Journal of BioScience and Biotechnology (JBB)
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p-ISSN: 1314-6238 / e-ISSN: 1314-6246
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Item Characterization of α-amylase and antimicrobial activity of Penicillium chrysogenum(Plovdiv University Press “Paisii Hilendarski”, 2024-07-10) Adeoyo, Olusegun R.; Adewumi, Olubunmi R.Fungi have been used to produce a number of bioactive compounds that include enzymes and antibacterial agents with unique characteristics in vitro. Penicillium chrysogenum is a mould with great potential for producing important secondary metabolites. This study aimed to characterize α-amylase and antimicrobial activity of P. chrysogenum. The organism was grown in a solid-state fermentation (SSF) medium using cassava peels as the only carbon source. The molecular mass of the amylase was estimated to be 59 kDa by combined results of Amicon Ultra-15 filter unit (Sigma) 50 kDa MWCO and polyacrylamide gel electrophoresis (PAGE). The optimum activity of 93.1 U/mL was noticed at 30oC and pH of 5.8. A stability assay revealed that the enzyme was stable at 40°C and 50°C by retaining 66% and 37% residual activities, respectively. Also, crude extract of P. chrysogenum was found to inhibit Pseudomonas aeruginosa and Staphylococcus aureus with minimum inhibitory concentrations (MIC) of 25 and 12.5 mg/mL respectively. Therefore, these properties make P. chrysogenum an interesting fungus that has the potential of being used to produce both antibacterial agents and α-amylase for the bio-economy.Item Bio-active components of Melaleuca alternifolia, Rosmarinus officinalis, Boswellia serrata essential oil as anti-diabetic therapeutics targeting α-amylase : In vitro α-amylase inhibition, antioxidant, binding interaction, and docking studies of predominant compounds(Plovdiv University Press “Paisii Hilendarski”, 2024-07-10) Sharma, Arun Dev; Chauhan, Amrita; Kaur, InderjeetTea tree essential oil (TEO), Rosemary Essential Oil (REO), and Guggul essential oil (GEO) (EOs) are priceless essential oils that have been linked to several biological activities, including antibacterial, antifungal, immunomodulatory, anticancer, and anti-inflammatory effects. α-amylase inhibition is a hopeful curative target against type-2 diabetes as it can downgrade fierce digestion and absorption of carbohydrates into absorbable monosaccharides. The purpose of the study is in silico molecular docking of principal components of TEO, REO, and GEO followed by in vitro validation of inhibition of α-amylase activity. For docking Cb-dock2 tool was utilized. Ligand-Protein 2-D interactions were also studied. From the perspective of human health, in silico ADMET pharmacoinformatic features (Physicochemical, Lipophilicity, Medicinal Chemistry, Druglikeness, Absorption, Water Solubility, Distribution, Metabolism, Pharmacokinetics, Excretion) have prospected. Using α-amylase, wet lab validation was carried out. 2, 2-Diphenyl-1-picryl hydrazyl (DPPH) radical inhibition assay was conducted to ascertain the antioxidant role of all EOs. Docking investigation demonstrated the effective binding of all the ligands with the α-amylase. The interaction results imply that the enzyme-ligand complexes form hydrogen, hydrophobic, and other interactions. In silico ADMET examination disclosed that all the ligand molecules have no toxic effect and acceptable absorption, as well. Further, TEO, REO, and GEO have dose-dependent inhibitory action against α-amylase. All EOs depicted good antioxidant potential. Kinetic analysis revealed that TEO, REO, and GEO competitively inhibited α-amylase. It was concluded that these substances can function as model molecules for the synthesis of novel anti-diabetic substances.Item Diversity of arbuscular mycorrhizal fungi under the rhizosphere soil of different cropping systems(2024-07-10) Zeleke, Girma; Dobo, Beyene; Asefa, FassilThis study has been conducted to evaluate the diversity of Arbuscular Mycorrhizal Fungi (AMF) under the rhizosphere soil of different monocrops in the grazing and croplands. A descriptive cross-sectional survey was employed to understand the rate of root colonization and diversity of AMF. 23 different types of AMF morphospecies in 12 AMF genera were isolated across 12 different types of land covers. The AMF under the rhizosphere soil of Eucalyptus tree has shown the highest biomass compared to all other land uses, with spore density of 1907.4±0.404 100g-1 of dry soil. The lowest AMF biomass has been recorded in the rhizosphere soil of the Mango tree, with a spore density of 260.1±0.121 100g-1 dry soil. The total root colonization (RLC) of the various land covers by AMF ranges from 11.15-85.41%. Finally, further study on the implication of agricultural inputs on the microbial community under different cropping systems is recommended.Item In vitro antimicrobial, antioxidant, antibiofilm, and quorum sensing inhibitory activities of Scandix pecten-veneris L., Artedia squamata L., Oenanthe pimpinelloides L.