Search results for: bacterial susceptibility
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1514

Search results for: bacterial susceptibility

1154 Effect of Bacillus subtilis Pb6 on Growth and Gut Microflora in Clostridium perfringens Challenged Broilers

Authors: A. Khalique, T. Naseem, N. Haque, Z. Rasool

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The objective of current study was to investigate the effect of Bacillus subtilis PB6 (CloSTAT) as a probiotic in broilers. The corn-soybean based diet was divided into four treatment groups; T1 (basal diet with no probiotic and no Clostridium perfringens); T2 (basal diet challenged with C. perfringens without probiotic); T3 (basal diet challenged with C. perfringens having 0.05% probiotic); T4 (basal diet challenged with C. perfringens having 0.1% probiotic). Every treatment group had four replicates with 24 birds each. Body weight and feed intake were measured on weekly basis, while ileal bacterial count was recorded on day-28 following Clostridium perfringens challenge. The 0.1% probiotic treatment showed 7.2% increase in average feed intake (P=0.05) and 8% increase in body weight compared to T2. In 0.1% treatment body weight was 5% higher than T3 (P=0.02). It was also observed that 0.1% treatment had improved feed conversion ratio (1.77) on 6th week. No effect of treatment was observed on mortality and ileal bacterial count. The current study indicated that 0.1% use of probiotic had positive response in C. perfringens challenged broilers.

Keywords: Bacillus subtilis PB6, antibiotic growth promoters, Clostridium perfringens, broilers

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1153 The Paleoenvironment and Paleoclimatological Variations during Aptian in North Central Tunisia

Authors: Houda Khaled, Frederic Boulvain, Fredj Chaabani

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This paper focuses on the sedimentological and mineralogical studies of Aptian series outcrops in the Serdj and Bellouta Mountain situated in north-central Tunisia. In the Serdj Mountain, the Aptian series is about 590 meters thick and it is defined by tow formations corresponding respectively to the Sidi Hamada formation (Barremian-Gragasian) and the Serdj formation (Middle Gragasian-Late Clansaysian). This later is consisting of five limestones sequences separated by marly levels limestones associated to some siltstones bed. The Bellouta section is especially composed of carbonate rocks and it is attributed to the Middle Gragasian - Late Clansaysian. These sections are studied in detail regarding lithology, micropaleontology, microfacies, magnetic susceptibility and mineralogical composition in order to provide new insights into the paleoenvironmental evolution and paleoclimatological implications during Aptian. The following facies associations representing different ramp palaeoenvironments have been identified: mudstone-wackestone outer ramp facies; skeletal grainstone-packstone mid-ramp facies, packstone-grainstone inner-ramp facies which include a variety of organisms such as ooliths, rudists ostracods associated to athor bioclats. The coastal facies is especially defined by a mudstone -wackestone texture coastal rich with miliolidea and orbitolines. The magnetic susceptibility (Xin) of all samples was compared with the lithological and microfacies variation. The MS curves show that the high values are correlated with the distal facies and the low values are registred in the coastal environment. The X-ray diffractometer analysis show the presence of kaolinite and illite.

Keywords: Aptian, Serdj formation, mineralogy, petrography

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1152 Phenotypical and Genotypical Assessment Techniques for Identification of Some Contagious Mastitis Pathogens

Authors: Ayman El Behiry, Rasha Nabil Zahran, Reda Tarabees, Eman Marzouk, Musaad Al-Dubaib

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Mastitis is one of the most economic disease affecting dairy cows worldwide. Its classic diagnosis using bacterial culture and biochemical findings is a difficult and prolonged method. In this research, using of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) permitted identification of different microorganisms with high accuracy and rapidity (only 24 hours for microbial growth and analysis). During the application of MALDI-TOF MS, one hundred twenty strains of Staphylococcus and Streptococcus species isolated from milk of cows affected by clinical and subclinical mastitis were identified, and the results were compared with those obtained by traditional methods as API and VITEK 2 Systems. 37 of totality 39 strains (~95%) of Staphylococcus aureus (S. aureus) were exactly detected by MALDI TOF MS and then confirmed by a nuc-based PCR technique, whereas accurate identification was observed in 100% (50 isolates) of the coagulase negative staphylococci (CNS) and Streptococcus agalactiae (31 isolates). In brief, our results demonstrated that MALDI-TOF MS is a fast and truthful technique which has the capability to replace conventional identification of several bacterial strains usually isolated in clinical laboratories of microbiology.

Keywords: identification, mastitis pathogens, mass spectral, phenotypical

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1151 Impact of Calcium Carbide Waste Dumpsites on Soil Chemical and Microbial Characteristics

Authors: C. E. Ihejirika, M. I. Nwachukwu, R. F. Njoku-Tony, O. C. Ihejirika, U. O. Enwereuzoh, E. O. Imo, D. C. Ashiegbu

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Disposal of industrial solid wastes in the environment is a major environmental challenge. This study investigated the effects of calcium carbide waste dumpsites on soil quality. Soil samples were collected with hand auger from three different dumpsites at varying depths and made into composite samples. Samples were subjected to standard analytical procedures. pH varied from 10.38 to 8.28, nitrate from 5.6mg/kg to 9.3mg/kg, phosphate from 8.8mg/kg to 12.3mg/kg, calcium carbide reduced from 10% to to 3%. Calcium carbide was absent in control soil samples. Bacterial counts from dumpsites ranged from 1.8 x 105cfu/g - 2.5 x 105cfu/g while fungal ranged from 0.8 x 103cfu/g - 1.4 x 103cfu/g. Bacterial isolates included Pseudomonas spp, Flavobacterium spp, and Achromobacter spp, while fungal isolates include Penicillium notatum, Aspergillus niger, and Rhizopus stolonifer. No organism was isolated from the dumpsites at soil depth of 0-15 cm, while there were isolates from other soil depths. Toxicity might be due to alkaline condition of the dumpsite. Calcium carbide might be bactericidal and fungicidal leading to cellular physiology, growth retardation, death, general loss of biodiversity and reduction of ecosystem processes. Detoxification of calcium carbide waste before disposal on soil might be the best option in management.

