are the important mutated variants of the BRAF proteins. The BRAF inhibitory anticancer agents are continuously developing, and sorafenib is a BRAF inhibitor that is under clinical use. The crystal structure of sorafenib bounded to wild type, and V599 is known, showing a similar interaction pattern in both the case. The mutated 599th residue, in both the case, is also found not interacting directly with the co-crystallized sorafenib molecule. However, the IC50 value of sorafenib was found extremely different in both the case, i.e., 22 nmol/L for wild and 38 nmol/L for V599E protein. Molecular docking study and MMGBSA binding energy results also revealed a significant difference in the binding pattern of sorafenib in both the case. Therefore, to explore the role of distinctively situated 599th residue, we have further conducted comprehensive computational studies. The molecular dynamics simulation, residue interaction network (RIN) analysis, and residue correlation study results revealed the importance of the 599th residue on the therapeutic outcome and overall dynamic of the BRAF protein. Therefore, although the position of 599th residue is very much distinctive from the ligand-binding cavity of BRAF, still it has exceptional control over the overall functional outcome of the protein. The insight obtained here may seem extremely important and guide us while designing ideal BRAF inhibitory anticancer molecules.

Keywords: BRAF, oncogenic, sorafenib, computational studies

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Authors: Rachel A. Brant, Grace E. Kenny, Paige A. Muñiz, Gerardo R. Camilo

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The social sweat bee, Halictus ligatus, has been documented to alter its phenology as a response to changes in temporal dynamics of resources. Furthermore, H. ligatus exhibits polyethism in natural environments as a consequence of the variation in resources. Yet, we do not know if or how H. ligatus responds to these variations in urban environments. As urban environments become much more widespread, and human population is expected to reach nine billion by 2050, it is crucial to distinguish how resources are allocated by bees in cities. We hypothesize that in urban regions, where floral availability varies with human activity, H. ligatus will exhibit polyethism in order to match the extremely localized spatial variability of resources. We predict that in an urban setting, where resources vary both spatially and temporally, the phenology of H. ligatus will alter in response to these fluctuations. This study was conducted in Saint Louis, Missouri, at fifteen sites each varying in size and management type (community garden, urban farm, prairie restoration). Bees were collected by hand netting from 2013-2016. Results suggest that the largest individuals, mostly gynes, occurred in lower income neighborhood community gardens in May and August. We used a model averaging procedure, based on information theoretical methods, to determine a best model for predicting bee size. Our results suggest that month and locality within the city are the best predictors of bee size. Halictus ligatus was observed to comply with the predictions of polyethism from 2013 to 2015. However, in 2016 there was an almost complete absence of the smallest worker castes. This is a significant deviation from what is expected under polyethism. This could be attributed to shifts in planting decisions, shifts in plant-pollinator matches, or local climatic conditions. Further research is needed to determine if this divergence from polyethism is a new strategy for the social sweat bee as climate continues to alter or a response to human dominated landscapes.