(2024-07-10) Ceylan, Özgür; Uğur, Aysel; Saraç, Nurdan; Arslan, Büşra EroğluPlants have been widely used as a pharmaceutical to treat diseases in many cultures for centuries. In vitro biological activities of some plants consumed as food and used in traditional medicine in Southwestern Anatolia have been reported. The present study focused on the antimicrobial, antibiofilm, antioxidant, and quorum sensing (QS) inhibitory activities of aqueous and ethanolic extracts of aerial parts of Scandix pecten-veneris L., Artedia squamata L., Oenanthe pimpinelloides L. The antimicrobial activity was evaluated against fifteen bacteria and one yeast using disc diffusion and microdilution methods. The aqueous and ethanol extracts of A. squamata showed the highest activity against Staphylococcus aureus MU 47 with 20 and 23 mm zone diameters, respectively. The ethanol extract of S. pecten-veneris inhibited Staphylococcus aureus ATCC 25923 at a 5 mg/mL MIC value. The aqueous extract of O. pimpinelloides inhibited biofilm formation of S. aureus MU 40, P. aeruginosa ATCC 27853, and C. albicans ATCC 10239 at MIC value. All extracts displayed violacein inhibition against Chromobacterium violaceum CV12472 in the concentration range of 1.25 mg/mL to 25 mg/mL. The aqueous extract of A. squamata showed valuable anti-quorum sensing (anti-QS) activity with a pigment inhibition zone of 17 mm on Chromobacterium violaceum CV026. The aqueous extract of S. pecten-veneris inhibited the swarming motility of Pseudomonas aeruginosa PA01 by 47.6%. The aqueous and ethanol extracts of A. squamata indicated the best antioxidant activity. It was proved that these edible plant extracts could be used as natural alternative inhibitors to control microbial pathogenesis and diseases.Item Co-immunoprecipitation based isolation and identification of Interacting Phytal and Rheumatoid arthritis subject’s serum globulins proteins(2024-07-10) Chunduri, Jayaparada Rao; Bhatt, Pragati; Desai, Rupal H.Phytalproteins have an immense role in plant’s growth and sustenance processes. They can be used as nutritional enhancers of foods we consume. Rheumatoid arthritis is an autoimmune disorder, prevalent in both sexes of humans and especially those above 50 years. Ayurveda suggests plant oils to reduce the pain and inflammation caused by Rheumatoid arthritis (RA). The current study is based on immunoprecipitation interactions between phytal proteins and gamma globulins of Rheumatoid arthritis subjects to assess their role in diagnostics or as immune modulators. A comparative study of proteins of Murraya koenigi, Basella alba (leaves), and Sapindus mukorossi (seed) has been considered for immunoassays. The protein-protein binding capability assessment has shown a positive response with Murraya koenigi phytal proteins than that of Basella alba. The globulins of Sapindus mukorossi seed proteins indicated more specific and prominent interactions than the previous two. The co-immunoprecipitation bands were subjected to LC/MS followed by Bioinformatic studies based on meta HawkDock and PDB sum. Based on the docking study’s scores, hydrogen bonds and salt bridges, bonding capability between 10 selected gamma globulin proteins and unique proteins, and 4 phytal proteins of Indian soapberry were established. Q3KNS1(sperm function), unique protein Q7Z351(associated with the immune system) and A0A024617 (Alpha-1-antitrypsin) proteins of human serum showed an interaction with A0A0U1XK40, A0A7H1CQR4 of Sapindicus. Studies of RuBISCo (A0A0U1XK40) of Indian soap nut protein and Q3KNS1(Patched domain-containing protein 3 that controls sperm development or sperm function and affects Rheumatoidism of muscles) interactions, can be considered for further studies.Item In vitro assay of antagonistic activities of endophytic fungi from Calabash tree leaves against pathogenic Fusarium oxysporum(2024-07-10) Sari, Noorkomala; Sari, Nukhak Nufita; Sari, Wijayanti PurnamaEndophytic fungi are microorganisms known as plant-associated fungi and are typically