Keywords: biodiversity, calcium-carbide, denitrification, toxicity

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1150 Synthesis of 3,4-Dihydro-1H-Quinoxalin-2-Ones and 1H‑Quinolin-2-Ones and Evaluation of Their Anti-Bacterial Activity

Authors: Ali Amiri, Arash Esfandiari, Elham Zarenezhad

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We report here an efficient and rapid method for the preparation of 3,4-dihydro-1H-quinoxalin-2-ones and 1H‑quinolin-2-ones that involves grinding of o-, m-, or p‑phenylenediamine and three dialkyl acetylenedicarboxylates using a pestle and mortar. This solvent-free approach requires only a few minutes of reaction time. This type of reaction is expected to be the most economical method since neither catalyst nor solvent is used. Finally, all synthesised compounds were screened for antimicrobial activity against two Gram-positive bacteria (Pseudomonas aeruginosa PTCC 1077, Escherichia coli PTCC1330) and two Gram-negative bacteria (Staphylococcus aureus PTCC 1133, Bacillus cereus PTCC 1015) and their activity. Compared with gentamycin and ampicillin as reference drugs for Gram-negative and Gram-positive bacteria, respectively. The minimum inhibitory concentration (MIC) of the synthesised compounds and reference drugs were determined by the microdilution method. Good antibacterial activity was observed for 3,4-dihydro-1H-quinoxalin-2-ones against all species of Gram-positive and Gram-negative bacteria, and1H‑quinolin-2-ones showed good antibacterial activity against two Gram-positive bacteria.

Keywords: quinolin, quinoxalin, anti-bacterial activity, minimum inhibitory concentration (MIC)

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1149 Temperature Susceptibility of Multigrade Bitumen Asphalt and an Approach to Account for Temperature Variation through Deep Pavements

Authors: Brody R. Clark, Chaminda Gallage, John Yeaman

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Multigrade bitumen asphalt is a quality asphalt product that is not utilised in many places globally. Multigrade bitumen is believed to be less sensitive to temperature, which gives it an advantage over conventional binders. Previous testing has shown that asphalt temperature changes greatly with depth, but currently the industry standard is to nominate a single temperature for design. For detailed design of asphalt roads, perhaps asphalt layers should be divided into nominal layer depths and different modulus and fatigue equations/values should be used to reflect the temperatures of each respective layer. A collaboration of previous laboratory testing conducted on multigrade bitumen asphalt beams under a range of temperatures and loading conditions was analysed. The samples tested included 0% or 15% recycled asphalt pavement (RAP) to determine what impact the recycled material has on the fatigue life and stiffness of the pavement. This paper investigated the temperature susceptibility of multigrade bitumen asphalt pavements compared to conventional binders by combining previous testing that included conducting a sweep of fatigue tests, developing complex modulus master curves for each mix and a study on how pavement temperature changes through pavement depth. This investigation found that the final design of the pavement is greatly affected by the nominated pavement temperature and respective material properties. This paper has outlined a potential revision to the current design approach for asphalt pavements and proposes that further investigation is needed into pavement temperature and its incorporation into design.

Keywords: asphalt, complex modulus, fatigue life, flexural stiffness, four point bending, multigrade bitumen, recycled asphalt pavement

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1148 Challenging Clinical Scenario of Blood Stream Candida Infections – An Indian Experience

Authors: P. Uma Devi, S. Sujith, K. Rahul, T. S. Dipu, V. Anil Kumar , Vidya Menon

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Introduction: Candida is an important cause of bloodstream infections (BSIs), causing significant mortality and morbidity. The epidemiology of Candida infection is also changing, mainly in relation to the number of episodes caused by species Candida non-albicans. However, in India, the true burden of candidemia is not clear. Thus, this study was conducted to evaluate the clinical characteristics, species distribution, antifungal susceptibility and outcome of candidemia at our hospital. Methodology: Between January 2012 and April 2014, adult patients with at least one positive blood culture for Candida species were identified through the microbiology laboratory database (for each patient only the first episode of candidemia was recorded). Patient data was collected by retrospective chart review of clinical characteristics including demographic data, risk factors; species distribution, resistance to antifungals and survival. Results: A total of 165 episodes of Candida BSI were identified, with 115 episodes occurring in adult patients. Most of the episodes occurred in males (69.6%). Nearly 82.6% patients were between 41 to 80 years and majority of the patients were in the intensive care unit (65.2%) at the time of diagnosis. On admission, 26.1% and 18.3% patients had pneumonia and urinary tract infection, respectively. Majority of the candidemia episodes were found in the general medicine department (23.5%) followed by gastrointestinal surgery (13.9%) and medical oncology & haematology (13%). Risk factors identified were prior hospitalization within one year (83.5%), antibiotic therapy within the last one month (64.3%), indwelling urinary catheter (63.5%), central venous catheter use (59.1%), diabetes mellitus (53%), severe sepsis (45.2%), mechanical ventilation (43.5%) and surgery (36.5%). C. tropicalis (30.4%) was the leading cause of infection followed by C. parapsilosis (28.7%) and C. albicans (13%). Other non-albicans species isolated included C. haemulonii (7.8%), C. glabrata (7%), C. famata (4.3%) and C. krusei (1.7%). Antifungal susceptibility to fluconazole was 87.9% (C. parapsilosis), 100% (C. tropicalis) and 93.3% (C. albicans). Mortality was noted in 51 patients (44.3%). Early mortality (within 7 days) was noted in 32 patients while late mortality (between 7 and 30 days) was noted in 19 patients. Conclusion: In recent years, candidemia has been flourishing in critically ill patients. Comparison of data from our own hospital from 2005 shows a doubling of the incidence. Rapid changes in the rate of infection, potential risk factors, and emergence of non-albicans Candida demand continued surveillance of this serious BSI. High index of suspicion and sensitive diagnostics are essential to improve outcomes in resource limited settings with emergence of non-albicans Candida.

Keywords: antifungal susceptibility, candida albicans, candidemia, non-albicans candida

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1147 Prevalence, Associated Risk Factors, and Bacterial Pathogens in Dairy Camels: A Review

Authors: Djeddi Khaled, Houssou Hind, Miloudi Abdelatif, Rabah Siham

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Camels play a vital role as multipurpose animals, providing milk meat and serving as a means of transportation. They serve as a financial reserve for pastoralists and hold significant cultural and social value. Camel milk, known for its exceptional nutritional properties, is considered a valuable substitute for human milk. However, udder infections, particularly mastitis, pose significant challenges to camel farming. Clinical and subclinical mastitis can lead to substantial economic losses. Mastitis, especially the subclinical form, is a persistent and prevalent condition affecting milk hygiene and quality in dairy camels. This review offers insights into the prevalence and risk factors associated with subclinical mastitis in camels. The prevalence of subclinical mastitis in dairy camels was found to range from 9.28% to 87.78%. Major pathogens responsible for camel mastitis include Staphylococcus aureus, Coagulase-negative Staphylococcus, Streptococcus agalactiae, Streptococcus dysgalactiae, Escherichia coli, Micrococcus spp, Pasteurella haemolytica and Corynebacterium spp. The study outlines key risk factors contributing to camel mastitis, emphasizing factors such as severe tick infestation, age, stage of lactation, parity, body condition score, skin lesion on the teats or udders, anti-suckling devices, previous history of the udder, conformation of the udder, breed, unhygienic milking practices, production system, amongst others have been reported to be important in the prevalence of subclinical mastitis. This comprehensive overview provides valuable insights into the multifaceted aspects of camel mastitis, encompassing prevalent bacterial pathogens and diverse risk factors. The findings underscore the importance of holistic management practices, emphasizing hygiene, health monitoring, and targeted interventions to ensure the well-being and productivity of camels in various agro-pastoral contexts.