Keywords: polyethism, urban environment, phenology, social sweat bee

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Authors: Jane E. Hill

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Many variables led to the changes in the insanity defense since the Wild Beast Standard of 1723 until the Federal Insanity Defense Reform Act of 1984. The insanity defense is used in criminal trials and argued that the defendant is ‘not guilty by reason of insanity’ because the individual was unable to distinguish right from wrong during the time they were breaking the law. The issue that surrounds whether or not to use the insanity defense in the criminal court depends on the mental state of the defendant at the time the criminal act was committed. This leads us to the question of did the defendant know right from wrong when they broke the law? In 1723, The Wild Beast Test stated that to be exempted from punishment the individual is totally deprived of their understanding and memory and doth not know what they are doing. The Wild Beast Test became the standard in England for over seventy-five years. In 1800, James Hadfield attempted to assassinate King George III. He only made the attempt because he was having delusional beliefs. The jury and the judge gave a verdict of not guilty. However, to legal confine him; the Criminal Lunatics Act was enacted. Individuals that were deemed as ‘criminal lunatics’ and were given a verdict of not guilty would be taken into custody and not be freed into society. In 1843, the M'Naghten test required that the individual did not know the quality or the wrongfulness of the offense at the time they committed the criminal act(s). Daniel M'Naghten was acquitted on grounds of insanity. The M'Naghten Test is still a modern concept of the insanity defense used in many courts today. The Irresistible Impulse Test was enacted in the United States in 1887. The Irresistible Impulse Test suggested that offenders that could not control their behavior while they were committing a criminal act were not deterrable by the criminal sanctions in place; therefore no purpose would be served by convicting the offender. Due to the criticisms of the latter two contentions, the federal District of Columbia Court of Appeals ruled in 1954 to adopt the ‘product test’ by Sir Isaac Ray for insanity. The Durham Rule also known as the ‘product test’, stated an individual is not criminally responsible if the unlawful act was the product of mental disease or defect. Therefore, the two questions that need to be asked and answered are (1) did the individual have a mental disease or defect at the time they broke the law? and (2) was the criminal act the product of their disease or defect? The Durham courts failed to clearly define ‘mental disease’ or ‘product.’ Therefore, trial courts had difficulty defining the meaning of the terms and the controversy continued until 1972 when the Durham rule was overturned in most places. Therefore, the American Law Institute combined the M'Naghten test with the irresistible impulse test and The United States Congress adopted an insanity test for the federal courts in 1984.

Keywords: insanity defense, psychology law, The Federal Insanity Defense Reform Act of 1984, The Wild Beast Standard in 1723

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Authors: Rahim Ada, Mustafa Harmankaya, Sadiye Ayse Celik

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The safflower seed contains about 25-40% solvent extract and 20-33% fiber. It is well known that dietary phospholipids lower serum cholesterol levels effectively. The nutrient composition of safflower seed changes depending on region, soil and genotypes. This research was made by using of six natural selected (A22, A29, A30, C12, E1, F4, G8, G12, J27) and three commercial (Remzibey, Dincer, Black Sun1) varieties of safflower genotypes. The research was conducted on field conditions for two years (2009 and 2010) in randomized complete block design with three replications in Konya-Turkey ecological conditions. Oil contents, mineral contents and their correlations were determined in the research. According to the results, oil content was ranged from 22.38% to 34.26%, while the minerals were in between the following values: 1469, 04-2068.07 mg kg^-1 for Ca, 7.24-11.71 mg kg^-1 for B, 13.29-17.41 mg kg^-1 for Cu, 51.00-79.35 mg kg^-1 for Fe, 3988-6638.34 mg kg^-1 for K, 1418.61-2306.06 mg kg^-1 for Mg, 11.37-17.76 mg kg^-1 for Mn, 4172.33-7059.58 mg kg^-1 for P and 32.60-59.00 mg kg^-1 for Zn. Correlation analysis that was made separately for the commercial varieties and wild lines showed that high level of oil content was negatively affected by all the investigated minerals except for K and Zn in the commercial varieties.

Keywords: safflower, oil, quality, mineral content

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Authors: Shagufta Gulraiz, Abu Saeed Hashmi, Muhammad Mohsin Javed

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Lysine required for poultry feed is imported in Pakistan to fulfil the desired dietary needs. Present study was designed to produce maximum lysine by utilizing cheap sources to save the foreign exchange. To achieve the goal of lysine production through fermentation, large scale production of lysine was carried out in 7.5 L stirred glass vessel fermenter with wild and mutant Brevibacterium flavum (B. flavum) using all pre-optimized conditions. The identification of produced lysine was carried out by TLC and amino acid analyzer. Toxicity evaluation of produced lysine was performed before feeding to broiler chicks. During biological trial concentrated fermented broth having 8% lysine was used in poultry rations as a source of Lysine for test birds. Fermenter scale studies showed that the maximum lysine (20.8 g/L) was produced at 250 rpm, 1.5 vvm aeration, 6.0% inoculum under controlled pH conditions after 56 h of fermentation with wild culture but mutant (BFENU2) gave maximum yield of lysine 36.3 g/L under optimized condition after 48 h. Amino acid profiling showed 1.826% Lysine in fermented broth by wild B. flavum and 2.644% by mutant strain (BFENU2). Toxicity evaluation report showed that the produced lysine is safe for consumption by broilers. Biological evaluation results showed that produced lysine was equally good as commercial lysine in terms of weight gain, feed intake and feed conversion ratio. A cheap and practical bioprocess of Lysine production was concluded, that can be exploited commercially in Pakistan to save foreign exchange.

