Search results for: pathogenic microorganisms

Authors: Ranjita Ghoshmoulick, Aswathi Madhavan, Subhavna Juneja, Prasenjit Sen, Jaydeep Bhattacharya

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Type 2 diabetes mellitus (T2DM) is a very complex and multifactorial metabolic disease where the blood sugar level goes up. One of the major consequence of this elevated blood sugar is the formation of AGE (Advance Glycation Endproducts), from a series of chemical or biochemical reactions. AGE are detrimental because it leads to severe pathogenic complications. They are a group of structurally diverse chemical compounds formed from nonenzymatic reactions between the free amino groups (-NH2) of proteins and carbonyl groups (>C=O) of reducing sugars. The reaction is known as Maillard Reaction. It starts with the formation of reversible schiff’s base linkage which after sometime rearranges itself to form Amadori Product along with dicarbonyl compounds. Amadori products are very unstable hence rearrangement goes on until stable products are formed. During the course of the reaction a lot of chemically unknown intermediates and reactive byproducts are formed that can be termed as Early Glycation Products. And when the reaction completes, structurally stable chemical compounds are formed which is termed as Advanced Glycation Endproducts. Though all glycation products have not been characterized well, some fluorescence compounds e.g pentosidine, Malondialdehyde (MDA) or carboxymethyllysine (CML) etc as AGE and α-dicarbonyls or oxoaldehydes such as 3-deoxyglucosone (3-DG) etc as the intermediates have been identified. In this work Gold NanoParticle (GNP) was used as an optical indicator of glycation products. To achieve faster glycation kinetics and high AGE accumulation, fructose was used instead of glucose. Hemoglobin A0 (HbA0) was fructosylated by in-vitro method. AGE formation was measured fluorimetrically by recording emission at 450nm upon excitation at 350nm. Thereafter this fructosylated HbA0 was fractionated by column chromatography. Fractionation separated the proteinaceous substance from the AGEs. Presence of protein part in the fractions was confirmed by measuring the intrinsic protein fluorescence and Bradford reaction. GNPs were synthesized using the templates of chromatographically separated fractions of fructosylated HbA0. Each fractions gave rise to GNPs of varying color, indicating the presence of distinct set of glycation products differing structurally and chemically. Clear solution appeared due to settling down of particles in some vials. The reactive groups of the intermediates kept the GNP formation mechanism on and did not lead to a stable particle formation till Day 10. Whereas SPR of GNP showed monotonous colour for the fractions collected in case of non fructosylated HbA0. Our findings accentuate the use of GNPs as a simple colorimetric sensing platform for the identification of intermediates of glycation reaction which could be implicated in the prognosis of the associated health risk due to T2DM and others.

Keywords: advance glycation endproducts, glycation, gold nano particle, sensor

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Authors: Thayse Marques Passos, Brid Quilty, Mary Pryce

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

Keywords: bacterial aggregation, photoinactivation, Pseudomonads, water disinfection

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Authors: N. Demertzis, J. L. Bowen

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Sepsis is a complex and rapidly evolving condition, resulting from uncontrolled prolonged activation of host immune system due to pathogenic insult. The aim of this study is the development of a multiplex electrochemical sensing platform, capable of detecting both pathogen associated and host immune markers to enable the rapid and definitive diagnosis of sepsis. A combination of aptamers and molecular imprinting approaches have been employed to generate sensing systems for lipopolysaccharide (LPS), c-reactive protein (CRP) and procalcitonin (PCT). Gold working electrodes were mechanically polished and electrochemically cleaned with 0.1 M sulphuric acid using cyclic voltammetry (CV). Following activation, a self-assembled monolayer (SAM) was generated, by incubating the electrodes with a thiolated anti-LPS aptamer / dithiodibutiric acid (DTBA) mixture (1:20). 3-aminophenylboronic acid (3-APBA) in combination with the anti-LPS aptamer was used for the development of the hybrid molecularly imprinted sensor (apta-MIP). Aptasensors, targeting PCT and CRP were also fabricated, following the same approach as in the case of LPS, with mercaptohexanol (MCH) replacing DTBA. In the case of the CRP aptasensor, the SAM was formed following incubation of a 1:1 aptamer: MCH mixture. However, in the case of PCT, the SAM was formed with the aptamer itself, with subsequent backfilling with 1 μM MCH. The binding performance of all systems has been evaluated using electrochemical impedance spectroscopy. The apta-MIP’s polymer thickness is controlled by varying the number of electropolymerisation cycles. In the ideal number of polymerisation cycles, the polymer must cover the electrode surface and create a binding pocket around LPS and its aptamer binding site. Less polymerisation cycles will create a hybrid system which resembles an aptasensor, while more cycles will be able to cover the complex and demonstrate a bulk polymer-like behaviour. Both aptasensor and apta-MIP were challenged with LPS and compared to conventional imprinted (absence of aptamer from the binding site, polymer formed in presence of LPS) and non-imprinted polymers (NIPS, absence of LPS whilst hybrid polymer is formed). A stable LPS aptasensor, capable of detecting down to 5 pg/ml of LPS was generated. The apparent Kd of the system was estimated at 17 pM, with a Bmax of approximately 50 pM. The aptasensor demonstrated high specificity to LPS. The apta-MIP demonstrated superior recognition properties with a limit of detection of 1 fg/ml and a Bmax of 100 pg/ml. The CRP and PCT aptasensors were both able to detect down to 5 pg/ml. Whilst full binding performance is currently being evaluated, there is none of the sensors demonstrate cross-reactivity towards LPS, CRP or PCT. In conclusion, stable aptasensors capable of detecting LPS, PCT and CRP at low concentrations have been generated. The realisation of a multiplex panel such as described herein, will effectively contribute to the rapid, personalised diagnosis of sepsis.

Keywords: aptamer, electrochemical impedance spectroscopy, molecularly imprinted polymers, sepsis

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Authors: Ahmed F. Ghanem, Marwa I. Wahba, Asmaa N. El-Dein, Mohamed A. EL-Raey, Ghada E.A. Awad

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Numerous attempts are being performed in order to formulate suitable packaging materials for meat products. However, to the best of our knowledge, the incorporation of free hibiscus extract or its microcapsules in the pure polyvinyl alcohol (PVA) matrix as packaging materials for meats is seldom reported. Therefore, this study aims at protection of the aqueous crude extract of hibiscus flowers utilizing spry drying encapsulation technique. Fourier transform infrared (FTIR), scanning electron microscope (SEM), and zetasizer results confirmed the successful formation of assembled capsules via strong interactions, spherical rough microparticles, and ~ 235 nm of particle size, respectively. Also, the obtained microcapsules enjoy high thermal stability, unlike the free extract. Then, the obtained spray-dried particles were incorporated into the casting solution of the pure PVA film with a concentration 10 wt. %. The segregated free-standing composite films were investigated, compared to the neat matrix, with several characterization techniques such as FTIR, SEM, thermal gravimetric analysis (TGA), mechanical tester, contact angle, water vapor permeability, and oxygen transmission. The results demonstrated variations in the physicochemical properties of the PVA film after the inclusion of the free and the extract microcapsules. Moreover, biological studies emphasized the biocidal potential of the hybrid films against microorganisms contaminating the meat. Specifically, the microcapsules imparted not only antimicrobial but also antioxidant activities to PVA. Application of the prepared films on the real meat samples displayed low bacterial growth with a slight increase in the pH over the storage time up to 10 days at 4 oC which further proved the meat safety. Moreover, the colors of the films did not significantly changed except after 21 days indicating the spoilage of the meat samples. No doubt, the dual-functional of prepared composite films pave the way towards combined active/smart food packaging applications. This would play a vital role in the food hygiene, including also quality control and assurance.