Keywords: bacterial pathogens, camel, mastitis, risk factors

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1146 Biological Hazards and Laboratory inflicted Infections in Sub-Saharan Africa

Authors: Godfrey Muiya Mukala

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This research looks at an array of fields in Sub-Saharan Africa comprising agriculture, food enterprises, medicine, organisms genetically modified, microbiology, and nanotechnology that can be gained from biotechnological research and development. Findings into dangerous organisms, mainly bacterial germs, rickettsia, fungi, parasites, or organisms that are genetically engineered, have immensely posed questions attributed to the biological danger they bring forth to human beings and the environment because of their uncertainties. In addition, the recurrence of previously managed diseases or the inception of new diseases are connected to biosafety challenges, especially in rural set-ups in low and middle-income countries. Notably, biotechnology laboratories are required to adopt biosafety measures to protect their workforce, community, environment, and ecosystem from unforeseen materials and organisms. Sensitization and inclusion of educational frameworks for laboratory workers are essential to acquiring a solid knowledge of harmful biological agents. This is in addition to human pathogenicity, susceptibility, and epidemiology to the biological data used in research and development. This article reviews and analyzes research intending to identify the proper implementation of universally accepted practices in laboratory safety and biological hazards. This research identifies ideal microbiological methods, adequate containment equipment, sufficient resources, safety barriers, specific training, and education of the laboratory workforce to decrease and contain biological hazards. Subsequently, knowledge of standardized microbiological techniques and processes, in addition to the employment of containment facilities, protective barriers, and equipment, is far-reaching in preventing occupational infections. Similarly, reduction of risks and prevention may be attained by training, education, and research on biohazards, pathogenicity, and epidemiology of the relevant microorganisms. In this technique, medical professionals in rural setups may adopt the knowledge acquired from the past to project possible concerns in the future.

Keywords: sub-saharan africa, biotechnology, laboratory, infections, health

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1145 Improved 3D Structure Prediction of Beta-Barrel Membrane Proteins by Using Evolutionary Coupling Constraints, Reduced State Space and an Empirical Potential Function

Authors: Wei Tian, Jie Liang, Hammad Naveed

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Beta-barrel membrane proteins are found in the outer membrane of gram-negative bacteria, mitochondria, and chloroplasts. They carry out diverse biological functions, including pore formation, membrane anchoring, enzyme activity, and bacterial virulence. In addition, beta-barrel membrane proteins increasingly serve as scaffolds for bacterial surface display and nanopore-based DNA sequencing. Due to difficulties in experimental structure determination, they are sparsely represented in the protein structure databank and computational methods can help to understand their biophysical principles. We have developed a novel computational method to predict the 3D structure of beta-barrel membrane proteins using evolutionary coupling (EC) constraints and a reduced state space. Combined with an empirical potential function, we can successfully predict strand register at > 80% accuracy for a set of 49 non-homologous proteins with known structures. This is a significant improvement from previous results using EC alone (44%) and using empirical potential function alone (73%). Our method is general and can be applied to genome-wide structural prediction.

Keywords: beta-barrel membrane proteins, structure prediction, evolutionary constraints, reduced state space

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1144 Evaluation of Antimicrobial Efficacy of Nanofluid Containing Carbon Nanotubes Functionalized with Antibiotic on Urinary Tract Infection

Authors: Erfan Rahimi, Hadi Bahari Far, Mojgan Shikhpour

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Background: Urinary tract infection is one of the most common nosocomial infections, especially among women. E. coli is one of the main causes of urinary tract infections and one of the most common antibiotics to fight this bacterium is ampicillin. As conventional antibiotics led to bacterial antibiotic resistance, modification of the pure drugs can address this issue. The aim of this study was to prepare nanofluids containing carbon nanotubes conjugated with ampicillin to improve drug performance and reduce antibiotic resistance. Methods: Multi-walled carbon nanotubes (MWCNTs) were activated with thionyl chloride by reflux system and nanofluids containing antibiotics were prepared by ultrasonic method. The properties of the prepared nano-drug were investigated by general element analysis, infrared spectroscopy, Raman spectroscopy, scanning electron microscopy and transmission electron microscopy. After the treatment of the desired strain with nanofluid, microbial studies were performed to evaluate the antibacterial effects and molecular studies were carried out to measure the expression of the resistance gene AcrAB. Result: We have shown that the antimicrobial effect of ampicillin-functionalized MWCNTs at low concentrations performed better than that of the conventional drug in both resistant and ATCC strains. Also, a decrease in antibiotic resistance of bacteria treated with ampicillin-functionalized MWCNTs compared to the pure drug was observed. Also, ampicillin-functionalized MWCNTs downregulated the expression of AcrAB in treated bacteria. Conclusion: Because carbon nanotubes are capable of destroying the bacterial wall, which provides antibiotic resistance features in bacteria, their usage in the form of nanofluids can make lower dosages (about three times less) than that of the pure drug more effective. Additionally, the expression of the bacterial resistance gene AcrAB decreased, thereby reducing antibiotic resistance and improving drug performance against bacteria.

