Search results for: cellulolytic bacteria

Authors: António Inês, Ana Guimarães, José Maria, Vânia Laranjo, Armando Venâncio, Luís Abrunhosa

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Lactic acid bacteria (LAB) play a key role in the biopreservation of a wide range of fermented food products, such as yogurt, cheese, fermented milks, meat, fish, vegetables (sauerkraut, olives and pickles), certain beer brands, wines and silage, allowing their safe consumption, which gave to these bacteria a GRAS (Generally Recognised as Safe) status. Besides that, the use of LAB in food and feed is a promising strategy to reduce the exposure to dietary mycotoxins, improving their shelf life and reducing health risks, given the unique mycotoxin decontaminating characteristic of some LAB. Mycotoxins present carcinogenic, mutagenic, teratogenic, neurotoxic and immunosuppressive effects over animals and Humans, being the most important ochratoxin A (OTA), aflatoxins (AFB1), trichothecenes, zearalenone (ZEA), fumonisin (FUM) and patulin. In a previous work of our group it was observed OTA biodegradation by some strains of Pediococcus parvulus isolated from Douro wines. So, the aim of this study was to enlarge the screening of the biodetoxification over more mycotoxins besides OTA, including AFB1, and ZEA. This ability was checked in a collection of LAB isolated from vegetable (wine, olives, fruits and silage) and animal (milk and dairy products, sausages) sources. All LAB strains were characterized phenotypically (Gram, catalase) and genotypically. Molecular characterisation of all LAB strains was performed using genomic fingerprinting by MSP-PCR with (GTG)5 and csM13 primers. The identification of the isolates was confirmed by 16S rDNA sequencing. To study the ability of LAB strains to degrade OTA, AFB1 and ZEA, a MRS broth medium was supplemented with 2.0 μg/mL of each mycotoxin. For each strain, 2 mL of MRS supplemented with the mycotoxins was inoculated in triplicate with 109 CFU/mL. The culture media and bacterial cells were extracted by the addition of an equal volume of acetonitrile/methanol/acetic acid (78:20:2 v/v/v) to the culture tubes. A 2 mL sample was then collected and filtered into a clean 2 mL vial using PP filters with 0.45 μm pores. The samples were preserved at 4 °C until HPLC analysis. Among LAB tested, 10 strains isolated from milk were able to eliminate AFB1, belonging to Lactobacillus casei (7), Lb. paracasei (1), Lb. plantarum (1) and 1 to Leuconostoc mesenteroides. Two strains of Enterococcus faecium and one of Ec. faecalis from sausage eliminated ZEA. Concerning to strains of vegetal origin, one Lb. plantarum isolated from elderberry fruit, one Lb. buchnerii and one Lb. parafarraginis both isolated from silage eliminated ZEA. Other 2 strains of Lb. plantarum from silage were able to degrade both ZEA and OTA, and 1 Lb. buchnerii showed activity over AFB1. These enzymatic activities were also verified genotypically through specific gene PCR and posteriorly confirmed by sequencing analysis. In conclusion, due the ability of some strains of LAB isolated from different sources to eliminate OTA, AFB1 and ZEA one can recognize their potential biotechnological application to reduce the health hazards associated with these mycotoxins. They may be suitable as silage inoculants or as feed additives or even in food industry.

Keywords: bio-detoxification, lactic acid bacteria, mycotoxins, food and feed

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Authors: A. Cheriguene, F. Chougrani

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A total of 120 wild lactic acid bacteria were isolated from goat’s milk collected from different areas in Western Algeria. The strains were screened for production and technological properties such as acid production, aminopeptidase activity, autolytic properties, antimicrobial activity, and exopolysaccharide production. In general most tested isolates showed a good biomass separation when collected by centrifugation; as for the production of the lactic acid, results revealed that our strains are weakly acidifying; nevertheless, lactococci showed a best acidifying activity compared to lactobacilli. Aminopeptidase activity was also weak in most strains; but, it was generally higher for lactobacilli compared to lactococci. Autolytic activity was generally higher for most strains, more particularly lactobacilli. Antimicrobial activity was detected in 50% of the isolates, particularly in lactobacilli where 80% of strains tested were able to inhibit the growth of other strains. The survey of the profile of the texture, the proteolysis as well as the development of the flavor in the Domiati cheese made on the basis of our isolated strains have been led during the ripening. The sensory assessment shows that the cheese salted in milk received the best scores in relation to cheese salted after drainage. Textural characteristics, such as hardness, cohesiveness, gumminess, and chewiness decreased in the two treatments during the 60 days of ripening. Otherwise, it has been noted that adhesiveness and adhesive force increased in the cheese salted in milk.

Keywords: lactic acid bacteria, technological properties, acidification, exopolysaccharide, bacteriocin, textural properties

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Authors: Harikrishna Yadav Nanganuru, Narasimhulu Korrapati

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Over the past many years, many sites in the world have been contaminated with heavy metals, which are the largest class of contaminants. Lead is one of the toxic heavy metals contaminated in the environment. Lead is not biodegradable, that’s why it is accumulated in the human body and impacts all the systems of the human body when it has been taken by humans. The accumulation of lead in the water environment has been showing adverse effects on the public health. So the removal of lead from the water environment by the biosorption process, which is emerged as a potential method for the lead removal, is an efficient approach. This work was focused to examine the removal of Lead [Pb (II)] ions from aqueous solution and effluent from battery industry. Lead contamination in water is a widespread problem throughout the world and mainly results from lead acid battery manufacturing effluent. In this work, isolated bacteria from wastewater of lead acid battery industry has been utilized for the removal of lead. First effluent from the lead acid battery industry was characterized by the inductively coupled plasma atomic emission spectrometry (ICP – AES). Then the bacteria was isolated from the effluent and used it’s immobilized dead mass for the biosorption of lead. Scanning electron microscopic (SEM) and Atomic force microscopy (AFM) studies clearly suggested that the Lead (Pb) was adsorbed efficiently. The adsorbed percentage of lead (II) from waste was 97.40 the concentration of lead (II) is measured by Atomic Absorption Spectroscopy (AAS). From the result of AAS it can be concluded that immobilized isolated dead mass was well efficient and useful for biosorption of lead contaminated waste water.

