Search results for: protein complementation assay

Authors: Yanhua Zhao, Yu Gou, Shu Feng, Dongdong Li, Chuanmin Tao

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The natural history of HBV infection could experience immune tolerant (IT), immune clearance (IC), HBeAg-negative inactive/quienscent carrier (ENQ), and HBeAg-negative hepatitis (ENH). As current biomarkers for discriminating these four phases have some weaknesses, additional serological indicators are needed. Hepatits B core-related antigen (HBcrAg) encoded with precore/core gene contains denatured HBeAg, HBV core antigen (HBcAg) and a 22KDa precore protein (p22cr), which was demonstrated to have a close association with natural history of hepatitis B infection, but no specific cutoff values and diagnostic parameters to evaluate the diagnostic efficacy. This study aimed to clarify the distribution of HBcrAg levels and evaluate its diagnostic performance during the natural history of infection from a Western Chinese perspective. 294 samples collected from treatment-naïve chronic hepatitis B (CHB) patients in different phases (IT=64; IC=72; ENQ=100, and ENH=58). We detected the HBcrAg values and analyzed the relationship between HBcrAg and HBV DNA. HBsAg and other clinical parameters were quantitatively tested. HBcrAg levels of four phases were 9.30 log U/mL, 8.80 log U/mL, 3.00 log U/mL, and 5.10 logU/mL, respectively (p < 0.0001). Receiver operating characteristic curve analysis demonstrated that the area under curves (AUCs) of HBcrAg and quantitative HBsAg at cutoff values of 9.25 log U/mL and 4.355 log IU/mL for distinguishing IT from IC phases were 0.704 and 0.694, with sensitivity 76.39% and 59.72%, specificity 53.13% and 79.69%, respectively. AUCs of HBcrAg and quantitative HBsAg at cutoff values of 4.15 log U/mlmL and 2.395 log IU/mlmL for discriminating between ENQ and ENH phases were 0.931 and 0.653, with sensitivity 87.93% and 84%, specificity 91.38% and 39%, respectively. Therefore, HBcrAg levels varied significantly among four natural phases of HBV infection. It had higher predictive performance than quantitative HBsAg for distinguishing between ENQ-patients and ENH-patients and similar performance with HBsAg for the discrimination between IT and IC phases, which indicated that HBcrAg could be a potential serological marker for CHB.

Keywords: chronic hepatitis B, hepatitis B core-related antigen, hepatitis B surface antigens, hepatitis B virus

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Authors: Sugandha Asthana, Geetika Vajpayee, Pratibha Kumari, Shanthy Sundaram

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An economically important disease of wheat in North Western region of India is Karnal Bunt caused by smut fungus Tilletia indica. This fungal pathogen spreads by air, soil and seed borne sporodia at the time of flowering, which ultimately leads to partial bunting of wheat kernels with fishy odor and taste to wheat flour. It has very serious effects due to quarantine measures which have to be applied for grain exports. Chemical fungicides such as mercurial compounds and Propiconazole applied to the control of Karnal bunt have been only partially successful. Considering the harmful effects of chemical fungicides on man as well as environment, many countries are developing biological control as the superior substitute to chemical control. Repeated use of fungicides can be responsible for the development of resistance in fungal pathogens against certain chemical compounds. The present investigation is based on the isolation and evaluation of antifungal properties of some isolated (from natural manure) and commercial bacterial strains against Tilletia indica. Total 23 bacterial isolates were obtained and antagonistic activity of all isolates and commercial bacterial strains (Bacillus subtilis MTCC8601, Bacillus pumilus MTCC 8743, Pseudomonas aeruginosa) were tested against T. indica by dual culture plate assay (pour plate and streak plate). Test for the production of antifungal volatile organic compounds (VOCs) by antagonistic bacteria was done by sealed plate method. Amongst all s1, s3, s5, and B. subtilis showed more than 80% inhibition. Production of extracellular hydrolytic enzymes such as protease, beta 1, 4 glucanase, HCN and ammonia was studied for confirmation of antifungal activity. s1, s3, s5 and B. subtilis were found to be the best for protease activity and s5 and B. subtilis for beta 1, 4 glucanase activity. Bacillus subtilis was significantly effective for HCN whereas s3, s5 and Bacillus subtilis for ammonia production. Isolates were identified as Pseudomonas aeruginosa (s1) and B. licheniformis (s3, s5) by various biochemical assays and confirmed by16s rRNA sequencing. Use of microorganisms or their secretions as biocontrol agents to avoid plant diseases is ecologically safe and may offer long term of protection to crop. The above study reports the promising effects of these strains in better pathogen free crop production and quality maintenance as well as prevention of the excessive use of synthetic fungicides.

Keywords: antagonistic, antifungal, biocontrol, Karnal bunt

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Authors: Sílvia Pereira Mello, Eliane Rodrigues, Maria de Lourdes Andrade, Marcelo Pereira, Giselle Dias, Jose Seixas Filho

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Frog meat is a highly digestible food and its use is recommended in diets aimed at fighting cholesterol, obesity, and arterial hypertension, as well as for treating gastrointestinal disorders. In this study, the soups were developed with frog meat in addition to other ingredients which did not present allergenic potential. The carcasses of the thawed frogs went through bleaching and deboning, and other ingredients (vegetables and condiments) were then added to the separated meat. After the process of cooking, the soups were cooled and later on frozen at -40° C for 3 hours and then taken to the LS 3000 B lyophilizer for 24 hours. The soups were submitted to microbiological analysis: enumeration of total coliforms and Bacillus cereus; identification of coagulase positive Staphylococcus; isolation and identification of Salmonella spp.; and physical-chemical analysis; application of micro-Kjeldahl method for protein, Soxhlet method for lipids, use of a heating chamber at 105ºC for moisture, incineration method (500-550°C) for ash, and Decagon's Pawkit equipment for determining water activity. Acceptance test was performed with 50 elderly people, all between 60 and 85 years of age. The degree of acceptance was demonstrated using a seven points structured hedonic scale in which the taster expressed their impression towards the product. Results of the microbiological analysis showed that all samples met the standards established by the National Health Surveillance Agency of Brazil (ANVISA). Results of the acceptance test indicated that all the soups were accepted considering overall impression and intended consumption. In addition to its excellent nutritional quality, the dehydrated soups made with frog meat are presented as a solution for consumers due to convenience in preparation, consumption and storage.

Keywords: bacteriological quality, lithobates catesbeianus, instant soup, proximate composition, sensory analysis

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Authors: O. I. Adeniran, M. A. Mogale

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Advanced glycation end-products (AGEs) have been implicated in the development and progression of vascular complications of diabetes mellitus and other age-related disease such as Alzheimer’s disease, heart diseases, stroke and limb amputation. The aim of the study was to determine the anti-glycation activity and AGE-cross-linking breaking ability of Sclerocarya birrea stem-bark extracts (SBSBETs). Hexane, ethyl acetate, methanol and water extracts of Sclerocarya birrea stem-bark and standard inhibitor, aminoguanidine (AG) were incubated with bovine serum albumin (BSA)-fructose mixture for 20 and 40 days. The amounts of total immunogenic AGEs (TIAGEs), fluorescent AGEs (FAGEs) and carboxymethyl lysine (CML) formed were determined and the percentage anti-glycation activity of each plant extract calculated. The ability of SBSBETs to break fructose-derived BSA-AGE-collagen cross-links was also investigated. All SBSBETs under investigation demonstrated less anti-glycation activity against TIAGE, FAGEs and CML than AG after 20 days incubation. After 40 days incubation, ethyl acetate, methanol and water SBSBETs demonstrated lower anti-glycation activity against TIAGEs than AG but exerted higher anti-glycation activity than AG against FAGEs. All SBSBETs except water demonstrated lower anti-glycation activity than AG against CML. With regard to the ability of SBSBETs to breakdown fructose-derived AGEs cross-links, the polar SBSBETs demonstrated higher ability to break AGE-cross-links than the non-polar ones. The results of this study may lead to the isolation of bio-active phyto-chemicals from SBSBETs that may be used for the prevention of vascular complication of diabetes.