Keywords: lysine, fermentation, broiler chicks, biological evaluation

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Authors: Neha Sahu, Awantika Singh, Brijesh Kumar, K. R. Arya

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Taraxacum officinale Weber or dandelion (Asteraceae) is an important Indian traditional herb used to treat liver detoxification, digestive problems, spleen, hepatic and kidney disorders, etc. The plant is well known to possess important phenolic and flavonoids to serve as a potential source of antioxidative and chemoprotective agents. Biosynthesis of bioactive compounds through in vitro cultures is a requisite for natural resource conservation and to provide an alternative source for pharmaceutical applications. Thus an efficient and reproducible protocol was developed for in vitro biosynthesis of bioactive antioxidative compounds from leaf derived callus and in vitro regenerated cultures of Taraxacum officinale using MS media fortified with various combinations of auxins and cytokinins. MS media containing 0.25 mg/l 2, 4-D (2, 4-Dichloro phenoxyacetic acid) with 0.05 mg/l 2-iP [N6-(2-Isopentenyl adenine)] was found as an effective combination for the establishment of callus with 92 % callus induction frequency. Moreover, 2.5 mg/l NAA (α-Naphthalene acetic acid) with 0.5 mg/l BAP (6-Benzyl aminopurine) and 1.5 mg/l NAA showed the optimal response for in vitro plant regeneration with 80 % regeneration frequency and rooting respectively. In vitro regenerated plantlets were further transferred to soil and acclimatized. Quantitative variability of accumulated bioactive compounds in cultures (in vitro callus, plantlets and acclimatized) were determined through UPLC-MS/MS (ultra-performance liquid chromatography-triple quadrupole-linear ion trap mass spectrometry) and compared with wild plants. The phytochemical determination of in vitro and wild grown samples showed the accumulation of 6 compounds. In in vitro callus cultures and regenerated plantlets, two major antioxidative compounds i.e. chlorogenic acid (14950.0 µg/g and 4086.67 µg/g) and umbelliferone (10400.00 µg/g and 2541.67 µg/g) were found respectively. Scopoletin was found to be highest in vitro regenerated plants (83.11 µg/g) as compared to wild plants (52.75 µg/g). Notably, scopoletin is not detected in callus and acclimatized plants, but quinic acid (6433.33 µg/g) and protocatechuic acid (92.33 µg/g) were accumulated at the highest level in acclimatized plants as compared to other samples. Wild grown plants contained highest content (948.33 µg/g) of flavonoid glycoside i.e. luteolin-7-O-glucoside. Our data suggests that in vitro callus and regenerated plants biosynthesized higher content of antioxidative compounds in controlled conditions when compared to wild grown plants. These standardized cultural conditions may be explored as a sustainable source of plant materials for enhanced production and adequate supply of oxidative polyphenols.

Keywords: anti-oxidative compounds, in vitro cultures, Taraxacum officinale, UPLC-MS/MS

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Authors: Andreea Halunga, Chris D. Orme, Takashi Yamagata