Keywords: biosorption, ICP-AES, lead (Pb), SEM

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Authors: Kristina Karapetyan, Flora Tkhruni, Tsovinar Balabekyan, Arevik Israyelyan, Tatyana Khachatryan

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The ability of the lactic acid bacteria (LAB) to prevent and cure a variety of diseases, their protective role against infections and colonization of pathogenic microorganisms in the digestive tract, has lead to the coining of the term probiotics or pro-life. LAB inhibiting the growth of pathogenic and food spoilage microorganisms, maintaining the nutritive quality and improving the shelf life of foods. They have also been used as flavor and texture producers. Enterococcus strains have been used for treatment of diseases such as diarrhea or antibiotic associated diarrhea, inflammatory pathologies that affect colon such as irritable bowel syndrome, or immune regulation, diarrhea caused by antibiotic treatments. The obtaining and investigation of biological properties of proteinoceous antibiotics, on the basis of probiotic LAB shown, that bacteriocins, metabiotics, and peptides of LAB represent bactericides have a broad range of activity and are excellent candidates for development of new prophylactic and therapeutic substances to complement or replace conventional antibiotics. The genotyping by 16S rRNA sequencing for LAB were used. Cell free culture broth (CFC) broth was purified by the Gel filtration method on the Sephadex Superfine G 25 resin. Antimicrobial activity was determined by spot-on-lawn method and expressed in arbitrary units (AU/ml). The diversity of multidrug-resistance (MDR) of pathogenic strains to antibiotics, most widely used for treatment of human diseases in the Republics of Armenia and Nagorno Karabakh were examined. It was shown, that difference of resistance of pathogens to antibiotics depends on their isolation sources. The influences of partially purified antimicrobial preparations (AMP), obtained from the different strains of Enterococcus genus on the growth of MDR pathogenic bacteria were investigated. It was shown, that bacteriocin containing partially purified preparations, obtained from different strains of Enterococcus faecium and durans species, possess bactericidal or bacteriostatic activity against antibiotic resistant intestinal, spoilage and food-borne pathogens such as Listeria monocytogenes, Staphylococcus aureus, E. coli, and Salmonella. Endemic strains of LAB, isolated from Matsoni made from donkey, buffalo and goat milk, shown broad spectrum of activity against food spoiling microorganisms, moulds and fungi, such as Salmonella sp., Esherichia coli, Aspergillus and Penicillium species. Highest activity against MDR pathogens shown bacteria, isolated from goat milk products. High stability of the investigated strains of the genus Enerococcus, isolated from samples of matsun from different regions of Nagorno-Karabakh (NKR) to the antibiotics was shown. The obtained data show high stability of the investigated different strains of the genus Enerococcus. The high genetic diversity in Enterococcus group suggests adaptations for specific mutations in different environments. Thus, endemic strains of LAB are able to produce bacteriocins with high and different inhibitory activity against broad spectrum of microorganisms isolated from different sources and belong to different taxonomic group. Prospect of the use of certain antimicrobial preparations against pathogenic strains is obvious. These AMP can be applied for long term use against different etiology antibiotic resistant pathogens for prevention or treatment of infectional diseases as an alternative to antibiotics.

Keywords: antimicrobial biopreparation, endemic lactic acid bacteria, intra-species diversity, multidrug resistance of pathogens

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Authors: Vladimíra Kňazovická, Mária Dovičičová, Miroslava Kačániová, Margita Čanigová

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The aim of the study was to test the storage of bee pollen at room temperature and in cold store, and to describe microorganisms originated from it. Fresh bee pollen originating in West Slovakia was collected in May 2010. It was tested for presence of particular microbial groups using dilution plating method, and divided into two parts with different storage (in cold store and at room temperature). Microbial analyses of pollen were repeated after one year of storage. Several bacterial strains were isolated and tested using Gram staining, for catalase and fructose-6-phosphate-phosphoketolase presence, and by rapid ID 32A (BioMérieux, France). Micromycetes were identified at genus level. Fresh pollen contained coliform bacteria, which were not detected after one year of storage in both ways. Total plate count (TPC) of aerobes and anaerobes and of yeasts in fresh bee pollen exceeded 5.00 log CFU/g. TPC of aerobes and anaerobes decreased below 2.00 log CFU/g after one year of storage in both ways. Count of yeasts decreased to 2.32 log CFU/g (at room temperature) and to 3.66 log CFU/g (in cold store). Microscopic filamentous fungi decreased from 3.41 log CFU/g (fresh bee pollen) to 1.13 log CFU/g (at room temperature) and to 1.89 log CFU/g (in cold store). In fresh bee pollen, 12 genera of micromycetes were identified in the following order according to their relative density: Penicillium > Mucor > Absidia > Cladosporium, Fusarium > Alternaria > Eurotium > Aspergillus, Rhizopus > Emericella > Arthrinium and Mycelium sterilium. After one year at room temperature, only three genera were detected in bee pollen (Penicillium > Aspergillus, Mucor) and after one year in cold store, seven genera were detected (Mucor > Penicillium, Emericella > Aspergillus, Absidia > Arthrinium, Eurotium). From the plates designated for anaerobes, eight colonies originating in fresh bee pollen were isolated. Among them, a single yeast isolate occurred. Other isolates were G+ bacteria, with a total of five rod shaped. In three out of these five, catalase was absent and fructose-6-phosphate-phosphoketolase was present. Bacterial isolates originating in fresh pollen belonged probably to genus Bifidobacterium or relative genera, but their identity was not confirmed unequivocally. In general, cold conditions are suitable for maintaining the natural properties of foodstuffs for a longer time. Slight decrease of microscopic fungal number and diversity was recorded in cold temperatures compared with storage at room temperature.

Keywords: bacteria, bee product, microscopic fungi, biosystems engineering

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Authors: Jarel Elgin Tolentino, Arby Denise Nera, Mary Rose Roco, Angela Vianca Aspa, Nikko Beltran, Else Dapat

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Drug resistance by cells has been the problem in the medical field for decades now. The use of medicinal plants as a source of creating powerful drugs has been nowadays recognized worldwide to treat such resistant diseases. In the present study, the potential for Diospyros philippinensis A. DC. to inhibit growth of both bacteria and cancer cell line was conducted. The leaf crude extracts were screened for the presence of phytochemicals and examined for potential bioactivities by employing several assays like Kirby-Bauer disc diffusion method, DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) assay and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) tetrazolium assay for the antibacterial, antioxidant and cytotoxic activities of the extract, respectively. Phytochemical test results of the extracts revealed the presence of alkaloids, flavonoids, saponins, phenols, quinones, cardiac glycosides, phlobatannins, carbohydrate, cardenolides and proteins. The leaf extracts were found to exhibit antibacterial activity against gram-positive bacteria, high antioxidant activity (99.22% ± 0.005) but did not show any sign of cytotoxicity towards HCT116 (ATCC CCL-247). The study therefore concludes that D. philippinensis A. DC. leaf extract can be a source of antibacterial and chemopreventive agents. This claim may be used as basis for future investigation.

Keywords: bioassay, medicinal plants, plant crude extracts, phytochemical screening

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Authors: Ubani Onyedikachi, Atagana Harrison Ifeanyichukwu, Thantsha Mapitsi Silvester

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Components of oil sludge (PAHs) are known cytotoxic, mutagenic and potentially carcinogenic compounds also bacteria and fungi have been found to degrade PAHs to innocuous compounds. This study is aimed at measuring the effect of pig, cow, horse and poultry manures on the degradation in selected PAHs present in oil sludge. Soil spiked with oil sludge was co-composted differently with each manure in a ratio of 2:1 (w/w) spiked soil: manure and wood-chips in a ratio of 2:1 (w/v) spiked soil: wood-chips. Control was set up similar as the one above but without manure. The mixtures were incubated for 10 months at room temperature. Compost piles were turned weekly and moisture level was maintained at between 50% and 70%. Moisture level, pH, temperature, CO2 evolution and oxygen consumption were measured monthly and the ash content at the end of experimentation. Highest temperature reached was 27.5 °C in all compost heaps, pH ranged from 5.5 to 7.8 and CO2 evolution was highest in poultry manure at 18.78μg/dwt/day. Microbial growth and activities were enhanced; bacteria identified were Bacillus, Arthrobacter and Staphylococcus species. Percentage reduction in PAHs was measured using automated soxhlet extractor with Dichloromethane coupled with gas chromatography/mass spectrometry (GC/MS). Results from PAH measurements showed reduction between 77% and 99%. Co-composting of spiked soils with animal manures enhanced the reduction in PAHs.