Keywords: advanced glycation end-products, anti-glycation, cross-link breaking, Sclerocarrya birrea

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Authors: Nimot Opeyemi Wahab

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Poor nutritional status during adolescence is a reflection of inadequate intake of nutrients. This can also be associated with a lack of consumption of diverse food. This study assessed the nutritional status and dietary diversity score (DDS) of in-school adolescents in Ibadan North, North East, and Ibadan South West Local Government Areas (LGA) of Oyo State, Nigeria. A cross-sectional study involving 3,510 in-school adolescents from the three LGA was conducted. Nutrient intake was measured using a validated 24-hour dietary recall, and the anthropometric measurement was also taken. Dietary diversity score (DDS) was assessed using the Individual Dietary Diversity Score (WDDS) of nine food groups. Participants were between 10-19years, and the mean age was 14.76±1.68, 15.32±1.77, and 15.45±1.62 in Ibadan North, Ibadan North East, and Ibadan South West, respectively. About 48% of the participants were male (47.9%), while 52.1% were female. BMI-for-age showed that 92.1%, 5.4%, 2.1%, and 0.4% of the participants were normal, underweight, overweight, and obese, respectively. The mean energy intake (143.193±695.98) of the female respondents was more than that of the male respondents (1406.86±767.41). The macronutrients intake (protein, carbohydrates, fiber, and fats) of the female participants was also found to be more than that of the male participants, with a non-significant difference of 0.336, 0.530, 0.234, and 0.069 (at p< 0.05). Out of all the vitamin intake, only vitamin C was found to be statistically different (p=0.038) at p<0.05 between the male and female respondents. Of all the mineral intake, only phosphorus showed a higher intake (575.20±362.12) among female respondents than the male respondents. The mean DDS of all participants was 4.59±0.939. The majority of the participants, 1183 (80.9%), were within the medium DDS category, 9.9% were low, while 1.5% were in the high category: of which males were 474 (71.5%) and females were 709 (88.6%). Participants from Ibadan North were 941(88.5%), and those from South West were 242(60.5%). A non-significant difference in the mean score of participants from the two locations (p=0.467) was also found. A negative correlation exists between DDS and BMI-for age (-0.11), DDS, and energy intake (-0.46) in Ibadan North and South West LGA. The nutritional status of in-school adolescents was normal, and DDS was within the medium category. Nutrition intervention regarding the consumption of diverse food is necessary among adolescents.

Keywords: nutritional status, dietary diversity, adolescents, nutrient intake

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Authors: Shazia Anwer Bukhari, Madiha Javeed Ghani, Muhammad Ibrahim Rajoka

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Objective: Biochemical, environmental, physical and genetic factors have a strong effect on the development of coronary disease (CAD). Plasma homocysteine (Hcy) level and DNA damage play a pivotal role in its development and progression. The aim of this study was to investigate the predictive strength of an oxidative stress, clinical biomarkers and total antioxidant status (TAS) in CAD patients to find the correlation of homocysteine, TOS and oxidative DNA damage with other clinical parameters. Methods: Sixty confirmed patients with CAD and 60 healthy individuals as control were included in this study. Different clinical and laboratory parameters were studied in blood samples obtained from patients and control subjects using commercially available biochemical kits and statistical software Results: As compared to healthy individuals, CAD patients had significantly higher concentrations of indices of oxidative stress: homocysteine (P=0.0001), total oxidative stress (TOS) (P=0.0001), serum cholesterol (P=0.04), low density lipoprotein cholesterol (LDL) (P=0.01), high density lipoprotein-cholesterol (HDL) (P=0.0001), and malondialdehyde (MDA) (P=0.001) than those of healthy individuals. Plasma homocysteine level and oxidative DNA damage were positively correlated with cholesterol, triglycerides, systolic blood pressure, urea, total protein and albumin (P values= 0.05). Both Hcy and oxidative DNA damage were negatively correlated with TAS and proteins. Conclusion: Coronary artery disease patients had a significant increase in homocysteine level and DNA damage due to increased oxidative stress. In conclusion, our study shows a significantly increase in lipid peroxidation, TOS, homocysteine and DNA damage in the erythrocytes of patients with CAD. A significant decrease level of HDL-C and TAS was observed only in CAD patients. Therefore these biomarkers may be useful diagnosis of patients with CAD and play an important role in the pathogenesis of CAD.

Keywords: antioxidants, coronary artery disease, DNA damage, homocysteine, oxidative stress, malondialdehyde, 8-Hydroxy-2’deoxyguanosine

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Authors: Carmiña L. Vargas-Zapata, María A. Conde-Sarmiento, Maria Consuelo Maestre-Vargas

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Calcium is an essential element for good growth and development of the organism, and its requirement is increased at school age. Low socio-economic populations of developing countries such as Colombia may have food deficiency of this mineral in schoolchildren that could be reflected in calcium biochemical indicators, bone alterations and anthropometric indicators. The objective of this investigation was to evaluate some calcium biochemical indicators in a group of schoolchildren of low socioeconomic level from Barranquilla city and to correlate with body mass index. 60 schoolchildren aged 7 to 15 years were selected from Jesus’s Heart Educational Institution in Barranquilla-Atlántico, apparently healthy, without suffering from infectious or gastrointestinal diseases, without habits of drinking alcohol or smoking another hallucinogenic substance and without taking supplementation with calcium in the last six months or another substance that compromises bone metabolism. The research was approved by the ethics committee at Universidad del Atlántico. The selected children were invited to donate a blood and urine sample in a fasting time of 12 hours, the serum was separated by centrifugation and frozen at ˗20 ℃ until analyzed and the same was done with the urine sample. On the day of the biological collections, the weight and height of the students were measured to determine the nutritional status by BMI using the WHO tables. Calcium concentrations in serum and urine (SCa, UCa), alkaline phosphatase activity total and of bone origin (SAPT, SBAP) and urinary creatinine (UCr) were determined by spectrophotometric methods using commercial kits. Osteocalcin and Cross-linked N-telopeptides of type I collagen (NTx-1) in serum were measured with an enzyme-linked inmunosorbent assay. For statistical analysis the Statgraphics software Centurium XVII was used. 63% (n = 38) and 37% (n = 22) of the participants were male and female, respectively. 78% (n = 47), 5% (n = 3) and 17% (n = 10) had a normal, malnutrition and high nutritional status, respectively. The averages of evaluated indicators levels were (mean ± SD): 9.50 ± 1.06 mg/dL for SCa; 181.3 ± 64.3 U/L for SAPT, 143.8 ± 73.9 U/L for SBAP; 9.0 ± 3.48 ng/mL for osteocalcin and 101.3 ± 12.8 ng/mL for NTx-1. UCa level was 12.8 ± 7.7 mg/dL that adjusted with creatinine ranged from 0.005 to 0.395 mg/mg. Considering serum calcium values, approximately 7% of school children were hypocalcemic, 16% hypercalcemic and 77% normocalcemic. The indicators evaluated did not correlate with the BMI. Low values ​​were observed in calcium urinary excretion and high in NTx-1, suggesting that mechanisms such as increase in renal retention of calcium and in bone remodeling may be contributing to calcium homeostasis.

Keywords: calcium, calcium biochemical, indicators, school children, low socioeconomic status

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Authors: Elif Tugce Aksun Tumerkan

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Although seafood is an important part of human diet and categorized highly traded food industry internationally, it is remain overlooked generally in the global food security aspect. Food product authentication is the main interest in the aim of both avoids commercial fraud and to consider the risks that might be harmful to human health safety. In recent years, with increasing consumer demand for regarding food content and it's transparency, there are some instrumental analyses emerging for determining food fraud depend on some analytical methodologies such as proteomic and metabolomics. While, fish and seafood consumed as fresh previously, within advanced technology, processed or packaged seafood consumption have increased. After processing or packaging seafood, morphological identification is impossible when some of the external features have been removed. The main fish and seafood quality-related issues are the authentications of seafood contents such as mislabelling products which may be contaminated and replacement partly or completely, by lower quality or cheaper ones. For all mentioned reasons, truthful consistent and easily applicable analytical methods are needed for assurance the correct labelling and verifying of seafood products. DNA-barcoding methods become popular robust that used in taxonomic research for endangered or cryptic species in recent years; they are used for determining food traceability also. In this review, when comparing the other proteomic and metabolic analysis, DNA-based methods are allowing a chance to identification all type of food even as raw, spiced and processed products. This privilege caused by DNA is a comparatively stable molecule than protein and other molecules. Furthermore showing variations in sequence based on different species and founding in all organisms, make DNA-based analysis more preferable. This review was performed to clarify the main advantages of using DNA-barcoding for determining seafood fraud among other techniques.