Keywords: PVA, hibiscus, extraction, encapsulation, active packaging, smart and intelligent packaging, meat spoilage

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Authors: Mukesh K. Dwivedi, Rajeshwar N. Srivastava, Amit K. Bhagat, Saloni Raj

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Introduction: Management of Pressure Ulcer (PU) is an ongoing clinical challenge particularly in traumatic paraplegia patients in developing countries where socio economic conditions often dictate treatment modalities. When negative pressure wound therapy (NPWT) was introduced, there were a series of devices (V.A.C., KCI, San Antonio, TX) manufactured. These devices for NPWT are costly and hard to afford by patients in developing countries like India. Considering this limitation, this study was planned to design an RCT to compare NPWT by an indigenized locally constructed NPD and conventional gauze dressing for the treatment of PU. Material and Methods: This RCT (CTRI/2014/09/0050) was conducted in the Department of Orthopaedic Surgery at King George’s Medical University (KGMU), India. Thirty-four (34) subjects of traumatic paraplegia having PU of stage 3 or 4, were enrolled and randomized in two treatment groups (NPWT Group & Conventional dressing group). The outcome measures of this study were surface area and depth of PU, exudates, microorganisms and matrix metalloproteinase-8 (MMP-8) during 0 to 9 weeks follow-ups. Levels of MMP-8 were analyzed in the tissues of PU at week 0, 3, 6 and week 9 by Enzyme Linked Immuno Sorbent Assay (ELISA). Results: Significantly reduced length of PU in NPWT group was observed at week 6 (p=0.04) which further reduced at week 9 (p=0.001) as compared to conventionally treated group. Similarly significant reduction of width and depth of PU was observed in NPWT at week 9 (p<0.05). The exudate became significantly (p=0.001) lower in NPWT group as compared with conventionally treated group from 6th to 9th week. Clearance and conversion of slough into red granulation tissue was significantly higher in NPWT group (p=0.001). At week 9, the wound culture was negative in all the subjects of NPWT group, while it was positive in 10 (41⋅6%) subjects of conventional group. Significantly lower level of MMP-8 was observed in subjects of NPWT group at week 6 (0.006**), and continually more reduction was observed at week 9 (<0.0001**) as compared to the conventional group. Conclusion: NPWT by locally constructed NPD is better wound care procedure for management of PU. Our device gave similar results as commercially available devices. Reduction of level of MMP-8 and increased rate of healing was achieved by negative pressure wound therapy (NPWT) as compared to conventional dressing.

Keywords: NPWT, NPD, MMP8, ELISA

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Authors: Carlos Y. Soto, Camila A. Lota, G. Astrid Garzón

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Bacterial biofilms cause an ongoing problem for food safety. They are formed when microorganisms aggregate to form a community that attaches to solid surfaces. Biofilms increase the resistance of pathogens to cleaning, disinfection and antibacterial products. This resistance gives rise to problems for human health, industry, and agriculture. At present, plant extracts rich in polyphenolics are being investigated as natural alternatives to degrade bacterial biofilms. The pomace of the tropical Berry Vaccinium meridionale S. contains high amounts of phenolic compounds. Therefore, in the current study, the antimicrobial and antibiofilm effects of extracts from the pomace of Vaccinium meridionale S. were tested on three foodborne pathogens: Enterohaemorrhagic Escherichia coli O157:H7 (ATCC®700728TM), Staphylococcus aureus subsp. aureus (ATCC® 6538TM), and Salmonella enterica serovar Enteritidis (ATCC® 13076TM). Microwave-assisted extraction was used to extract polyphenols with aqueous methanol (80% v/v) at a solid to solvent ratio of 1:10 (w/v) for 20 min. The magnetic stirring was set at 400 rpm, and the microwave power was adjusted to 400 W. The antimicrobial effect of the extract was assessed by determining the half maximal inhibitory concentration (IC50) against the three food poisoning pathogens at concentrations ranging from 50 to 2,850 μg gallic acid equivalents (GAE)/mL of the extract. Biofilm inhibition was assessed using a crystal violet assay applying the same range of concentration. Three replications of the experiments were carried out, and all analyses were run in triplicate. IC50 values were determined using the GraphPad Prism8® program. Significant differences (P<0.05) among means were identified using one-factor analysis of variance (ANOVA) and the post-hoc least significant difference (LSD) test using the Statgraphics plus program, version 2.1.There was significant difference among the mean IC50 values for the tested bacteria. The IC50 for S. aureus was 48 ± 9 μg GAE/mL, followed by 123 ± 49 μg GAE/mL for Salmonella and 376 ± 32 μg GAE/mL for E. coli. The percent inhibition of the extract on biofilm formation was significantly higher for S. aureus (85.8  0.3), followed by E. coli (74.5  1.0) and Salmonella (53.6  9.7). These findings suggest that polyphenolic extracts obtained from the pomace of V. meridionale S. might be used as natural antimicrobial and anti-biofilm natural agents, effective against S. aureus, E. coli and Salmonella enterica.