Keywords: urinary tract infection, antibiotic resistance, carbon nanotube, nanofluid

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1143 A Prospective Study on the Pattern of Antibiotics Use and Prevalence of Multidrug Resistant Escherichia Coli in Poultry Chickens and Its Correlation with Urinary Tract Infection

Authors: Stelvin Sebastian, Andriya Annie Tom, Joyalanna Babu, Merin Joshy

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Introduction: The worldwide increase in the use of antibiotics in poultry and livestock industry to treat and prevent bacterial diseases and as growth promoters in feeds has led to the problem of development of antibiotic resistance both in animals and human population. Aim: To study the pattern of antibiotic use and prevalence of multidrug-resistant Escherichia coli in poultry chickens in selected farms in Muvattupuzha and to compare the spread of multidrug-resistant bacteria from poultry environment to UTI patients. Methodology: Two farms from each of 6 localities in Muvattupuzha were selected. A questionnaire on the pattern of antibiotic use and various farming practices were surveyed from farms. From each farm, 60samples of fresh fecal matter, litter from inside, litter from the outside shed, agricultural soil and control soil were collected, and antimicrobial susceptibility testing of E. coli was done. Antibiogram of UTI patients was collected from the secondary care hospital included in the study, and those were compared with resistance patterns of poultry samples. Results: From survey response antibiotics such as ofloxacin, enrofloxacin, levofloxacin, ciprofloxacin, colistin, ceftriaxone, neomycin, cephalexin, and oxytetracycline were used for treatment and prevention of infections in poultry. 31of 48 samples (51.66%) showed E. coli growth. 7 of 15 antibiotics (46.6%) showed resistance. Ampicillin, amoxicillin, meropenem, tetracycline showed 100% resistance to all samples. Statistical analysis confirmed similar resistance pattern in the poultry environment and UTI patients for antibiotics such as ampicillin, amoxicillin, amikacin, and ofloxacin. Conclusion: E. coli were resistant not only to extended-spectrum beta-lactams but also to carbapenems, which may be disseminated to the environment where litter was used as manure. This may due to irrational use of antibiotics in chicken or from their use in poultry feed as growth promoters. The study concludes the presence of multidrug-resistant E.coli in poultry and its spread to environment and humans, which may cause potentially serious implications for human health.

Keywords: multidrug resistance, escherichia coli, urinary tract infection, poultry

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1142 Molecular Detection of Staphylococcus aureus in the Pork Chain Supply and the Potential Anti-Staphylococcal Activity of Natural Compounds

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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1141 Unconventional Strategies for Combating Multidrug Resistant Bacterial Biofilms

Authors: Soheir Mohamed Fathey

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Biofilms are complex biological communities which are hard to be eliminated by conventional antibiotic administration and implemented in eighty percent of humans infections. Green remedies have been used for centuries and have shown obvious effects in hindering and combating microbial biofilm infections. Nowadays, there has been a growth in the number of researches on the anti-biofilm performance of natural agents such as plant essential oil (EOs) and propolis. In this study, we investigated the antibiofilm performance of various natural agents, including four essential oils (EOs), cinnamon (Cinnamomum cassia), tea tree (Melaleuca alternifolia), and clove (Syzygium aromaticum), as well as propolis versus the biofilm of both Gram-positive pathogenic bacterium Staphylococcus aureus and Gram-negative pathogenic bacterium Pseudomonas aeruginosa which are major human and animal pathogens rendering a high risk due to their biofilm development ability. The antibiofilm activity of the tested agents was evaluated by crystal violet staining assay and detected by scanning electron and fluorescent microscopy. Antibiofilm performance declared a potent effect of the tested products versus the tested bacterial biofilms.

Keywords: biofilm, essential oils, electron microscopy, fluorescent

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1140 Biodegradation of Chlorpyrifos in Real Wastewater by Acromobacter xylosoxidans SRK5 Immobilized in Calcium Alginate

Authors: Saira Khalid, Imran Hashmi

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Agrochemical industries produce huge amount of wastewater containing pesticides and other harmful residues. Environmental regulations make it compulsory to bring pesticides to a minimum level before releasing wastewater from industrial units.The present study was designed with the objective to investigate biodegradation of CP in real wastewater using bacterial cells immobilized in calcium alginate. Bacterial strain identified as Acromobacter xylosoxidans SRK5 (KT013092) using 16S rRNA nucleotide sequence analysis was used. SRK5 was immobilized in calcium alginate to make calcium alginate microspheres (CAMs). Real wastewater from industry having 50 mg L⁻¹ of CP was inoculated with free cells or CAMs and incubated for 96 h at 37˚C. CP removal efficiency with CAMs was 98% after 72 h of incubation, and no lag phase was observed. With free cells, 12h of lag phase was observed. After 96 h of incubation 87% of CP removal was observed when inoculated with free cells. No adsorption was observed on vacant CAMs. Phytotoxicity assay demonstrated considerable loss in toxicity. Almost complete COD removal was achieved at 96 h with CAMs. Study suggests the use of immobilized cells of SRK5 for bioaugmentation of industrial wastewater for CP degradation instead of free cells.

Keywords: biodegradation, chlorpyrifos, immobilization, wastewater

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1139 Health Monitoring of Primates in a Conservation Unit in Brazil

Authors: Elisângela de Albuquerque Sobreira Borovoski, Ricardo Willian Borovoski

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Microbiological infections acquired by animals pose a risk to public health. In public health, monitoring the health of primates is linked to the risk of transmission of zoonoses through scratches, bites and contact with biological samples. The project was approved by the Ethics Committee on the Use of Animals Protocol No. 170/2019. It was authorized by ICMBio Protocol No. 52117-1. The study was carried out in the period 2019-2022 in the municipality of Anápolis. Iron and galvanized wire traps were used and the animals were anesthetized with 4.4mg/kg zolethyl intramuscularly and saliva was collected through swabs. Fifty-three capuchin monkeys were captured from the Onofre Quinan Environmental Park in Anápolis-Goiás for health monitoring purposes. In the laboratory, the samples were deposited on the agar surface and seeded by exhaustion to obtain isolated colonies. These colonies were analyzed according to morphocolonial characteristics. Morphometric characterization and biochemical tests for bacterial identification were performed. A total of 861 bacterial samples were isolated, nine of which were strict anaerobic bacteria of the genus Peptostreptococcus. Previous and constant knowledge of the prevalence of pathogenic agents in biological samples is essential to be prepared to act in pandemic situations.

Keywords: Brazil, microbiology, monkeys, public health

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1138 Evaluation on Heat and Drought Tolerance Capacity of Chickpea

Authors: Derya Yucel, Nigar Angın, Dürdane Mart, Meltem Turkeri, Volkan Catalkaya, Celal Yucel

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Chickpea (Cicer arietinum L.) is one of the important legumes widely grown for dietery proteins in semi-arid Mediteranean climatic conditions. To evaluate the genetic diversity with improved heat and drought tolerance capacity in chickpea, thirty-four selected chickpea genotypes were tested under different field-growing conditions (rainfed winter sowing, irrigated-late sowing and rainfed-late sowing) in 2015 growing season. A factorial experiment in randomized complete block design with 3 reps was conducted at the Eastern Mediterranean Research Institute Adana, Turkey. Based on grain yields under different growing conditions, several indices were calculated to identify economically higher-yielding chickpea genotypes with greater heat and drought tolerance capacity. Average across chickpea genotypes, the values of tolerance index, mean productivity, yield index, yield stability index, stress tolerance index, stress susceptibility index, and geometric mean productivity were ranged between 1.1 to 218, 38 to 202, 0.3 to 1.7, 0.2 to 1, 0.1 to 1.2, 0.02 to 1.4, and 36 to 170 for drought stress and 3 to 54, 23 to 118, 0.3 to 1.7, 0.4 to 0.9, 0.2 to 2, 0.2to 2.3, and 23 to 118 for heat stress, respectively. There were highly significant differences observed among the tested chickpea genotypes response to drought and heat stresses. Among the chickpea genotypes, the Aksu, Arda, Çakır, F4 09 (X 05 TH 21-16189), FLIP 03-108 were identified with a higher drought and heat tolerance capacity. Based on our field studies, it is suggested that the drought and heat tolerance indicators of plants can be used by breeders to select stress-resistant economically productive chickpea genotypes suitable to grow under Mediteranean climatic conditions.