found asymptomatically inside plant tissue. By directly generating secondary metabolites, endophytic fungi help their host plants grow and become more resistant to plant diseases. Furthermore, they can also biosynthesize bioactive compounds used for antimicrobials, which were previously believed to be produced only by the host plant. Medicinal plants as host plants for endophytic fungi which produce secondary metabolites and their ability as biological agents against pathogens. Calabash tree is a medicinal plant that contains active compounds that function as antifungal and antibacterial. This study aims to determine the antagonistic activity of endophytic fungi from calabash tree leaves against F. oxysporum Fos, the pathogen that causes fusarium wilt in shallot plants. Twelve endophytic fungi were obtained from isolation from the leaves of the calabash tree, namely EnM6C22, EnM11P2, EnM6A2, EnM5H21, EnM10H22, EnM12P, EnM11P, EnM111, EnM5H22, EnM9Pt22, EnM5H1 and EnM5K. Five of the twelve endophytic fungi of calabash tree leaves have been identified, as Fusarium sp., Rhizoctonia sp., Pestalotia sp., Colletotrichum sp., and Bipolaris sp. with inhibition percentages of 36.43%, 32.19%, 25.41%, 32.19%, and 27.53% respectively at 7 days after inoculation.Item Exploring the Impact of Hesperidin on Probiotic Properties of Lactobacillus acidophilus LA-5 and Its Synbiotic Interaction with Colon Cancer Cells(2024-07-10) Celebioglu, Hasan U.; Evin, Emre; Bulut, Sena Davran; Derebasi, Buse N.; Adali, OrhanProbiotics are live microorganisms that provide health benefits to the host when taken in adequate amounts. Hesperidin is a phenolic compound found in banana fruit, lemon fruit, and lemon peel. It exhibits anti-inflammatory, anti-oxidative, and anti-cancer activities. Although these compounds are present in the gastrointestinal tract (GIT), their synbiotic interaction on the GIT is not well understood. This study aims to investigate the effects of hesperidin on the probiotic properties of Lactobacillus acidophilus LA-5 (LA-5) and the impact of their synbiotic interaction on the cancer cell line HT-29. Results showed that hesperidin did not negatively affect the auto-aggregation, adhesion, and antioxidant capacity of LA-5. LA-5 grown with hesperidin exhibited greater resistance to pepsin and bile salt compared to LA-5 alone. Furthermore, in vitro cancer studies indicated that bacteria grown in the presence of hesperidin may increase caspase-3 activity in HT-29 cells and thus induce apoptosis in this way. This study suggests that hesperidin may enhance and contribute to the probiotic properties of LA-5. Consequently, LA-5 grown with hesperidin may exert more beneficial effects on the host.Item Water and surfaces as potential sources of pathogenic bacteria in Harare hospitals’ intensive care units(2024-07-10) Dururu, Mathias; Muchesa, Petros; Chidzwondo, Farisai; Mangoyi, RumbidzaiWater and contaminated hospital surfaces have been reported as major causes of Hospital acquirеd (nosocomial) infections (HAIs), a significant global safety problem for both patients and medical staff. However, the frequency of Intensive Care Units (ICU)-HAIs has been reported to be at least 2-3 times greater in low- to middle-income countries, particularly in Zimbabwe, than in high-income countries. Thus, this study aimed at determining the presence of pathogenic bacteria in water and surfaces from selected high-risk areas within the intensive care units of three hospitals (A, B, and C) in Harare, Zimbabwe. Water and surface samples were collected and processed within 24 hours of collection. The water samples were tested for the presence of microbial indicators using standard methodology of microbial plating. The surface swabs were also investigated using microbial plating. The organisms that were found to be present in surface samples were then identified using biochemical tests and 16S polymerase chain reaction (16S PCR). The results showed no bacteria present in any of the water samples tested during the study. However, using morphological characteristics, six different types of bacteria were identified in surface samples. The bacteria identified to be present on surfaces were Escherichia coli, Salmonella, Shigella, Staphylococcus, Pseudomonas, and Streptococcus. Hospital A surfaces may be the major cause of some bacterial infection outbreaks, hence, there is a need for better ways of reducing contamination of surfaces in the intensive care units.Item Estimation of some heavy metals and biochemical