Keywords: antibiofilm, antimicrobial, E. coli, S. aureus, salmonella, IC50, pomace, V. meridionale

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Authors: Govinda Pathak

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In modern agriculture, the sustainable enhancement of crop productivity while minimizing environmental impacts remains a paramount challenge. Plant Growth Promoting Rhizobacteria (PGPR) have emerged as a promising solution to address this challenge. The rhizosphere, the dynamic interface between plant roots and soil, hosts intricate microbial interactions crucial for plant health and nutrient acquisition. PGPR, a subset of rhizospheric microorganisms, exhibit multifaceted beneficial effects on plants. Their abilities to stimulate growth, confer stress tolerance, enhance nutrient availability, and suppress pathogens make them invaluable contributors to sustainable agriculture. This work examines the pivotal role of free living nitrogen fixer in optimizing agricultural practices. We delve into the intricate mechanisms underlying PGPR-mediated plant-microbe interactions, encompassing quorum sensing, root exudate modulation, and signaling molecule exchange. Furthermore, we explore the diverse strategies employed by PGPR to enhance plant resilience against abiotic stresses such as drought, salinity, and metal toxicity. Additionally, we highlight the role of PGPR in augmenting nutrient acquisition and soil fertility through mechanisms such as nitrogen fixation, phosphorus solubilization, and mineral mobilization. Furthermore, we discuss the potential of PGPR in minimizing the reliance on chemical fertilizers and pesticides, thereby contributing to environmentally friendly agriculture. However, harnessing the full potential of PGPR requires a comprehensive understanding of their interactions with host plants and the surrounding microbial community. We also address challenges associated with PGPR application, including formulation, compatibility, and field efficacy. As the quest for sustainable agriculture intensifies, harnessing the remarkable attributes of PGPR offers a holistic approach to propel agricultural productivity while maintaining ecological balance. This work underscores the promising prospect of free living nitrogen fixer as a panacea for addressing critical agricultural challenges regarding chemical urea in an era of sustainable and resilient food production.

Keywords: PGPR, nitrogen fixer, quorum sensing, Rhizobacteria, pesticides

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Authors: Shalini TV, Gangadharan GG, Sriranjini S Jaideep, ASN Seshasayee, Awadhesh Pandit

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Introduction: Prakriti (body-mind constitution of an individual) is a conventional, customized and unique understanding of which is essential for the personalized medicine described in Ayurveda, Indian System of Medicine. Based on the Doshas( functional, bio humoral unit in the body), individuals are categorized into three major Prakriti- Vata, Pitta, and Kapha. The human gut microbiome hosts plenty of highly diverse and metabolically active microorganisms, mainly dominated by the bacteria, which are known to influence the physiology of an individual. Few researches have shown the correlation between the Prakriti and the biochemical parameters. In this study, an attempt was made to explore any correlation between the Prakriti (phenotype of an individual) with the Genetic makeup of the gut microbiome in healthy individuals. Materials and methods: 270 multi-ethnic, healthy volunteers of both sex with the age group between 18 to 40 years, with no history of antibiotics in the last 6 months were recruited into three groups of Vata, Pitta, and Kapha. The Prakriti of the individual was determined using Ayusoft, a software designed by CDAC, Pune, India. The volunteers were subjected to initial screening for the assessment of their height, weight, Body Mass Index, Vital signs and Blood investigations to ensure they are healthy. The stool and saliva samples of the recruited volunteers were collected as per the standard operating procedure developed, and the bacterial DNA was isolated using Qiagen kits. The extracted DNA was subjected to 16s rRNA sequencing using the Illumina kits. The sequencing libraries are targeting the variable V3 and V4 regions of the 16s rRNA gene. Paired sequencing was done on the MiSeq system and data were analyzed using the CLC Genomics workbench 11. Results: The 16s rRNA sequencing of the V3 and V4 regions showed a diverse pattern in both the oral and stool microbial DNA. The study did not reveal any specific pattern of bacterial flora amongst the Prakriti. All the p-values were more than the effective alpha values for all OTUs in both the buccal cavity and stool samples. Therefore, there was no observed significant enrichment of an OTU in the patient samples from either the buccal cavity or stool samples. Conclusion: In healthy volunteers of multi-ethnicity, due to the influence of the various factors, the correlation between the Prakriti and the gut microbiome was not seen.

Keywords: gut microbiome, ayurveda Prakriti, sequencing, multi-ethnic urban population

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Authors: Doris Lacej, Eni Bushi

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Cleaning and disinfection procedures are essential for maintaining the cleanliness status of the pharmaceutical manufacturing environment particularly of the cleanrooms and sterile unit area. The Good Manufacturing Practice (GMP) Annex 1 recommendation highly requires the implementation of the standard and validated cleaning and disinfection protocols. However, environmental monitoring has shown that even a validated cleaning method with certified agents may result in the presence of atypical microorganisms’ colony that exceeds GMP limits for a specific cleanroom area. In response to this issue, this case study aims to arrive at the root cause of the microbial contamination observed in the sterile production environment in Profarma pharmaceutical industry in Albania through applying a problem-solving practical approach that ensures the appropriate sterility grade. The guidelines and literature emphasize the importance of several factors in the prevention of possible microbial contamination occurring in cleanrooms, grade A and C. These factors are integrated into a practical framework, to identify the root cause of the presence of Aspergillus Niger colony in the sterile production environment in Profarma pharmaceutical industry in Albania. In addition, the application of a semi-automatic disinfecting system such as H2O2 FOG into sterile grade A and grade C cleanrooms has been an effective solution in eliminating the atypical colony of Aspergillus Niger. Selecting the appropriate detergents and disinfectants at the right concentration, frequency, and combination; the presence of updated and standardized guidelines for cleaning and disinfection as well as continuous training of operators on these practices in accordance with the updated GMP guidelines are some of the identified factors that influence the success of achieving sterility grade. However, to ensure environmental sustainability it is important to be prepared for identifying the source of contamination and making the appropriate decision. The proposed case-based practical approach may help pharmaceutical companies to achieve sterile production and cleanliness environmental sustainability in challenging situations. Apart from the integration of valid agents and standardized cleaning and disinfection protocols according to GMP Annex 1, pharmaceutical companies must be careful and investigate the source and all the steps that can influence the results of an abnormal situation. Subsequently apart from identifying the root cause it is important to solve the problem with a successful alternative approach.

Keywords: cleanrooms, disinfectants, environmental monitoring, GMP Annex 1

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Authors: Yemane Tadesse Gebreslassie, Fisseha Guesh Gebremeskel

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Nanotechnology has made remarkable advancements in recent years, revolutionizing various scientific fields, industries, and research institutions through the utilization of metal and metal oxide nanoparticles. Among these nanoparticles, copper oxide nanoparticles (CuO NPs) have garnered significant attention due to their versatile properties and wide-range applications, particularly, as effective antimicrobial and anticancer agents. CuO NPs can be synthesized using different methods, including physical, chemical, and biological approaches. However, conventional chemical and physical approaches are expensive, resource-intensive, and involve the use of hazardous chemicals, which can pose risks to human health and the environment. In contrast, biological synthesis provides a sustainable and cost-effective alternative by eliminating chemical pollutants and allowing for the production of CuO NPs of tailored sizes and shapes. This comprehensive review focused on the green synthesis of CuO NPs using various biological resources, such as plants, microorganisms, and other biological derivatives. Current knowledge and recent trends in green synthesis methods for CuO NPs are discussed, with a specific emphasis on their biomedical applications, particularly in combating cancer and microbial infections. This review highlights the significant potential of CuO NPs in addressing these diseases. By capitalizing on the advantages of biological synthesis, such as environmental safety and the ability to customize nanoparticle characteristics, CuO NPs have emerged as promising therapeutic agents for a wide range of conditions. This review presents compelling findings, demonstrating the remarkable achievements of biologically synthesized CuO NPs as therapeutic agents. Their unique properties and mechanisms enable effective combating against cancer cells and various harmful microbial infections. CuO NPs exhibit potent anticancer activity through diverse mechanisms, including induction of apoptosis, inhibition of angiogenesis, and modulation of signaling pathways. Additionally, their antimicrobial activity manifests through various mechanisms, such as disrupting microbial membranes, generating reactive oxygen species, and interfering with microbial enzymes. This review offers valuable insights into the substantial potential of biologically synthesized CuO NPs as an alternative approach for future therapeutic interventions against cancer and microbial infections.