Keywords: irrigation, rainfed, stress susceptibility, tolerance indice

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1137 Comparison of Rumen Microbial Analysis Pipelines Based on 16s rRNA Gene Sequencing

Authors: Xiaoxing Ye

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To investigate complex rumen microbial communities, 16S ribosomal RNA (rRNA) sequencing is widely used. Here, we evaluated the impact of bioinformatics pipelines on the observation of OTUs and taxonomic classification of 750 cattle rumen microbial samples by comparing three commonly used pipelines (LotuS, UPARSE, and QIIME) with Usearch. In LotuS-based analyses, 189 archaeal and 3894 bacterial OTUs were observed. The observed OTUs for the Usearch analysis were significantly larger than the LotuS results. We discovered 1495 OTUs for archaea and 92665 OTUs for bacteria using Usearch analysis. In addition, taxonomic assignments were made for the rumen microbial samples. All pipelines had consistent taxonomic annotations from the phylum to the genus level. A difference in relative abundance was calculated for all microbial levels, including Bacteroidetes (QIIME: 72.2%, Usearch: 74.09%), Firmicutes (QIIME: 18.3%, Usearch: 20.20%) for the bacterial phylum, Methanobacteriales (QIIME: 64.2%, Usearch: 45.7%) for the archaeal class, Methanobacteriaceae (QIIME: 35%, Usearch: 45.7%) and Methanomassiliicoccaceae (QIIME: 35%, Usearch: 31.13%) for archaeal family. However, the most prevalent archaeal class varied between these two annotation pipelines. The Thermoplasmata was the top class according to the QIIME annotation, whereas Methanobacteria was the top class according to Usearch.

Keywords: cattle rumen, rumen microbial, 16S rRNA gene sequencing, bioinformatics pipeline

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1136 Effect of Tooth Bleaching Agents on Enamel Demineralisation

Authors: Najlaa Yousef Qusti, Steven J. Brookes, Paul A. Brunton

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Background: Tooth discoloration can be an aesthetic problem, and tooth whitening using carbamide peroxide bleaching agents are a popular treatment option. However, there are concerns about possible adverse effects such as demineralisation of the bleached enamel; however, the cause of this demineralisation is unclear. Introduction: Teeth can become stained or discoloured over time. Tooth whitening is an aesthetic solution for tooth discoloration. Bleaching solutions of 10% carbamide peroxide (CP) have become the standard agent used in dentist-prescribed and home-applied ’vital bleaching techniques’. These materials release hydrogen peroxide (H₂O₂), the active whitening agent. However, there is controversy in the literature regarding the effect of bleaching agents on enamel integrity and enamel mineral content. The purpose of this study was to establish if carbamide peroxide bleaching agents affect the acid solubility of enamel (i.e., make teeth more prone to demineralisation). Materials and Methods: Twelve human premolar teeth were sectioned longitudinally along the midline and varnished to leave the natural enamel surface exposed. The baseline behavior of each tooth half in relation to its demineralisation in acid was established by sequential exposure to 4 vials containing 1ml of 10mM acetic acid (1 minute/vial). This was followed by exposure to 10% CP for 8 hours. After washing in distilled water, the tooth half was sequentially exposed to 4 further vials containing acid to test if the acid susceptibility of the enamel had been affected. The corresponding tooth half acted as a control and was exposed to distilled water instead of CP. The mineral loss was determined by measuring [Ca²⁺] and [PO₄³⁻] released in each vial using a calcium ion-selective electrode and the phosphomolybdenum blue method, respectively. The effect of bleaching on the tooth surfaces was also examined using SEM. Results: Exposure to carbamide peroxide did not significantly alter the susceptibility of enamel to acid attack, and SEM of the enamel surface revealed a slight alteration in surface appearance. SEM images of the control enamel surface showed a flat enamel surface with some shallow pits, whereas the bleached enamel appeared with an increase in surface porosity and some areas of mild erosion. Conclusions: Exposure to H₂O₂ equivalent to 10% CP does not significantly increase subsequent acid susceptibility of enamel as determined by Ca²⁺ release from the enamel surface. The effects of bleaching on mineral loss were indistinguishable from distilled water in the experimental system used. However, some surface differences were observed by SEM. The phosphomolybdenum blue method for phosphate is compromised by peroxide bleaching agents due to their oxidising properties. However, the Ca²⁺ electrode is unaffected by oxidising agents and can be used to determine the mineral loss in the presence of peroxides.

Keywords: bleaching, carbamide peroxide, demineralisation, teeth whitening

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1135 Repeated Reuse of Insulin Injection Syringes and Incidence of Bacterial Contamination among Diabetic Patients in Jimma University Specialized Hospital, Jimma, Ethiopia

Authors: Muluneh Ademe, Zeleke Mekonnen

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Objective: to determine the level of bacterial contamination of reused insulin syringes among diabetic patients. Method: A facility based cross-sectional study was conducted among diabetic patients. Data on socio-demographic variables, history of injection syringe reuse, and frequency of reuse of syringes were collected using predesigned questionnaire. Finally, the samples from the syringes were cultured according to standard microbiological techniques. Result: Eighteen diabetic patients at Jimma University Hospital participated. A total of 83.3% of participants reused a single injection syringe for >30 consecutive injections, while 16.7% reused for >30 injections. Our results showed 22.2% of syringes were contaminated with methicillin-resistant Staphylococcus aures. Conclusion: We conclude reuse of syringe is associated with microbial contamination. The findings that 4/18 syringes being contaminated with bacteria is an alarming situation. A mechanism should be designed for patients to get injection syringes with affordable price. If reusing is not avoidable, reducing number of injections per a single syringe and avoiding needle touching with hand or other non-sterile material may be an alternative to reduce the risk of contamination.