variables in blood and soil samples from Kirkuk city, Iraq(2024-07-10) Beebany, Shakhawan; Yousif, Heshu; Hussain, KameranThis study is focused on the determination of some heavy metals levels (Se, Pb and Cd) with the study of biochemical parameters in blood samples, such as superoxide dismutases (SOD), catalases (CAT), glutathione peroxidases (GPX), glutathione reductases (GRX), vitamin C and vitamin. The samples were collected from 50 individuals at four distinct fuel stations in Kirkuk city in Iraq. To obtain the practical data, atomic absorption spectrometer, UV spectrophotometer and high performance liquid chromatography were used. The results indicated remarkable differences in the concentrations of Se, Pb and Cd in the analyzed samples compared to the control group. These Levels were lower in fuel station workers, whereas and the Pb and Cd concentrations were higher in their blood. Additionally, the results of chemical variables showed a decline in the antioxidant concentrations (SOD, GPx, Vit C, Vit E). In terms of oxidative stress (MDA), there was a significant increase in its concentration among the fuel station workers. In addition, the concentration of toxic heavy metals (Pb, Cd) in the soil of fuel stations was measured using the graphite furnace atomic absorption spectrometer. The Pb and Cd levels in the soil were within the permissible limits, indicating no adverse impact of on the soil.Item Biosocial characteristics of patients with paranoid schizophrenia(Plovdiv University Press “Paisii Hilendarski”, 2016-04-26) Panayotova, Gergana K.; Chengeliyska, Vesela D.; Ivanova, Evgeniya N.Schizophrenia is known as a complex disorder that combines both genetic and environmental factors. Different genes have been tested as candidates for association with schizophrenia and different environmental factors have been examined in many studies on epidemiology of schizophrenia. Specific environmental factors, such as nonspecific stress, mental and physical abuse, maternal diet during pregnancy, drug use, living in an urban setting, migration, seasonal effects on birth and exposure to infections, have been discussed as possible risk for schizophrenia. The present preliminary study is focused on the relations between biological and social characteristics of patients with paranoid schizophrenia with different cognitive levels, emotional and creative styles. Descriptive statistics, the Student’s t-test and SPSS software, were used to analyse the relations mentioned. Differences between sexes and these concerning age of individuals (risk level of inheritance, ABO blood group distribution, triggering factors, aggressive behavior, single or multiple suicide attempts, levels of education and creative talents) were indicated and discussed. The study identifies important trends and discuses essential biosocial relations in context of the knowledge for schizophrenia in Bulgaria. Future comparative investigations, including genetic markers and psychogenetic approaches, should be used in complex, in order to characterize the reasons for developing paranoid schizophrenia and the possible relations between biological, psychological and social factors better.Item Sensitivity of watermelon variety Bojura to mutant agents 60Co and EMS(Plovdiv University Press “Paisii Hilendarski”, 2016-04-26) Velkov, Nikolay; Tomlekova, Nasya; Sarsu, FatmaA study on the sensitivity of watermelon variety Bojura to mutagenic agents was carried out in 2013-2014. The goal was to establish effective doses for mutagenic treatment of dry seeds with 60Co gamma rays (80, 100, 200, 250, 350 and 450 Gy) and swollen seeds with water for 24 hours were treated with ethyl methanesulfonate (EMS) at a concentration of 2%. Dominant mutations were not observed in the M1 generation. Morphological changes in 14 of 1395 M2 plants were observed. Phenotypic variations changes were the colour of the seed coat, chlorophyll disorders of cotyledons, leaves, petals, and alterations of the location of the fruit set in the central stem. Visible changes of the morphological characteristics of the fruit were not observed. The doses induced certain morphological changes, however, higher doses or combined gamma rays 60Co and EMS treatments would induce mutations more efficiently. Subsequent experiments are required to obtain mutants with changes that affect flowers and fruits. The results are important for increasing mutation efficiency in watermelon breeding.Item Production of therapeutic proteins through plant tissue and cell