Keywords: copper oxide nanoparticles, green synthesis, nanotechnology, microbial infection

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Authors: Tatyane Lopes de Freitas, Antonio Diogo S. Vieira, Susana Marta Isay Saad, Maria Ines Genovese

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The fruit processing industries generate a large volume of residues to produce juices, pulps, and jams. These residues, or by-products, consisting of peels, seeds, and pulps, are routinely discarded. Fruits are rich in bioactive compounds, including polyphenols, which have positive effects on health. Dry residues from two fruits, acerola (M. emarginata D. C.) and orange (C. sinensis), were characterized in relation to contents of ascorbic acid, minerals, total dietary fibers, moisture, ash, lipids, proteins, and carbohydrates, and also high performance liquid chromatographic profile of flavonoids, total polyphenols and proanthocyanidins contents, and antioxidant capacity by three different methods (Ferric reducing antioxidant power assay-FRAP, Oxygen Radical Absorbance Capacity-ORAC, 1,1-diphenyl-2-picrylhydrazil (DPPH) radical scavenging activity). Acerola by-products presented the highest acid ascorbic content (605 mg/100 g), and better antioxidant capacity than orange by-products. The dry residues from acerola demonstrated high contents of proanthocyanidins (617 µg CE/g) and total polyphenols (2525 mg gallic acid equivalents - GAE/100 g). Both presented high total dietary fiber (above 60%) and protein contents (acerola: 10.4%; orange: 9.9%), and reduced fat content (acerola: 1.6%; orange: 2.6%). Both residues showed high levels of potassium, calcium, and magnesium, and were considered sources of these minerals. With acerola by-product, four formulations of probiotics fermented milks were produced: F0 (without the addition of acerola residue (AR)), F2 (2% AR), F5 (5% AR) and F10 (10% AR). The physicochemical characteristics of the fermented milks throughout of storage were investigated, as well as the impact of in vitro simulated gastrointestinal conditions on flavonoids and probiotics. The microorganisms analyzed maintained their populations around 8 log CFU/g during storage. After the gastric phase of the simulated digestion, the populations decreased, and after the enteric phase, no colonies were detected. On the other hand, the flavonoids increased after the gastric phase, maintaining or suffering small decrease after enteric phase. Acerola by-products powder is a valuable ingredient to be used in functional foods because is rich in vitamin C, fibers and flavonoids. These flavonoids appear to be highly resistant to the acids and salts of digestion.

Keywords: acerola, orange, by-products, fermented milk

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Authors: S. D. N. K. Bathige, G. I. Godahewa, Navaneethaiyer Umasuthan, Jehee Lee

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Kunitz-type serine protease inhibitors (KTIs) are identified in various organisms including animals, plants and microbes. These proteins shared single or multiple Kunitz inhibitory domains link together or associated with other types of domains. Characteristic Kunitz type domain composed of around 60 amino acid residues with six conserved cysteine residues to stabilize by three disulfide bridges. KTIs are involved in various physiological processes, such as ion channel blocking, blood coagulation, fibrinolysis and inflammation. In this study, two Kunitz-type domain containing protein was identified from rock bream database and designated as RbKunitz. The coding sequence of RbKunitz encoded for 507 amino acids with 56.2 kDa theoretical molecular mass and 5.7 isoelectric point (pI). There are several functional domains including MANEC superfamily domain, PKD superfamily domain, and LDLa domain were predicted in addition to the two characteristic Kunitz domain. Moreover, trypsin interaction sites were also identified in Kunitz domain. Homology analysis revealed that RbKunitz shared highest identity (77.6%) with Takifugu rubripes. Completely conserved 28 cysteine residues were recognized, when comparison of RbKunitz with other orthologs from different taxonomical groups. These structural evidences indicate the rigidity of RbKunitz folding structure to achieve the proper function. The phylogenetic tree was constructed using neighbor-joining method and exhibited that the KTIs from fish and non-fish has been evolved in separately. Rock bream was clustered with Takifugu rubripes. The SYBR Green qPCR was performed to quantify the RbKunitz transcripts in different tissues and challenged tissues. The mRNA transcripts of RbKunitz were detected in all tissues (muscle, spleen, head kidney, blood, heart, skin, liver, intestine, kidney and gills) analyzed and highest transcripts level was detected in gill tissues. Temporal transcription profile of RbKunitz in rock bream blood tissues was analyzed upon LPS (lipopolysaccharide), Poly I:C (Polyinosinic:polycytidylic acid) and Edwardsiella tarda challenge to understand the immune responses of this gene. Compare to the unchallenged control RbKunitz exhibited strong up-regulation at 24 h post injection (p.i.) after LPS and E. tarda injection. Comparatively robust expression of RbKunits was observed at 3 h p.i. upon Poly I:C challenge. Taken together all these data indicate that RbKunitz may involve into to immune responses upon pathogenic stress, in order to protect the rock bream.

Keywords: Kunitz-type, rock bream, immune response, serine protease inhibitor

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Authors: Neveen AlQasas, Daniel Johnson

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Membranes for water treatment are an established technology that attracts great attention due to its simplicity and cost effectiveness. However, membranes in operation suffer from the adverse effect of membrane fouling. Bio-fouling is a phenomenon that occurs at the water-membrane interface, and is a dynamic process that is initiated by the adsorption of dissolved organic material, including biomacromolecules, on the membrane surface. After initiation, attachment of microorganisms occurs, followed by biofilm growth. The biofilm blocks the pores of the membrane and consequently results in reducing the water flux. Moreover, the presence of a fouling layer can have a substantial impact on the membrane separation properties. Understanding the mechanism of the initiation phase of biofouling is a key point in eliminating the biofouling on membrane surfaces. The adhesion and attachment of different fouling materials is affected by the surface properties of the membrane materials. Therefore, surface properties of different polymeric materials had been studied in terms of their surface energies and Hansen solubility parameters (HSP). The difference between the combined HSP parameters (HSP distance) allows prediction of the affinity of two materials to each other. The possibilities of measuring the HSP of different polymer films via surface measurements, such as contact angle has been thoroughly investigated. Knowing the HSP of a membrane material and the HSP of a specific foulant, facilitate the estimation of the HSP distance between the two, and therefore the strength of attachment to the surface. Contact angle measurements using fourteen different solvents on five different polymeric films were carried out using the sessile drop method. Solvents were ranked as good or bad solvents using different ranking method and ranking was used to calculate the HSP of each polymeric film. Results clearly indicate the absence of a direct relation between contact angle values of each film and the HSP distance between each polymer film and the solvents used. Therefore, estimating HSP via contact angle alone is not sufficient. However, it was found if the surface tensions and viscosities of the used solvents are taken in to the account in the analysis of the contact angle values, a prediction of the HSP from contact angle measurements is possible. This was carried out via training of a neural network model. The trained neural network model has three inputs, contact angle value, surface tension and viscosity of solvent used. The model is able to predict the HSP distance between the used solvent and the tested polymer (material). The HSP distance prediction is further used to estimate the total and individual HSP parameters of each tested material. The results showed an accuracy of about 90% for all the five studied films