Keywords: DNA-barcoding, genetic analysis, food fraud, mislabelling, packaged seafood

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Authors: Hany M. Fayed, Rehab F. Abdel-Rahman, Alyaa F. Hessin, Hanan A. Ogaly, Gihan F. Asaad, Abeer A. A. Salama, Sahar Abdelrahman, Mahmoud S. Arbid, Marwan Abd Elbaset Mohamed

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Liver fibrosis is a serious global health problem that occurs as a result of a variety of chronic liver disorders. Apigenin, a flavonoid found in many plants, has several pharmacological properties. The aim of this study was to evaluate the antifibrotic efficacy of apigenin (APG) against experimentally induced hepatic fibrosis in rats via using thioacetamide (TAA) and to explore the possible underlying mechanisms. TAA (100 mg/kg, i.p.) was given three times each week for two weeks to induce liver fibrosis. After TAA injections, APG was given orally (5 and 10 mg/kg) daily for two weeks. Biochemical, molecular, histological and immunohistochemical analyses were performed on blood and liver tissue samples. The functioning of the liver, oxidative stress, inflammation, and liver fibrosis indicators were all evaluated. The findings showed that TAA markedly increased the activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), as well as the levels of malondialdehyde (MDA), focal adhesion kinase (FAK), hypoxia-inducible factor-1 (HIF-1), nuclear factor-κB (NF-κB), transforming growth factor-beta (TGF-β), tumor necrosis factor-alpha (TNF-α) and interleukin-1β (IL-1β) with a reduction in albumin, total protein, A/G ratio, GSH content and interleukin-10 (IL-10). Moreover, TAA elevated the content of collagen I, α -smooth muscle actin (α-SMA), and hydroxyproline in the liver. The treatment with APG in a dose-dependent manner has obviously prevented these alterations and amended the harmful effects induced by TAA. The histopathological and immunohistochemical observations supported this biochemical evidence. The higher dose of APG produced the most significant antifibrotic effect. As a result of these data, APG appears to be a promising antifibrotic drug and could be used as a new herbal medication or dietary supplement in the future for the treatment of liver fibrosis. This effect might be related to the inhibition of the HIF-1/FAK signaling pathway.

Keywords: apigenin, FAK, HIF-1, liver fibrosis, rat, thioacetamide

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Authors: Rajendra Jangde, Deependra Singh

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Present work was based with objective to release of the antimicrobial and debriding agent in sustained manner at the wound surface. In order to provide a long-lasting antimicrobial action and moist environment on wound space, Biocompatible moist system was developed for complete healing. In the present study, a biocompatible moist system of PVA-gelatin hydrogel was developed capable of carrying multiple drugs- Quercetin and Cabopol in controlled manner for effective and complete wound healing. Carbopol and Quercetin were prepared by thin film hydration techniques and optimized system was incorporated in PVA-Gelatin slurry. PVA-Gelatin hydrogels were prepared by freeze thaw method. The prepared dispersion was casted into films to prepare multiphase hydrogel system and characterized by in vitro and in vivo studies. Results revealed the uniform dispersion of microspheres in a three-dimensional matrix of the PVA-Gelatin hydrogel observed at different magnifications. The in vitro release data showed typical biphasic release pattern, i.e., a burst release followed by a slower sustained release for 5 days. Prepared system was found to be stable under both normal and accelerated conditions. Histopathological study showed significant (p<0.05) increase in fibroblast cells, collagen fibres and blood vessels formation. All parameters such as wound contraction, tensile strength, histopathological and biochemical parameters- hydroxyproline content, protein level, etc. were observed significant (p<0.05) in comparison to control group. Present results suggest an accelerated re-epithelialization under moist wound environment with delivery of multiple drugs effective at different stages of wound healing cascade with minimum disturbance of wound bed.

Keywords: multiphase hydrogel, optimization quercetin, wound healing

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Authors: Liliane Samara Ferreira Leite, Francys Kley Vieira Moreira, Luiz Henrique Capparelli Mattoso

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The increasing environmental concern regarding the plastic pollution worldwide has stimulated the development of low-cost biodegradable materials. Proteins are renewable feedstocks that could be used to produce biodegradable plastics. Gelatin, for example, is a cheap film-forming protein extracted from animal skin and connective tissues of Brazilian Livestock residues; thus it has a good potential in low-cost biodegradable plastic production. However, gelatin plastics are limited in terms of mechanical and barrier properties. Cellulose nanocrystals (CNC) are efficient nanofillers that have been used to extend physical properties of polymers. This work was aimed at evaluating the reinforcing efficiency of CNC on gelatin films. Specifically, we have employed the continuous casting as the processing method for obtaining the gelatin/CNC bionanocomposites. This required a first rheological study for assessing the effect of gelatin-CNC and CNC-CNC interactions on the colloidal state of the aqueous bionanocomposite formulations. CNC were isolated from eucalyptus pulp by sulfuric acid hydrolysis (65 wt%) at 55 °C for 30 min. Gelatin was solubilized in ultra-pure water at 85°C for 20 min and then mixed with glycerol at 20 wt.% and CNC at 0.5 wt%, 1.0 wt% and 2.5 wt%. Rotational measurements were performed to determine linear viscosity (η) of bionanocomposite solutions, which increased with increasing CNC content. At 2.5 wt% CNC, η increased by 118% regarding the neat gelatin solution, which was ascribed to percolation CNC network formation. Storage modulus (G’) and loss modulus (G″) further determined by oscillatory tests revealed that a gel-like behavior was dominant in the bionanocomposite solutions (G’ > G’’) over a broad range of temperature (20 – 85 °C), particularly at 2.5 wt% CNC. These results confirm effective interactions in the aqueous gelatin-CNC bionanocomposites that could substantially increase the physical properties of the gelatin plastics. Tensile tests are underway to confirm this hypothesis. The authors would like to thank the Fapesp (process n 2016/03080-3) for support.

Keywords: bionanocomposites, cellulose nanocrystals, gelatin, viscoelastic characterization

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Authors: S. A. N. Rashani, S. Pigera, P. N. J. Fernando, S. Jayawickema, M. A. Niriella, A. P. De Silva

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Dietary supplement (DS) consumption is drastically trending among the young athlete generation in developing countries. Many athletes try to fulfill their nutrition requirements using dietary supplements without knowing their effects on health and performance. This study aimed to assess the DS usage patterns of university athletes in Sri Lanka. A self-administered questionnaire was employed to collect data from state university students representing a university team, and a sample of 200 respondents was selected based on a stratified random sampling technique. Incomplete questionnaires were omitted from the analysis. The data were analyzed using IBM SPSS statistics for Windows version 25. The level of significance was set at p<0.05 in the data analysis. The prevalence of DS was 48.2% (n= 94), with no significant association between gender and DS intake. Protein (15.9%), vitamin (14.9%), sports drinks (12.8%), and creatine (8.2%) were the most consumed DS by students. Weightlifting (85.0%), football (62.5%), rugby (57.7%), and wrestling (40.9%) players showed higher DS usage among other sports. Coaches were reported as the most frequent person who was advised to use DS (43.0%). Students who won interuniversity games showed significantly low DS intake (p = 0.002) compared to others. Interestingly, DS use was significantly affected by the season of use (p = 0.000), pointing out that during competition and training seasons (62.4%) was the most frequent use. The pharmacy (27.0%) was the commonest place to buy DS. Students who used nutrient-dense meal plans during the training and competition period still showed a 61.0% tendency to consume DS. Most claimed reason to use DS was to increase energy and strength (29.0%). A majority reported that they used DS for less than one month (35.5%), while the second-highest duration was over three years (17.2%). Considering body mass index (BMI), healthy weight students showed 71.0% DS prevalence. DS prevalence was moderate among Sri Lankan university students, highlighting that the highest DS use was during competition and training seasons. Moreover, it emphasizes the need for nutrition and anti-doping counseling in the Sri Lankan university system.