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This paper proposes a heteroskedasticity-robust Breusch-Pagan test of the null hypothesis of zero cross-section (or contemporaneous) correlation in linear panel-data models, without necessarily assuming independence of the cross-sections. The procedure allows for either fixed, strictly exogenous and/or lagged dependent regressor variables, as well as quite general forms of both non-normality and heteroskedasticity in the error distribution. The asymptotic validity of the test procedure is predicated on the number of time series observations, T, being large relative to the number of cross-section units, N, in that: (i) either N is fixed as T→∞; or, (ii) N²/T→0, as both T and N diverge, jointly, to infinity. Given this, it is not expected that asymptotic theory would provide an adequate guide to finite sample performance when T/N is "small". Because of this, we also propose and establish asymptotic validity of, a number of wild bootstrap schemes designed to provide improved inference when T/N is small. Across a variety of experimental designs, a Monte Carlo study suggests that the predictions from asymptotic theory do, in fact, provide a good guide to the finite sample behaviour of the test when T is large relative to N. However, when T and N are of similar orders of magnitude, discrepancies between the nominal and empirical significance levels occur as predicted by the first-order asymptotic analysis. On the other hand, for all the experimental designs, the proposed wild bootstrap approximations do improve agreement between nominal and empirical significance levels, when T/N is small, with a recursive-design wild bootstrap scheme performing best, in general, and providing quite close agreement between the nominal and empirical significance levels of the test even when T and N are of similar size. Moreover, in comparison with the wild bootstrap "version" of the original Breusch-Pagan test our experiments indicate that the corresponding version of the heteroskedasticity-robust Breusch-Pagan test appears reliable. As an illustration, the proposed tests are applied to a dynamic growth model for a panel of 20 OECD countries.

Keywords: cross-section correlation, time-series heteroskedasticity, dynamic panel data, heteroskedasticity robust Breusch-Pagan test

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Authors: Siyamthanda Makhwabe

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Tourism development and spatial planning within the broader spectrum of the Eastern Cape needs to be strategically integrated to give effectiveness to development planning within the province. Tourism was severely affected and limited by policies of the previous regime. Tourism development in the Eastern Cape has been identified as one of the underdeveloped sectors that have the potential to improve the province’s local economic development trajectory The proposed study reviews literature on tourism development in an urban/rural and regional context in the Eastern Cape province. The proposed study will therefore offer an in-depth literature review on issues pertaining to spatial planning, land use management policies and tourism development within the Eastern Cape using the scoping review method. The intention of the proposed study is to identify synergies between the intertwined municipalities within the Wild Coast region in order to create a tourism belt that would yield benefit from Coffee Bay to East London.

Keywords: development, Eastern Cape, policies, spatial planning, tourism

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Authors: Muhammad Naeem, Amina Zubari, Abdus Salam, Syed Ali Ayub Bukhari, Naveed Ahmad Khan

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Seventy three wild Labeo bata of different body sizes, ranging from 8.20-16.00 cm total length and 7.4-86.19 g body weight, were studied for the analysis of body composition parameters (Water content, ash content, fat content, protein content) in relation to body size and condition factor. Mean percentage is found as for water 77.71 %, ash 3.42 %, fat 2.20 % and protein content 16.65 % in whole wet body weight. Highly significant positive correlations were observed between condition factor and body weight (r = 0.243). Protein contents, organic content and ash (% wet body weight) increase with increasing percent water contents for Labeo bata while these constituents (% dry body weight) and fat contents (% wet and dry body weight) have no influence on percent water. It was observed that variations in the body constituents have no association to body weight or length.

Keywords: Labeo bata, body size, body composition, condition factor

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Authors: Michal Kielbik, Izabela Szulc-Kielbik, Anna Brzostek, Jaroslaw Dziadek, Magdalena Klink