Keywords: diabetes mellitus, Ethiopia, subcutaneous insulin injection, syringe reuse

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1134 Antimicrobial Activity of Fatty Acid Salts against Microbes for Food Safety

Authors: Aya Tanaka, Mariko Era, Manami Masuda, Yui Okuno, Takayoshi Kawahara, Takahide Kanyama, Hiroshi Morita

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Objectives— Fungi and bacteria are present in a wide range of natural environments. They are breed in the foods such as vegetables and fruit, causing corruption and deterioration of these foods in some cases. Furthermore, some species of fungi and bacteria are known to cause food intoxication or allergic reactions in some individuals. To prevent fungal and bacterial contamination, various fungicides and bactericidal have been developed that inhibit fungal and bacterial growth. Fungicides and bactericides must show high antifungal and antibacterial activity, sustainable activity, and a high degree of safety. Therefore, we focused on the fatty acid salt which is the main component of soap. We focused on especially C10K and C12K. This study aimed to find the effectiveness of the fatty acid salt as antimicrobial agents for food safety. Materials and Methods— Cladosporium cladosporioides NBRC 30314, Penicillium pinophilum NBRC 6345, Aspergillus oryzae (Akita Konno store), Rhizopus oryzae NBRC 4716, Fusarium oxysporum NBRC 31631, Escherichia coli NBRC 3972, Bacillus subtilis NBRC 3335, Staphylococcus aureus NBRC 12732, Pseudomonas aenuginosa NBRC 13275 and Serratia marcescens NBRC 102204 were chosen as tested fungi and bacteria. Hartmannella vermiformis NBRC 50599 and Acanthamoeba castellanii NBRC 30010 were chosen as tested amoeba. Nine fatty acid salts including potassium caprate (C10K) and laurate (C12K) at 350 mM and pH 10.5 were used as antifungal activity. The spore suspension of each fungus (3.0×10⁴ spores/mL) or the bacterial suspension (3.0×10⁵ or 3.0×10⁶ or 3.0×10⁷ CFU/mL) was mixed with each of the fatty acid salts (final concentration of 175 mM). Samples were counted at 0, 10, 60, and 180 min by plating (100 µL) on potato dextrose agar or nutrient agar. Fungal and bacterial colonies were counted after incubation for 1 or 2 days at 30 °C. Results— C10K was antifungal activity of 4 log-unit incubated time for 10 min against fungi other than A. oryzae. C12K was antifungal activity of 4 log-unit incubated time for 10 min against fungi other than P. pinophilum and A. oryzae. C10K and C12K did not show high anti-yeast activity. C10K was antibacterial activity of 6 or 7 log-unit incubated time for 10 min against bacteria other than B. subtilis. C12K was antibacterial activity of 5 to 7 log-unit incubated time for 10 min against bacteria other than S. marcescens. C12K was anti-amoeba activity of 4 log-unit incubated time for 10 min against H. vermiformis. These results suggest C10K and C12K have potential in the field of food safety.

Keywords: food safety, microbes, antimicrobial, fatty acid salts

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1133 Microbial Evaluation of Geophagic and Cosmetic Clays from Southern and Western Nigeria: Potential Natural Nanomaterials

Authors: Bisi-Johnson, Mary A., Hamzart A. Oyelade, Kehinde A. Adediran, Saheed A. Akinola

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Geophagic and cosmetic clays are among potential nano-material which occur naturally and are of various forms. The use of these nano-clays is a common practice in both rural and urban areas mostly due to tradition and medicinal reasons. These naturally occurring materials can be valuable sources of nano-material by serving as nano-composites. The need to ascertain the safety of these materials is the motivation for this research. Physical Characterization based on the hue value and microbiological qualities of the nano-clays were carried out. The Microbial analysis of the clay samples showed considerable contamination with both bacteria and fungi with fungal contaminants taking the lead. This observation may not be unlikely due to the ability of fungi species to survive harsher growth conditions than bacteria. 'Atike pupa' showed no bacterial growth. The clay with the largest bacterial count was Calabash chalk (Igbanke), while that with the highest fungal count was 'Eko grey'. The most commonly isolated bacteria in this study were Clostridium spp. and Corynebacterium spp. while fungi included Aspergillus spp. These results are an indication of the need to subject these clay materials to treatments such as heating before consumption or topical usage thereby ascertaining their safety.

Keywords: nano-material, clay, microorganism, quality

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1132 Genetics of Atopic Dermatitis: Role of Cytokines Genes Polymorphisms

Authors: Ghaleb Bin Huraib, Fahad Al Harthi, Misbahul Arfin, Abdulrahman Al-Asmari

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Atopic dermatitis (AD), also known as atopic eczema, is a chronic inflammatory skin disease characterized by severe itching and recurrent relapsing eczema-like skin lesions, affecting up to 20% of children and 10% of adults in industrialized countries. AD is a complex multifactorial disease, and its exact etiology and pathogenesis have not been fully elucidated. The aim of this study was to investigate the impact of gene polymorphisms of T helper cell subtype Th1 and Th2 cytokines, interferon-gamma (IFN-γ), interleukin-6 (IL-6) and transforming growth factor (TGF)-β1on AD susceptibility in a Saudi cohort. One hundred four unrelated patients with AD and 195 healthy controls were genotyped for IFN-γ (874A/T), IL-6 (174G/C) and TGF-β1 (509C/T) polymorphisms using ARMS-PCR and PCR-RFLP technique. The frequency of genotypes AA and AT of IFN-γ (874A/T) differed significantly among patients and controls (P 0.001). The genotype AT was increased while genotype AA was decreased in AD patients as compared to controls. AD patients also had higher frequency of T containing genotypes (AT+TT) than controls (P = 0.001). The frequencies of allele T and A were statistically different in patients and controls (P = 0.04). The frequencies of genotype GG and allele G of IL-6 (174G/C) were significantly higher while genotype GC and allele C were lower in AD patients than controls. There was no significant difference in the frequencies of alleles and genotypes of TGF-β1 (509C/T) polymorphism between patient and control groups. These results showed that susceptibility to AD is influenced by presence or absence of genotypes of IFN-γ (874A/T) and IL-6 (174G/C) polymorphisms. It is concluded that T-allele and T-containing genotypes (AT+TT) of IFN-γ (874A/T) and G-allele and GG genotype ofIL-6 (174G/C) polymorphisms are susceptible to AD in Saudis.On the other hand, the TGF-β1 (509C/T) polymorphism may not be associated with AD risk in Saudi population however further studies with large sample size are required to confirm these findings.