Keywords: surface characterization, hansen solubility parameter estimation, contact angle measurements, artificial neural network model, surface measurements

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Authors: R. Hurtado-Ortiz, D. Clermont, M. Schüngel, C. Bizet, D. Smith, E. Stackebrandt

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Microbiological resources and their derivatives are the essential raw material for the advancement of human health, agro-food, food security, biotechnology, research and development in all life sciences. Microbial resources, and their genetic and metabolic products, are utilised in many areas such as production of healthy and functional food, identification of new antimicrobials against emerging and resistant pathogens, fighting agricultural disease, identifying novel energy sources on the basis of microbial biomass and screening for new active molecules for the bio-industries. The complexity of public collections, distribution and use of living biological material (not only living but also DNA, services, training, consultation, etc.) and service offer, demands the coordination and sharing of policies, processes and procedures. The Microbial Resource Research Infrastructure (MIRRI) is an initiative within the European Strategy Forum Infrastructures (ESFRI), bring together 16 partners including 13 European public microbial culture collections and biological resource centres (BRCs), supported by several European and non-European associated partners. The objective of MIRRI is to support innovation in microbiology by provision of a one-stop shop for well-characterized microbial resources and high quality services on a not-for-profit basis for biotechnology in support of microbiological research. In addition, MIRRI contributes to the structuring of microbial resources capacity both at the national and European levels. This will facilitate access to microorganisms for biotechnology for the enhancement of the bio-economy in Europe. MIRRI will overcome the fragmentation of access to current resources and services, develop harmonised strategies for delivery of associated information, ensure bio-security and other regulatory conditions to bring access and promote the uptake of these resources into European research. Data mining of the landscape of current information is needed to discover potential and drive innovation, to ensure the uptake of high quality microbial resources into research. MIRRI is in its Preparatory Phase focusing on governance and structure including technical, legal governance and financial issues. MIRRI will help the Biological Resources Centres to work more closely with policy makers, stakeholders, funders and researchers, to deliver resources and services needed for innovation.

Keywords: culture collections, microbiology, infrastructure, microbial resources, biotechnology

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Authors: Yemane Tadesse Gebreslassie, Fisseha Guesh Gebremeskel

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Nanotechnology has made remarkable advancements in recent years, revolutionizing various scientific fields, industries, and research institutions through the utilization of metal and metal oxide nanoparticles. Among these nanoparticles, copper oxide nanoparticles (CuO NPs) have garnered significant attention due to their versatile properties and wide-range applications, particularly, as effective antimicrobial and anticancer agents. CuO NPs can be synthesized using different methods, including physical, chemical, and biological approaches. However, conventional chemical and physical approaches are expensive, resource-intensive, and involve the use of hazardous chemicals, which can pose risks to human health and the environment. In contrast, biological synthesis provides a sustainable and cost-effective alternative by eliminating chemical pollutants and allowing for the production of CuO NPs of tailored sizes and shapes. This comprehensive review focused on the green synthesis of CuO NPs using various biological resources, such as plants, microorganisms, and other biological derivatives. Current knowledge and recent trends in green synthesis methods for CuO NPs are discussed, with a specific emphasis on their biomedical applications, particularly in combating cancer and microbial infections. This review highlights the significant potential of CuO NPs in addressing these diseases. By capitalizing on the advantages of biological synthesis, such as environmental safety and the ability to customize nanoparticle characteristics, CuO NPs have emerged as promising therapeutic agents for a wide range of conditions. This review presents compelling findings, demonstrating the remarkable achievements of biologically synthesized CuO NPs as therapeutic agents. Their unique properties and mechanisms enable effective combating against cancer cells and various harmful microbial infections. CuO NPs exhibit potent anticancer activity through diverse mechanisms, including induction of apoptosis, inhibition of angiogenesis, and modulation of signaling pathways. Additionally, their antimicrobial activity manifests through various mechanisms, such as disrupting microbial membranes, generating reactive oxygen species, and interfering with microbial enzymes. This review offers valuable insights into the substantial potential of biologically synthesized CuO NPs as an alternative approach for future therapeutic interventions against cancer and microbial infections.

Keywords: biological synthesis, copper oxide nanoparticles, microbial infection, nanotechnology

Procedia PDF Downloads 62

Authors: Fatemeh Elmi, Zahra Etemadifar

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Improvement of soil mechanical properties is crucial before its use in construction, as the low mechanical strength and unstable structure of soil in many parts of the world can lead to the destruction of engineering infrastructure, resulting in financial and human losses. Although, conventional methods, such as chemical injection, are often utilized to enhance soil strength and stiffness, they are generally expensive, require heavy machinery, and cause significant environmental effects due to chemical usage, and also disrupt urban infrastructure. Moreover, they are not suitable for treating large volume of soil. Recently, an alternative method to improve various soil properties, including strength, hardness, and permeability, has received much attention: the application of biological methods. One of the most widely used is biocementation, which is based on the microbial precipitation of calcium carbonte crystalls using ureolytic bacteria However, there are still limitations to its large-scale use that need to be resolved before it can be commercialized. These issues have not received enough attention in prior research. One limitation of MICP (microbially induced calcium carbonate precipitation) is that microorganisms cannot operate effectively in harsh and variable environments, unlike the controlled conditions of a laboratory. Another limitation of applying this technique on a large scale is the high cost of producing a substantial amount of bacterial culture and reagents required for soil treatment. Therefore, the purpose of the present study was to investigate soil improvement using the biocementation activity of poly-extremophile, calcium carbonate crystal- producing bacterial strain, Bhargavaea cecembensis N1, in whey as an inexpensive medium. This strain was isolated and molecularly identified from sandy soils in our previous research, and its 16S rRNA gene sequences was deposited in the NCBI Gene Bank with an accession number MK420385. This strain exhibited a high level of urease activity (8.16 U/ml) and produced a large amount of calcium carbonate (4.1 mg/ ml). It was able to improve the soil by increasing the compressive strength up to 205 kPa and reducing permeability by 36%, with 20% of the improvement attributable of calcium carbonate production. This was achieved using this strain in a whey culture medium. This strain can be an eco-friendly and economical alternative to conventional methods in soil stabilization, and other MICP related applications.