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The goal of many research groups in the world is to find new components that are important for survival of mycobacteria in the host cells. Mycobacterium tuberculosis (Mtb) possesses a number of enzymes degrading cholesterol that are considered to be an important factor for its survival and persistence in host macrophages. One of them - cholesterol oxidase (ChoD), although not being essential for cholesterol degradation, is discussed as a virulence compound, however its involvement in macrophages’ response to Mtb is still not sufficiently determined. The recognition of tubercle bacilli antigens by pathogen recognition receptors is crucial for the initiation of the host innate immune response. An important receptor that has been implicated in the recognition and/or uptake of Mtb is Toll-like receptor type 2 (TLR2). Engagement of TLR2 results in the activation and phosphorylation of intracellular signaling proteins including IRAK-1 and -4, TRAF-6, which in turn leads to the activation of target kinases and transcription factors responsible for bactericidal and pro-inflammatory response of macrophages. The aim of these studies was a detailed clarification of the role of Mtb cholesterol oxidase as a virulence factor affecting the TLR2 signaling pathway in human macrophages. As human macrophages the THP-1 differentiated cells were applied. The virulent wild-type Mtb strain (H37Rv), its mutant lacking a functional copy of gene encoding cholesterol oxidase (∆choD), as well as complimented strain (∆choD–choD) were used. We tested the impact of Mtb strains on the expression of TLR2-depended signaling proteins (mRNA level, cytosolic level and phosphorylation status). The cytokine and bactericidal response of THP-1 derived macrophages infected with Mtb strains in relation to TLR2 signaling pathway dependence was also determined. We found that during the 24-hours of infection process the wild-type and complemented Mtb significantly reduced the cytosolic level and phosphorylation status of IRAK-4 and TRAF-6 proteins in macrophages, that was not observed in the case of ΔchoD mutant. Decreasement of TLR2-dependent signaling proteins, induced by wild-type Mtb, was not dependent on the activity of proteasome. Blocking of TLR2 expression, before infection, effectively prevented the induced by wild-type strain reduction of cytosolic level and phosphorylation of IRAK-4. None of the strains affected the surface expression of TLR2. The mRNA level of IRAK-4 and TRAF-6 genes were significantly increased in macrophages 24 hours post-infection with either of tested strains. However, the impact of wild-type Mtb strain on both examined genes was significantly stronger than its ΔchoD mutant. We also found that wild-type strain stimulated macrophages to release high amount of immunosuppressive IL-10, accompanied by low amount of pro-inflammatory IL-8 and bactericidal nitric oxide in comparison to mutant lacking cholesterol oxidase. The influence of wild-type Mtb on this type of macrophages' response strongly dependent on fully active IRAK-1 and IRAK-4 signaling proteins. In conclusion, Mtb using cholesterol oxidase causes the over-activation of TLR2 signaling proteins leading to the reduction of their cytosolic level and activity resulting in the modulation of macrophages response to allow its intracellular survival. Supported by grant: 2014/15/B/NZ6/01565, National Science Center, Poland

Keywords: Mycobacterium tuberculosis, cholesterol oxidase, macrophages, TLR2-dependent signaling pathway

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Authors: Hasan Hüseyin Doğan, Murad Aydın Şanda

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Element contents were analyzed in twelve wild Amanita taxa [A. caesarea (Scop.) Pers., A. citrina (Schaeff.) Pers., A. excelsa (Fr.) Bertill., A. franchetii (Boud.) Fayod, A. gemmata (Fr.) Bertill., A. mairei Foley, A. muscaria (L.) Lam., A. pantherina (DC.) Krombh., A. phalloides (Fr.) Link, A. rubescens Pers., A. vaginata (Bull.) Lam., and A. verna (Bull.) Lam.] from Marmara Region of Turkey by ICP-AES equipment. The element uptake levels were observed at different amounts in each Amanita species. The highest Pb and P concentrations were determined as 15.11 and 0.861 mg.kg-1 in A. caesarea. Fe, Co, As, Sr, Ca, Mg, Al and Na concentrations were determined as 0.832, 4.56, 15.6, 18.9, 0.44, 0.253 and 0.190 mg.kg-1 in A. gemmata respectively. A. muscaria has highest Mo, Th, Sb, V, Cr, and B concentrations as 1.45, 1.17, 1.06, 44, 75, and 7 mg.kg-1 respectively, whereas A. rubescens has highest Zn, Ba, K, S, and Se as 430.6, 65.7, 5.47, 1.16, 11.5 mg.kg-1 respectively. A. muscaria has highest Hg concentrations as 5855 µg.kg-1.The highest Mn concentration were found in A. pantherina with 1176 mg.kg-1, the highest Cd were found in A. phalloides as 10.77 mg.kg-1. In contrast to A. verna has highest Ag and Au content as 77728 and 192 µg.kg-1. Although A. citrina has only the highest Ni content as 75.9 mg.kg-1and A. vaginata has Cu content as 67.04 mg.kg-1 on the other hand A. phalloides has highest Cd concentrations as 10.77 mg.kg-1.

