Search results for: genus Pagrus
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 339

Search results for: genus Pagrus

69 Clostridium thermocellum DBT-IOC-C19, A Potential CBP Isolate for Ethanol Production

Authors: Nisha Singh, Munish Puri, Collin Barrow, Deepak Tuli, Anshu S. Mathur

Abstract:

The biological conversion of lignocellulosic biomass to ethanol is a promising strategy to solve the present global crisis of exhausting fossil fuels. The existing bioethanol production technologies have cost constraints due to the involvement of mandate pretreatment and extensive enzyme production steps. A unique process configuration known as consolidated bioprocessing (CBP) is believed to be a potential cost-effective process due to its efficient integration of enzyme production, saccharification, and fermentation into one step. Due to several favorable reasons like single step conversion, no need of adding exogenous enzymes and facilitated product recovery, CBP has gained the attention of researchers worldwide. However, there are several technical and economic barriers which need to be overcome for making consolidated bioprocessing a commercially viable process. Finding a natural candidate CBP organism is critically important and thermophilic anaerobes are preferred microorganisms. The thermophilic anaerobes that can represent CBP mainly belong to genus Clostridium, Caldicellulosiruptor, Thermoanaerobacter, Thermoanaero bacterium, and Geobacillus etc. Amongst them, Clostridium thermocellum has received increased attention as a high utility CBP candidate due to its highest growth rate on crystalline cellulose, the presence of highly efficient cellulosome system and ability to produce ethanol directly from cellulose. Recently with the availability of genetic and molecular tools aiding the metabolic engineering of Clostridium thermocellum have further facilitated the viability of commercial CBP process. With this view, we have specifically screened cellulolytic and xylanolytic thermophilic anaerobic ethanol producing bacteria, from unexplored hot spring/s in India. One of the isolates is a potential CBP organism identified as a new strain of Clostridium thermocellum. This strain has shown superior avicel and xylan degradation under unoptimized conditions compared to reported wild type strains of Clostridium thermocellum and produced more than 50 mM ethanol in 72 hours from 1 % avicel at 60°C. Besides, this strain shows good ethanol tolerance and growth on both hexose and pentose sugars. Hence, with further optimization this new strain could be developed as a potential CBP microbe.

Keywords: Clostridium thermocellum, consolidated bioprocessing, ethanol, thermophilic anaerobes

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68 Variability of the Arbuscular Mycorrhizal Fungi Communities Associated with Wild Agraz Plants (Vaccinium meridionale Swartz) in the Colombian Andes

Authors: Gabriel Roveda-Hoyos, Margarita Ramirez-Gomez, Adrian Perez, Diana Paola Serralde

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The objective of this study was to determine the variability of arbuscular mycorrhizal fungi (HFMA) communities associated with wild agraz plants (Vaccinium meridionale Swartz) in the Colombian Andes. This species is one of the most promising fruits within the genus Vaccinium because of the high content of anthocyanins and antioxidants in its fruits, and like other species of the Ericaceae family, it depends on the association with HFM for its development in the natural environment. In this study, the presence of mycorrhizae in wild communities of V. meridionale was evaluated, and their relationship with the edaphic and climatic conditions of the study area was analyzed. Sampling was conducted in the rural area of the municipalities of Raquira, and Chiquinquira, Chia, and Tabio in the departments of Cundinamarca and Boyaca, Colombia. Seven sites were selected, and in each site, 5 plants were randomly selected, root and soil samples were taken from each plant in the rhizosphere zone for the quantification of colonization and the presence of spores. The samples were collected on different soils, taxonomic orders Entisols, Inceptisols, and Alfisols, located at altitudes between 2,600 and 3,000 above sea level in the Eastern Cordillera of Colombia. The physicochemical characteristics of the soil were compared with the density of spores and the percentage of presence of mycorrhizae in the roots and variables with the morphometric and physiological characteristics of the plants. Four types of mutual associations were found: arbuscular mycorrhizae, ectendomycorrhiza, ericoid mycorrhizae, and endophytic septate fungi. The main results obtained show a predominance of spores of the genera Glomus and Acaulsopora, in most of the soils analyzed. The spore density of Glomeromycete fungi in the soil varied considerably between the different sites; it was higher ( > 50 spores/g of dry soil) in soil samples with lower bulk density and higher content of organic matter; in these soils a higher cation exchange capacity was found, as well as of nitrogen, calcium, magnesium, manganese and zinc concentration. It can be concluded that Vaccinium meridionale is able to establish in a natural way, association with HFMA.

Keywords: Ericaceae, Arbuscular mycorrhizae, Andes, soils, Glomus sp.

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67 Isolation, Identification and Measurement of Cottonseed Oil Gossypol in the Treatment of Drug-Resistant Cutaneous Leishmaniasis

Authors: Sara Taghdisi, Mehrosadat Mirmohammadi, Mostafa Mokhtarian, Mohammad Hossein Pazandeh

Abstract:

Leishmaniasis is one of the 10 most important diseases of the World Health Organization with health problems in more than 90 countries. Over one billion people are at risk of these diseases on almost every continent. The present human study was performed to evaluate the therapeutic effect of cotton plant on cutaneous leishmaniasis leision. firstly, the cotton seeds were cleaned and grinded to smaller particles. In the second step, the seeds were oiled by cold press method. In order to separate bioactive compound, after saponification of the oil, its gossypol was hydrolyzed and crystalized. finally, the therapeutic effect of Cottonseed Oil on cutaneous leishmaniasis was investigated. In the current project, Gossypol was extracted with a liquid-liquid extraction method in 120 minutes in the presence of Phosphoric acid from the cotton seed oil of Golestan beach varieties, then got crystallized in darkness using Acetic acid and isolated as Gossypol Acetic acid. The efficiency of the extracted crystal was obtained at 1.28±0.12. the cotton plant could be efficient in the treatment of Cutaneous leishmaniasis. This double-blind randomized controlled clinical trial was performed on 88 cases of leishmaniasis wounds. Patients were randomly divided into two groups of 44 cases. two groups received conventional treatment. In addition to the usual treatment (glucantime), the first group received cottonseed oil and the control group received placebo. The results of the present study showed that the surface of lesion before the intervention and in the first to fourth weeks after the intervention was not significantly different between the two groups (P-value> 0.05). But the surface of lesion in the Intervention group in the eighth and twelfth weeks was lower than the control group (P-value <0.05). This study showed that the improvement of leishmaniasis lesion using topical cotton plant mark in the eighth and twelfth weeks after the intervention was significantly more than the control group. Considering the most common chemical drugs for Cutaneous leishmaniasis treatment are sodium stibogluconate, and meglumine antimonate, which not only have relatively many side effects, but also some species of the Leishmania genus have become resistant to them. Therefore, a plant base bioactive compound such as cottonseed oil can be useful whit fewer side effects.

Keywords: cottonseed oil, crystallization, gossypol, leishmaniasis

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66 Population Dynamics of Cyprinid Fish Species (Mahseer: Tor Species) and Its Conservation in Yamuna River of Garhwal Region, India

Authors: Davendra Singh Malik

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India is one of the mega-biodiversity countries in the world and contributing about 11.72% of global fish diversity. The Yamuna river is the longest tributary of Ganga river ecosystem, providing a natural habitat for existing fish diversity of Himalayan region of Indian subcontinent. The several hydropower dams and barrages have been constructed on different locations of major rivers in Garhwal region. These dams have caused a major ecological threat to change existing fresh water ecosystems altering water flows, interrupting ecological connectivity, fragmenting habitats and native riverine fish species. Mahseer fishes (Indian carp) of the genus Tor, are large cyprinids endemic to continental Asia popularly known as ‘Game or sport fishes’ have continued to be decimated by fragmented natural habitats due to damming the water flow in riverine system and categorized as threatened fishes of India. The fresh water fish diversity as 24 fish species were recorded from Yamuna river. The present fish catch data has revealed that mahseer fishes (Tor tor and Tor putitora) were contributed about 32.5 %, 25.6 % and 18.2 % in upper, middle and lower riverine stretches of Yaumna river. The length range of mahseer (360-450mm) recorded as dominant size of catch composition. The CPUE (catch per unit effort) of mahseer fishes also indicated about a sharp decline of fish biomass, changing growth pattern, sex ratio and maturity stages of fishes. Only 12.5 – 14.8 % mahseer female brooders have showed only maturity phases in breeding months. The fecundity of mature mahseer female fish brooders ranged from 2500-4500 no. of ova during breeding months. The present status of mahseer fishery has attributed to the over exploitative nature in Yamuna river. The mahseer population is shrinking continuously in down streams of Yamuna river due to cumulative effects of various ecological stress. Mahseer conservation programme have implemented as 'in situ fish conservation' for enhancement of viable population size of mahseer species and restore the genetic loss of mahseer fish germplasm in Yamuna river of Garhwal Himalayan region.