Keywords: biocementation, Bhargavaea cecembensis, soil improvement, whey culture medium

Procedia PDF Downloads 54

Authors: Akinola Helen Olufunke, Soetan Olatunbosun Jonathan, Adeleye Oludamola

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Introduction: Antibiotic resistance has grown significantly, which is a major cause for concern. There have not been many significant developments in antibiotics over the past few decades, and practically all of the ones that are currently in use are losing effectiveness against pathogenic germs. Researchers are starting to focus more on the physiologically active compounds found in plants, particularly phytobiotics in poultry production. Consumption of chicken products is among the greatest in the country, but numerous nations, including Nigeria, use excessive amounts of necessary antibiotics in poultry farming, endangering the safety of such goods (through antimicrobial residues). Drug resistance has become a widespread issue as a result of the risky use of antibiotics in the chicken production industry. In order to replace antibiotics, biotic or natural products like phytobiotics (also known as botanicals or phytogenics) have drawn a lot of interest. Phytobiotics or their components are thought to be a relatively recent category of natural herbs that have acquired acceptance and favor among chicken farmers. The addition of several phytobiotic additions to poultry feed has demonstrated its capacity to improve both the broiler and layer populations' productivity. Design: Experimental research design and cross-sectional study was carried out at every 300 purposively selected farming household in the six-geopolitical zone in Nigeria. Data Analysis: A semi-structured questionnaire was administered to each farmer, and quantitative data were analyzed using Statistical Package for Social Science (SPSS) while the Chi-square test was used to analyze factors mitigating the use of Phytobiotics. Result: The result shows that the benefits associated with the use of phytobiotics are contributed to growth promotion in chickens and enhancement of productive performance of broiler and layer, which could be attributed to their antioxidant activity. The result further revealed that factors mitigating the use of phytobiotics were lack of knowledge in the use of phytobiotics, overdose or underdose usage, and seasonal availability of the phytobiotics. Others are the educational level of the farmers, intrinsic motivation, income poultry farming experience, price of phytobiotics based additives feeds, and intensity of extension agents in visiting them. Conclusion: The difficulties associated with using phytobiotics in chicken farms limit their willingness to boost productivity. The study found that most farmers were ignorant, which prevented them from handling this notion and turning their poultry into a viable enterprise while also allowing them to be creative. They believed that packing phytobiotics-based additive feed was expensive, and lastly, the seasonal availability of some phytobiotics. Recommendation: Further research in phytobiotics use in Nigeria should be carried out in order to establish its efficiency, safety, and awareness.

Keywords: mitigating, antibiotics, phytobiotics, poultry farming

Procedia PDF Downloads 171

Authors: Ayodele A. Otaiku

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Purpose: Nigeria has become the largest importer of rice in Africa and second in the world, 2015. Investigate interactions of organic rice farming on soil quality and health from bio-waste converted to biofertilizer and its environmental impact on rice crop. Methodology: Bio-wastes, poultry waste, organic agriculture wastes, wood ash mixed with microbial inoculant organisms called OBD-Plus microbes (broad spectrum) composted in anaerobic digester to OBD-biofertilizer (2010 - 2012) uses microbes to build humus and other stable carbons. Two field experiments were carried out at Badeggi, Niger state in 2011 and 2012 to evaluate the response of lowland rice production using biofertilizer. The experimental field was laid out in a strip-plot design with five treatments and three replications and at twenty-one day old seedlings of FARO 44 and FARO 52 rice varieties were transplanted. Plots without fertiliser application served as control. Findings: The highest rice grain yield increase of 4.4 t/ha over the control in 2012 against the Nigeria average of lowland rice grain yields of 1.5 t/ha. The utilization of OBD-Biofertilizer can decrease the use of chemical nitrogen fertilizer, prevent the depletion of soil organic matter and reduce environmental pollution. Increasing the floodwater productivity and optimizing the recycling of nutrients cum grazer populations and disease by biocontrols microbes present in the OBD-Biofertilizer. Organic matter in the soil improves by 58% and C/N 15 (2011) and 13.35 (2012). Implications: OBD- Biofertilizer produce plant growth hormones such as indole acetic acid (IAA), glomalin related soil protein and extracellular enzymes as phosphatases that promote soil health and quality. Conclusion: Microorganisms can enhance nutrients use efficiency by increasing root surface area e.g., mycorrhizal, fungi, promoting other beneficial symbioses of the host plant and microbial interactions resulting to increase in soil organic matter. By 2030, climate change is projected to depress cereal production in Africa by 2 to 3 percent. Improved seeds and increased fertilizer use should more than compensate, but this factor will still weigh heavily on efforts to make progress.

Keywords: OBD-plus microbial consortia, OBD-biofertilizer, rice production, soil quality, sustainable agriculture

Procedia PDF Downloads 269

Authors: Andrea M. Rivas-Castillo, Marlenne Gómez-Ramirez, Isela Rodríguez-Pozos, Norma G. Rojas-Avelizapa

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Spent catalysts are considered as hazardous residues of major concern, mainly due to the simultaneous presence of several metals in elevated concentrations. Although hydrometallurgical, pyrometallurgical and chelating agent methods are available to remove and recover some metals contained in spent catalysts; these procedures generate potentially hazardous wastes and the emission of harmful gases. Thus, biotechnological treatments are currently gaining importance to avoid the negative impacts of chemical technologies. To this end, diverse microorganisms have been used to assess the removal of metals from spent catalysts, comprising bacteria, archaea and fungi, whose resistance and metal uptake capabilities differ depending on the microorganism tested. Acidophilic sulfur oxidizing bacteria have been used to investigate the biotreatment and extraction of valuable metals from spent catalysts, namely Acidithiobacillus thiooxidans and Acidithiobacillus ferroxidans, as they present the ability to produce leaching agents such as sulfuric acid and sulfur oxidation intermediates. In the present work, the ability of A. thiooxidans DSM 26636 for the bioleaching of metals contained in five different spent catalysts was assessed by growing the culture in modified Starkey mineral medium (with elemental sulfur at 1%, w/v), and 1% (w/v) pulp density of each residue for up to 21 days at 30 °C and 150 rpm. Sulfur-oxidizing activity was periodically evaluated by determining sulfate concentration in the supernatants according to the NMX-k-436-1977 method. The production of sulfuric acid was assessed in the supernatants as well, by a titration procedure using NaOH 0.5 M with bromothymol blue as acid-base indicator, and by measuring pH using a digital potentiometer. On the other hand, Inductively Coupled Plasma - Optical Emission Spectrometry was used to analyze metal removal from the five different spent catalysts by A. thiooxidans DSM 26636. Results obtained show that, as could be expected, sulfuric acid production is directly related to the diminish of pH, and also to highest metal removal efficiencies. It was observed that Al and Fe are recurrently removed from refinery spent catalysts regardless of their origin and previous usage, although these removals may vary from 9.5 ± 2.2 to 439 ± 3.9 mg/kg for Al, and from 7.13 ± 0.31 to 368.4 ± 47.8 mg/kg for Fe, depending on the spent catalyst proven. Besides, bioleaching of metals like Mg, Ni, and Si was also obtained from automotive spent catalysts, which removals were of up to 66 ± 2.2, 6.2±0.07, and 100±2.4, respectively. Hence, the data presented here exhibit the potential of A. thiooxidans DSM 26636 for the simultaneous bioleaching of metals contained in spent catalysts from diverse provenance.