Keywords: animal manures, bioremediation, co-composting, oil refinery sludge, PAHs

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Authors: Rasha F. Sharaf

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Treatment of dental caries in young children is considered a great challenge for all dentists, especially with uncooperative children. Recently non-invasive techniques have been highlighted as they alleviate the need for local anesthesia and other painful procedures during management of carious teeth and, at the same time, increase the success rate of the treatment done. Silver Diamine Fluoride (SDF) is one of the most effective cariostatic materials that arrest the progression of carious lesions and aid in remineralizing the demineralized tooth structure. Both fluoride and silver ions proved to have an antibacterial action and aid in the precipitation of an insoluble layer that prevents further decay. At the same time, Moringa proved to have an effective antibacterial action against different types of bacteria, therefore, it can be used as a non-invasive technique for the management of caries in children. One of the important theories for the control of caries is by depriving the cariogenic bacteria from nutrients causing their starvation and death, which can be achieved by applying stainless steel crown on primary molars with carious lesions which are not involving the pulp, and this technique is known as Hall technique. The success rate of the Hall technique can be increased by arresting the carious lesion using either SDF or Moringa and gaining the benefit of their antibacterial action. Multiple clinical cases with 1 year follow up will be presented, comparing different treatment options, and using various materials and techniques for non-invasive and non-painful management of carious primary teeth.

Keywords: SDF, hall technique, carious primary teeth, moringa extract

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Authors: Cheng-Chih Tsai, Yu-Hsuan Liu, Cheng-Ying Ho, Chun-Chin Huang

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The aim of this study evaluated the in vitro and in vivo antimicrobial activity of selected lactic acid bacteria (LAB) against Uropathogenic Escherichia coli (UPEC) for prevention and amelioration of UTIs. We screened LAB strains with antimicrobial effects on UPEC using a well-diffusion assay, bacterial adherence to the uroepithelium cell line SV-HUC-1 (BCRC 60358), and a coculture inhibition assay. The results showed that the 7 LAB strains (Lactobacillus paracasei, L. salivarius, two Pediococcus pentosaceus strains, two L. plantarum strains, and L. crispatus) and the fermented probiotic products produced by these multi-LAB strains exhibited potent zones of inhibition against UPEC. Moreover, the LAB strains and probiotic products adhered strongly to the uroepithelium SV-HUC-1 cell line. The growth of UPEC strains was also markedly inhibited after co-culture with the LAB strains and probiotic products in human urine. In addition, the enhanced levels of IL-6, IL-8 and lactic acid dehydrogenase were significantly decreased by treatments with the LAB strains and probiotic products in UPEC-induced SV-HUC-1 cells. Furthermore, oral administration of probiotic products reduced the number of viable UPEC in the urine of UPEC-challenged BALB/c mice. Taken together, this study demonstrates that probiotic supplementation may be useful as an adjuvant therapy for the treatment of bacterial-induced urinary tract infections.

Keywords: lactic acid bacterium, SV-HUC-1 uroepithelium, urinary tract infection, uropathogenic Escherichia coli, BALB/c mice

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

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

Keywords: antimicrobial, antioxidant, cytotoxicity, ethnobotanical, fruits

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Authors: Hamed Ahari, Behrouz Akbari Adreghani, Vadood Razavilar, Amirali Anvar, Sima Moradi, Hourieh Shalchi

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Considering the ever increasing population and industrialization of the developmental trend of humankind's life, we are no longer able to detect the toxins produced in food products using the traditional techniques. This is due to the fact that the isolation time for food products is not cost-effective and even in most of the cases, the precision in the practical techniques like the bacterial cultivation and other techniques suffer from operator errors or the errors of the mixtures used. Hence with the advent of nanotechnology, the design of selective and smart sensors is one of the greatest industrial revelations of the quality control of food products that in few minutes time, and with a very high precision can identify the volume and toxicity of the bacteria. Methods and Materials: In this technique, based on the bacterial antibody connection to nanoparticle, a sensor was used. In this part of the research, as the basis for absorption for the recognition of bacterial toxin, medium sized silica nanoparticles of 10 nanometer in form of solid powder were utilized with Notrino brand. Then the suspension produced from agent-linked nanosilica which was connected to bacterial antibody was positioned near the samples of distilled water, which were contaminated with Staphylococcus aureus bacterial toxin with the density of 10-3, so that in case any toxin exists in the sample, a connection between toxin antigen and antibody would be formed. Finally, the light absorption related to the connection of antigen to the particle attached antibody was measured using spectrophotometry. The gene of 23S rRNA that is conserved in all Staphylococcus spp., also used as control. The accuracy of the test was monitored by using serial dilution (l0-6) of overnight cell culture of Staphylococcus spp., bacteria (OD600: 0.02 = 107 cell). It showed that the sensitivity of PCR is 10 bacteria per ml of cells within few hours. Result: The results indicate that the sensor detects up to 10-4 density. Additionally, the sensitivity of the sensors was examined after 60 days, the sensor by the 56 days had confirmatory results and started to decrease after those time periods. Conclusions: Comparing practical nano biosensory to conventional methods like that culture and biotechnology methods(such as polymerase chain reaction) is accuracy, sensitiveness and being unique. In the other way, they reduce the time from the hours to the 30 minutes.

Keywords: exotoxin, nanobiosensor, recognition, Staphylococcus aureus

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Authors: Khaled Khleifat, Muayyad Abboud, Amjad Khleifat, Humodi Saeed

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In this study, Pseudomonas aeruginosa was isolated from infected burn patients and characterized by standard biochemical tests. The in vitro copper uptake was compared between this isolated pathogenic strain and two non-pathogenic control strains of Gram positive bacteria Bacillusthuringiensis strain Israelisas well as Gram negative bacteria Enterobacter aerogenes. Maximum copper uptake of 470 ppm/g biomass was obtained by P. aeruginosa strain, while the control strains B. thuringiensis andEnterobacter aerogenes had copper uptake of 350 and 383 ppm/g biomass, respectively. However, the lowest copper uptake (60 ppm/g biomass) was observed with another control the saprophytic strain Pseudomonas (Shewanella) putrefaciens. A further investigation regarding the effect of copper toxicity on bacterial growth, gave an MIC score of 600 ppm for P. aeruginosa strain compared to 460 and 300 ppm for the two Gram positive and Gram negative control strains, respectively. In tandem with these in vitro findings, blood analysis on burn patients infected with P. aeruginosa has indicated a selective decrease of copper (hypocupremia) and ceruloplasmin plasma levels. The iron metabolism was also affected by this copper deprivation leading to a similar decrease in plasma levels of PCV, iron, total iron binding capacity, and transferrin. All these hematological changes were significantly different (P < 0.05) from the matched group of non-infected burn patients. The observed hypocupremia in infected burn patients was attributed to demanding scavenger ability by P. aeruginosa strain for the copper of plasma.