Keywords: conservation practice, population dynamics, tor fish species, Yamuna River

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65 Bioactivities and Phytochemical Studies of Acrocarpus fraxinifolius Bark Wight and Arn

Authors: H. M. El-Rafie, A. H. Abou Zeid, R. S. Mohammed, A. A. Sleem

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Acrocarpus is a genus of flowering plants in the legume family Fabaceae which considered as a large and economically important family. This study aimed to investigate the phytoconstituents of the petroleum ether extract (PEE) of Acrocarpus fraxinofolius bark by Gas chromatography coupled with mass spectrometry (GC/MS) analysis of its fractions (fatty acid and unsaponifiable matter). Concerning this, identification of 52 compounds constituting 97.03 % of the total composition of the unsaponifiable matter fraction. Cycloeucalenol was found to be the major compound representing 32.52% followed by 4a, 14a-dimethyl-A8~24(28)-ergostadien (26.50%) and ß-sitosterol(13.74%), furthermore Gas liquid chromatography (GLC) analysis of the sterol fraction revealed the identification of cholesterol (7.22 %), campesterol (13.30 %), stigmasterol (10.00 %) and β - sitosterol (69.48 %). Meanwhile, the identification of 33 fatty acids representing 90.71% of the total fatty acid constituents. Methyl-9,12-octadecadienoate (40.39%) followed by methyl hexadecanoate (23.64%) were found to be the major compounds. On the other hand, column chromatography and Thin layer chromatography (TLC) fractionation of PEE separate the triterpenoid: 21β-hydroxylup-20(29)-en-3-one and β- amyrin which were structurally identified by spectroscopic analysis (NMR, MS and IR). PEE has been biologically evaluated for 1: management of diabetes in alloxan induced diabetic rats 2: cytotoxic activity against four human tumor cell lines (Cervix carcinoma cell line[HELA], Breast carcinoma cell line [MCF7], Liver carcinoma cell line[HEPG2] and Colon carcinoma cell line[HCT-116] 3: hepatoprotective activity against CCl4-induced hepatotoxicity in rats and the activity was studied by assaying the serum marker enzymes like AST, ALT, and ALP. Concerning this, the anti-diabetic activity exhibited by 100mg of PEE extract was 74.38% relative to metformin (100% potency). It also showed a significant anti-proliferative activity against MCF-7 (IC50= 2.35µg), Hela(IC50=3.85µg) and HEPG-2 (IC50= 9.54µg) compared with Doxorubicin as reference drug. The hepatoprotective activity was evidenced by significant decrease in liver function enzymes, i.e. AST, ALT and ALP by (29.18%, 28.26%, and 34.11%, respectively using silymarin as the reference drug, compared to their concentration levels in an untreated group with liver damage induced by CCl₄. This study was performed for the first time on the bark of this species.

Keywords: Acrocarpus fraxinofolius, antidiabetic, cytotoxic, hepatoprotective

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64 Reconsidering the Palaeo-Environmental Reconstruction of the Wet Zone of Sri Lanka: A Zooarchaeological Perspective

Authors: Kelum N. Manamendra-Arachchi, Kalangi Rodrigo

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Bones, teeth, and shells have been acknowledged over the last two centuries as evidence of chronology, Palaeo-environment, and human activity. Faunal traces are valid evidence of past situations because they have properties that have not changed over long periods of time. Sri Lanka has been known as an Island, which has a diverse variation of prehistoric occupation among ecological zones. Defining the Paleoecology of the past societies has been an archaeological thought developed in the 1960s. It is mainly concerned with the reconstruction from available geological and biological evidence of past biota, populations, communities, landscapes, environments, and ecosystems. Sri Lanka has dealt with this subject and considerable research has been already undertaken. The fossil and material record of Sri Lanka’s Wet Zone tropical forests continues from c. 38,000–34,000 ybp. This early and persistent human fossil, technical, and cultural florescence, as well as a collection of well-preserved tropical-forest rock shelters with associated ' on-site ' Palaeoenvironmental records, makes Sri Lanka a central and unusual case study to determine the extent and strength of early human tropical forest encounters. Excavations carried out in prehistoric caves in the low country wet zone has shown that in the last 50,000 years, the temperature in the lowland rainforests has not exceeded 5 degrees. Based on Semnopithecus Priam (Gray Langur) remains unearned from wet zone prehistoric caves, it has been argued that periods of momentous climate changes during the LGM and Terminal Pleistocene/Early Holocene boundary, with a recognizable preference for semi-open ‘Intermediate’ rainforest or edges. Continuous Genus Acavus and Oligospira occupation along with uninterrupted horizontal pervasive of Canarium sp. (‘kekuna’ nut) have proven that temperatures in the lowland rain forests have not changed by at least 5 oC over the last 50,000 years. Site Catchment or Territorial analysis cannot be no longer defensible, due to time-distance based factors as well as optimal foraging theory failed as a consequences of prehistoric people were aware of the decrease in cost-benefit ratio and located sites, and generally played out a settlement strategy that minimized the ratio of energy expanded to energy produced.

Keywords: palaeo-environment, prehistory, palaeo-ecology, zooarchaeology

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63 Response of Planktonic and Aggregated Bacterial Cells to Water Disinfection with Photodynamic Inactivation

Authors: Thayse Marques Passos, Brid Quilty, Mary Pryce

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The interest in developing alternative techniques to obtain safe water, free from pathogens and hazardous substances, is growing in recent times. The photodynamic inactivation of microorganisms (PDI) is a promising ecologically-friendly and multi-target approach for water disinfection. It uses visible light as an energy source combined with a photosensitiser (PS) to transfer energy/electrons to a substrate or molecular oxygen generating reactive oxygen species, which cause cidal effects towards cells. PDI has mainly been used in clinical studies and investigations on its application to disinfect water is relatively recent. The majority of studies use planktonic cells. However, in their natural environments, bacteria quite often do not occur as freely suspended cells (planktonic) but in cell aggregates that are either freely floating or attached to surfaces as biofilms. Microbes can form aggregates and biofilms as a strategy to protect them from environmental stress. As aggregates, bacteria have a better metabolic function, they communicate more efficiently, and they are more resistant to biocide compounds than their planktonic forms. Among the bacteria that are able to form aggregates are members of the genus Pseudomonas, they are a very diverse group widely distributed in the environment. Pseudomonas species can form aggregates/biofilms in water and can cause particular problems in water distribution systems. The aim of this study was to evaluate the effectiveness of photodynamic inactivation in killing a range of planktonic cells including Escherichia coli DSM 1103, Staphylococcus aureus DSM 799, Shigella sonnei DSM 5570, Salmonella enterica and Pseudomonas putida DSM 6125, and aggregating cells of Pseudomonas fluorescens DSM 50090, Pseudomonas aeruginosa PAO1. The experiments were performed in glass Petri dishes, containing the bacterial suspension and the photosensitiser, irradiated with a multi-LED (wavelengths 430nm and 660nm) for different time intervals. The responses of the cells were monitored using the pour plate technique and confocal microscopy. The study showed that bacteria belonging to Pseudomonads group tend to be more tolerant to PDI. While E. coli, S. aureus, S. sonnei and S. enterica required a dosage ranging from 39.47 J/cm2 to 59.21 J/cm2 for a 5 log reduction, Pseudomonads needed a dosage ranging from 78.94 to 118.42 J/cm2, a higher dose being required when the cells aggregated.

Keywords: bacterial aggregation, photoinactivation, Pseudomonads, water disinfection

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62 Optimization of Culture Conditions of Paecilomyces Tenuipes, Entomopathogenic Fungi Inoculated into the Silkworm Larva, Bombyx Mori

Authors: Sung-Hee Nam, Kwang-Gill Lee, You-Young Jo, HaeYong Kweon

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Entomopathogenic fungi is a Cordyceps species that is isolated from dead silkworm and cicada. Fungi on cicadas were described in old Chinese medicinal books and From ancient times, vegetable wasps and plant worms were widely known to have active substance and have been studied for pharmacological use. Among many fungi belonging to the genus Cordyceps, Cordyceps sinensis have been demonstrated to yield natural products possessing various biological activities and many bioactive components. Generally, It is commonly used to replenish the kidney and soothe the lung, and for the treatment of fatigue. Due to their commercial and economic importance, the demand for Cordyceps has been rapidly increased. However, a supply of Cordyceps specimen could not meet the increasing demand because of their sole dependence on field collection and habitat destruction. Because it is difficult to obtain many insect hosts in nature and the edibility of host insect needs to be verified in a pharmacological aspect. Recently, this setback was overcome that P. tenuipes was able to be cultivated in a large scale using silkworm as host. Pharmacological effects of P. tenuipes cultured on silkworm such as strengthening immune function, anti-fatigue, anti-tumor activity and controlling liver etc have been proved. They are widely commercialized. In this study, we attempted to establish a method for stable growth inhibition of P. tenuipes on silkworm hosts and an optimal condition for synnemata formation. To determine optimum culturing conditions, temperature and light conditions were varied. The length and number of synnemata was highest at 25℃ temperature and 100~300 lux illumination. On an average, the synnemata of wild P. tenuipes measures 70 ㎜ in length and 20 in number; those of the cultured strain were relatively shorter and more in number. The number of synnemata may have increased as a result of inoculating the host with highly concentrated conidia, while the length may have decreased due to limited nutrition per individual. It is not able that changes in light illumination cause morphological variations in the synnemata. However, regulation of only light and temperature could not produce stromata like perithecia, asci, and ascospores. Yamanaka reported that although a complete fruiting body can be produced under optimal culture conditions, it should be regarded as synnemata because it does not develop into an ascoma bearing ascospores.