Keywords: athlete, dietary, supplements, university

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Authors: Wasim Sajjad, Manzoor Ahmad, Sundas Qadir, Muhammad Rafiq, Fariha Hasan, Richard Tehan, Kerry L. McPhail, Aamer Ali Shah

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Aim: This study aims to investigate the possible radiation protective role of a compatible solute in the tolerance of radio-halophilic bacterium against stresses, like desiccation and exposure to ionizing radiation. Methods and Results: Nine different radio-resistant bacteria were isolated from desert soil, where strain WMA-LM19 was chosen for detailed studies on the basis of its high tolerance for ultraviolet radiation among all these isolates. 16S rRNA gene sequencing indicated that the bacterium was closely related to Stenotrophomonas sp. (KT008383). A bacterial milking strategy was applied for extraction of intracellular compatible solutes in 70% (v/v) ethanol, which were purified by high-performance liquid chromatography (HPLC). The compound was characterized as ectoine by 1H and 13C nuclear magnetic resonance (NMR), and mass spectrometry (MS). Ectoine demonstrated more efficient preventive activity (54.80%) to erythrocyte membranes and also inhibited oxidative damage to proteins and lipids in comparison to the standard ascorbic acid. Furthermore, a high level of ectoine-mediated protection of bovine serum albumin against ionizing radiation (1500-2000 Jm-2) was observed, as indicated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Conclusion: The results indicated that ectoine can be used as a potential mitigator and radio-protective agent to overcome radiation- and salinity-mediated oxidative damage in extreme environments. Significance and Impact of the Study: This study shows that ectoine from radio-halophiles can be used as a potential source in topical creams as sunscreen. The investigation of ectoine as UV protectant also changes the prospective that radiation resistance is specific only to molecular adaptation.

Keywords: ectoine, anti-oxidant, stenotrophomonas sp., ultraviolet radiation

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Authors: Ying Li, Rui Hu, Shizhen Chen, Xin Zhou, Yunhuang Yang

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Prostate cancer is one of the leading causes of cancer-related death among men. Prostate-specific antigen (PSA), a specific product of prostatic epithelial cells, is an important indicator of prostate cancer. Though PSA is not a specific serum biomarker for the screening of prostate cancer, it is recognized as an indicator for prostate cancer recurrence and response to therapy for patient’s post-prostatectomy. Since radical prostatectomy eliminates the source of PSA production, serum PSA levels fall below 50 pg/mL, and may be below the detection limit of clinical immunoassays (current clinical immunoassay lower limit of detection is around 10 pg/mL). Many clinical studies have shown that intervention at low PSA levels was able to improve patient outcomes significantly. Therefore, ultra-sensitive and precise assays that can accurately quantify extremely low levels of PSA (below 1-10 pg/mL) will facilitate the assessment of patients for the possibility of early adjuvant or salvage treatment. Currently, the commercially available ultra-sensitive ELISA kit (not used clinically) can only reach a detection limit of 3-10 pg/mL. Other platforms developed by different research groups could achieve a detection limit as low as 0.33 pg/mL, but they relied on sophisticated instruments to get the final readout. Herein we report a microfluidic platform for point-of-care (POC) detection of PSA with a detection limit of 0.5 pg/mL and without the assistance of any equipment. This platform is based on a previously reported volumetric-bar-chart chip (V-Chip), which applies platinum nanoparticles (PtNPs) as the ELISA probe to convert the biomarker concentration to the volume of oxygen gas that further pushes the red ink to form a visualized bar-chart. The length of each bar is used to quantify the biomarker concentration of each sample. We devised a long reading channel V-Chip (LV-Chip) in this work to achieve a wide detection window. In addition, LV-Chip employed a unique enzyme-free ELISA probe that enriched PtNPs significantly and owned 500-fold enhanced catalytic ability over that of previous V-Chip, resulting in a significantly improved detection limit. LV-Chip is able to complete a PSA assay for five samples in 20 min. The device was applied to detect PSA in 50 patient serum samples, and the on-chip results demonstrated good correlation with conventional immunoassay. In addition, the PSA levels in finger-prick whole blood samples from healthy volunteers were successfully measured on the device. This completely stand-alone LV-Chip platform enables convenient POC testing for patient follow-up in the physician’s office and is also useful in resource-constrained settings.

Keywords: point-of-care detection, microfluidics, PSA, ultra-sensitive

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Authors: Valeria Velasco, Ana M. Bonilla, José L. Vergara, Alcides Lofa, Jorge Campos, Pedro Rojas-García

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Staphylococcus aureus is both commensal bacterium and opportunistic pathogen that can cause different diseases in humans and can rapidly develop antimicrobial resistance. Since this bacterium has the ability to colonize the nares and skin of humans and animals, there is a risk of contamination of food in different steps of the food chain supply. Emerging strains have been detected in food-producing animals and meat, such as methicillin-resistant S. aureus (MRSA). The aim of this study was to determine the prevalence and oxacillin susceptibility of S. aureus in the pork chain supply in Chile and to suggest some natural antimicrobials for control. A total of 487 samples were collected from pigs (n=332), carcasses (n=85), and retail pork meat (n=70). Presumptive S. aureus colonies were isolated by selective enrichment and culture media. The confirmation was carried out by biochemical testing (Api® Staph) and molecular technique PCR (detection of nuc and mecA genes, associated with S. aureus and methicillin resistance, respectively). The oxacillin (β-lactam antibiotic that replaced methicillin) susceptibility was assessed by minimum inhibitory concentration (MIC) using the Epsilometer test (Etest). A preliminary assay was carried out to test thymol, carvacrol, oregano essential oil (Origanum vulgare L.), Maqui or Chilean wineberry extract (Aristotelia chilensis (Mol.) Stuntz) as anti-staphylococcal agents using the disc diffusion method at different concentrations. The overall prevalence of S. aureus in the pork chain supply reached 33.9%. A higher prevalence of S. aureus was determined in carcasses (56.5%) than in pigs (28.3%) and pork meat (32.9%) (P ≤ 0.05). The prevalence of S. aureus in pigs sampled at farms (40.6%) was higher than in pigs sampled at slaughterhouses (23.3%) (P ≤ 0.05). The contamination of no packaged meat with S. aureus (43.1%) was higher than in packaged meat (5.3%) (P ≤ 0.05). The mecA gene was not detected in S. aureus strains isolated in this study. Two S. aureus strains exhibited oxacillin resistance (MIC ≥ 4µg/mL). Anti-staphylococcal activity was detected in solutions of thymol, carvacrol, and oregano essential oil at all concentrations tested. No anti-staphylococcal activity was detected in Maqui extract. Finally, S. aureus is present in the pork chain supply in Chile. Although the mecA gene was not detected, oxacillin resistance was found in S. aureus and could be attributed to another resistance mechanism. Thymol, carvacrol, and oregano essential oil could be used as anti-staphylococcal agents at low concentrations. Research project Fondecyt No. 11140379.