Keywords: pseudomonas, Cu uptake, burn patients, biosorption

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Authors: E. Bartkiene, M. Ruzauskas, V. Lele, P. Zavistanaviciute, J. Bernatoniene, V. Jakstas, L. Ivanauskas, D. Zadeike, D. Klupsaite, P. Viskelis, J. Bendoraitiene, V. Navikaite-Snipaitiene, G. Juodeikiene

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In this study, antimicrobial nutraceuticals; gummy candies (GC) from bovine colostrum (BC), essential oils (EOs), probiotic lactic acid bacteria (PLAB), and their combinations, were developed. For antimicrobial GC preparation, heteropolysaccharide (agar) was used. The antimicrobial properties of EOs (Eugenia caryophyllata, Thymus vulgaris, Citrus reticulata L., Citrus paradisi L.), BC, L. paracasei LUHS244, L. plantarum LUHS135, and their combinations against pathogenic bacteria strains (Streptococcus mutans, Enterococcus faecalis, Staphylococcus aureus, Salmonella enterica, Escherichia coli, Proteus mirabilis, and Pseudomonas aeruginosa) were evaluated. The highest antimicrobial properties by EO’s (Eugenia caryophyllata and Thymus vulgaris) were established. The optimal ingredients composition for antimicrobial GC preparation was established, which incorporate the BC fermented with L. paracasei LUHS244 in combination with Thymus vulgaris or Eugenia caryophyllata. These ingredients showed high inhibition properties of all tested pathogenic strains (except Pseudomonas aeruginosa). Antimicrobial GC formula consisting of thyme EO (up to 0.2%) and fermented BC (up to 3%), and for taste masking, mandarin or grapefruit EOs (up to 0.2%) was used. Developed GC high overall acceptability and antimicrobial properties, thus, antimicrobial GC could be a preferred form of nutraceuticals. This study was fulfilled with the support of the LSMU-KTU joint project.

Keywords: antimicrobial activity, bovine colostrum, essential oil, gummy candy, probiotic

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Authors: Saifalarab A. Mohmmed, Morgana E. Vianna, Jonathan C. Knowles

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The issue of apical periodontitis has received considerable critical attention. Bacteria is integrated into communities, attached to surfaces and consequently form biofilm. The biofilm structure provides bacteria with a series protection skills against, antimicrobial agents and enhances pathogenicity (e.g. apical periodontitis). Sodium hypochlorite (NaOCl) has become the irrigant of choice for elimination of bacteria from the root canal system based on its antimicrobial findings. The aim of the study was to investigate the effect of different agitation techniques on the efficacy of 2.5% NaOCl to eliminate the biofilm from the surface of the lateral canal using the residual biofilm, and removal rate of biofilm as outcome measures. The effect of canal complexity (lateral canal) on the efficacy of the irrigation procedure was also assessed. Forty root canal models (n = 10 per group) were manufactured using 3D printing and resin materials. Each model consisted of two halves of an 18 mm length root canal with apical size 30 and taper 0.06, and a lateral canal of 3 mm length, 0.3 mm diameter located at 3 mm from the apical terminus. E. faecalis biofilms were grown on the apical 3 mm and lateral canal of the models for 10 days in Brain Heart Infusion broth. Biofilms were stained using crystal violet for visualisation. The model halves were reassembled, attached to an apparatus and tested under a fluorescence microscope. Syringe and needle irrigation protocol was performed using 9 mL of 2.5% NaOCl irrigant for 60 seconds. The irrigant was either left stagnant in the canal or activated for 30 seconds using manual (gutta-percha), sonic and ultrasonic methods. Images were then captured every second using an external camera. The percentages of residual biofilm were measured using image analysis software. The data were analysed using generalised linear mixed models. The greatest removal was associated with the ultrasonic group (66.76%) followed by sonic (45.49%), manual (43.97%), and passive irrigation group (control) (38.67%) respectively. No marked reduction in the efficiency of NaOCl to remove biofilm was found between the simple and complex anatomy models (p = 0.098). The removal efficacy of NaOCl on the biofilm was limited to the 1 mm level of the lateral canal. The agitation of NaOCl results in better penetration of the irrigant into the lateral canals. Ultrasonic agitation of NaOCl improved the removal of bacterial biofilm.

Keywords: 3D printing, biofilm, root canal irrigation, sodium hypochlorite

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Authors: Eusèbe Gnonlonfoun, Xavier Framboisier, Michel Fick, Emmanuel Rondags

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Pathogenic fungi represent a generic problem for cereals, including barley, as they can produce a number of thermostable toxic metabolites such as mycotoxins that contaminate plants and food products, leading to serious health issues for humans and animals and causing significant losses in global food production. In addition, mycotoxins represent a significant technological concern for the malting and brewing industries, as they may affect the quality and safety of raw materials (barley and malt) and final products (beer). Moreover, this situation is worsening due to the highly variable climatic conditions that favor microbial development and the societal desire to reduce the use of phytosanitary products, including fungicides. In this complex environmental, regulatory and economic context for the French barley-malt-beer industry, this project aims to develop an innovative biocontrol process by using technological bacteria, isolated from infection-resistant barley cultures, that are able to reduce the development of spoilage fungi and the associated mycotoxin production. The experimental approach consists of i) coculturing bacterial and pathogenic fungal strains in solid and liquid media to access the growth kinetics of these microorganisms and to evaluate the impact of these bacteria on fungal growth and mycotoxin production; then ii) the results will be used to carry out a micro-malting process in order to develop the aforementioned process, and iii) the technological and sanitary properties of the generated barley malts will finally be evaluated in order to validate the biocontrol process developed. The process is expected to make it possible to guarantee, with controlled costs, an irreproachable hygienic and technological quality of the malt, despite the increasingly complex and variable conditions for barley production. Thus, the results will not only make it possible to maintain the dominant world position of the French barley-malt chain but will also allow it to conquer emerging markets, mainly in Africa and Asia. The use of this process will also contribute to the reduction of the use of phytosanitary products in the field for barley production while reducing the level of contamination of malting plant effluents. Its environmental impact would therefore be significant, especially considering that barley is the fourth most-produced cereal in the world.