Keywords: atopic dermatitis, interferon-γ, interleukin-6, transforming growth factor-β1, polymorphism

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1131 Multi-Index Performance Investigation of Rubberized Reclaimed Asphalt Mixture

Authors: Ling Xu, Giuseppe Loprencipe, Antonio D'Andrea

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Asphalt pavement with recycled and sustainable materials has become the most commonly adopted strategy for road construction, including reclaimed asphalt pavement (RAP) and crumb rubber (CR) from waste tires. However, the adhesion and cohesion characteristics of rubberized reclaimed asphalt pavement were still ambiguous, resulting in deteriorated adhesion behavior and life performance. This research investigated the effect of bonding characteristics on rutting resistance and moisture susceptibility of rubberized reclaimed asphalt pavement in terms of two RAP sources with different oxidation levels and two tire rubber with different particle sizes. Firstly, the binder bond strength (BBS) test and bonding failure distinguishment were conducted to analyze the surface behaviors of binder-aggregate interaction. Then, the compatibility and penetration grade of rubberized RAP binder were evaluated by rotational viscosity test and penetration test, respectively. Hamburg wheel track (HWT) test with high-temperature viscoelastic deformation analysis was adopted, which illustrated the rutting resistance. Additionally, a water boiling test was employed to evaluate the moisture susceptibility of the mixture and the texture features were characterized with the statistical parameters of image colors. Finally, the colloid structure model of rubberized RAP binder with surface interaction was proposed, and statistical analysis was established to release the correlation among various indexes. This study concluded that the gel-phase colloid structure and molecular diffusion of the free light fraction would affect the surface interpretation with aggregate, determining the bonding characteristic of rubberized RAP asphalt.

Keywords: bonding characteristics, reclaimed asphalt pavement, rubberized asphalt, sustainable material

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1130 Antimicrobial Activity of Ilex paraguariensis Sub-Fractions after Liquid-Liquid Partitioning

Authors: Sabah El-Sawalhi, Elie Fayad, Roula M. Abdel-Massih

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Ilex paraguariensis (Yerba Mate) is a medium to large tree commonly consumed by South Americans. Its leaves and stems are associated with different biological activities. The purpose of this study was to evaluate the antibacterial activity of Yerba Mate against Gram-positive and Gram-negative bacterial strains and its action against some resistant bacteria with different resistance profiles. Yerba Mate aqueous extracts were prepared at 70°C for 2 hrs, and the microdilution method was used to determine the minimum inhibitory concentration (MIC). Gram-positive bacteria exhibited a stronger antibacterial activity (MIC ranged between 0.468 mg/mL and 15 mg/mL) than Gram-negative bacteria. Yerba Mate was also extracted with acetone: water (1:1) and then further sub-fractionated with hexane, chloroform, and ethyl acetate. MIC values against Staphylococcus aureus ranged from 0.78 to 2.5 mg/ml for the chloroform fraction, from 1.56 to 3.75 mg/ml for the ethyl acetate fraction, and 0.78 to 1.87 mg/ml for the water fraction. The water fraction also exhibited antibacterial activity against Salmonella species (MIC ranged from 1.56 mg/ml to 3.12 mg/ml). The water fraction exhibited the highest antibacterial activity among all the fractions obtained. More studies are needed to determine the molecule or molecules responsible for this activity.

Keywords: antibacterial activity, bacterial resistance, minimum inhibitory concentration, yerba mate

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1129 Antimicrobial Peptide Produced by Lactococcus garvieae with a Broad Inhibition Spectrum

Authors: Hai Chi, Ibrahim Mehmeti, Kirill Ovchinnikov, Hegle Holo, Ingolf F. Nes, Dzung B. Diep

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By using a panel of multiple indicator strains of different bacterial species and genera, we screened a large collection of bacterial isolates (over 1800 isolates) derived from raw milk, for bacteriocin producers with broad inhibition spectra (BIS). Fourteen isolates with BIS were identified, and by 16S rDNA sequencing they were found to belong to Lactococcus garvieae (10 isolates) and Enterococcus feacalis (4 isolates). Further analysis of the ten L. garvieae isolates revealed that they were very similar, if not identical, to each other in metabolic and genetic terms: they had the same fermentation profile on different types of sugars, repetitive sequence-based PCR (rep-PCR) DNA pattern as well as they all had the same inhibition profile towards over 50 isolates of different species. The bacteriocin activity from one of the L. garvieae isolates was assessed further. The bacteriocin which was termed garvicin KS, was found to be heatstable and proteinase-labile and its inhibition spectrum contained many distantly related genera of Firmicutes, comprising most lactic acid bacteria (LAB) as well as problematic species of Bacillus, Listeria, Streptococcus and Staphylococcus and their antibiotic resistant derivatives (e.g. VRE, MRSA). Taken together, the results indicate that this is a potent bacteriocin from L. garvieae and that its very broad inhibition spectrum can be a very useful property for use in food preservation as well as in infection treatments caused by gram-positive pathogens and their antibiotic-derivatives.

Keywords: bacteriocin, lactic acid bacteria, Lactococcus garvieae, antibiotics resistance

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1128 The Comparison Study of Human Microbiome in Chronic Rhinosinusitis between Adults and Children

Authors: Il Ho Park, Joong Seob Lee, Sung Hun Kang, Jae-Min Shin, Il Seok Park, Seok Min Hong, Seok Jin Hong

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Introduction: The human microbiota is the aggregate of microorganisms, and the bacterial microbiome of the human digestive tract contributes to both health and disease. In health, bacteria are key components in the development of mucosal barrier function and in innate and adaptive immune responses, and they also work to suppress the establishment of pathogens. In human upper airway, the sinonasal microbiota might play an important role in chronic rhinosinusitis (CRS). The purpose of this study is to investigate the human upper airway microbiome in CRS patients and to compare the sinonasal microbiome of adults with children. Materials and methods: A total of 19 samples from 19 patients (Group1; 9 CRS in children, aged 5 to 14 years versus Group 2; 10 CRS in adults aged 21 to 59 years) were examined. Swabs were collected from the middle meatus and/or anterior ethmoid region under general anesthesia during endoscopic sinus surgery or tonsillectomy. After DNA extraction from swab samples, we analysed bacterial microbiome consortia using 16s rRNA gene sequencing approach (the Illumina MiSeq platform). Results: In this study, relatively abundance of the six bacterial phyla and tremendous genus and species found in substantial amounts in the individual sinus swab samples, include Corynebacterium, Hemophilus, Moraxella, and Streptococcus species. Anaerobes like Fusobacterium and Bacteroides were abundantly present in the children group, Bacteroides and Propionibacterium were present in adults group. In genus, Haemophilus was the most common CRS microbiome in children and Corynebacterium was the most common CRS microbiome in adults. Conclusions: Our results show the diversity of human upper airway microbiome, and the findings will suggest that CRS is a polymicrobial infection. The Corynebacterium and Hemophilus may live as commensals on mucosal surfaces of sinus in the upper respiratory tract. The further study will be needed for analysis of microbiome-human interactions in upper airway and CRS.