Keywords: bioleaching, metal removal, spent catalysts, Acidithiobacillus thiooxidans

Procedia PDF Downloads 140

Authors: Maneesha Pahlani, Najmin Sultana

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A hospital is one of the most suitable places for acquiring an infection because it harbors a high population of virulent strains of microorganisms that may be resistant to antibiotics, especially the prevalence of Methicillin-Resistant Staphylococcus Aureus (MRSA) infections. The hospital-acquired infection has become a global challenge. In developed countries, healthcare-associated infections occur in 5-15% of hospitalized clients, affecting 9-37% of those admitted to intensive care units (ICU). A non-experimental descriptive study was conducted among 50 nursing officers working in a selected hospital in Bangalore to assess the nursing officers’ level of knowledge regarding the prevention and management of MRSA infections and to associate the pre-test knowledge mean scores of nursing officers with selected socio-demographic variables. Data was collected using a structured questionnaire consisting of socio-demographic data and a structured questionnaire on knowledge regarding the prevention and management of MRSA infections. The data was analyzed in terms of frequencies and percentages for the analysis of demographic variables and computing chi-square to determine the association between knowledge means scores and selected demographic variables. The study findings revealed that the nursing officer had an overall good level of knowledge (63.05%) regarding the prevention and management of MRSA infections, and there is no significant association found between the level of knowledge mean scores for prevention and management of MRSA infection with the selected socio-demographic variables. However, the categorization of knowledge items showed that the nursing officer must thoroughly receive education on correct guidance and information regarding MRSA infection control policy, including measures and practices on hygiene precautions and information regarding antibiotic resistance for effective nursing care to patients with MRSA infections. The conclusions drawn from the study findings showed that it is necessary that the nursing officer thoroughly receive education on correct guidance and information regarding MRSA infection control policy, including measures and practices on hygiene precautions and information regarding antibiotic resistance to provide effective nursing care to patients with MRSA infection as they constantly care for the patient who can be at risk for multi-drug resistance organisms to reduce the risk of MRSA infection in hospital care settings as well community settings.

Keywords: MRSA, nursing officers, knowledge, preventive and management

Procedia PDF Downloads 69

Authors: Najmin Sultana, Maneesha Pahlani

Abstract:

A hospital is one of the most suitable places for acquiring an infection because it harbors a high population of virulent strains of microorganisms that may be resistant to antibiotics, especially the prevalence of Methicillin-Resistant Staphylococcus Aureus (MRSA) infections. The hospital-acquired infection has become a global challenge. In developed countries, healthcare-associated infections occur in 5-15% of hospitalized clients, affecting 9-37% of those admitted to intensive care units (ICU). A non-experimental descriptive study was conducted among 50 nursing officers working in a selected hospital in bengaluru to assess the nursing officers’ level of knowledge regarding the prevention and management of MRSA infections and to associate the pre-test knowledge mean scores of nursing officers with selected socio-demographic variables. Data was collected using a structured questionnaire consisting of socio-demographic data and a structured questionnaire on knowledge regarding the prevention and management of MRSA infections. The data was analyzed in terms of frequencies and percentages for the analysis of demographic variables and computing chi-square to determine the association between knowledge means scores and selected demographic variables. The study findings revealed that the nursing officer had an overall good level of knowledge (63.05%) regarding the prevention and management of MRSA infections, and there is no significant association found between the level of knowledge mean scores for prevention and management of MRSA infection with the selected socio-demographic variables. However, the categorization of knowledge items showed that the nursing officer must thoroughly receive education on correct guidance and information regarding MRSA infection control policy, including measures and practices on hygiene precautions and information regarding antibiotic resistance for effective nursing care to patients with MRSA infections. The conclusions drawn from the study findings showed that it is necessary that the nursing officer thoroughly receive education on correct guidance and information regarding MRSA infection control policy, including measures and practices on hygiene precautions and information regarding antibiotic resistance to provide effective nursing care to patients with MRSA infection as they constantly care for the patient who can be at risk for multi-drug resistance organisms to reduce the risk of MRSA infection in hospital care settings as well community settings.

Keywords: MRSA, knowledge, nursing officers', prevention and management

Procedia PDF Downloads 63

Authors: Kanika Gupta, Ashok Kumar

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Biofilms are dense, highly hydrated cell clusters that are irreversibly attached to a substratum, to an interface or to each other, and are embedded in a self-produced gelatinous matrix composed of extracellular polymeric substances. Research in biofilm field has become very significant, as biofilm has shown high mechanical resilience and resistance to antibiotic treatment and constituted as a significant problem in both healthcare and other industry related to microorganisms. The massive information both stated and hidden in the biofilm literature are growing exponentially therefore it is not possible for researchers and practitioners to automatically extract and relate information from different written resources. So, the current work proposes and discusses the use of text mining techniques for the extraction of information from biofilm literature corpora containing 34306 documents. It is very difficult and expensive to obtain annotated material for biomedical literature as the literature is unstructured i.e. free-text. Therefore, we considered unsupervised approach, where no annotated training is necessary and using this approach we developed a system that will classify the text on the basis of growth and development, drug effects, radiation effects, classification and physiology of biofilms. For this, a two-step structure was used where the first step is to extract keywords from the biofilm literature using a metathesaurus and standard natural language processing tools like Rapid Miner_v5.3 and the second step is to discover relations between the genes extracted from the whole set of biofilm literature using pubmed.mineR_v1.0.11. We used unsupervised approach, which is the machine learning task of inferring a function to describe hidden structure from 'unlabeled' data, in the above-extracted datasets to develop classifiers using WinPython-64 bit_v3.5.4.0Qt5 and R studio_v0.99.467 packages which will automatically classify the text by using the mentioned sets. The developed classifiers were tested on a large data set of biofilm literature which showed that the unsupervised approach proposed is promising as well as suited for a semi-automatic labeling of the extracted relations. The entire information was stored in the relational database which was hosted locally on the server. The generated biofilm vocabulary and genes relations will be significant for researchers dealing with biofilm research, making their search easy and efficient as the keywords and genes could be directly mapped with the documents used for database development.