Keywords: barley, pathogenic fungi, mycotoxins, malting, bacterial biocontrol

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Authors: Atinderpal Kaur, Shweta Dang

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Our aim is to develop a nanoemulsion-based delivery system containing polyphenon 60 (P60) and ciprofloxacin (Cipro) for intravaginal delivery to treat urinary tract infection. In the present study Polyphenon 60 (P60) and ciprofloxacin (Cipro) were loaded in a single nano emulsion (NE) system via ultra-sonication technique and characterized for particle size, in vitro release and antibacterial efficacy against Bcl-2 level Escherichia coli bacteria. To determine in vivo pharmacokinetic parameters and intravaginal transportation of NE, gamma scintigraphy and biodistribution study was conducted by radiolabelling NE with technetium pertechnetate (99mTc). The preliminary antibacterial investigation showed synergy between these compounds with FICindex of 0.42. The developed formulation showed zeta potential +55.3 and particle size of 151.7 nm, with PDI of 0.196. The in vitro release percentage of P60 at the end of 7th hours was 94.8 ± 0.9 % whereas the release for Cipro was 75.1± 0.15 % in simulated vaginal media. MBC was identified and the findings demonstrated that in both ESBL (Extended Spectrum β- lactamase) and MBL (Metallo β- lactamase) cultures the P60+Cipro NE showed inhibition of growth of all the isolates at 2 mg/ml dilutions. The percentage per gram of radiolabelled drug was found (3.50±0.26) and (3.81±0.30) in kidney and urinary bladder, respectively at 3 h. From the findings, it was concluded that the developed P60+Cipro NE was transported efficiently throughout the target organs, had long duration of action and high biocompatibility via intravaginal administration as compared to oral administration.

Keywords: ciprofloxacin, gamma scintigraphy, intravaginal drug delivery, Polyphenon 60

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Authors: Shivankar Agrawal, Sunil Kumar Deshmukh, Colin Barrow, Alok Adholeya

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Millions of people worldwide are affected by infectious diseases caused by bacteria and fungi. Skin and skin structure infections (SSSI) represent a significant category of infectious disease. Unexpectedly, many pathogens have developed resistance towards current antibiotics and over the time this problem has become more and more serious. All these new problems necessitate the continuous search for novel and alternative antibiotics and antifungals. The aim of our research is the screening of extracts of marine fungi for their antibacterial activity against bacteria causing skin and wound infection in humans. A total of 40 marine samples were collected from west coast and Andaman Island of India and 35 morphologically different marine fungi were isolated using natural sea water medium. Among 35 marine fungi, eight isolates exhibited significant antimicrobial activity against human pathogens. In the course of systematic screening program for bioactive marine fungi, strain 'D5' was found to be most potent strain with MIC value of 1 mg/mL, which was morphologically identified as Simplicillium lamellicola. The effects of the most active crude extracts against their susceptible test microorganisms were also investigated by SEM analysis. Purification and characterization of crude extracts for identification of active lead molecule is under process. The results of diversity and antimicrobial activity have increased the scope of finding industrially important marine fungi from Indian marine environments and these organisms could be vital sources for the discovery of pharmaceutically useful molecules.

Keywords: antimicrobial activity, antibiotic, marine fungi, skin infections

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Authors: Claudio Lamilla, Misael Riquelme, Victoria Saez, Fernanda Sepulveda, Monica Pavez, Leticia Barrientos

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Biosurfactants are compounds synthesized by microorganisms that show various chemical structures, including glycolipids, lipopeptides, polysaccharide-protein complex, phospholipids, and fatty acids. These molecules have attracted attention in recent years due to the amphipathic nature of these compounds, which allows their application in various activities related to emulsification, foaming, detergency, wetting, dispersion and solubilization of hydrophobic compounds. Microorganisms that produce biosurfactants are ubiquitous, not only present in water, soil, and sediments but in extreme conditions of pH, salinity or temperature such as those present in Antarctic ecosystems. Due to this, it is of interest to study biosurfactants producing bacterial strains isolated from Antarctic environments, with the potential to be used in various biotechnological processes. The objective of this research was to characterize biosurfactants produced by bacterial strains isolated from Antarctic environments, with potential use in biotechnological processes for the cleaning of sites contaminated with hydrocarbons. The samples were collected from soils and sediments in the South Shetland Islands and the Antarctic Peninsula, during the Antarctic Research Expedition INACH 2016, from both pristine and human occupied areas (influenced). The bacteria isolation was performed from solid R2A, M1 and LB media. The selection of strains producing biosurfactants was done by hemolysis test on blood agar plates (5%) and blue agar (CTAB). From 280 isolates, it was determined that 10 bacterial strains produced biosurfactants after stimulation with different carbon sources. 16S rDNA taxonomic markers, using the universal primers 27F-1492R, were used to identify these bacterias. Biosurfactants production was carried out in 250 ml flasks using Bushnell Hass liquid culture medium enriched with different carbon sources (olive oil, glucose, glycerol, and hexadecane) during seven days under constant stirring at 20°C. Each cell-free supernatant was characterized by physicochemical parameters including drop collapse, emulsification and oil displacement, as well as stability at different temperatures, salinity, and pH. In addition, the surface tension of each supernatant was quantified using a tensiometer. The strains with the highest activity were selected, and the production of biosurfactants was stimulated in six liters of culture medium. Biosurfactants were extracted from the supernatants with chloroform methanol (2:1). These biosurfactants were tested against crude oil and motor oil, to evaluate their displacement activity (detergency). The characterization by physicochemical properties of 10 supernatants showed that 80% of them produced the drop collapse, 60% had stability at different temperatures, and 90% had detergency activity in motor and olive oil. The biosurfactants obtained from two bacterial strains showed a high activity of dispersion of crude oil and motor oil with halos superior to 10 cm. We can conclude that bacteria isolated from Antarctic soils and sediments provide biological material of high quality for the production of biosurfactants, with potential applications in the biotechnological industry, especially in hydrocarbons -contaminated areas such as petroleum.

Keywords: antarctic, bacteria, biosurfactants, hydrocarbons

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Authors: K. Rasool

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Sulfate-reducing bacteria (SRBs) induced biofilm formation is a global industrial concern due to its role in the development of microbial-induced corrosion (MIC). Herein, we have developed a biodegradable chitosan/lignosulfonate nanocomposite (CS@LS) as an efficient green biocide for the inhibition of SRBs biofilms. We investigated in detail the inhibition mechanism of SRBs by CS@LS in seawater media. Stable CS@LS-1:1 with 150–200 nm average size and zeta potential of + 34.25 mV was synthesized. The biocidal performance of CS@LS was evaluated by sulfate reduction profiles coupled with analysis of extracted extracellular polymeric substances (EPS) and lactate dehydrogenase (LDH) release assays. As the nanocomposite concentration was increased from 50 to 500 µg/mL, the specific sulfate reduction rate (SSRR) decreased from 0.278 to 0.036 g-sulfate/g-VSS*day showing a relative sulfate reduction inhibition of 86.64% as compared to that of control. Similarly, the specific organic uptake rate (SOUR) decreased from 0.082 to 0.039 0.036 g-TOC/g-VSS*day giving a relative co-substrate oxidation inhibition of 52.19% as compared to that of control. The SRBs spiked with 500 µg/mL CS@LS showed a reduction in cell viability to 1.5 × 106 MPN/mL. To assess the biosafety of the nanocomposite on the marine biota, the 72-hours acute toxicity assays using the zebrafish embryo model revealed that the LC50 for the CS@LS was 103.3 µg/mL. Thus, CS@LS can be classified as environmentally friendly. The nanocomposite showed long-term stability and excellent antibacterial properties against SRBs growth and is thus potentially useful for combating the problems of biofilm growth in harsh marine and aquatic environments.