Keywords: antimicrobials, mecA gen, nuc gen, oxacillin susceptibility, pork meat

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Authors: A. Natsir, M. Nadir, S. Syahrir, A. Mujnisa

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In tropical countries such as in Indonesia, rice straw plays an important role in fulfilling the needs of feed for ruminant, especially during the dry season in which the availability of forage is very limited. However, the main problem of using rice straw as a feedstuff is low digestibility due to the existence of the links between lignin and cellulose or hemicellulose, and imbalance of its minerals content. One alternative to solve this problem is by application of biodecomposer (BS) derived from rumen bacterial of the ruminant. This study was designed to assess the effects of BS application on the changes of the chemical composition of rice straw. Four adults local buffalo raised under typical feeding conditions were used as a source of inoculum for BS development. The animal was fed for a month with a diet consisted of rice straw and elephant grass before taking rumen fluid samples. Samples of rumen fluid were inoculated in the carboxymethyl cellulose (CMC) media under anaerobic condition for 48 hours at 37°C. The mixture of CMC media and microbes are ready to be used as a biodecomposer following incubation of the mixture under anaerobic condition for 7 days at 45°C. The effectiveness of BS then assessed by applying the BS on the straw according to completely randomized design consisted of four treatments and three replication. One hundred g of ground coarse rice straw was used as the substrate. The BS was applied to the rice straw substrate with the following composition: Rice straw without BS (P0), rice straw + 5% BS (P1), rice straw +10% BS (P2), and rice straw + 15% BS. The mixture of rice straw and BS then fermented under anaerobic for four weeks. Following the fermentation, the chemical composition of rice straw was evaluated. The results indicated that the crude protein content of rice straw significantly increased (P < 0.05) as the level of BS increased. On the other hand, the concentration of crude fiber of the rice straw was significantly decreased (P < 0.05) as the level of BS increased. Other nutrients such as minerals did not change (P > 0.05) due to the treatments. In conclusion, application of BS developed from rumen bacterial of buffalo has a promising prospect to be used as a biological agent to improve the quality of rice straw as feeding for ruminant.

Keywords: biodecomposer, local buffalo, rumen microbial, chemical composition

Procedia PDF Downloads 164

Authors: S. E. Mohmad-Saberi, W. Song, N. Oliver, M. Adrian, T.C. Hsu, A. Darbyshire

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Biodegradable polyurethane (PU) has emerged as a potential material to promote repair and regeneration of damaged/diseased tissues in heart valve regeneration due to its excellent biomechanical profile. Understanding the effects of sterilization on their properties is vital since they are more sensitive and more critical of porous structures compared to bulk ones. In this study, the effects of low concentration of hydrogen peroxide (H₂O₂) solution sterilization has been investigated to determine whether the procedure would be efficient and non-destructive to porous three-dimensional (3D) elastomeric nanocomposite, polyhedral oligomeric silsesquioxane-terminated poly (ethylene-diethylene glycol succinate-sebacate) urea-urethane (POSS-EDSS-PU) scaffold. All the samples were tested for sterility following sterilization using phosphate buffer saline (PBS) as control and 5 % v/v H₂O₂ solution. The samples were incubated in tryptic soy broth for the cultivation of microorganisms under agitation at 37˚C for 72 hours. The effects of the 5 % v/v H₂O₂ solution sterilization were evaluated in terms of morphology, chemical and mechanical properties using scanning electron microscopy (SEM), Fourier transform infrared (FTIR) and tensile tester apparatus. Toxicity effects of the 5 % v/v H₂O₂ solution decontamination were studied by in vitro cytotoxicity test, where the cellular responses of human dermal fibroblast (HDF) were examined. A clear, uncontaminated broth using 5 % v/v H₂O₂ solution method indicated efficient sterilization after 3 days, while the non-sterilized control shows clouding broth indicated contamination. The morphology of 3D POSS-EDSS-PU scaffold appeared to have similar morphology after sterilization with 5 % v/v H₂O₂ solution regarding of pore size and surface. FTIR results show that the sterilized samples and non-sterilized control share the same spectra pattern, confirming no significant alterations over the surface chemistry. For the mechanical properties of the H₂O₂ solution-treated scaffolds, the tensile strain was not significantly decreased, however, become significantly stiffer after the sterilization. No cytotoxic effects were observed after the 5 % v/v H₂O₂ solution sterilization as confirmed by cell viability assessed by Alamar Blue assay. The results suggest that low concentration of 5 % v/v hydrogen peroxide solution can be used as an alternative method for sterilizing biodegradable 3D porous scaffold with micro/nano-architecture without structural deformation. This study provides the understanding of the sterilization effects on biomechanical profile and cell proliferation of 3D POSS-EDSS-PU scaffolds.

Keywords: biodegradable, hydrogen peroxide solution, POSS-EDSS-PU, sterilization

Procedia PDF Downloads 130

Authors: Yik Sin Chan, Bee Ling Chuah, Wei Quan Chan, Ri Jin Cheng, Yan Hang Oon, Kong Soo Khoo, Nam Weng Sit

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Traditionally, medicinal plants have been used to treat different kinds of ailments including infectious diseases. They serve as a good source of lead compounds for the development of new and safer anti-infective agents. This study aimed to investigate the antimicrobial potential of the leaves of three medicinal plants, namely Catunaregam spinosa (Rubiaceae; Mountain pomegranate), Houttuynia cordata (Saururaceae; "fishy-smell herb") and Rhapis excelsa (Arecaceae; “broadleaf lady palm”). The leaves extracts were obtained by sequential extraction using hexane, chloroform, ethyl acetate, ethanol, methanol and water. The antibacterial and antifungal activities were assessed using a colorimetric broth microdilution method against a panel of human pathogenic bacteria (Gram-positive: Bacillus cereus and Staphylococcus aureus; Gram-negative: Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa) and fungi (yeasts: Candida albicans, Candida parapsilosis and Cryptococcus neoformans; Moulds: Aspergillus fumigatus and Trichophyton mentagrophytes) respectively; while antiviral activity was evaluated against the Chikungunya virus on monkey kidney epithelial (Vero) cells by neutral red uptake assay. All the plant extracts showed bacteriostatic activity, however, only 72% of the extracts (13/18) were found to have bactericidal activity. The lowest minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were given by the hexane extract of C. spinosa against S. aureus with the values of 0.16 and 0.31 mg/mL respectively. All the extracts also possessed fungistatic activity. Only the hexane, chloroform and ethyl acetate extracts of H. cordata exerted inhibitory activity against A. fumigatus, giving the lowest fungal susceptibility index of 16.7%. In contrast, only 61% of the extracts (11/18) showed fungicidal activity. The ethanol extract of R. excelsa exhibited the strongest fungicidal activity against C. albicans, C. parapsilosis and T. mentagrophytes with minimum fungicidal concentration (MFC) values of 0.04–0.08 mg/mL, in addition to its methanol extract against T. mentagrophytes (MFC=0.02 mg/mL). For anti-Chikungunya virus activity, only chloroform and ethyl acetate extracts of R. excelsa showed significant antiviral activity with 50% effective concentrations (EC50) of 29.9 and 78.1 g/mL respectively. Extracts of R. excelsa warrant further investigations into their active principles responsible for antifungal and antiviral properties.

Keywords: bactericidal, Chikungunya virus, extraction, fungicidal

Procedia PDF Downloads 370

Authors: Christelle E. Chua, Alicia L. Ho

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The development of a panel of differential gene expression signatures has been of interest in the field of biomarker discovery for radiation exposure. In the absence of the availability of exposed human subjects, lymphocyte cell lines have often been used as a surrogate to human whole blood, when performing ex vivo irradiation studies. The extent of variation between different lymphocyte cell lines is currently unclear, especially with regard to the expression of extracellular miRNA. This study compares the expression profile of extracellular miRNA isolated from different lymphocyte cell lines. It also compares the profile of miRNA obtained when different exosome isolation kits are used. Lymphocyte cell lines were created using lymphocytes isolated from healthy adult males of similar racial descent (Chinese American and Chinese Singaporean) and immortalised with Epstein-Barr virus. The cell lines were cultured in exosome-free cell culture media for 72h and the cell culture supernatant was removed for exosome isolation. Two exosome isolation kits were used. Total exosome isolation reagent (TEIR, ThermoFisher) is a polyethylene glycol (PEG)-based exosome precipitation kit, while ExoSpin (ES, Cell Guidance Systems) is a PEG-based exosome precipitation kit that includes an additional size exclusion chromatography step. miRNA from the isolated exosomes were isolated using miRNEASY minikit (Qiagen) and analysed using nCounter miRNA assay (Nanostring). Principal component analysis (PCA) results suggested that the overall extracellular miRNA expression profile differed between the lymphocyte cell line originating from the Chinese American donor and the cell line originating from the Chinese Singaporean donor. As the gender, age and racial origins of both donors are similar, this may suggest that there are other genetic or epigenetic differences that account for the variation in extracellular miRNA gene expression in lymphocyte cell lines. However, statistical analysis showed that only 3 miRNA genes had a fold difference > 2 at p < 0.05, suggesting that the differences may not be of that great a significance as to impact overall conclusions drawn from different cell lines. Subsequent analysis using cell lines from other donors will give further insight into the reproducibility of results when difference cell lines are used. PCA results also suggested that the method of exosome isolation impacted the expression profile. 107 miRNA had a fold difference > 2 at p < 0.05. This suggests that the inclusion of an additional size exclusion chromatography step altered the subset of the extracellular vesicles that were isolated. In conclusion, these results suggest that extracellular miRNA can be isolated and analysed from exosomes derived from lymphocyte cell lines. However, care must be taken in the choice of cell line and method of exosome isolation used.