Keywords: amanita, element, macrofungi, Turkey

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Authors: Dorothy Kagazi

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The African elephant (Loxodonta africana) remains one of the most crop-damaging species around Kibale National Park, western Uganda. Elephant crop raiding deprives communities of food and incomes, consequently impacting livelihoods, attitude, and support for conservation. It also attracts an aggressive reaction from local communities including the retaliatory killing of a species that is already endangered and listed under Appendix I of the Convention on Endangered Species of Flora and Fauna (CITES). In order to mitigate against elephant crop raiding and minimize conflict, a number of interventions were devised by the government of Uganda such as physical guarding, scare-shooting, excavation of trenches, growing of unpalatable crops and fire lighting all of which have over the years been implemented around the park. These generated varying degrees of effectiveness but largely never solved the problem of elephants crossing into communities to destroy food and shelter which had a negative effect onto sustainable tourism of the communities who often resorted to killing these animals and hence contributing the falling numbers of these animals. It was until government discovered that there are far more effective ways of deterring these animals from crossing to communities that it commissioned a study to deploy the African honeybee (Apis mellifera scutellata) as a deterrent against elephant crop raiding and income enhancement for local people around the park. These efforts led to a number of projects around Kibale National Park where communities were facilitated to keep bees for human-elephant conflict mitigation and rural income enhancement through the sale of honey. These projects have registered tremendous success in reducing crop damage, enhance rural incomes, influence positive attitude change and ultimately secure community support for elephant and park conservation which is a clear manifestation of sustainable tourism development in the area. To address the issue of sustainability, the project was aligned with four major objectives that contributed to the overall goal of maintaining the areas around the parks and the national park itself in such a manner that it remains viable over an infinite period. Among these included determining deterrence effects of bees against elephant crop raiding, assessing the contribution of beekeeping towards rural income enhancement, determining the impact of community involvement of park conservation and management among others. The project deployed 500 improved hives by placing them at specific and previously identified and mapped out elephant crossing points along the park boundary. A control site was established without any intervention to facilitate comparison of findings and data was collected on elephant raiding frequency, patterns, honey harvested, and community attitude towards the park. A socio-economic assessment was also undertaken to ascertain the contribution of beekeeping to incomes and attitude change. In conclusion, human-wildlife conflicts have disturbed conservation and sustainable tourism development efforts. Such success stories like the beekeeping strategy should hence be extensively discussed and widely shared as a conservation technique for sustainable tourism.

Keywords: bees, communities, conservation, elephants

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Authors: Nausheen Irshad

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Human-wildlife conflict (HWC) exists in both developed and developing countries though it is more serious in developing nations. Knowledge of species ecology and species sensitivity to anthropogenic pressures is an important prerequisite for conservation/management. Therefore, three districts (Poonch, Bagh, and Muzaffarabad) of Azad Jammu and Kashmir were selected to highlight the wildlife hunting practices from January 2015 to November 2018. The study area was thoroughly explored to recover dead animals. Moreover, the local community was investigated (questionnaire survey) to catch on motives of killing. The results showed HWC mainly arises due to feeding habits of wild animals as some are frugivorous (small Indian civet and small Kashmir flying squirrel) who damaged human cultivated fruit trees. Besides, Indian crested porcupine and wild boar act as serious crop pests. The feeding upon domestic animals (common leopard) and poultry (Asiatic Jackal and Red fox) were also reported as factors of conflict. Hence numerous wild animals and birds (N=120) were found killed by natives in revenge. Despite protected status in Pakistan, the killed mammals belonged to categories of critically endangered (Panthera pardus) and near threatened (Viverricula indica) species. The important birds include critically endangered (Falco peregrines) and endangered (Lophura leucomelanos) species. It was found that mammals were primarily killed due to HWC (60%) followed by recreation (20%) and trade (15%) Whereas, the foremost hunting reasons for birds are recreation (50%), food (25%) and trade (25%). The drastic hunting/killing of the species needs our immediate attention. This unwarranted killing must be stopped forthwith otherwise these animals become extinct.