Keywords: microbiome, upper airway, chronic rhinosinusitis, adult and children

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1127 Assessment of cellulase and xylanase Production by chryseobacterium sp. Isolated from Decaying Biomass in Alice, Eastern Cape, South Africa

Authors: A. Nkohla, U. Nwodo, L. V. Mabinya, A. I. Okoh

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A potential source for low-cost production of value added products is the utilization of lignocellulosic materials. However, the huddle needing breaching would be the dismantlement of the complex lignocellulosic structure as to free sugar base therein. the current lignocellosic material treatment process is expensive and not eco-friendly hence, the advocacy for enzyme based technique which is both cheap and eco-friendly is highly imperative. Consequently, this study aimed at the screening of cellulose and xylan degrading bacterial strain isolated from decaying sawdust samples. This isolate showed high activity for cellulase and xylanase when grown on carboxymethyl cellulose and birtchwood xylan as the sole carbon source respectively. The 16S rDNA nucleotide sequence of the isolate showed 98% similarity with that of Chryseobacterium taichungense thus, it was identified as a Chryseobacterium sp. Optimum culture conditions for cellulase and xylanase production were medium pH 6, incubation temperature of 25 °C at 50 rpm and medium pH 6, incubation temperature of 25 °C at 150 rpm respectively. The high enzyme activity obtained from this bacterial strain portends it as a good candidate for industrial use in the degradation of complex biomass for value added products.

Keywords: lignocellulosic material, chryseobacterium sp., submerged fermentation, cellulase, xylanase

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1126 Antifungal Susceptibility of Saprolegnia parasitica Isolated from Rainbow Trout and Its Host Pathogen Interaction in Zebrafish Disease Model

Authors: Sangyeop Shin, D. C. M. Kulatunga, S. H. S. Dananjaya, Chamilani Nikapitiya, Jehee Lee, Mahanama De Zoysa

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Saprolegniasis is one of the most devastating fungal diseases in freshwater fish which is caused by species in the genus Saprolegnia including Saprolegnia parasitica. In this study, we isolated the strain of S. parasitica from diseased rainbow trout in Korea. Morphological and molecular based identification confirmed that isolated fungi belong to the member of S. parasitica, supported by its typical fungal features including cotton-like whitish mycelium, zoospores (primary and secondary) and phylogenetic analysis with internal transcribed spacer (ITS) region. Pathogenicity of isolated S. parasitica was developed in embryo, larvae, juvenile and adult zebrafish as a disease model. Up regulation of host genes encoding ZfTnf-α, Zfc-Rel, ZfIl-12, ZfLyz-c, Zfβ-def, and ZfHsp-70 was identified in zebrafish larvae after experimental challenge of S. parasitica showing the host immune responses against the S. parasitica. Survival of the juveniles upon fungal infection might be due to the increased immune protection in the host. Investigation of antifungal susceptibility of S. parasitica with natural lawsone (2-hydroxy-1,4-naphthoquinone) revealed the minimum inhibitory concentration (MIC) and percentage inhibition of radial growth (PIRG %) as 200 µg/mL and 31.8%, respectively. Lawsone was able to change the membrane permeability, and cause irreversible damage and disintegration to the cellular membranes of S. parasitica which might have effect on fungi growth inhibition. Moreover, the mycelium exposed to lawsone (MIC level) changed the transcriptional responses of S. parasitica genes. Overall results indicate that lawsone could be a potential and novel anti-S. parasitica agent for controlling S. parasitica infection.

Keywords: host-pathogen interactions, lawsone, rainbow trout, Saprolegnia parasitica, Saprolegniasis, zebrafish

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1125 Microbial Load, Prevalence and Antibiotic Resistance of Microflora Isolated from the Ghanaian Paper Currency Note: A Potential Health Threat

Authors: Simon Nyarko

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This study examined the microbial flora contamination of the Ghanaian paper currency notes and antibiotic resistance in Ejura Municipal, Ashanti Region, Ghana. This is a descriptive cross-sectional study designed to assess the profile of microflora contamination of the Ghanaian paper currency notes and antibiotic-resistant in the Ejura Municipality. The research was conducted in Ejura, a town in the Ejura Sekyeredumase Municipal of the Ashanti region of Ghana. 70 paper currency notes which were freshly collected from the bank, consisting of 15 pieces of GH ¢1, GH ¢2, and GH ¢5, 10 pieces of GH ¢10 and GH ¢20, and 5 pieces of GH ¢50, were randomly sampled from people by exchanging their money in usage with those freshly secured from the bank. The surfaces of each GH¢ note were gently swabbed and sent to the lab immediately in sterile Zip Bags and sealed, and tenfold serial dilution was inoculated on plate count agar (PCA), MacConkey agar (MCA), mannitol salt agar (MSA), and deoxycholate citrate agar (DCA). For bacterial identification, the study used appropriate laboratory and biochemical tests. The data was analyzed using SPSS-IBM version 20.0. It was found that 95.2 % of the 70 GH¢ notes tested positive for one or more bacterial isolates. On each GH¢ note, mean counts on PCA ranged from 3.0 cfu/ml ×105 to 4.8 cfu/ml ×105. Of 124 bacteria isolated. 36 (29.03 %), 32 (25.81%), 16 (12.90 %), 20 (16.13%), 13 (10.48 %), and 7 (5.66 %) were from GH¢1, GH¢2, GH¢10, GH¢5, GH¢20, and GH¢50, respectively. Bacterial isolates were Escherichia coli (25.81%), Staphylococcus aureus (18.55%), coagulase-negative Staphylococcus (15.32%), Klebsiella species (12.10%), Salmonella species (9.68%), Shigella species (8.06%), Pseudomonas aeruginosa (7.26%), and Proteus species (3.23%). Meat shops, commercial drivers, canteens, grocery stores, and vegetable shops contributed 25.81 %, 20.16 %, 19.35 %, 17.74 %, and 16.94 % of GH¢ notes, respectively. There was 100% resistance of the isolates to Erythromycin (ERY), and Cotrimoxazole (COT). Amikacin (AMK) was the most effective among the antibiotics as 75% of the isolates were susceptible to it. This study has demonstrated that the Ghanaian paper currency notes are heavily contaminated with potentially pathogenic bacteria that are highly resistant to the most widely used antibiotics and are a threat to public health.

Keywords: microflora, antibiotic resistance, staphylococcus aureus, culture media, multi-drug resistance

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