Keywords: green biocides, chitosan/lignosulfonate nanocomposite, SRBs, toxicity

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Authors: Sadia Roshan, Kulsoom Sughra, Shazia Shamas, Shamaila Irum, Haleema Sadia

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To evaluate synergistic medicinal effects of Withania somnifera (Ashwaghandha) and Murraya koenigii (Curry pata) in vitro. Antimicrobial activity was determined using the disc diffusion method against five bacterial and two fungal strains. The antioxidant activity was evaluated by the DPPH assay. The antidiabetic activity was evaluated by alpha-glucosidase inhibition assay and alpha-amylase inhibition assay. Synergistic antibacterial activity was observed against all the strains of bacteria, either Gram-positive or Gram-negative and fungi under study conditions. The maximum antibacterial activity was displayed by combined extract against E. coli i.e. 26±0.4mm. Maximum antifungal activity was shown by combined extract against Aspergillus niger, i.e., 17.3±0.5mm. The antioxidant activity of the combined extract was also significant. Alpha-glucosidase inhibition and alpha-amylase inhibition assays also showed synergism. Results indicate that Withania somnifera and Murraya koengii have medicinal properties. The combined extract of both plants is more potent than their individual extracts, suggesting that these can work in synergism. The research suggests that different plant extracts could be used in combination to increase their medicinal activities by many folds, thus giving an insight into future use of herbal medication.

Keywords: withania somnifera, murraya koenigii, antimicrobial activity, gram-positive bacetria, gram-negative bacteria

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Authors: Kayla Youhanaie, Kenneth Dott, Greg Gillis-Smith

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Access to clean water is a global challenge that affects nearly one-third of the world’s population. A lack of safe drinking water negatively affects a person’s health, safety, and economic status. However, many regions of the world that face this clean water challenge also have high solar energy potential. To address this worldwide issue and utilize available resources, a solar-powered water purification device was developed that could be implemented in communities around the world that lack access to potable water. The device uses ozone to destroy water-borne pathogens and sand filtration to filter out particulates from the water. To select the best method for this application, a quantitative energy efficiency comparison of three water purification methods was conducted: heat, UV light, and ozone. After constructing an initial prototype, the efficacy of the device was tested using agar petri dishes to test for bacteria growth in treated water samples at various time intervals after applying the device to contaminated water. The results demonstrated that the water purification device successfully removed all bacteria and particulates from the water within three minutes, making it safe for human consumption. These results, as well as the proposed design that utilizes widely available resources in target communities, suggest that the device is a sustainable solution to address the global water crisis and could improve the quality of life for millions of people worldwide.

Keywords: clean water, solar powered water purification, ozonation, sand filtration, global water crisis

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Authors: Harikrishna Yadav Nanganuru, Narasimhulu Korrapati

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The demand for the lead (Pb) in the battery industry has been growing for last twenty years. On an average about 2.35 million tons of lead is used in the battery industry. According to the survey of supply and demand battery industry is using 75% of lead produced every year. Due to the increase in battery scrap, secondary lead production has been increasing in this decade. Europe and USA together account for 75% of the world’s secondary lead production. The effluent from used battery scrap consists of high concentrations of lead. Unauthorized disposal of spent batteries, which contain intolerable concentration of lead, into landfills or municipal water canals causes release of Pb into the environment. Lead is one of the toxic heavy metals that have large damaging effects on the human health. Due to its persistence and toxicity, the presence of Pb in drinking water is considered as a special concern. Accumulation of Pb in the human body for long period of time can result in the malfunctioning of some organs. Many technologies have been developed for the removal of lead using microorganisms. In this paper, effluent was taken from the spent battery scrap and was characterized by inductively coupled plasma atomic emission spectrometer. Microorganisms play an important role in removal of lead from the contaminated sites. So, the bacteria were isolated from the effluent. Optimum conditions for the microbial growth and applied for the lead removal. These bacterial cells were immobilized and used for the removal of Pb from the known concentration of metal solution. Scanning electron microscopic (SEM) studies were shown that the Pb was efficiently adsorbed by the immobilized bacteria. From the results of Atomic Absorption Spectroscopy (AAS), 83.40 percentage of Pb was removed in a batch culture.

Keywords: adsorption, effluent, immobilization, lead (Pb)

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Authors: Muhammad Tariq, Muhammad Waseem, Muhammad Hidayat Rasool

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Isolation and characterization of chromium tolerant Staphylococcus aureus from industrial wastewater and their potential use to bioremediate environmental chromium. Objectives: Chromium with its great economic importance in industrial use is major metal pollutant of the environment. Chromium are used in different industries for various applications such as textile, dyeing and pigmentation, wood preservation, manufacturing pulp and paper, chrome plating, steel and tanning. The release of untreated chromium in industrial effluents causes serious threat to environment and human health, therefore, the current study designed to isolate chromium tolerant Staphylococcus aureus for removal of chromium prior to their final discharge into the environment due to its cost effective and beneficial advantage over physical and chemical methods. Methods: Wastewater samples were collected from discharge point of different industries. Heavy metal analysis by atomic absorption spectrophotometer and microbiological analysis such as total viable count, total coliform, fecal coliform and Escherichia coli were conducted. Staphylococcus aureus was identified through gram’s staining, biomeriux vitek 2 microbial identification system and 16S rRNA gene amplification by polymerase chain reaction. Optimum growth conditions with respect to temperature, pH, salt concentrations and effect of chromium on the growth of bacteria, resistance to other heavy metal ions, minimum inhibitory concentration and chromium uptake ability of Staphylococcus aureus strain K1 was determined by spectrophotometer. Antibiotic sensitivity pattern was also determined by disc diffusion method. Furthermore, chromium uptake ability was confirmed by Fourier transform infrared spectroscopy (FTIR) and scanning electron microscope equipped with Oxford Energy Dipersive X-ray (EDX) micro analysis system. Results: The results presented that optimum temperature was 35ᵒC, pH was 8.0 and salt concentration was 0.5% for growth of Staphylococcus aureus K1. The maximum uptake ability of chromium by bacteria was 20mM than other heavy metal ions. The antibiotic sensitivity pattern revealed that Staphylococcus aureus was vancomycin and methicillin sensitive. Non hemolytic activity on blood agar and negative coagulase reaction showed that it was non-pathogenic. Furthermore, the growth of bacteria decreases in the presence of chromium and maximum chromium uptake by bacteria observed at optimum growth conditions. Fourier transform infrared spectroscopy (FTIR), scanning electron microscope (SEM) and Energy dispersive X-ray (EDX) analysis confirmed the presence of chromium uptake by Staphylococcus aureus K1. Conclusion: The study revealed that Staphylococcus aureus K1 have the potential to bio-remediate chromium toxicity from wastewater. Gradually, this biological treatment becomes more important due to its advantage over physical and chemical methods to protect environment and human health.