Keywords: Azad Jammu and Kashmir, anthropogenic pressures, endangered species, human-wildlife conflicts

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Authors: Chen Zhang, Fengxue Xin, Jianzhong He

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A unique Clostridium beijerinckii species strain BGS1 was obtained from grass land samples, which is capable of producing 8.43g/L butanol and 3.21 isopropanol from 60g/L glucose while generating 4.68g/L volatile fatty acids (VFAs) from 30g/L xylan. The concentration of isopropanol produced by culture BGS1 is ~15% higher than previously reported wild-type Clostridium beijerinckii under similar conditions. Compared to traditional Acetone-Butanol-Ethanol (ABE) fermentation species, culture BGS1 only generates negligible amount of ethanol and acetone, but produces butanol and isopropanol as biosolvent end-products which are pure alcohols and more economical than ABE. More importantly, culture BGS1 can consume acetone to produce isopropanol, e.g., 1.84g/L isopropanol from 0.81g/L acetone in 60g/L glucose medium containing 6.15g/L acetone. The analysis of BGS1 draft genome annotated by RAST server demonstrates that no ethanol production is caused by the lack of pyruvate decarboxylase gene – related to ethanol production. In addition, an alcohol dehydrogenase (adhe gene) was found in BGS1 which could be a potential gene responsible for isopropanol-generation. This is the first report on Isopropanol-Butanol (IB) fermentation by wild-type Clostridium strain and its application for isopropanol and butanol production.

Keywords: acetone conversion, butanol, clostridium, isopropanol

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Authors: Weaam Aldeeban, Majd Aljamali, Lama A. Youssef

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Background & Objective: Warfarin is considered a problematic drug due to its narrow therapeutic window and wide inter-individual response variations, which are attributed to demographic, environmental, and genetic factors, particularly single nucleotide polymorphism (SNPs) in the genes encoding VKORC1 and CYP2C9 involved in warfarin's mechanism of action and metabolism, respectively. CYP2C9*2rs1799853 and CYP2C9*3rs1057910 alleles are linked to reduced enzyme activity, as carriers of either or both alleles are classified as moderate or slow metabolizers, and therefore exhibit higher sensitivity of warfarin compared with wild type (CYP2C9*1*1). Our study aimed to assess the frequency of *1, *2, and *3 alleles in the CYP2C9 gene in a cohort of Syrian patients receiving a maintenance dose of warfarin for different indications, the impact of genotypes on warfarin dosing, and the frequency of adverse effects (i.e., bleedings). Subjects & Methods: This retrospective cohort study encompassed 94 patients treated with warfarin. Patients’ genotypes were identified by sequencing the polymerase chain reaction (PCR) specific products of the gene encoding CYP2C9, and the effects on warfarin therapeutic outcomes were investigated. Results: Sequencing revealed that 43.6% of the study population has the *2 and/or *3 SNPs. The mean weekly maintenance dose of warfarin was 37.42 ± 15.5 mg for patients with the wild-type allele (CYP2C9*1*1), whereas patients with one or both variants (*2 and/or *3) demanded a significantly lower dose (28.59 ±11.58 mg) of warfarin, (P= 0.015). A higher percentage (40.7%) of patients with allele *2 and/or *3 experienced hemorrhagic accidents compared with only 17.9% of patients with the wild type *1*1, (P = 0.04). Conclusions: Our study proves an association between *2 and *3 genotypes and higher sensitivity to warfarin and a tendency to bleed, which necessitates lowering the dose. These findings emphasize the significance of CYP2C9 genotyping prior to commencing warfarin therapy in order to achieve optimal and faster dose control and to ensure effectiveness and safety.