culture(Plovdiv University Press “Paisii Hilendarski”, 2016-04-27) Gharelo, Reza S.; Oliaei, Ebrahim D.; Bandehagh, Ali; Khodadadi, Ehsaneh; Noparvar, Pouya M.Nowadays, pharmaceutical recombinant protein is increasingly used in treatment of many diseases such as hepatitis, anemia, diabetes and cancer. Different protein expression systems have been used for the expression of recombinant proteins in which each of them face obstacles that make utilizing them as comprehensive expression system in order to express wide variety of proteins difficult. Plant cell as a eukaryotic expression system have many advantages compared to other hosts. They are very “safe” and significantly decrease concerns about the contamination of recombinant proteins with human pathogens. In addition to this, plants as eukaryotic expression system perform proper post-translational modification, in case of eukaryotic proteins, and appropriate folding resulting in right function in biological environments. Therefore, the production of pharmaceutical protein through plant cells can be absolutely promising approach. In this review, the production of pharmaceutical protein in plant cells, advantages and disadvantages, offered methods and techniques for developing recombinant protein yields, and affective factors on the whole process of pharmaceutical protein expression in the molecular level will be reviewed.Item The effects of different sizes of insoluble grit on growth performance and carcass traits in broiler chickens(Plovdiv University Press “Paisii Hilendarski”, 2016-04-27) Moghaddam, Abbas Ali Rajabiyan; Ebrahimnezhad, Yahya; Teli, Abdol-Ahad ShaddelThe present study was conducted to investigate the effects of different sizes of insoluble grit on growth performance and carcass traits in broiler chickens. A total of 200 broilers (Ross 308), 10 days old, were randomly allotted to five experimental equal groups with four replicates of 10 chickens (five male and five female) and fed with basal diet + ground wheat (without grit); basal diet + whole wheat (without grit) and basal diet contain 1.5% grit of diet with sizes of 2, 3 and 4 mm. Growth performance (evaluated through weight gain, feed intake and feed conversion ratio) was determined on day 24 and 42. Also, carcass traits (relative weights of carcass, breast, thigh, liver, heart, gizzard and intestine) and intestine length were assessed on day 42. Weight gains and feed conversion ratio were significantly improved in broilers added with grit 2 mm compared to the control group (p<0.05), whereas; carcass traits were not significantly altered. These data suggest that grit with size of 2 mm improve growth performance in broiler chickens.Item Genetic analysis of Apis mellifera macedonica (type rodopica) populations selectively reared for purposive production of honey bee queens in Bulgaria(Plovdiv University Press “Paisii Hilendarski”, 2016-04-27) Georgieva, Vida H.; Petrov, Plamen P.; Petkov, Nikolay G.; Ivanova, Evgeniya N.The genetic polymorphism in selectively reared in Bulgaria, local honey bee populations of Apis mellifera macedonica subspecies (type rodopica), has been studied, using analysis of six enzyme systems (MDH-1, ME, EST-3, ALP, PGM and HK) corresponding to 6 loci. Totally 458 worker bees from 12 bee breeding bases for artificially inseminated queens were used for this study. All these stations are part of the National Bee Breeding Association which officially implements a National Program for sustainable beekeeping in Bulgaria. All of the six loci were found to be polymorphic. Only EST-3 locus was established as fixed in one of the investigated populations. Polymorphism with three alleles was ascertained for MDH, ME, ALP, PGM and HK loci and with five alleles for EST-3 locus. The most common alleles in almost all of the populations were MDH-1 100, ME 100, EST-3 100, PGM 100 and HK 100. Two private alleles (frequency < 0.05) were found for two of the populations. The calculated level of polymorphism was 88.33% in only one of the populations and 100% - in all others. The observed and expected heterozygosities (Ho and He) ranged from 0.157 to 0.250 and from 0.206 to 0.272, respectively. The estimated mean FST value from allozyme data was 0.035. On the bases of the allele frequencies of the studied allozyme loci the Nei’s (1972) genetic distance was estimated. It ranged between 0.002 and 0.060 among the populations studied.Item Effect of house type on growth performance, litter quality and incidence of foot lesions in broiler chickens