Keywords: wastewater, staphylococcus, chromium, bioremediation

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Authors: Blessing C. Nwachukwu, Michael O. Taiwo, Wasiu A. Abibu, Isaac O. Ayodeji

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Milk is an important source of micro and macronutrients for humans. Soft Cheese (Wara) is an example of a by-product of milk. In addition, water is considered as one of the most vital resources in cattle farms. Due to the high consumption rate of milk and soft cheese and the traditional techniques involved in their production in Nigeria, there was a need for a microbiological assessment which will be of utmost public health importance. The study thus investigated microbial risk assessments associated with consumption of milk and soft cheese (Wara). It also investigated common pathogens present in dairy water in farms and antibiotic sensitivity profiling for implicated pathogens were conducted. Samples were collected from three different Fulani dairy herds in Ido local government, Ibadan, Oyo State, Nigeria and subjected to microbiological evaluation and antimicrobial susceptibility testing. Aspergillus flavus was the only isolated fungal isolate from Wara while Staphylococcus aureus, Vibro cholera, Hafnia alvei, Proteus mirabilis, Escherishia coli, Psuedomonas aeuroginosa, Citrobacter freundii, and Klebsiella pneumonia were the bacteria genera isolated from Wara, dairy milk and dairy drinking water. Bacterial counts from Wara from the three selected farms A, B and C were 3.5×105 CFU/ml, 4.0×105 CFU/ml and 5.3×105 CFU/ml respectively while the fungal count was 3CFU/100µl. The total bacteria count from dairy milk from the three selected farms A, B and C were Farms 2.0 ×105 CFU/ml, 3.5 × 105 CFU/ml and 6.5 × 105 CFU/ml respectively. 1.4×105 CFU/ml, 1.9×105 CFU/ml and 4.9×105 CFU/ml were the recorded bacterial counts from dairy water from farms A, B and C respectively. The highest antimicrobial resistance of 100% was recorded in Wara with Enrofloxacin, Gentamycin, Cefatriaxone and Colistin. The highest antimicrobial susceptibility of 100% was recorded in Raw milk with Enrofloxacin and Gentamicin. Highest antimicrobial intermediate response of 100% was recorded in Raw milk with Streptomycin. The study revealed that most of the cheeses sold at Ido local Government are contaminated with pathogens. Further research is needed on standardizing the production method to prevent pathogens from gaining access. The presence of bacteria in raw milk indicated contamination due to poor handling and unhygienic practices. Thus, drinking unpasteurized milk is hazardous as it increases the risk of zoonoses. Also, the Provision of quality drinking water is crucial for optimum productivity of dairy. Health education programs aiming at increasing awareness of the importance of clean water for animal health will be helpful.

Keywords: dairy, raw milk, soft cheese, Wara

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Authors: Solomon Gebremariam, Mulugeta Naizigi, Aregawi Haileselassie

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Background: Knowledge of the pattern of bacterial isolates and their antimicrobial susceptibility is crucial for guiding empirical treatment and infection prevention and control measures. Objective: The aim of this study was to analyze the pattern of bacterial isolates and their susceptibility patterns from various specimens. Methods: Retrospectively, a total of 1067 microbiological culture results that were isolated, characterized, and identified by standard microbiological methods and whose antibiotic susceptibility was determined using CLSI guidelines between 2017 and 2019 were retrieved and analyzed. Data were entered and analyzed using the Stata release 10.1 statistical package. Result: The positivity rate of culture was 26.04% (419/1609). The most common bacteria isolated were S. aureus 23.8% (94), E. coli 15.1% (60), Klebsiella pneumonia 14.1% (56), Pseudomonas aeruginosa 8.5% (34), and CONS 7.3% (29). S. aureus and CONS showed a high (58.1% - 96.2%) rate of resistance to most antibiotics tested. They were less resistant to Vancomycin which is 18.6% (13/70) and 11.8% (2/17), respectively. Similarly, the resistance of E. coli, Klebsella pneumonia, and Pseudomonas aeruginosa was high (69.4% - 100%) to most antibiotics. They were less resistant to Ciprofloxacilin, which is 41.1% (23/56), 19.2% (10/52), and 16.1% (5/31), respectively. Conclusion: This study has shown that there is a high rate of antibiotic resistance among bacterial isolates in this hospital. A combination of Vancomycin and Ciprofloxacin should be considered in the choice of antibiotics for empirical treatment of suspected infections due to S. aureus, CONS, E. coli, Klebsiella pneumonia, Pseudomonas such as in infections within hospital setup.

Keywords: antimicrobial, resistance, bacteria, hospital

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Authors: Shubham Sharma, Swarna Jaiswal, Brendan Duffy, Amit Jaiswal

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The key features of any active packaging film are its biodegradability and antimicrobial properties. Biological macromolecules such as polyphenols (ferulic acid (FA) and tannic acids (TA)) are naturally found in plants such as grapes, berries, and tea. In this study, antimicrobial activity screening of several polyphenols was carried out by using minimal inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against two strains of gram-negative bacteria - Salmonella typhimurium, Escherichia coli, and two-gram positive strains - Staphylococcus aureus and Listeria monocytogenes. FA and TA had shown strong antibacterial activity at the low concentration against both gram-positive and gram-negative bacteria. The selected polyphenols FA and TA were incorporated at various concentrations (1%, 5%, and 10% w/w) in the poly(lactide) – poly (butylene adipate-co-terephthalate) (PLA-PBAT) composite film by using the solvent casting method. The effect of TA and FA incorporation in the packaging was characterized based on morphological, optical, color, mechanical, thermal, and antimicrobial properties. The thickness of the FA composite film was increased by 1.5 – 7.2%, while for TA composite film, it increased by 0.018 – 1.6%. FA and TA (10 wt%) composite film had shown approximately 65% - 66% increase in the UV barrier property. As the FA and TA concentration increases from 1% - 10% (w/w), the TS value increases by 1.98 and 1.80 times, respectively. The water contact angle of the film was observed to decrease significantly with the increase in the FA and TA content in the composite film. FA has shown more significant increase in antimicrobial activity than TA in the composite film against Listeria monocytogenes and E. coli. The FA and TA composite film has the potential for its application as an active food packaging.

Keywords: active packaging, biodegradable film, polyphenols, UV barrier, tensile strength

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Authors: Yonas Lamore, Abebe Beyene, Samuel Fekadu, Moa Megersa

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Unaffordable construction cost of conventional water treatment plant and distribution system in most developing countries makes difficult to provide safe and adequate water for all households, especially for the rural setup. Water treatment at the source can be the best alternative. Solar disinfection is one alternative among point of use treatments. In this study, aqua lens, photovoltaic box and glass bottle were used subsequent to plant coagulants to evaluate microbial reduction potentials. Laboratory- and field-based experiments were conducted from May to August 2016. The Escherichia coli, total coliforms and heterotrophic plate counts were used as indicator organisms. The result indicated that aqua lens (AL), photovoltaic box (PV) and glass bottle (GB) have high inactivation rate subsequently almost for all indicator organisms in short solar exposure time. Total coliforms were inactivated in AL (SD = 15.8 °C, R2 = 0.92) followed by PV inactivation temperature associa- tion (SD = 11.6 C, R2 = 0.90), and the GB concentrator was inactivated (SD = 10.9 °C, R2 = 0.70) at turbidity level of 3.41 NTU. As the study indicated, aqua lens coupled with Moringa oleifera coagulant can be an effective with minimum cost for household water treatment system. The study also concludes heterotrophic bacteria were more resistant than other types of bacteria in SODIS with similar exposure time.