Keywords: paecilomyces tenuipes, entomopathogenic fungi, silkworm larva, bombyx mori

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61 Determination of Identification and Antibiotic Resistance Rates of Serratia marcescens and Providencia Spp. from Various Clinical Specimens by Using Both the Conventional and Automated (VITEK2) Methods

Authors: Recep Keşli, Gülşah Aşık, Cengiz Demir, Onur Türkyılmaz

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Objective: Serratia species are identified as aerobic, motile Gram negative rods. The species Serratia marcescens (S. marcescens) causes both opportunistic and nosocomial infections. The genus Providencia is Gram-negative bacilli and includes urease-producing that is responsible for a wide range of human infections. Although most Providencia infections involve the urinary tract, they are also associated with gastroenteritis, wound infections, and bacteremia. The aim of this study was evaluate the antimicrobial resistance rates of S. marcescens and Providencia spp. strains which had been isolated from various clinical materials obtained from different patients who belongs to intensive care units (ICU) and inpatient clinics. Methods: A total of 35 S. marcescens and Providencia spp. strains isolated from various clinical samples admitted to Medical Microbiology Laboratory, ANS Research and Practice Hospital, Afyon Kocatepe University between October 2013 and September 2015 were included in the study. Identification of the bacteria was determined by conventional methods and VITEK 2 system (bio-Merieux, Marcy l’etoile, France) was used additionally. Antibacterial resistance tests were performed by using Kirby Bauer disc (Oxoid, Hampshire, England) diffusion method following the recommendations of CLSI. Results: The distribution of clinical samples were as follows: upper and lower respiratory tract samples 26, 74.2 % wound specimen 6, 17.1 % blood cultures 3, 8.5%. Of the 35 S. marcescens and Providencia spp. strains; 28, 80% were isolated from clinical samples sent from ICU. The resistance rates of S. marcescens strains against trimethoprim-sulfamethoxazole, piperacillin-tazobactam, imipenem, gentamicin, ciprofloxacin, ceftazidime, cefepime and amikacin were found to be 8.5 %, 22.8 %, 11.4 %, 2.8 %, 17.1 %, 40 %, 28.5 % and 5.7 % respectively. Resistance rates of Providencia spp. strains against trimethoprim-sulfamethoxazole, piperacillin-tazobactam, imipenem, gentamicin, ciprofloxacin, ceftazidime, cefepime and amikacin were found to be 10.2 %, 33,3 %, 18.7 %, 8.7 %, 13.2 %, 38.6 %, 26.7%, and 11.8 % respectively. Conclusion: S. marcescens is usually resistant to ampicillin, amoxicillin, amoxicillin/clavulanate, ampicillin/sulbactam, cefuroxime, cephamycins, nitrofurantoin, and colistin. The most effective antibiotic on the total of S. marcescens strains was found to be gentamicin 2.8 %, of the totally tested strains the highest resistance rate found against to ceftazidime 40 %. The lowest and highest resistance rates were found against gentamiycin and ceftazidime with the rates of 8.7 % and 38.6 % for Providencia spp.

Keywords: Serratia marcescens, Providencia spp., antibiotic resistance, intensive care unit

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60 Chemical Composition of Volatiles Emitted from Ziziphus jujuba Miller Collected during Different Growth Stages

Authors: Rose Vanessa Bandeira Reidel, Bernardo Melai, Pier Luigi Cioni, Luisa Pistelli

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Ziziphus jujuba Miller is a common species of the Ziziphus genus (Rhamnaceae family) native to the tropics and subtropics known for its edible fruits, fresh consumed or used in healthy food, as flavoring and sweetener. Many phytochemicals and biological activities are described for this species. In this work, the aroma profiles emitted in vivo by whole fresh organs (leaf, bud flower, flower, green and red fruits) were analyzed separately by mean of solid phase micro-extraction (SPME) coupled with gas chromatography mass spectrometry (GC-MS). The emitted volatiles from different plant parts were analysed using Supelco SPME device coated with polydimethylsiloxane (PDMS, 100µm). Fresh plant material was introduced separately into a glass conical flask and allowed to equilibrate for 20 min. After the equilibration time, the fibre was exposed to the headspace for 15 min at room temperature, the fibre was re-inserted into the needle and transferred to the injector of the CG and CG-MS system, where the fibre was desorbed. All the data were submitted to multivariate statistical analysis, evidencing many differences amongst the selected plant parts and their developmental stages. A total of 144 compounds were identified corresponding to 94.6-99.4% of the whole aroma profile of jujube samples. Sesquiterpene hydrocarbons were the main chemical class of compounds in leaves also present in similar percentage in flowers and bud flowers where (E, E)-α-farnesene was the main constituent in all cited plant parts. This behavior can be due to a protection mechanism against pathogens and herbivores as well as resistance to abiotic factors. The aroma of green fruits was characterized by high amount of perillene while the red fruits release a volatile blend mainly constituted by different monoterpenes. The terpenoid emission of flesh fruits has important function in the interaction with animals including attraction of seed dispersers and it is related to a good quality of fruits. This study provides for the first time the chemical composition of the volatile emission from different Ziziphus jujuba organs. The SPME analyses of the collected samples showed different patterns of emission and can contribute to understand their ecological interactions and fruit production management.

Keywords: Rhamnaceae, aroma profile, jujube organs, HS-SPME, GC-MS

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59 Investigation of Azol Resistance in Aspergillosis Caused by Gradient Test and Agar Plaque Methods

Authors: Zeynep Yazgan, Gökhan Aygün, Reyhan Çalışkan

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Objective: Invasive fungal infections are a serious threat in terms of morbidity and mortality, especially in immunocompromised patients. The most frequently isolated agents are Aspergillus genus fungi, and sensitivity to azoles, which are the first choice in treatment, decreases. In our study, we aimed to investigate the use of the agar plate screening method as a fast, easy, and practical method in determining azole resistance in Aspergillus spp. species. Methods: Our study was conducted with 125 Aspergillus spp. isolates produced from various clinical samples. Aspergillus spp. isolates were identified by conventional methods and azole resistance was determined by gradient test and agar plate screening method. Broth microdilution method was applied to resistant isolates, and CypA-L98H and CypA-M220 mutations in the cyp51A gene were investigated. Results: In our study, 55 A. fumigatus complex (44%), 42 A. flavus (33.6%), 6 A. terreus (5%), 4 A. niger (3%) and 18 Aspergillus spp. (14%) were identified. With the gradient test method, resistance to VOR and POS was detected in 1 (1.8%) of A.fumigatus isolates, and resistance to ITR was detected in 3 (5.45%). With the agar plate method, 1 of the A.fumigatus isolates (1.8%) had VOR, ITR, POS, 1 of the A.terreus isolates (16.7%) had VOR, 1 of the A.niger isolates (25%) had ITR. Resistance to VOR and POS was detected in 2 Aspergillus spp. isolates (11%), and resistance to ITR was detected in 1 (5.6%). Sensitivity and specificity were determined as 100% for VOR and POS in A. fumigatus species, 33.3% and 100% for ITR, respectively, 100% for ITR in A. flavus species, and 100% for ITR and POS in A. terreus species. By broth microdilution method in 7 isolates in which resistance was detected by gradient test and/or agar plate screening method; 1 A.fumigatus resistant to ITR, VOR, POS, 2 A.fumigatus resistant to ITR, 2 Aspergillus spp. ITR, VOR, POS MICs were determined as 2µg/ml and 8µg/ml, 8µg/ml and >32µg/ml, 0.5µg/ml and 4µg/ml, respectively. CypA-L98H mutations were detected in 5 of these isolates, CypA-M220 mutations were detected in 6, and no mutation was detected in 1. CypA-L98H and CypA-M220 mutations were detected in 1 isolate for which resistance was not detected. Conclusion: The need for rapid antifungal susceptibility screening tests is increasing in the treatment of aspergillosis. Although the sensitivity of the agar plate method was determined to be 33.3% for A.fumigatus ITR in our study, its sensitivity and specificity were determined to be 100% for ITR, VOR, and POS in other species. The low sensitivity value detected for A.fumigatus showed that agar plate drug concentrations should be updated in accordance with the latest regulations of EUCAST guidelines. The CypA-L98H and CypA-M220 mutations detected in our study suggested that the distribution of azole resistance-related mutations in different regions in our country should be investigated. In conclusion, it is thought that the agar plate method, which can be easily applied to detect azole resistance, is a fast and practical method in routine use and can contribute to both the determination of effective treatment strategies and the generation of epidemiological data.

Keywords: Aspergillus, agar plate, azole resistance, cyp51A, cypA-L98H, cypA-M220

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58 Spatial Variability of Phyotoplankton Assemblages during the Intermonsoon in Baler Bay, Outer and Inner Casiguran Sound, Aurora, Fronting Philipine Rise

Authors: Aime P. Lampad-Dela Pena, Rhodora V. Azanza, Cesar L. Villanoy, Ephrime B. Metillo, Aletta T. Yniguez

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Phytoplankton community changes in relation to environmental parameters were compared between and within, the three interconnected basins. Phytoplankton samples were collected from thirteen stations of Baler Bay and Casiguran Sound, Aurora last May 2013 by filtering 10 L buckets of surface water and 5 L Niskin samples at 20 meters and at 30 to 40 meters depths through a 20um sieve. Duplicate samples per station were preserved, counted, and identified up to genus level, in order to determine the horizontal and vertical spatial variation of different phytoplankton functional groups during the summer ebb and flood flow. Baler Bay, Outer and Inner Casiguran Sound had a total of 89 genera from four phytoplankton groups: Diatom (62), Dinoflagellate (25), Silicoflagellate (1) and Cyanobacteria (1). Non-toxic diatom Chaetoceros spp. bloom (averaged 2.0 x 105 to 2.73 x 106 cells L⁻¹) co-existed with Bacteriastrum spp. at surface waters in Inner and Outer Casiguran. Pseudonitzschia spp. (1.73 x 106 cells L⁻¹) bloomed at bottom waters of the innermost embayment near Casiguran mangrove estuary. Cyanobacteria Trichodesmium spp. significantly increased during ebb tide at the mid-water layers (20 meters depth) in the three basins (ranged from 6, 900 to 15, 125 filaments L⁻¹), forming another bloom. Gonyaulax spp. - dominated dinoflagellate did not significantly change with depth across the three basins. Overall, diatoms and dinoflagellates community assemblages significantly changed between sites (p < 0.001) while diatoms and cyanobacteria varied within Casiguran outer and inner sites (p < 0.001) only. Tidal fluctuations significantly affected dinoflagellates and diatom groups (p < 0.001) in inner and baler sites. Chlorophyll significantly varied between (KW, p < 0.001) and within each basins (KW, p < 0.05), no tidal influence, with the highest value at inner Casiguran and at deeper waters indicating deep chlorophyll maxima. Aurora’s distinct shelf morphology favoring counterclockwise circulation pattern, advective transport, and continuous stratification of the water column could basically affect the phytoplankton assemblages and water quality of Baler Bay and Casiguran inner and outer basins. Observed spatial phytoplankton community changes with multi-species diatom and cyanobacteria bloom at different water layers of the three inter-connected embayments would be vital for any environmental management initiatives in Aurora.