Keywords: biomarker, extracellular miRNA, isolation methods, lymphocyte cell line

Procedia PDF Downloads 166

Authors: Yan Li, Qingxiang Meng, Bo Zhou, Zhenming Zhou

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The objective of this study was to compare the effects of ensiled mulberry leaves (EML), and sun-dried mulberry fruit pomace (SMFP) on fecal bacterial communities in Simmental crossbred finishing steers fed the following 3 diets: a standard TMR diet, standard diet containing EML and standard diet containing SMFP, and the diets had similar protein and energy levels. Bacterial communities in the fecal content were analyzed using Illumina Miseq sequencing of the V4 region of the 16S rRNA gene amplification. Quantitative real-time PCR was used to detect the selected bacterial species in the feces. Most of the sequences were assigned to phyla Firmicutes (56.67%) and Bacteroidetes(35.90%), followed by Proteobacteria(1.86%), Verrucomicrobia(1.80%) and Tenericutes(1.37%). And the predominant genera included the 5-7N15 (5.91%), CF231 (2.49%), Oscillospira (2.33%), Paludibacter (1.23%) and Akkermansia(1.11%). As for the treatments, no significant differences were observed in Firmicutes (p = 0.28), Bacteroidetes (p = 0.63), Proteobacteria (p = 0.46), Verrucomicrobia (p = 0.17) and Tenericutes (p = 0.75). On the genus level, classified genera with high abundance (more than 0.1%) mainly came from two phyla: Bacteroidetes and Firmicutes. Also no differences were observed in most genera level, 5-7N15 (p = 0.21), CF231 (p = 0.62), Oscillospira (p = 0.9), Paludibacter (p = 0.33) and Akkermansia (p = 0.37), except that rc4-4 were lower in the CON and SMFP groups compared to the EML animals (p = 0.02). Additionally, there were no differences in richness estimate and diversity indices (p > 0.16), and treatments had no significant effect on most selected bacterial species in the fecal (p > 0.06), except that Ruminococcus albus were higher in the EML group (p < 0.01) and Streptococcus bovis were lower in the CON group (p < 0.01). In conclusion, diets supplemented with EML and SMFP have little influence on fecal bacterial community composition in finishing steers.

Keywords: fecal bacteria community composition, sequencing, ensiled mulberry leaves (EML), sun-dried mulberry fruit pomace (SMFP)

Procedia PDF Downloads 287

Authors: Mariam G. Eshak, Ahmed Abbas, M. I. El-Sabry, M. M. Mashaly

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This investigation was conducted to determine the physiological response and evaluate the expression of inflammatory and cell death genes and DNA damage induced by endotoxic shock in laying hens. Endotoxic shock was induced by a single intravenous injection of 107 Escherichia coli (E. coli,) colony/hen. In the present study, 240 forty-week-old laying hens (H&N) were randomly assigned into 2 groups with 3 replicates of 40 birds each. Hens were reared in battery cages with wire floors in an open-sided housing system under natural conditions. Housing and general management practices were similar for all groups. At 42-wk of age, 45 hens from the first group (15 replicate) were infected with E. coli, while the same number of hens from the second group was injected with saline and served as a control. Heat shock protein-70 (HSP-70) expression, plasma corticosterone concentration, body temperature, and the gene expression of bax, caspase-3 activity, P38, Interlukin-1β (Il-1β), and tumor necrosis factor alpha (TNF-α) genes and DNA damage in the brain and liver were measured. Hens treated with E. coli showed significant (P≤0.05) increase of body temperature by 1.2 ᴼC and plasma corticosterone by 3 folds compared to the controls. Further, hens injected with E.Coli showed markedly over-expression of HSP-70 and increase DNA damage in brain and liver. These results were synchronized with activating cell death program since our data showed significant (P≤0.05) high expression of bax and caspase-3 activity genes in the brain and liver. These results were related to remarkable over-inflammation gene expression of P38, IL-1β, and TNF-α in brain and liver. In conclusion, our results indicate that endotoxic shock induces inflammatory physiological response and triggers cell death program by promoting P38, IL-1β, and TNF-α gene expression in the brain and liver.

Keywords: chicken, DNA damage, Escherichia coli, gene expression, inflammation

Procedia PDF Downloads 317

Authors: Afroza Parvin, Md. Mahmudul Hasan, Md. Rokunozzaman, Papon Debnath

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The effluents of textile industries have considerable amounts of heavy metals, causing potential microbial metal loads if discharged into the environment without treatment. Aim: In this present study, both lactose and non-lactose fermenting bacterial isolates were isolated from textile industrial effluents of a specific region of Bangladesh, named Savar, to compare and understand the load of heavy metals in these microorganisms determining the effects of heavy metal resistance properties on antibiotic resistance. Methods: Five different textile industrial canals of Savar were selected, and effluent samples were collected in 2016 between June to August. Total bacterial colony (TBC) was counted for day 1 to day 5 for 10-6 dilution of samples to 10-10 dilution. All the isolates were isolated and selected using 4 differential media, and tested for the determination of minimum inhibitory concentration (MIC) of heavy metals and antibiotic susceptibility test with plate assay method and modified Kirby-Bauer disc diffusion method, respectively. To detect the combined effect of heavy metals and antibiotics, a binary exposure experiment was performed, and to understand the plasmid profiling plasmid DNA was extracted by alkaline lysis method of some selective isolates. Results: Most of the cases, the colony forming units (CFU) per plate for 50 ul diluted sample were uncountable at 10-6 dilution, however, countable for 10-10 dilution and it didn’t vary much from canal to canal. A total of 50 Shigella, 50 Salmonella, and 100 E.coli (Escherichia coli) like bacterial isolates were selected for this study where the MIC was less than or equal to 0.6 mM for 100% Shigella and Salmonella like isolates, however, only 3% E. coli like isolates had the same MIC for nickel (Ni). The MIC for chromium (Cr) was less than or equal to 2.0 mM for 16% Shigella, 20% Salmonella, and 17% E. coli like isolates. Around 60% of both Shigella and Salmonella, but only 20% of E.coli like isolates had a MIC of less than or equal to 1.2 mM for lead (Pb). The most prevalent resistant pattern for azithromycin (AZM) for Shigella and Salmonella like isolates was found 38% and 48%, respectively; however, for E.coli like isolates, the highest pattern (36%) was found for sulfamethoxazole-trimethoprim (SXT). In the binary exposure experiment, antibiotic zone of inhibition was mostly increased in the presence of heavy metals for all types of isolates. The highest sized plasmid was found 21 Kb and 14 Kb for lactose and non-lactose fermenting isolates, respectively. Conclusion: Microbial resistance to antibiotics and metal ions, has potential health hazards because these traits are generally associated with transmissible plasmids. Microorganisms resistant to antibiotics and tolerant to metals appear as a result of exposure to metal-contaminated environments.