Keywords: warfarin, CYP2C9, polymorphisms, Syrian, hemorrhage

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Authors: Zewdneh Zana Zate

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The study focuses on the evolutionary and ecological genomics of both wild and cultivated Coffea arabica L. at its center of origin, Ethiopia, aiming to uncover how this vital species may withstand future climate changes. Utilizing bioclimatic models, we project the future distribution of Arabica under varied climate scenarios for 2050 and 2080, identifying potential conservation zones and immediate risk areas. Through whole-genome resequencing of accessions from Ethiopian gene banks, this research assesses genetic diversity and divergence between wild and cultivated populations. It explores relationships, demographic histories, and potential hybridization events among Coffea arabica accessions to better understand the species' origins and its connection to parental species. This genomic analysis also seeks to detect signs of natural or artificial selection across populations. Integrating these genomic discoveries with ecological data, the study evaluates the current and future ecological and genomic vulnerabilities of wild Coffea arabica, emphasizing necessary adaptations for survival. We have identified key genomic regions linked to environmental stress tolerance, which could be crucial for breeding more resilient Arabica varieties. Additionally, our ecological modeling predicted a contraction of suitable habitats, urging immediate conservation actions in identified key areas. This research not only elucidates the evolutionary history and adaptive strategies of Arabica but also informs conservation priorities and breeding strategies to enhance resilience to climate change. By synthesizing genomic and ecological insights, we provide a robust framework for developing effective management strategies aimed at sustaining Coffea arabica, a species of profound global importance, in its native habitat under evolving climatic conditions.

Keywords: coffea arabica, climate change adaptation, conservation strategies, genomic resilience

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Authors: Apurva Gupta, Surendra Singh

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Spirulina spp., as a promising source of many commercially valuable products, is grown photo autotrophically in open ponds and raceways on a large scale. However, the economic exploitation in an open system seems to have been limited because of lack of multiple stress-tolerant strains. The present study aims to isolate a stable stress tolerant mutant of Spirulina platensis with improved growth rate and enhanced potential to produce its commercially valuable bioactive compounds. N-methyl-n'-nitro-n-nitrosoguanidine (NTG) at 250 μg/mL (concentration permitted 1% survival) was employed for chemical mutagenesis to generate random mutants and screened against NaCl. In a preliminary experiment, wild type S. platensis was treated with NaCl concentrations from 0.5-1.5 M to calculate its LC₅₀. Mutagenized colonies were then screened for tolerance at 0.8 M NaCl (LC₅₀), and the surviving colonies were designated as NaCl tolerant mutants of S. platensis. The mutant cells exhibited 1.5 times improved growth against NaCl stress as compared to the wild type strain in control conditions. This might be due to the ability of the mutant cells to protect its metabolic machinery against inhibitory effects of salt stress. Salt stress is known to adversely affect the rate of photosynthesis in cyanobacteria by causing degradation of the pigments. Interestingly, the mutant cells were able to protect its photosynthetic machinery and exhibited 4.23 and 1.72 times enhanced accumulation of Chl a and phycobiliproteins, respectively, which resulted in enhanced rate of photosynthesis (2.43 times) and respiration (1.38 times) against salt stress. Phycocyanin production in mutant cells was observed to enhance by 1.63 fold. Nitrogen metabolism plays a vital role in conferring halotolerance to cyanobacterial cells by influx of nitrate and efflux of Na+ ions from the cell. The NaCl tolerant mutant cells took up 2.29 times more nitrate as compared to the wild type and efficiently reduce it. Nitrate reductase and nitrite reductase activity in the mutant cells also improved by 2.45 and 2.31 times, respectively against salt stress. From these preliminary results, it could be deduced that enhanced nitrogen uptake and its efficient reduction might be a reason for adaptive and halotolerant behavior of the S. platensis mutant cells. Also, the NaCl tolerant mutant of S. platensis with significant improved growth and phycocyanin accumulation compared to the wild type can be commercially promising.

Keywords: chemical mutagenesis, NaCl tolerant mutant, nitrogen metabolism, photosynthetic machinery, phycocyanin

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