Keywords: aurora fronting Philippines Rise, intermonsoon, multi-species diatom bloom, spatial variability

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57 Automatic Identification of Aquatic Insects Based on Deep Learning and Computer Vision

Authors: Predrag Simović, Katarina Stojanović, Milena Radenković, Dimitrija Savić Zdravković, Aleksandar Milosavljević, Bratislav Predić, Milenka Božanić, Ana Petrović, Djuradj Milošević

Abstract:

Mayflies (Ephemeroptera), stoneflies (Plecoptera), and caddisflies (Trichoptera) (collectively referred to as EPT) are key participants in most freshwater habitats and often exhibit high diversity. Moreover, their presence and relative abundance are used in freshwater ecological and biomonitoring studies. Current methods for freshwater ecosystem biomonitoring follow a traditional approach of taxa monitoring based on morphological characters, which is time-consuming, and often generates data sets with low taxonomic resolution and unverifiable identification precision. To assist in solving identification problems and contribute to the knowledge of the distribution of many species, there was a need to develop alternative approaches in macroinvertebrate sample identification. Here, we establish an automatic machine-based identification approach for EPT taxa (Insect) using deep Convolutional Neural Networks (CNNs) and computer vision to increase the efficiency and taxonomic resolution in biomonitoring. The 5 550 specimens were collected from freshwater ecosystems of Serbia, and the deep model was built upon 90 EPT taxa. The protocol for obtaining images included the following stages: taxonomic identification by human experts and DNA barcoding validation, mounting the larvae, and photographing the dorsal side using a stereomicroscope and camera (16 650 individuals). The most informative image regions (the dorsal segments of individuals) for the decision-making process in the deep learning model were visualized using Gradient Weighted Class Activation Mapping (Grad-CAM). After training the artificial neural network, a CNN model was then built that was able to classify the 90 EPT taxa into their respective taxonomic categories automatically with 98.7%. Our approach offers a straightforward and efficient solution for routine monitoring programs, focusing on key biotic descriptors, such as EPT taxa. In addition, this application provides a streamlined solution that not only saves time, reduces equipment and expert requirements but also significantly enhances reliability and information content. The identification of the EPT larvae is difficult because of the variation of morphological features even within a single genus or the close resemblance of several species, and therefore, future research should focus on increasing the number of entities (species) in the model.

Keywords: convolutional neural networks, DNA barcoding, EPT taxa, biomonitoring

Procedia PDF Downloads 51
56 Adaptative Metabolism of Lactic Acid Bacteria during Brewers' Spent Grain Fermentation

Authors: M. Acin-Albiac, P. Filannino, R. Coda, Carlo G. Rizzello, M. Gobbetti, R. Di Cagno

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Demand for smart management of large amounts of agro-food by-products has become an area of major environmental and economic importance worldwide. Brewers' spent grain (BSG), the most abundant by-product generated in the beer-brewing process, represents an example of valuable raw material and source of health-promoting compounds. To the date, the valorization of BSG as a food ingredient has been limited due to poor technological and sensory properties. Tailored bioprocessing through lactic acid bacteria (LAB) fermentation is a versatile and sustainable means for the exploitation of food industry by-products. Indigestible carbohydrates (e.g., hemicelluloses and celluloses), high phenolic content, and mostly lignin make of BSG a hostile environment for microbial survival. Hence, the selection of tailored starters is required for successful fermentation. Our study investigated the metabolic strategies of Leuconostoc pseudomesenteroides and Lactobacillus plantarum strains to exploit BSG as a food ingredient. Two distinctive BSG samples from different breweries (Italian IT- and Finish FL-BSG) were microbially and chemically characterized. Growth kinetics, organic acid profiles, and the evolution of phenolic profiles during the fermentation in two BSG model media were determined. The results were further complemented with gene expression targeting genes involved in the degradation cellulose, hemicelluloses building blocks, and the metabolism of anti-nutritional factors. Overall, the results were LAB genus dependent showing distinctive metabolic capabilities. Leuc. pseudomesenteroides DSM 20193 may degrade BSG xylans while sucrose metabolism could be furtherly exploited for extracellular polymeric substances (EPS) production to enhance BSG pro-technological properties. Although L. plantarum strains may follow the same metabolic strategies during BSG fermentation, the mode of action to pursue such strategies was strain-dependent. L. plantarum PU1 showed a great preference for β-galactans compared to strain WCFS1, while the preference for arabinose occurred at different metabolic phases. Phenolic compounds profiling highlighted a novel metabolic route for lignin metabolism. These findings will allow an improvement of understanding of how lactic acid bacteria transform BSG into economically valuable food ingredients.

Keywords: brewery by-product valorization, metabolism of plant phenolics, metabolism of lactic acid bacteria, gene expression

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55 Comparision of Neutrophil Response to Curvularia, Bipolaris and Aspergillus Species

Authors: Eszter J. Tóth, Alexandra Hoffmann, Csaba Vágvölgyi, Tamás Papp

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Members of the genera Curvularia and Bipolaris are closely related melanin producing filamentous fungi; both of them have the teleomorph states in genus Cochliobolus. While Bipolaris species infect only plants and may cause serious agriculture damages, some Curvularia species was recovered from opportunistic human infections. The human pathogenic species typically cause phaeohyphomycoses, i.e. mould infections caused by melanised fungi, which can manifest as invasive mycoses with frequent involvement of the central nervous system in immunocompromised patients or as local infections (e.g. keratitis, sinusitis, and cutaneous lesions) in immunocompetent people. Although their plant-fungal interactions have been intensively studied, there is only little information available about the human pathogenic feature of these fungi. The aim of this study was to investigate the neutrophil granulocytes’ response to hyphal forms of Curvularia and Bipolaris in comparison with the response to Aspergillus. In the present study Curvularia lunata SZMC 23759 and Aspergillus fumigatus SZMC 23245 both isolated from human eye infection, and Bipolaris zeicola BRIP 19582b isolated from plant leaf were examined. Neutrophils were isolated from heparinised venous blood of healthy donors with dextran sedimentation followed by centrifugation over Ficoll and hypotonic lysis of erythrocytes. Viability and purity of the cells were checked with trypan blue and Wright staining, respectively. Infection of neutrophils was carried out with germinated conidia in a ratio of 5:1. Production of hydrogen peroxide, superoxide anion, and nitrogen monoxide was measured both intracellularly and extracellularly in response to the germinated spores with or without the supernatant and after serum treatment. ROS and NOS production of neutrophils in interaction with the three fungi were compared. It is already known that Aspergillus species induce ROS production of neutrophils only after serum treatment. Although, in case of C. lunata, serum opsonisation also induced an intensive production of reactive species, lower level of production was measured in the lack of serum as well. After interaction with the plant pathogenic B. zeicola, amount of reactive species found to be similar with and without serum treatment. The presence of germination supernatant decreased the reactive species production in case of each fungus. Interaction with Curvularia, Bipolaris and Aspergillus species induced different response of neutrophils. It seems that recognition of C. lunata and B. zeicola is independent of serum opsonisation, albeit it increases the level of the produced reactive species in response for C. lunata. The study was supported by the grant LP2016-8/2016.

Keywords: Curvularia, neutrophils, NOS, ROS, serum opsonisation

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54 Evaluation of Antarctic Bacteria as Potential Producers of Cellulolytic Enzymes of Industrial Interest

Authors: Claudio Lamilla, Andrés Santos, Vicente Llanquinao, Jocelyn Hermosilla, Leticia Barrientos

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The industry in general is very interested in improving and optimizing industrial processes in order to reduce the costs involved in obtaining raw materials and production. Thus, an interesting and cost-effective alternative is the incorporation of bioactive metabolites in such processes, being an example of this enzymes which catalyze efficiently a large number of enzymatic reactions of industrial and biotechnological interest. In the search for new sources of these active metabolites, Antarctica is one of the least explored places on our planet where the most drastic cold conditions, salinity, UVA-UVB and liquid water available are present, features that have shaped all life in this very harsh environment, especially bacteria that live in different Antarctic ecosystems, which have had to develop different strategies to adapt to these conditions, producing unique biochemical strategies. In this work the production of cellulolytic enzymes of seven bacterial strains isolated from marine sediments at different sites in the Antarctic was evaluated. Isolation of the strains was performed using serial dilutions in the culture medium at M115°C. The identification of the strains was performed using universal primers (27F and 1492R). The enzyme activity assays were performed on R2A medium, carboxy methyl cellulose (CMC)was added as substrate. Degradation of the substrate was revealed by adding Lugol. The results show that four of the tested strains produce enzymes which degrade CMC substrate. The molecular identifications, showed that these bacteria belong to the genus Streptomyces and Pseudoalteromonas, being Streptomyces strain who showed the highest activity. Only some bacteria in marine sediments have the ability to produce these enzymes, perhaps due to their greater adaptability to degrade at temperatures bordering zero degrees Celsius, some algae that are abundant in this environment and have cellulose as the main structure. The discovery of new enzymes adapted to cold is of great industrial interest, especially for paper, textiles, detergents, biofuels, food and agriculture. These enzymes represent 8% of industrial demand worldwide and is expected to increase their demand in the coming years. Mainly in the paper and food industry are required in extraction processes starch, protein and juices, as well as the animal feed industry where treating vegetables and grains helps improve the nutritional value of the food, all this clearly puts Antarctic microorganisms and their enzymes specifically as a potential contribution to industry and the novel biotechnological applications.