Keywords: antibiotics, effluents, heavy metals, minimum inhibitory concentration, resistance

Procedia PDF Downloads 280

Authors: Aylin Hasbay Büyükkaragöz, Zehra Büyüktuncer, Tuğçe Nur Balcı, Nevin Ergun

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Purpose: Nutritional supplement usage is widespread among athletes all over the world. However, the usage among disabled athletes is not well-known. This study aimed to evaluate dietary supplement use in disabled athletes, their motivation for consuming supplements, sources of information, and their side effect. Methods: A total of 75 Turkish National Team's disabled athletes (38 female, 37 male) from 5 sport branches (soccer, weight lifting, shooting, table tennis and basketball), aged 13- 55 years, were participated in the study. Nutritional supplement usage was inquired using a questionnaire by a dietitian at their preparation camps. Results: A total of 22.7% of the athletes (18.4% and 27% of, respectively females and males) used some type of dietary supplements. Protein (35.3%), amino acid (29.4%), carnitine (29.4%), creatine (23.5%) and glucosamine (23.5%) were mostly preferred nutritional supplements by all athletes. The most common supplements use was obtained among weightlifters (71.4%), followed by the athletes of soccer (23.5%), table tennis (15.4%), and basketball (6.7%). No nutritional supplement usage was observed among shooters. Total of 41.2% consumers declared more than one reason for taking nutritional supplements. The main motivation for supplement usage was improving athletic performance (63.5%). Other reasons were weight loss, weight gain, muscle development, health protection and nutritional support. Athletes were more likely to get recommendation about nutritional supplement usage from team coaches (48.9%). Of 35.6% athletes reported that they made their own decision about using supplements. Other information sources were health professional, family member, friend and sale manager of sport retail store. Only 3 of 17 athletes reported side effects which were increased urine output, weight gain, loss of appetite and intestinal gas. Conclusions: Nutritional supplement usage was not common among disabled athletes. However, getting information from incompetent sources is disquieting. Considering their health problems, accurate information from competent sources should be provided to disabled athletes. Moreover, long term effects of nutritional supplements among disabled athletes should be examined in further studies.

Keywords: disabled athletes, ergogenic aid, nutritional supplement, vitamin supplementation

Procedia PDF Downloads 292

Authors: M. Naimi, M. B. Khaled

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The aim of the present work was to test in vitro inhibition of food pathogens and spoilage bacteria by crude bacteriocins from autochthonous lactic acid bacteria. Thirty autochthonous lactic acid bacteria isolated previously, belonging to the genera: Lactobacillus, Carnobacterium, Lactococcus, Vagococcus, Streptococcus, and Pediococcus, have been screened by an agar spot test and a well diffusion assay against Gram-positive and Gram-negative harmful bacteria: Bacillus cereus, Bacillus subtilis ATCC 6633, Escherichia coli ATCC 8739, Salmonella typhimurium ATCC 14028, Staphylococcus aureus ATCC 6538, and Pseudomonas aeruginosa under conditions means to reduce lactic acid and hydrogen peroxide effect to select bacteria with high bacteriocinogenic potential. Furthermore, crude bacteriocins semiquantification and heat sensitivity to different temperatures (80, 95, 110°C, and 121°C) were performed. Another exploratory test concerning the response of St. aureus ATCC 6538 to the presence of crude bacteriocins was realized. It has been observed by the agar spot test that fifteen candidates were active toward Gram-positive targets strains. The secondary screening demonstrated an antagonistic activity oriented only against St. aureus ATCC 6538, leading to the selection of five isolates: Lm14, Lm21, Lm23, Lm24, and Lm25 with a larger inhibition zone compared to the others. The ANOVA statistical analysis reveals a small variation of repeatability: Lm21: 0.56%, Lm23: 0%, Lm25: 1.67%, Lm14: 1.88%, Lm24: 2.14%. Conversely, slight variation was reported in terms of inhibition diameters: 9.58± 0.40, 9.83± 0.46, and 10.16± 0.24 8.5 ± 0.40 10 mm for, Lm21, Lm23, Lm25, Lm14and Lm24, indicating that the observed potential showed a heterogeneous distribution (BMS = 0.383, WMS = 0.117). The repeatability coefficient calculated displayed 7.35%. As for the bacteriocins semiquantification, the five samples exhibited production amounts about 4.16 for Lm21, Lm23, Lm25 and 2.08 AU/ml for Lm14, Lm24. Concerning the sensitivity the crude bacteriocins were fully insensitive to heat inactivation, until 121°C, they preserved the same inhibition diameter. As to, kinetic of growth , the µmax showed reductions in pathogens load for Lm21, Lm23, Lm25, Lm14, Lm24 of about 42.92%, 84.12%, 88.55%, 54.95%, 29.97% in the second trails. Inversely, this pathogen growth after five hours displayed differences of 79.45%, 12.64%, 11.82%, 87.88%, 85.66% in the second trails, compared to the control. This study showed potential inhibition to the growth of this food pathogen, suggesting the possibility to improve the hygienic food quality.

Keywords: exploratory test, lactic acid bacteria, crude bacteriocins, spoilage, pathogens

Procedia PDF Downloads 184

Authors: Dwi Arisandi Wijaya, Ernawati Arifin Giri-Rachman, Neni Nurainy

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Typhoid fever disease can be prevented by using a polysaccharide-based vaccine Vi which is a virulence factor of S.typhi. To produce high yield Vi polysaccharide from bacteria, it is important to know the biosynthesis of Vi polysaccharide and the regulators involved. In the In vivo condition, S. typhi faces different osmolarity, and the bacterial two-component system OmpR-EnvZ, regulate by up and down Capsular Vi polysaccharide biosynthesis. A high yielded Vi Polysaccharide strain, S. typhi strain C6524 used to study the effect of media osmolarity on Vi polysaccharide biosynthesis and the osmoregulation pattern of S. typhi strain C6524. The methods were performed by grown S. typhi strain C6524 grown on medium with 50 mM, 100 mM, and 150 mM osmolarity with the batch system. Vi polysaccharide concentration was measured by ELISA method. For further investigation of the osmoregulation pattern of strain C6524, the osmoregulator gene, OmpR, has been isolated and sequenced using the specific primer of the OmpR gene. Nucleotide sequence analysis is done with BLAST and Lallign. Amino Acid sequence analysis is done with Prosite and Multiple Sequence Alignment. The results of cultivation showed the average content of polysaccharide Vi for 50 mM, 100 mM, and 150 mM osmolarities 11.49 μg/mL, 12.06 μg/mL, and 14.53 μg/mL respectively. Analysis using Anova stated that the osmolarity treatment of 150 mM significantly affects Vi content. Analysis of nucleotide sequences shows 100% identity between S. typhi strain C6524 and Ty2. Analysis of amino acid sequences shows that the OmpR response regulator protein of the C6524 strain also has a α4-β5-α5 motif which is important for the regulatory activation system when phosphorylation occurs by domain kinase. This indicates that the regulator osmolarity response of S. typhi strain C6524 has no difference with the response regulator owned by S. typhi strain Ty2. A high Vi response rate in the 150 mM osmolarity treatment requires further research for RcsB-RcsC, another two-component system involved in Vi Biosynthesis.

Keywords: osmoregulator, OmpR, Salmonella, Vi polysaccharide

Procedia PDF Downloads 162

Authors: Amriana Hifizah, Philip Edward Vercoe, Graeme Bruce Martin, Teuku Reza Ferasy, Muhammad Hambal

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Many native forage plant species have been used in Indonesia as feed for ruminants. However, less information is available about how these plants affect productivity, let alone methane emissions. In the province of Aceh, where the traditional practice is to feed local forages to small ruminants, the farmers are not satisfied with the productivity of their livestock, and they attribute this problem to poor availability and too few options for good quality forages. Forage quality is reduced by high environmental temperatures which increase the amount of lignification. In addition to reducing productivity, these factors also increase enteric methane production. A preliminary survey about potential forage species was completed in three different districts, two of low elevation and one of high elevation: Syiah Kuala (05°30’5.08” N to 095°24’7.35” E), elevation 29 m MSL; Kajhu (05°32’34.6” N to 095°21’17.7” E), elevation 30 m MSL; Lembah Seulawah (05°28'06.4" N to 095°43' 14.2" E), elevation 254 m MSL. Information about local plants was collected in a semi-structured interview with scientists, government field officers and local farmers, in the city of Banda Aceh and in those three districts. The outcome was a list 40 species that could be useful, of which 21 were selected for further study. The selection process was based on several criteria: high availability, high protein content, low toxicity, and evidence of secondary metabolites (eg, history of medicinal plants for both human and animals). For some of the selected medicinal plants, there is experimental evidence of effects on methane production during rumen fermentation. Subsequently, the selected forages were tested for their effects on rumen fermentation in vitro, using batch culture. The data produced will be used to identify forages with the potential to reduce CH4 emissions. These candidates will then be assessed for their benefits (fermentability and productivity) and potential deleterious side-effects.