reared in varying stocking density(Plovdiv University Press “Paisii Hilendarski”, 2016-04-26) Farhadi, Danial; Hosseini, Seyed Majid; Dezfuli, Bahareh TaheriTo evaluate the effect of different stocking density on broiler performance two repeated experiments were conducted in a conventional and an environmentally controlled house using a total of 10,232 broiler chickens. In each experiment, a total of 5,116 one-day old Ross 308 broiler chickens were randomly distributed to 16 floor pens arranged within each house at density of 16, 18, 20 or 22 birds/m2. Results showed that the broiler chickens grown in environmentally controlled house had greater (p<0.05) weight gain, kg body weight (BW)/m2, production efficiency index (PEI) and spleen relative weight at day 42 and also had lower (p<0.05) feed conversion ratio and mortality rate during 21 to 42 and 1 to 42 days of age than those reared in conventional house. Weight gain, feed intake and PEI decreased (p<0.05) and kg BW/m2 and carcass yield increased (p<0.05), when broiler density increased more than 20 birds/m2. House type and stocking density had no effect on relative weights of liver, abdominal fat, bursa of Fabricius, and litter pH and ammonia emission. However, higher litter moisture in conventional house led to a greater (p<0.05) incidence of foot pad lesions and hock burns, which intensified with increased density. In conclusion, broiler chickens reared in environmentally controlled house had superior performance, higher liability, and lower litter moisture content and foot lesions. Moreover, broiler rearing at the density of 22 birds/m2 adversely affected growth performance and foot quality, despite the greater kg BW/m2 compared to broilers grown at lower densities.Item Evaluation of genetic divergence and heritability in pea (Pisum sativum L.)(Plovdiv University Press “Paisii Hilendarski”, 2016-01-15) Georgieva, Natalia; Nikolova, Ivelina; Kosev, ValentinAn experiment on genetic evaluation of five genotypes of forage pea (Glyans, Svit, Kamerton, Modus, Pleven 4) was conducted during 2012-2014 period. Analysis of variance showed significant differences among genotypes for the traits pod width, seeds per plant, seed weight per plant and 1000 seed weight. The estimates of genetic parameters of five varieties of Pisum sativum L. indicated a good amount of genetic variation in the experimental materials under investigation. Moderate phenotypic and genotypic coefficients of variation were observed for most of traits except pod length and pod width. For the traits studied seeds per plant, seed weight per plant and plant height were found high heritability along with high genetic gain indicating preponderance of additive effects. Therefore, selection programme based on these characters would be more effective in improving yield parameters of forage pea. The seed yield was positively and significantly correlated with 1000 seed weight and pod stem, which suggested the possibilities of improving seed yield by simultaneous improvement of these traits.Item Development of microsatellite markers and their correlation with morphological and chemical markers in Withania somnifera (L.) Dunal(Plovdiv University Press “Paisii Hilendarski”, 2016-01-15) Khanna, Punit Kumar; Chandra, Ratna; Kumar, Arun; Dogra, Nidhi; Gupta, Heena; Gupta, Gourav; Verma, VijeshwarThe genetic variation and relationships among 14 Withania accessions were evaluated using morphological, chemical and Simple Sequence Repeat (SSR) markers. Wild accessions are more robust and better performing in morphological and chemical metabolite accumulation than cultivated one. The results revealed that out of fourteen, four primers showed distinct polymorphism, indicating the robust nature of microsatellites in revealing polymorphism. The banding pattern was recorded in the form of 0-1 data sheet which was analyzed using unweighted pair group method with arithmetic mean (UPGMA) based on Jaccard's similarity coefficient. The cluster analysis showed higher level of genetic variation among the accessions. Similarity coefficients ranged from 0.125 to 1. The dendrogram revealed 3 major distinct clusters. Higher range of similarity values for related genotypes using simple sequence repeats (SSR) provides greater confidence for the assessment of genetic diversity and relationships. The polymorphism information content (PIC) value for the SSR loci ranged from 0.0 to 0.40. Higher PIC values were associated with higher level of polymorphism. Results of this study showed a high