Keywords: antarctic, bacteria, biotechnological, cellulolytic enzymes

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53 Isolation and Selection of Strains Perspective for Sewage Sludge Processing

Authors: A. Zh. Aupova, A. Ulankyzy, A. Sarsenova, A. Kussayin, Sh. Turarbek, N. Moldagulova, A. Kurmanbayev

Abstract:

One of the methods of organic waste bioconversion into environmentally-friendly fertilizer is composting. Microorganisms that produce hydrolytic enzymes play a significant role in accelerating the process of organic waste composting. We studied the enzymatic potential (amylase, protease, cellulase, lipase, urease activity) of bacteria isolated from the sewage sludge of Nur-Sultan, Rudny, and Fort-Shevchenko cities, the dacha soil of Nur-Sultan city, and freshly cut grass from the dacha for processing organic waste and identifying active strains. Microorganism isolation was carried out by the cultures enrichment method on liquid nutrient media, followed by inoculating on different solid media to isolate individual colonies. As a result, sixty-one microorganisms were isolated, three of which were thermophiles (DS1, DS2, and DS3). The highest number of isolates, twenty-one and eighteen, were isolated from sewage sludge of Nur-Sultan and Rudny cities, respectively. Ten isolates were isolated from the wastewater of the sewage treatment plant in Fort-Shevchenko. From the dacha soil of Nur-Sultan city and freshly cut grass - 9 and 5 isolates were revealed, respectively. The lipolytic, proteolytic, amylolytic, cellulolytic, ureolytic, and oil-oxidizing activities of isolates were studied. According to the results of experiments, starch hydrolysis (amylolytic activity) was found in 2 isolates - CB2/2, and CB2/1. Three isolates - CB2, CB2/1, and CB1/1 were selected for the highest ability to break down casein. Among isolated 61 bacterial cultures, three isolates could break down fats - CB3, CBG1/1, and IL3. Seven strains had cellulolytic activity - DS1, DS2, IL3, IL5, P2, P5, and P3. Six isolates rapidly decomposed urea. Isolate P1 could break down casein and cellulose. Isolate DS3 was a thermophile and had cellulolytic activity. Thus, based on the conducted studies, 15 isolates were selected as a potential for sewage sludge composting - CB2, CB3, CB1/1, CB2/2, CBG1/1, CB2/1, DS1, DS2, DS3, IL3, IL5, P1, P2, P5, P3. Selected strains were identified on a mass spectrometer (Maldi-TOF). The isolate - CB 3 was referred to the genus Rhodococcus rhodochrous; two isolates CB2 and CB1 / 1 - to Bacillus cereus, CB 2/2 - to Cryseobacterium arachidis, CBG 1/1 - to Pseudoxanthomonas sp., CB2/1 - to Bacillus megaterium, DS1 - to Pediococcus acidilactici, DS2 - to Paenibacillus residui, DS3 - to Brevibacillus invocatus, three strains IL3, P5, P3 - to Enterobacter cloacae, two strains IL5, P2 - to Ochrobactrum intermedium, and P1 - Bacillus lichenoformis. Hence, 60 isolates were isolated from the wastewater of the cities of Nur-Sultan, Rudny, Fort-Shevchenko, the dacha soil of Nur-Sultan city, and freshly cut grass from the dacha. Based on the highest enzymatic activity, 15 active isolates were selected and identified. These strains may become the candidates for bio preparation for sewage sludge processing.

Keywords: sewage sludge, composting, bacteria, enzymatic activity

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52 Variation of Carbon Isotope Ratio (δ13C) and Leaf-Productivity Traits in Aquilaria Species (Thymelaeceae)

Authors: Arlene López-Sampson, Tony Page, Betsy Jackes

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Aquilaria genus produces a highly valuable fragrant oleoresin known as agarwood. Agarwood forms in a few trees in the wild as a response to injure or pathogen attack. The resin is used in perfume and incense industry and medicine. Cultivation of Aquilaria species as a sustainable source of the resin is now a common strategy. Physiological traits are frequently used as a proxy of crop and tree productivity. Aquilaria species growing in Queensland, Australia were studied to investigate relationship between leaf-productivity traits with tree growth. Specifically, 28 trees, representing 12 plus trees and 16 trees from yield plots, were selected to conduct carbon isotope analysis (δ13C) and monitor six leaf attributes. Trees were grouped on four diametric classes (diameter at 150 mm above ground level) ensuring the variability in growth of the whole population was sampled. Model averaging technique based on the Akaike’s information criterion (AIC) was computed to identify whether leaf traits could assist in diameter prediction. Carbon isotope values were correlated with height classes and leaf traits to determine any relationship. In average four leaves per shoot were recorded. Approximately one new leaf per week is produced by a shoot. Rate of leaf expansion was estimated in 1.45 mm day-1. There were no statistical differences between diametric classes and leaf expansion rate and number of new leaves per week (p > 0.05). Range of δ13C values in leaves of Aquilaria species was from -25.5 ‰ to -31 ‰ with an average of -28.4 ‰ (± 1.5 ‰). Only 39% of the variability in height can be explained by δ13C in leaf. Leaf δ13C and nitrogen content values were positively correlated. This relationship implies that leaves with higher photosynthetic capacities also had lower intercellular carbon dioxide concentrations (ci/ca) and less depleted values of 13C. Most of the predictor variables have a weak correlation with diameter (D). However, analysis of the 95% confidence of best-ranked regression models indicated that the predictors that could likely explain growth in Aquilaria species are petiole length (PeLen), values of δ13C (true13C) and δ15N (true15N), leaf area (LA), specific leaf area (SLA) and number of new leaf produced per week (NL.week). The model constructed with PeLen, true13C, true15N, LA, SLA and NL.week could explain 45% (R2 0.4573) of the variability in D. The leaf traits studied gave a better understanding of the leaf attributes that could assist in the selection of high-productivity trees in Aquilaria.

Keywords: 13C, petiole length, specific leaf area, tree growth

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51 Cellulolytic and Xylanolytic Enzymes from Mycelial Fungi

Authors: T. Sadunishvili, L. Kutateladze, T. Urushadze, R. Khvedelidze, N. Zakariashvili, M. Jobava, G. Kvesitadze

Abstract:

Multiple repeated soil-climatic zones in Georgia determines the diversity of microorganisms. Hundreds of microscopic fungi of different genera have been isolated from different ecological niches, including some extreme environments. Biosynthetic ability of microscopic fungi has been studied. Trichoderma ressei, representative of the Ascomycetes secrete cellulolytic and xylanolytic enzymes that act in synergy to hydrolyze polysaccharide polymers to glucose, xylose and arabinose, which can be fermented to biofuels. The other mesophilic strains producing cellulases are Allesheria terrestris, Chaetomium thermophile, Fusarium oxysporium, Piptoporus betulinus, Penicillium echinulatum, P. purpurogenum, Aspergillus niger, A. wentii, A. versicolor, A. fumigatus etc. In the majority of the cases the cellulases produced by strains of genus Aspergillus usually have high β-glucosidase activity and average endoglucanases levels (with some exceptions), whereas strains representing Trichoderma have high endo enzyme and low β-glucosidase, and hence has limited efficiency in cellulose hydrolysis. Six producers of stable cellulases and xylanases from mesophilic and thermophilic fungi have been selected. By optimization of submerged cultivation conditions, high activities of cellulases and xylanases were obtained. For enzymes purification, their sedimentation by organic solvents such as ethyl alcohol, acetone, isopropanol and by ammonium sulphate in different ratios have been carried out. Best results were obtained with precipitation by ethyl alcohol (1:3.5) and ammonium sulphate. The yields of enzyme according to cellulase activities were 80-85% in both cases. Cellulase activity of enzyme preparation obtained from the strain Trichoderma viride X 33 is 126 U/g, from the strain Penicillium canescence D 85–185U/g and from the strain Sporotrichum pulverulentum T 5-0 110 U/g. Cellulase activity of enzyme preparation obtained from the strain Aspergillus sp. Av10 is 120 U/g, xylanase activity of enzyme preparation obtained from the strain Aspergillus niger A 7-5–1155U/g and from the strain Aspergillus niger Aj 38-1250 U/g. Optimum pH and temperature of operation and thermostability, of the enzyme preparations, were established. The efficiency of hydrolyses of different agricultural residues by the microscopic fungi cellulases has been studied. The glucose yield from the residues as a result of enzymatic hydrolysis is highly determined by the ratio of enzyme to substrate, pH, temperature, and duration of the process. Hydrolysis efficiency was significantly increased as a result of different pretreatment of the residues by different methods. Acknowledgement: The Study was supported by the ISTC project G-2117, funded by Korea.