Keywords: batch culture, forage, methane, rumen

Procedia PDF Downloads 300

Authors: Saima Sharif Nilla, Mahmudur Rahman Khan, Anisur Rahman Khan, Ghulam Mustafa1

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The microbial quality of Indian white shrimp (Peneaus indicus) from Bagerhat, Khulna and Satkhira of southwest Bangladesh was assessed where the parameters varied with different sources and the quality was found to be poor for Satkhira shrimp samples. Shrimp samples in fresh condition were collected to perform the microbial assessment and 10 pathogenic isolates for antibiotic sensitivity test to 12 antibiotics. The results show that total bacterial count of all the samples were beyond the acceptable limit 105 cfu/g. In case of total coliform and E. coli density, no substantial difference (p<0.5) was found between the different shrimp samples from different districts and also high quantity of TC exceeding the limit (>102 cfu/g) proves the poor quality of shrimp. The FC abundance found in shrimps of Bagerhat and Satkhira was similar and significantly higher (p<0.5) than that of Khulna samples. No significant difference (p<0.5) was found among the high density of Salmonella-Shigella, Vibrio spp., and Staphylococcus spp. of the shrimp samples from the source places. In case of antibiotic sensitivity patterns, all of them were resistant to ampicillin, Penicillin and sensitive to kanamycin. Most of the isolates were frequently sensitive to ciprofloxacin and streptomycin in the sensitivity test. In case of nutritional composition, no significant difference (t-test, p<0.05) was found among protein, lipid, moisture and ash contents of shrimp samples. The findings prove that shrimp under this study was more or less contaminated and samples from Satkhira were highly privileged with food borne pathogens which confirmed the unhygienic condition of the shrimp farms as well as the presence of antibiotic resistance bacteria in shrimp fish supposed to threat food safety and deteriorate the export quality.

Keywords: food borne pathogens, satkhira, penaeus indicus, antibiotic sensitivity, southwest Bangladesh, food safety

Procedia PDF Downloads 678

Authors: Abrha Reta, Ajebu Nurfeta, Genet Mengistu, Mohammed Beyan

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Fodder trees such as Ziziphus spina-christi have the potential to enhance the utilization of natural grazing resources and also to mitigate seasonal feed shortages. The experiment was conducted with the objective of evaluating the effect of supplementing Ziziphus spina-christi leaf meal (ZSCLM) to natural pasture hay on feed intake, body weight gain, digestibility, and carcass characteristics of Tigray highland sheep. A randomized complete block design was employed with 5 blocks based on initial body weight, and sheep were randomly assigned to five treatments. Treatments were: 100g concentrate mix + ad libtum natural pasture hay (T1), T1+ 100g ZSCLM (T2), T1 + 200g ZSCLM (T3), T1 + 300g ZSCLM (T4), and T1 + 400g ZSCLM (T5) on dry matter (DM) basis. Dry matter intake was greater (P<0.05) in sheep on T5 compared to T3 and T1, while the total DM intake among T2, T4, and T5 were similar. Crude protein and metabolizable energy intake differed (P<0.05) among treatments with highest and lowest values in T5 and T1, respectively. Average daily gain was higher (P<0.05) in sheep kept on T2, T3, and T4 diets than T1. Higher (P<0.05) DM digestibility was found in T4 and T5 than T1. The highest (P<0.05) OM and CP digestibility was observed in sheep fed T3, T4, and T5 diets. Rib eye muscle area was higher (P<0.05) for T4 than T1 and T2. Dressing percentage was similar (P>0.05) among treatments. The current study indicated that supplementation of Tigray highland sheep with 200g air-dried Ziziphus spina-christi leaf meal leaves with 100g of concentrate mixture in their diet significantly increased feed intake and apparent digestibility, body weight gain, hot carcass weight, and rib eye muscle area by improving feed conversion efficiency.

Keywords: body weight, carcass, digestibility, and ziziphus spina-christi leaf meal

Procedia PDF Downloads 71

Authors: Abdolrasoul Daneshjoo

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The purpose of this research was to study the effect of aerobic exercises with 75% heart beats on the amount of urea, uric acid and creatine in blood of Iranian soccer national U-23 players. 27 players were selected according to the following demographic specifications: age: 21.4±1.60 years old; weight: 68±9.4 kg; height: 174.2±8.6 cm. Urea, uric acid and creatine in blood are considered as dependent variations where as 40 minutes running on a track with maximum 75% heart beats are independent variations. Heart beat and blood pressure in rest time, age, height, and weight are considered as the controlled variations. Maximum heart beats are recorded under maximum exercises (8 minutes and 150-250 watt energy) on ergo meter. Then, in order to determine independent variations, 75% maximum heart beats are considered for each player. Blood is taken twice (before and after determining independence variation). Moreover, the players are given a few instructions to be fulfilled 24 hours before the main exercises. Laboratory analysis method for blood urea sample is deacetyl ammoniom, for uric acid Karvy test and for creatine pyric acid. 'T' formula is applied for analyzing statistical data in dependent groups with degree of freedom 7 (d.f=7) urea and uric acid contain P>0.01 and P>0.05 for creatine. 1. Aerobic exercise can effect on the concentration of urea of blood as well as uric acid and creatine in blood serum and increase the amount of them. 2. Urea of blood serum increases from 26.75±2.59 to 28.9±2.67 (25%) with 40 minutes running and 75% heart beat. 3. Aerobic exercise causes uric acid increase 12.5% from 5.7±0.52 (before exercise) to 6.1±0.71 (after exercise). Creatine of blood serum increases from 1.36±0.27 (before exercise) to 1.85±0.49 (after exercise). We came to this result that during aerobic exercise catabolism of protein substrate increases. Moreover, augmentation of urea, uric acid and creatine in blood serum as metabolic poisons causes disorder in kidney. Also, tendons and joints are affected by these poisons. Appropriate diet and exercise can prevent production of these poisons resulted from heavy exercise.

Keywords: aerobic exercise, urea, uric acid, creatine, blood, soccer national players

Procedia PDF Downloads 506

Authors: Yahia Massinissa, Yahia Mouloud

Abstract:

Cystic fibrosis (CF), the most common lethal genetic disease in the Europe population, is caused by mutations in the transmembrane conductance regulator gene (CFTR). It affects most organs including an epithelial tissue, base of hydroelectrolytic transepithelial transport, notably that aerial ways, the pancreas, the biliary ways, the intestine, sweat glands and the genital tractus. The gene whose anomalies are responsible of the cystic fibrosis codes for a protein Cl channel named CFTR (cystic fibrosis transmembrane conductance regulator) that exercises multiple functions in the cell, one of the most important in control of sodium and chlorine through epithelia. The deficient function translates itself notably by an abnormal production of viscous secretion that obstructs the execrator channels of this target organ: one observes then a dilatation, an inflammation and an atrophy of these organs. It also translates itself by an increase of the concentration in sodium and in chloride in sweat, to the basis of the sweat test. In order to do a phenotypical and genotypical diagnosis at a part of the Algerian population, our survey has been carried on 16 patients with evocative symptoms of the cystic fibrosis at that the clinical context has been confirmed by a sweat test. However, anomalies of the CFTR gene have been determined by electrophoresis in gel of polyacrylamide of the PCR products (polymerase chain reaction), after enzymatic digestion, then visualized to the ultraviolet (UV) after action of the ethidium bromide. All mutations detected at the time of our survey have already been identified at patients attained by this pathology in other populations of the world. However, the important number of found mutation with regard to the one of the studied patients testifies that the origin of this big clinical variability that characterizes the illness in the consequences of an enormous diversity of molecular defects of the CFTR gene.

Keywords: cystic fibrosis, CFTR gene, polymorphism, algerian population, sweat test, genotypical diagnosis

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