Keywords: acrylic glass, aqua lens, moringa olifera, photovoltaic box, solar disinfection, water treatment

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Authors: Faozia A. S. Abuhtana, Yahia S. Abujnah, Said O. Gnann

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Ultra High Temperature (UHT) processed milk is widely distributed and preferred in numerous countries all over the world due its relatively high quality and long shelf life. Because of the notable high consumption rate of UHT in Libya in addition to negligible studies related to such product on the local level, this study was designed to assess the shelf life of locally produced as well as imported reconstituted sterilized whole milk samples marketed in Tripoli, Libya . Four locally produced vs. three imported brands were used in this study. All samples were stored at room temperature (25± 2C ) for 8 month long period, and subjected to physical, chemical, microbiological and sensory tests. These tests included : measurement of pH, specific gravity, percent acidity, and determination of fat, protein and melamine content. Microbiological tests included total aerobic count, total psychotropic bacteria, total spore forming bacteria and total coliform counts. Results indicated no detection of microbial growth of any type during the study period, in addition to no detection of melamine in all samples. On the other hand, a gradual decline in pH accompanied with gradual increase in % acidity of both locally produced and imported samples was observed. Such changes in both pH and % acidity reached their lowest and highest values respectively during the 24th week of storage. For instance pH values were (6.40, 6.55, 6.55, 6.15) and (6.30, 6.50, 6.20) for local and imported brands respectively. On the other hand, % acidity reached (0.185, 0181, 0170, 0183) and (0180, 0.180, 0.171) at the 24th week for local and imported brands respectively. Similar pattern of decline was also observed in specific gravity, fat and protein content in some local and imported samples especially at later stages of the study. In both cases, some of the recorded pH values, % acidity, sp. gravity and fat content were in violation of the accepted limits set by Libyan standard no. 356 for sterilized milk. Such changes in pH, % acidity and other UHT sterilized milk constituents during storage were coincided with a gradual decrease in the degree of acceptance of the stored milk samples of both types as shown by sensory scores recorded by the panelists. In either case degree of acceptance was significantly low at late stages of storage and most milk samples became relatively unacceptable after the 18th and 20th week for both untrained and trained panelists respectively.

Keywords: UHT milk, shelf life, quality, gravity, bacteria

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Authors: Mary Ghobrial, Sahar Wefky

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Marine macroalgae have proven to be rich source of bioactive compounds with biomedical potential, not only for human but also for veterinary medicine. Emergence of microbial disease in aquaculture industries implies serious loses. Usage of commercial antibiotics for fish disease treatment produces undesirable side effects. Marine organisms are a rich source of structurally novel biologically active metabolites. Competition for space and nutrients led to the evolution of antimicrobial defense strategies in the aquatic environment. The interest in marine organisms as a potential and promising source of pharmaceutical agents has increased in the last years. Many bioactive and pharmacologically active substances have been isolated from microalgae. Compounds with antibacterial, antifungal and antiviral activities have been also detected in green, brown and red algae. Selected species of marine benthic algae belonging to the Phaeophyta and Rhodophyta, collected from different coastal areas of Alexandria (Egypt), were investigated for their antibacterial and antifungal, activities. Macroalgae samples were collected during low tide from the Alexandria Mediterranean coast. Samples were air dried under shade at room temperature. The dry algae were ground, using electric mixer grinder. They were soaked in 10 ml of each of the solvents acetone, ethanol, methanol and hexane. Antimicrobial activity was evaluated using well-cut diffusion technique In vitro screening of organic solvent extracts from the marine macroalgae Laurencia pinnatifida, Pterocladia capillaceae, Stepopodium zonale, Halopteris scoparia and Sargassum hystrix, showed specific activity in inhibiting the growth of five virulent strains of bacteria pathogenic to fish Pseudomonas fluorescens, Aeromonas hydrophila, Vibrio anguillarum, V. tandara, Escherichia coli and two fungi Aspergillus flavus and A. niger. Results showed that, acetone and ethanol extracts of all test macroalgae exhibited antibacterial activity, while acetone extract of the brown Sargassum hystrix displayed the highest antifungal activity. The extracts of seaweeds inhibited bacteria more strongly than fungi and species of the Rhodophyta showed the greatest activity against the bacteria rather than fungi tested. The gas liquid chromatography coupled with mass spectrometry detection technique allows good qualitative and quantitative analysis of the fractionated extracts with high sensitivity to the smaller amounts of components. Results indicated that, the main common component in the acetone extracts of L. pinnatifida and P. capillacea is 4-hydroxy-4-methyl2-pentanone representing 64.38 and 58.60%. Thus, the extracts derived from the red macroalgae were more efficient than those obtained from the brown macroalgae in combating bacterial pathogens rather than pathogenic fungi. The most preferred species over all was the red Laurencia pinnatifida. In conclusion, the present study provides the potential of red and brown macroalgae extracts for development of anti-pathogenic agents for use in fish aquaculture.

Keywords: bacteria, fungi, extracts, solvents

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Authors: Nesrine Benarous, Hassiba Bougueria, Nabila Moussa Slimane, Aouatef Cherouana

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Crystal polymorphism is important for the synthesis of more potent and bioactive pharmaceutical compounds, including their different properties, such as packing arrangement and conformation. In fact, polymorphism plays a vital role in drug development. Different parameters affect the crystallization and give their degree of freedom. Severalproperties affected polymorphism, like kinetics, thermodynamics, spectroscopy, and mechanical property. Various techniques are used for characterizing polymorphs, are crystallography, morphology, phase transitions, molecular motion, and chemical environment. In this work, crystal structures of two polymorphs (I and II) of the Schiff base (SB) title compound were prepared by condensation reaction. The crystal structures of both polymorphs were determined by single X-ray analysis. The two polymorphs crystallize in two different space groups: P21/c for I and Pbca for II. The dihedral angles between the two phenyl rings are 4.81º for I and 82.27º for II. Both crystal structures are built on the basis of moderate and weak hydrogen bonds, 𝜋-stacking, and halogen⋯halogeninteractions. On the other hand, Hirshfeld surface (HS) analysis indicates that the most important contributions to the crystal packing for the two polymorphs are from Cl⋯H/H⋯Cl, H⋯H, and N⋯H/H⋯N contacts. These are followed by C⋯H/H⋯C for compound I and C⋯C and by C⋯H/H⋯C contacts for compound II. Afterwards, the in vitro antibacterial activity revealed that the SB have been found effective against G- bacteria Klebsiella pneumonia andG+ bacteria Staphylococcus aureuswith MIC value of14.37μg/mL. Moreover, the SBexhibited moderate toxicity against Brine Shrimp with LC50 value of 44.19μg/mL.

Keywords: polymorph, crystal structure, hirshfeld surface analysis, in vitro antibacterial activity, toxicity

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