Keywords: cellulase, xylanase, microscopic fungi, enzymatic hydrolysis

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50 Genetic Diversity Analysis in Ecological Populations of Persian Walnut

Authors: Masoud Sheidai, Fahimeh Koohdar, Hashem Sharifi

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Juglans regia (L.) commonly known as Persian walnut of the genus Juglans L. (Juglandaceae) is one of the most important cultivated plant species due to its high-quality wood and edible nuts. The genetic diversity analysis is essential for conservation and management of tree species. Persian walnut is native from South-Eastern Europe to North-Western China through Tibet, Nepal, Northern India, Pakistan, and Iran. The species like Persian walnut, which has a wide range of geographical distribution, should harbor extensive genetic variability to adapt to environmental fluctuations they face. We aimed to study the population genetic structure of seven Persian walnut populations including three wild and four cultivated populations by using ISSR (Inter simple sequence repeats) and SRAP (Sequence related amplified polymorphism) molecular markers. We also aimed to compare the genetic variability revealed by ISSR neutral multilocus marker and rDNA ITS sequences. The studied populations differed in morphological features as the samples in each population were clustered together and were separate from the other populations. Three wild populations studied were placed close to each other. The mantel test after 5000 times permutation performed between geographical distance and morphological distance in Persian walnut populations produced significant correlation (r = 0.48, P = 0.002). Therefore, as the populations become farther apart, they become more divergent in morphological features. ISSR analysis produced 47 bands/ loci, while we obtained 15 SRAP bands. Gst and other differentiation statistics determined for these loci revealed that most of the ISSR and SRAP loci have very good discrimination power and can differentiate the studied populations. AMOVA performed for these loci produced a significant difference (< 0.05) supporting the above-said result. AMOVA produced significant genetic difference based on ISSR data among the studied populations (PhiPT = 0.52, P = 0.001). AMOVA revealed that 53% of the total variability is due to among population genetic difference, while 47% is due to within population genetic variability. The results showed that both multilocus molecular markers and ITS sequences can differentiate Persian walnut populations. The studied populations differed genetically and showed isolation by distance (IBD). ITS sequence based MP and Bayesian phylogenetic trees revealed that Iranian walnut cultivars form a distinct clade separated from the cultivars studied from elsewhere. Almost all clades obtained have high bootstrap value. The results indicated that a combination of multilpcus and sequencing molecular markers can be used in genetic differentiation of Persian walnut.

Keywords: genetic diversity, population, molecular markers, genetic difference

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49 Coprophagus Beetles (Scarabaeidae: Coleoptera) of Buxa Tiger Reserve, West Bengal, India

Authors: Subhankar Kumar Sarkar

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Scarab beetles composing the family Scarabaeidae is one of the largest families in the order Coleoptera. The family is comprised of 11 subfamilies. Of these, the subfamily Scarabaeinae includes 13 tribes globally. Indian species are however considered within 2 tribes Scarabaeini and Coprini. Scarab beetles under this subfamily also known as Coprophagus beetles play an indispensable role in forestry and agriculture. Both adults and larvae of these beetles do a remarkable job of carrying excrement into the soil thus enriching the soil to a great extent. Eastern and North Eastern states of India are heavily rich in diversity of organisms as this region exhibits the tropical rain forests of the eastern Himalayas, which exhibits one of the 18 biodiversity hotspots of the world and one of the three of India. Buxa Tiger Reserve located in Dooars between latitudes 26°30” to 26°55” North & longitudes 89°20” to 89°35” East is one such fine example of rain forests of the eastern Himalayas. Despite this, the subfamily is poorly known, particularly from this part of the globe and demands serious revisionary studies. It is with this background; the attempt is being made to assess the Scarabaeinae fauna of the forest. Both extensive and intensive surveys were conducted in different beats under different ranges of Buxa Tiger Reserve. For collection sweep nets, bush beating and collection in inverted umbrella, hand picking techniques were used. Several pit fall traps were laid in the collection localities of the Reserve to trap ground dwelling scarabs. Dung of various animals was also examined to make collections. In the evening hours UV light, trap was used to collect nocturnal beetles. The collected samples were studied under Stereozoom Binocular Microscopes Zeiss SV6, SV11 and Olympus SZ 30. The faunistic investigation of the forest revealed in the recognition of 19 species under 6 genera distributed over 2 tribes. Of these Heliocopris tyrannus Thomson, 1859 was recorded new from the Country, while Catharsius javanus Lansberge, 1886, Copris corpulentus Gillet, 1910, C. doriae Harold, 1877 and C. sarpedon Harold, 1868 from the state. 4 species are recorded as endemic to India. The forest is dominated by the members of the Genus Onthophagus, of which Onthophagus (Colobonthophagus) dama (Fabricius, 1798) is represented by highest number of individuals. Their seasonal distribution is most during Premonsoon followed by Monsoon and Postmonsoon. Zoogeographically all the recorded species are of oriental distribution.

Keywords: buxa tiger reserve, diversity, India, new records, scarabaeinae, scarabaeidae

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48 The Evolution of Man through Cranial and Dental Remains: A Literature Review

Authors: Rishana Bilimoria

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Darwin’s insightful anthropological theory on the evolution drove mankind’s understanding of our existence in the natural world. Scientists consider analysis of dental and craniofacial remains to be pivotal in uncovering facts about our evolutionary journey. The resilient mineral content of enamel and dentine allow cranial and dental remains to be preserved for millions of years, making it an excellent resource not only in anthropology but other fields of research including forensic dentistry. This literature review aims to chronologically approach each ancestral species, reviewing Australopithecus, Paranthropus, Homo Habilis, Homo Rudolfensis, Homo Erectus, Homo Neanderthalis, and finally Homo Sapiens. Studies included in the review assess the features of cranio-dental remains that are of evolutionary importance, such as microstructure, microwear, morphology, and jaw biomechanics. The article discusses the plethora of analysis techniques employed to study dental remains including carbon dating, dental topography, confocal imaging, DPI scanning and light microscopy, in addition to microwear study and analysis of features such as coronal and root morphology, mandibular corpus shape, craniofacial anatomy and microstructure. Furthermore, results from these studies provide insight into the diet, lifestyle and consequently, ecological surroundings of each species. We can correlate dental fossil evidence with wider theories on pivotal global events, to help us contextualize each species in space and time. Examples include dietary adaptation during the period of global cooling converting the landscape of Africa from forest to grassland. Global migration ‘out of Africa’ can be demonstrated by enamel thickness variation, cranial vault variation over time demonstrates accommodation to larger brain sizes, and dental wear patterns can place the commencement of lithic technology in history. Conclusions from this literature review show that dental evidence plays a major role in painting a phenotypic and all rounded picture of species of the Homo genus, in particular, analysis of coronal morphology through carbon dating and dental wear analysis. With regards to analysis technique, whilst studies require larger sample sizes, this could be unrealistic since there are limitations in ability to retrieve fossil data. We cannot deny the reliability of carbon dating; however, there is certainly scope for the use of more recent techniques, and further evidence of their success is required.

Keywords: cranio-facial, dental remains, evolution, hominids

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47 Sceletium Tortuosum: A review on its Phytochemistry, Pharmacokinetics, Biological and Clinical Activities

Authors: Tomi Lois Olatunji, Frances Siebert, Ademola Emmanuel Adetunji, Brian Harvey, Johane Gericke, Josias Hamman, Frank Van Der Kooy

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Ethnopharmacological relevance: Sceletium tortuosum (L.) N.E.Br, the most sought after and widely researched species in the genus Sceletium is a succulent forb endemic to South Africa. Traditionally, this medicinal plant is mainly masticated or smoked and used for the relief of toothache, abdominal pain, and as a mood-elevator, analgesic, hypnotic, anxiolytic, thirst and hunger suppressant, and for its intoxicating/euphoric effects. Sceletium tortuosum is currently of widespread scientific interest due to its clinical potential in treating anxiety and depression, relieving stress in healthy individuals, and enhancing cognitive functions. These pharmacological actions are attributed to its phytochemical constituents referred to as mesembrine-type alkaloids. Aim of the review: The aim of this review was to comprehensively summarize and critically evaluate recent research advances on the phytochemistry, pharmacokinetics, biological and clinical activities of the medicinal plant S. tortuosum. Additionally, current ongoing research and future perspectives are also discussed. Methods: All relevant scientific articles, books, MSc and Ph.D. dissertations on botany, behavioral pharmacology, traditional uses, and phytochemistry of S. tortuosum were retrieved from different databases (including Science Direct, PubMed, Google Scholar, Scopus and Web of Science). For pharmacokinetics and pharmacological effects of S. tortuosum, the focus fell on relevant publications published between 2009 and 2021. Results: Twenty-five alkaloids belonging to four structural classes viz: mesembrine, Sceletium A4, joubertiamine, and tortuosamine, have been identified from S. tortuosum, of which the mesembrine class is predominant. The crude extracts and commercially available standardized extracts of S. tortuosum have displayed a wide spectrum of biological activities (e.g. antimalarial, anti-oxidant, immunomodulatory, anti-HIV, neuroprotection, enhancement of cognitive function) in in vitro or in vivo studies. This plant has not yet been studied in a clinical population, but has potential for enhancing cognitive function, and managing anxiety and depression. Conclusion: As an important South African medicinal plant, S. tortuosum has garnered many research advances on its phytochemistry and biological activities over the last decade. These scientific studies have shown that S. tortuosum has various bioactivities. The findings have further established the link between the phytochemistry and pharmacological application, and support the traditional use of S. tortuosum in the indigenous medicine of South Africa.