degree of variation among analyzed accessions, indicating an important source of genetic diversity that can be used in future breeding programs.Item Humoral autoimmune response against specific collagen type II epitopes in Bulgarian patients with rheumatoid arthritis(Plovdiv University Press “Paisii Hilendarski”, 2016-01-15) Batsalova, Tsvetelina; Ivanova, Petya; Antova, Plamka; Dzhambazov, BalikCollagen type II (CII) is a strong candidate autoantigen for rheumatoid arthritis (RA) pathogenesis. CII is the main structural protein of synovial cartilage and it is attacked by both antibodies and T-cells during RA disease course. Experiments with mouse models have identified an immunodominant T-cell epitope from CII as well as several epitopes that are recognized by the majority of CII-specific autoantibodies. It has been shown that some epitope-specific anti-CII antibodies are arthritogenic and are associated with development of chronic arthritis. In addition, the immunodominant CII epitopes could be posttranslationally modified and these modified epitopes might be involved in induction and/or perpetuation of autoimmune humoral response and arthritic pathology. The aim of the present study was to evaluate the CII epitope-specific humoral response in a subgroup of Bulgarian patients with rheumatoid arthritis. Our results demonstrate that RA patients have significantly increased levels of anti-CII antibodies compared to healthy individuals and patients with other type of autoimmune disease. The majority of anti-CII antibodies in Bulgarian patients are directed against the U1 and J1 conserved epitopes. We show that D8 epitope-specific antibodies react to the triple-helical structure of the epitope and thus recognize both the native and the posttranslationally citrullinated D8. This is the first article presenting an evaluation of CII-specific humoral autoimmune response in Bulgarian patients with rheumatoid arthritis.Item Production, purification and characterization of thermostable α-amylase from soil isolate Bacillus sp. strain B-10(Plovdiv University Press “Paisii Hilendarski”, 2016-01-03) Singh, Ravindra Nath; Bahuguna, Ashutosh; Chauhan, Peeyush; Sharma, Vijay Kumar; Kaur, Satinder; Singh, Santosh Kumar; Khan, AmirA bacterial strain B-10 that produces α-amylase was isolated from compost and kitchen waste receiving agricultural soil. Based on microbiological and biochemical tests the isolate B-10 was identified as Bacillus sp. Alpha-amylase produced by this isolate was purified by (NH4)2SO4 precipitation and DEAE cellulose ion-exchange chromatography showing 15.91 and 48.21 fold purification, respectively. SDS-PAGE of the purified enzyme confirmed the purification and monomeric nature of the enzyme. The purified α-amylase showed maximum activity at pH 7 and temperature 50°C. The enzyme was significantly active in the temperature range of 30-60°C for the studied period of 2 h. During the incubation of purified enzyme at pH ranging from 5 to 10 for 24 h the maximum stability was observed at pH 7 followed by pH 8, whereas at extreme pH, the stability was very poor. Km and Vmax were found to be 1.4 mg/mL and 6.2 U/mL, respectively.Item Optimized production of extracellular proteases by Bacillus subtilis from degraded abattoir waste(Plovdiv University Press “Paisii Hilendarski”, 2015-12-07) Badhe, Pallavi; Joshi, Manasi; Adivarekar, RavindraProteases are ubiquitous in occurrence and are found in all living organisms. These are essential for cell growth and differentiation. The extracellular proteases are of a high commercial value and find multiple applications in various industrial sectors. The present study describes the screening of protease producing bacteria from a hitherto unexplored source i.e. degraded waste from abattoir. Three isolates were found namely yellow, white and orange coloured bacteria. Amongst them, white colored colony was found to be more suitable for protease production. The morphological, cultural, biochemical and 16S rRNA confirmed that the isolate was Bacillus subtilis. Physical and chemical parameters were optimized for maximum protease production and optimum temperature and pH was found to be 40ºC at pH 14. Glucose as a carbon source and yeast extract as a nitrogen source further stimulated the production process giving maximum protease activity to be 20.74 U/ml and 20.67 U/ml. The applications of protease in detergent and solvent industry were tested and it was revealed that the purified enzyme can be used as an additive in detergent industry.