Keywords: Aizoaceae, Mesembrine, Serotonin, Sceletium tortuosum, Zembrin®, psychoactive, antidepressant

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46 The One, the Many, and the Doctrine of Divine Simplicity: Variations on Simplicity in Essentialist and Existentialist Metaphysics

Authors: Mark Wiebe

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One of the tasks contemporary analytic philosophers have focused on (e.g., Wolterstorff, Alston, Plantinga, Hasker, and Crisp) is the analysis of certain medieval metaphysical frameworks. This growing body of scholarship has helped clarify and prevent distorted readings of medieval and ancient writers. However, as scholars like Dolezal, Duby, and Brower have pointed out, these analyses have been incomplete or inaccurate in some instances, e.g., with regard to analogical speech or the doctrine of divine simplicity (DDS). Additionally, contributors to this work frequently express opposing claims or fail to note substantial differences between ancient and medieval thinkers. This is the case regarding the comparison between Thomas Aquinas and others. Anton Pegis and Étienne Gilson have argued along this line that Thomas’ metaphysical framework represents a fundamental shift. Gilson describes Thomas’ metaphysics as a turn from a form of “essentialism” to “existentialism.” One should argue that this shift distinguishes Thomas from many Analytic philosophers as well as from other classical defenders of the DDS. Moreover, many of the objections Analytic Philosophers make against Thomas presume the same metaphysical principles undergirding the above-mentioned form of essentialism. This weakens their force against Thomas’ positions. In order to demonstrate these claims, it will be helpful to consider Thomas’ metaphysical outlook alongside that of two other prominent figures: Augustine and Ockham. One area of their thinking which brings their differences to the surface has to do with how each relates to Platonic and Neo-Platonic thought. More specifically, it is illuminating to consider whether and how each distinguishes or conceives essence and existence. It is also useful to see how each approaches the Platonic conflicts between essence and individuality, unity and intelligibility. In both of these areas, Thomas stands out from Augustine and Ockham. Although Augustine and Ockham diverge in many ways, both ultimately identify being with particularity and pit particularity against both unity and intelligibility. Contrastingly, Thomas argues that being is distinct from and prior to essence. Being (i.e., Being in itself) rather than essence or form must therefore serve as the ground and ultimate principle for the existence of everything in which being and essence are distinct. Additionally, since change, movement, and addition improve and give definition to finite being, multitude and distinction are, therefore, principles of being rather than non-being. Consequently, each creature imitates and participates in God’s perfect Being in its own way; the perfection of each genus exists pre-eminently in God without being at odds with God’s simplicity, God has knowledge, power, and will, and these and the many other terms assigned to God refer truly to the being of God without being either meaningless or synonymous. The existentialist outlook at work in these claims distinguishes Thomas in a noteworthy way from his contemporaries and predecessors as much as it does from many of the analytic philosophers who have objected to his thought. This suggests that at least these kinds of objections do not apply to Thomas’ thought.

Keywords: theology, philosophy of religion, metaphysics, philosophy

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45 A Study of the Prevalence of Trichinellosis in Domestic and Wild Animals for the Region of Sofia, Bulgaria

Authors: Valeria Dilcheva, Svetlozara Petkova, Ivelin Vladov

Abstract:

Nemathodes of the genus Trichinella are zoonotic parasites with a cosmopolitan distribution. More than 100 species of mammals, birds and reptiles are involved in the natural cycle of this nematode. At present, T. spiralis, T. pseudospiralis, and T. britovi have been found in Bulgaria. The existence of natural wildlife and domestic reservoirs of Trichinella spp. can be a serious threat to human health. Three trichinella isolates caused human trichinella infection outbreaks from three regions of Sofia City Province were used for the research: sample No. 1 - Ratus norvegicus, sample No. 2 – domestic pig (Sus scrofa domestica), sample No. 3 - domestic pig (Sus scrofa domestica). Trichinella larvae of the studied species were isolated via digestive method (pepsin, hydrochloric acid, water) at 37ºC by standard procedure and were determined by gender (male and female) based on their morphological characteristics. As a reference trichinella species were used: T. spiralis, T. pseudospiralis, T. nativa and T. britovi. Single male and female larvae of the three isolates were crossed with single male and female larvae of the reference trichinella species as well as reciprocally. As a result of cross-breeding, offspring of muscular larvae with T. spiralis and T. britovi were obtained, while in experiments with T. pseudospiralis and T. nativa, trichinella larvae were not found in the laboratory mice. The results obtained in the control groups indicate that the trichinella larvae used from the isolates and the four trichinella species are infective. Also, the infective ability of the F1 offspring from the successful cross-breeding between isolates and reference species was investigated. Through the data obtained in the experiment was found that isolates No. 1 and No. 2 belong to the species T. spiralis, and isolate No. 3 belongs to the species T. britovi. The results were confirmed by PCR and real-time PCR analysis. Thus the presence and circulation of the species T. spiralis and T. britovi in Bulgaria was confirmed. Probably the rodents (rats) are involved in the distribution of T. spiralis in urban environment. The species T. britovi found in a domestic pig speaks of some contact with wild animals for which T. britovi is characteristic. The probable reason is that a large number of farmers in Bulgaria practice the free-range breeding of domestic pigs. Part of the farmers also used as food for domestic pigs waste products from the game (foxes, jackals, bears, wolves) and probably thus the infection was obtained. The distribution range of trichinella species in Bulgaria is not strictly outlined. It is believed that T. spiralis is most common in domestic animals and T. britovi and T. pseudospiralis are characteristic of wildlife. To answer the question whether wild and synanthropic animals are infected with the same or different trichinella species, which species predominate in nature and what their distribution among different hosts is, further research is required.

Keywords: cross-breeding, Sofia, trichinellosis, Trichinella britovi, Trichinella spiralis

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44 Value Adding of Waste Biomass of Capsicum and Chilli Crops for Medical and Health Supplement Industries

Authors: Mursleen Yasin, Sunil Panchal, Michelle Mak, Zhonghua Chen

Abstract:

“The use of agricultural and horticultural waste to obtain beneficial products. Thus reduce its environmental impact and help the general population.” Every year 20 billion dollars of food is wasted in the world. All the energy, resources, nutrients and metabolites are lost to the landfills as well. On farm production losses are a main issue in agriculture. Almost 25% vegetables never leave the farm because they are not considered perfect for supermarkets and treated as waste material along with the rest of the plant parts. For capsicums, this waste is 56% of the total crop. Capsicum genus is enriched with a group of compounds called capsaicinoids which are a source of spiciness of these fruits. Capsaicin and dihydrocapsaicin are the major members comprising almost 90% of this group. The major production and accumulation site is the non-edible part of fruit i.e., placenta. Other parts of the plant, like stem, leaves, pericarp and seeds, also contain these pungent compounds. Capsaicinoids are enriched with properties like analgesic, antioxidants, anti-inflammatory, antibacterial, anti-virulence anti-carcinogenic, chemo preventive, chemotherapeutic, antidiabetic etc. They are also effective in treating problems related to gastrointestinal tract, lowering cholesterol and triglycerides in obesity. The aim of the study is to develop a standardised technique for capsaicinoids extraction and to identify better nutrient treatment for fruit and capsaicinoids yield. For research 3 capsicum and 2 chilli varieties were grown in a high-tech glass house facility in Sydney, Australia. Plants were treated with three levels of nutrient treatments i.e., EC 1.8, EC 2.8 and EC 3.8 in order to check its effect on fruit yield and capsaicinoids concentration. Solvent extraction procedure is used with 75% ethanol to extract these secondary metabolites. Physiological, post-harvest and waste biomass measurement and metabolomic analysis are also performed. The results showed that EC 2.8 gave the better fruit yield of capsicums, and those fruits have the higher capsaicinoids concentration. For chillies, higher EC levels had better results than lower treatment. The UHPLC analysis is done to quantify the compounds, and a decrease in capsaicin concentration is observed with the crop maturation. The outcome of this project is a sustainable technique for extraction of capsaicinoids which can easily be adopted by farmers. In this way, farmers can help in value adding of waste by extracting and selling capsaicinoids to nutraceutical and pharmaceutical industries and also earn some secondary income from the 56% waste of capsicum crop.

Keywords: capsaicinoids, plant waste, capsicum, solvent extraction, waste biomass

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43 Eucalyptus camaldulensis Leaves Attacked by the Gall Wasp Leptocybe invasa: A Phyto-Volatile Constituents Study

Authors: Maged El-Sayed Mohamed

Abstract:

Eucalyptus camaldulensis is one on the most well-known species of the genus Eucalyptus in the Middle east, its importance relay on the high production of its unique volatile constituents which exhibits many medicinal and pharmacological activities. The gall-forming wasp (Leptocybe invasa) has recently come into sight as the main pest attacking E. camaldulensis and causing severe injury. The wasp lays its eggs in the petiole and midrib of leaves and stems of young shoots of E. camaldulensis, which leads to gall formation. Gall formation by L. invasa damages growing shoot and leaves of Eucalyptus, resulting in abscission of leaves and drying. AIM: This study is an attempt to investigate the effect of the gall wasp (Leptocybe invasa) attack on the volatile constitutes of E. camaldulensis. This could help in the control of this wasp through stimulating plant defenses or production of a new allelochemicals or insecticide. The study of volatile constitutes of Eucalyptus before and after attack by the wasp can help the re-use and recycle of the infected Eucalyptus trees for new pharmacological and medicinal activities. Methodology: The fresh gall wasp-attacked and healthy leaves (100 g each) were cut and immediately subjected to hydrodistillation using Clevenger-type apparatus for 3 hours. The volatile fractions isolated were analyzed using Gas chromatography/mass spectrometry (GC/MS). Kovat’s retention indices (RI) were calculated with respect to a set of co-injected standard hydrocarbons (C10-C28). Compounds were identified by comparing their spectral data and retention indices with Wiley Registry of Mass Spectral Data 10th edition (April 2013), NIST 11 Mass Spectral Library (NIST11/2011/EPA/NIH) and literature data. Results: Fifty-nine components representing 89.13 and 88.60% of the total volatile fraction content respectively were quantitatively analyzed. Twenty-six major compounds at an average concentration greater than 0.1 ± 0.02% have been used for the statistical comparison. From those major components, twenty-one were found in both the attacked and healthy Eucalyptus leaves’ fractions in different concentration and five components, mono terpene p-Mentha-2-4(8) diene and the sesquiterpenes δ-elemene, β-elemene, E-caryophyllene and Bicyclogermacrene, were unique and only produced in the attacked-leaves’ fraction. CONCLUSION: Newly produced components or those commonly found in the volatile fraction and changed in concentration could represent a part of the plant defense mechanisms or might be an element of the plant allelopathic and communication mechanisms. Identification of the components of the gall wasp-damaged leaves can help in their recycling for different physiological, pharmacological and medicinal uses.

Keywords: Eucalyptus camaldulensis, eucalyptus recycling, gall wasp, Leptocybe invasa, plant defense mechanisms, Terpene fraction

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42 Oxidative Stability of Corn Oil Supplemented with Natural Antioxidants from Cypriot Salvia fruticosa Extracts

Authors: Zoi Konsoula

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Vegetable oils, which are rich in polyunsaturated fatty acids, are susceptible to oxidative deterioration. The lipid oxidation of oils results in the production of rancid odors and unpleasant flavors as well as the reduction of their nutritional quality and safety. Traditionally, synthetic antioxidants are employed for their retardation or prevention of oxidative deterioration of oils. However, these compounds are suspected to pose health hazards. Consequently, recently there has been a growing interest in the use of natural antioxidants of plant origin for improving the oxidative stability of vegetable oils. The genus Salvia (sage) is well known for its antioxidant activity. In the Cypriot flora Salvia fruticosa is the most distributed indigenous Salvia species. In the present study, extracts were prepared from S. fruticosa aerial parts using various solvents and their antioxidant activity was evaluated by the 1,1-diphenyl-2-picrylhydrazine (DPPH) radical scavenging and Ferric Reducing Antioxidant Power (FRAP) method. Moreover, the antioxidant efficacy of all extracts was assessed using corn oil as the oxidation substrate, which was subjected to accelerated aging (60 °C, 30 days). The progress of lipid oxidation was monitored by the determination of the peroxide, p-aniside, conjugated dienes and trienes value according to the official AOCS methods. Synthetic antioxidants (butylated hydroxytoluene-BHT and butylated hydroxyanisole-BHA) were employed at their legal limit (200 ppm) as reference. Finally, the total phenolic (TPC) and flavonoid content (TFC) of the prepared extracts was measured by the Folin-Ciocalteu and aluminum-flavonoid complex method, respectively. The results of the present study revealed that although all sage extracts prepared from S. fruticosa exhibited antioxidant activity, the highest antioxidant capacity was recorded in the methanolic extract, followed by the non-toxic, food grade ethanol. Furthermore, a positive correlation between the antioxidant potency and the TPC of extracts was observed in all cases. Interestingly, sage extracts prevented lipid oxidation in corn oil at all concentrations tested, however, the magnitude of stabilization was dose dependent. More specifically, results from the different oxidation parameters were in agreement with each other and indicated that the protection offered by the various extracts depended on their TPC. Among the extracts, the methanolic extract was more potent in inhibiting oxidative deterioration. Finally, both methanolic and ethanolic sage extracts at a concentration of 1000 ppm exerted a stabilizing effect comparable to that of the reference synthetic antioxidants. Based on the results of the present study, sage extracts could be used for minimizing or preventing lipid oxidation in oils and, thus, prolonging their shelf-life. In particular, given that the use of dietary alcohol, such as ethanol, is preferable than methanol in food applications, the ethanolic extract prepared from S. fruticosa could be used as an alternative natural antioxidant.

Keywords: antioxidant activity, corn oil, oxidative deterioration, sage

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41 Predictive Pathogen Biology: Genome-Based Prediction of Pathogenic Potential and Countermeasures Targets

Authors: Debjit Ray

Abstract:

Horizontal gene transfer (HGT) and recombination leads to the emergence of bacterial antibiotic resistance and pathogenic traits. HGT events can be identified by comparing a large number of fully sequenced genomes across a species or genus, define the phylogenetic range of HGT, and find potential sources of new resistance genes. In-depth comparative phylogenomics can also identify subtle genome or plasmid structural changes or mutations associated with phenotypic changes. Comparative phylogenomics requires that accurately sequenced, complete and properly annotated genomes of the organism. Assembling closed genomes requires additional mate-pair reads or “long read” sequencing data to accompany short-read paired-end data. To bring down the cost and time required of producing assembled genomes and annotating genome features that inform drug resistance and pathogenicity, we are analyzing the performance for genome assembly of data from the Illumina NextSeq, which has faster throughput than the Illumina HiSeq (~1-2 days versus ~1 week), and shorter reads (150bp paired-end versus 300bp paired end) but higher capacity (150-400M reads per run versus ~5-15M) compared to the Illumina MiSeq. Bioinformatics improvements are also needed to make rapid, routine production of complete genomes a reality. Modern assemblers such as SPAdes 3.6.0 running on a standard Linux blade are capable in a few hours of converting mixes of reads from different library preps into high-quality assemblies with only a few gaps. Remaining breaks in scaffolds are generally due to repeats (e.g., rRNA genes) are addressed by our software for gap closure techniques, that avoid custom PCR or targeted sequencing. Our goal is to improve the understanding of emergence of pathogenesis using sequencing, comparative genomics, and machine learning analysis of ~1000 pathogen genomes. Machine learning algorithms will be used to digest the diverse features (change in virulence genes, recombination, horizontal gene transfer, patient diagnostics). Temporal data and evolutionary models can thus determine whether the origin of a particular isolate is likely to have been from the environment (could it have evolved from previous isolates). It can be useful for comparing differences in virulence along or across the tree. More intriguing, it can test whether there is a direction to virulence strength. This would open new avenues in the prediction of uncharacterized clinical bugs and multidrug resistance evolution and pathogen emergence.

Keywords: genomics, pathogens, genome assembly, superbugs

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40 The Gut Microbiome in Cirrhosis and Hepatocellular Carcinoma: Characterization of Disease-Related Microbial Signature and the Possible Impact of Life Style and Nutrition

Authors: Lena Lapidot, Amir Amnon, Rita Nosenko, Veitsman Ella, Cohen-Ezra Oranit, Davidov Yana, Segev Shlomo, Koren Omry, Safran Michal, Ben-Ari Ziv

Abstract:

Introduction: Hepatocellular carcinoma (HCC) is one of the leading causes of cancer related mortality worldwide. Liver Cirrhosis is the main predisposing risk factor for the development of HCC. The factor(s) influencing disease progression from Cirrhosis to HCC remain unknown. Gut microbiota has recently emerged as a major player in different liver diseases, however its association with HCC is still a mystery. Moreover, there might be an important association between the gut microbiota, nutrition, life style and the progression of Cirrhosis and HCC. The aim of our study was to characterize the gut microbial signature in association with life style and nutrition of patients with Cirrhosis, HCC-Cirrhosis and healthy controls. Design: Stool samples were collected from 95 individuals (30 patients with HCC, 38 patients with Cirrhosis and 27 age, gender and BMI-matched healthy volunteers). All participants answered lifestyle and Food Frequency Questionnaires. 16S rRNA sequencing of fecal DNA was performed (MiSeq Illumina). Results: There was a significant decrease in alpha diversity in patients with Cirrhosis (qvalue=0.033) and in patients with HCC-Cirrhosis (qvalue=0.032) compared to healthy controls. The microbiota of patients with HCC-cirrhosis compared to patients with Cirrhosis, was characterized by a significant overrepresentation of Clostridium (pvalue=0.024) and CF231 (pvalue=0.010) and lower expression of Alphaproteobacteria (pvalue=0.039) and Verrucomicrobia (pvalue=0.036) in several taxonomic levels: Verrucomicrobiae, Verrucomicrobiales, Verrucomicrobiaceae and the genus Akkermansia (pvalue=0.039). Furthermore, we performed an analysis of predicted metabolic pathways (Kegg level 2) that resulted in a significant decrease in the diversity of metabolic pathways in patients with HCC-Cirrhosis (qvalue=0.015) compared to controls, one of which was amino acid metabolism. Furthermore, investigating the life style and nutrition habits of patients with HCC-Cirrhosis, we found significant correlations between intake of artificial sweeteners and Verrucomicrobia (qvalue=0.12), High sugar intake and Synergistetes (qvalue=0.021) and High BMI and the pathogen Campylobacter (qvalue=0.066). Furthermore, overweight in patients with HCC-Cirrhosis modified bacterial diversity (qvalue=0.023) and composition (qvalue=0.033). Conclusions: To the best of the our knowledge, we present the first report of the gut microbial composition in patients with HCC-Cirrhosis, compared with Cirrhotic patients and healthy controls. We have demonstrated in our study that there are significant differences in the gut microbiome of patients with HCC-cirrhosis compared to Cirrhotic patients and healthy controls. Our findings are even more pronounced because the significantly increased bacteria Clostridium and CF231 in HCC-Cirrhosis weren't influenced by diet and lifestyle, implying this change is due to the development of HCC. Further studies are needed to confirm these findings and assess causality.

Keywords: Cirrhosis, Hepatocellular carcinoma, life style, liver disease, microbiome, nutrition

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