Keywords: biofilms literature, classifiers development, text mining, unsupervised learning approach, unstructured data, relational database

Procedia PDF Downloads 170

Authors: Z. C. Dlamini, R. L. S. Langa, A. I. Okoh, O. A. Aiyegoro

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The indiscriminate usage of antibiotics in swine production have consequential outcomes; such as development of bacterial resistance to prophylactic antibiotics and possibility of antibiotic residues in animal products. The use of probiotics appears to be the most effective procedure with positive metabolic nutritional implications. The aim of this study was to investigate the efficacy of probiotic bacteria (Lactobacillus reuteri ZJ625, Lactobacillus reuteri VB4, Lactobacillus salivarius ZJ614 and Streptococcus salivarius NBRC13956) administered as direct-fed microorganisms in weaned piglets. 45 weaned piglets blocked by weight were dived into 5 treatments groups: diet with antibiotic, diet with no-antibiotic and no probiotic, and diet with probiotic and diet with combination of probiotics. Piglets performance was monitored during the trials. Faecal and Ileum samples were collected for microbial count analysis. Blood samples were collected from pigs at the end of the trial, for analysis of haematological, biochemical and IgG stimulation. The data was analysed by Split-Plot ANOVA using SAS statistically software (SAS 9.3) (2003). The difference was observed between treatments for daily weight and feed conversion ratio. No difference was observed in analysis of faecal samples in regards with bacterial counts, difference was observed in ileums samples with enteric bacteria colony forming unit being lower in P2 treatment group as compared with lactic acid and total bacteria. With exception of globulin and albumin, biochemistry blood parameters were not affected, likewise for haematology, only basophils and segmented neutrophils were differed by having higher concentration in NC treatment group as compared with other treatment groups. Moreover, in IgG stimulation analysis, difference was also observed, with P2 treatment group having high concentration of IgG in P2 treatment group as compared to other groups. The results of this study suggest that probiotics have a beneficial effect on growth performances, blood parameters and IgG stimulation of pigs, most effective when they are administered in synergy form. This means that it is most likely that these probiotics will offer a significant benefit in pig farming by reducing risk of morbidity and mortality and produce quality meat that is more affordable to poorer communities, and thereby enhance South African pig industry’s economy. In addition, these results indicate that there is still more research need to be done on probiotics in regards with, i.e. dosage, shelf life and mechanism of action.

Keywords: antibiotics, biochemistry, haematology, IgG-stimulation, microbial count, probiotics

Procedia PDF Downloads 301

Authors: Ahmed F. Ghanem, Marwa I. Wahba, Asmaa N. El-Dein, Mohamed A. EL-Raey, Ghada E. A. Awad

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Numerous attempts are being performed in order to formulate suitable packaging materials for the meat products. However, to the best of our knowledge, the incorporation of the free hibiscus extract or its microcapsules in the pure polyvinyl alcohol (PVA) matrix as packaging materials for the meats is seldom reported. Therefore, this study aims at the protection of the aqueous crude extract of the hibiscus flowers utilizing the spry drying encapsulation technique. Results of the Fourier transform infrared (FTIR), the scanning electron microscope (SEM), and the particle size analyzer confirmed the successful formation of the assembled capsules via strong interactions, the spherical rough microparticles, and the particle size of ~ 235 nm, respectively. Also, the obtained microcapsules enjoy higher thermal stability than the free extract. Then, the obtained spray-dried particles were incorporated into the casting solution of the pure PVA film with a concentration of 10 wt. %. The segregated free-standing composite films were investigated, compared to the neat matrix, with several characterization techniques such as FTIR, SEM, thermal gravimetric analysis (TGA), mechanical tester, contact angle, water vapor permeability, and oxygen transmission. The results demonstrated variations in the physicochemical properties of the PVA film after the inclusion of the free and the extract microcapsules. Moreover, biological studies emphasized the biocidal potential of the hybrid films against the microorganisms contaminating the meat. Specifically, the microcapsules imparted not only antimicrobial but also antioxidant activities to the PVA matrix. Application of the prepared films on the real meat samples displayed a low bacterial growth with a slight increase in the pH over the storage time which continued up to 10 days at 4 oC, as further evidence to the meat safety. Moreover, the colors of the films did not significantly changed except after 21 days indicating the spoilage of the meat samples. No doubt, the dual-functional of the prepared composite films pave the way towards combined active and smart food packaging applications. This would play a vital role in the food hygiene, including also the quality control and the assurance.

Keywords: PVA, hibiscus, extraction, encapsulation, active packaging, smart and intelligent packaging, meat spoilage

Procedia PDF Downloads 90

Authors: W. S. Thulasitha, N. Umasuthan, G. I. Godahewa, Jehee Lee

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In fish, innate immune defense is the first immune response against microbial pathogens which consists of several antimicrobial components. Galectins are one of the carbohydrate binding lectins that have the ability to identify pathogen by recognition of pathogen associated molecular patterns. Galectins play a vital role in the regulation of innate and adaptive immune responses. Rock bream Oplegnathus fasciatus is one of the most important cultured species in Korea and Japan. Considering the losses due to microbial pathogens, present study was carried out to understand the molecular and functional characteristics of a galectin in normal and pathogenic conditions, which could help to establish an understanding about immunological components of rock bream. Complete cDNA of rock bream galectin like protein B (rbGal like B) was identified from the cDNA library, and the in silico analysis was carried out using bioinformatic tools. Genomic structure was derived from the BAC library by sequencing a specific clone and using Spidey. Full length of rbGal like B (contig14775) cDNA containing 517 nucleotides was identified from the cDNA library which comprised of 435 bp in the open reading frame encoding a deduced protein composed of 145 amino acids. The molecular mass of putative protein was predicted as 16.14 kDa with an isoelectric point of 8.55. A characteristic conserved galactose binding domain was located from 12 to 145 amino acids. Genomic structure of rbGal like B consisted of 4 exons and 3 introns. Moreover, pairwise alignment showed that rock bream rbGal like B shares highest similarity (95.9 %) and identity (91 %) with Takifugu rubripes galectin related protein B like and lowest similarity (55.5 %) and identity (32.4 %) with Homo sapiens. Multiple sequence alignment demonstrated that the galectin related protein B was conserved among vertebrates. A phylogenetic analysis revealed that rbGal like B protein clustered together with other fish homologs in fish clade. It showed closer evolutionary link with Takifugu rubripes. Tissue distribution and expression patterns of rbGal like B upon immune challenges were performed using qRT-PCR assays. Among all tested tissues, level of rbGal like B expression was significantly high in gill tissue followed by kidney, intestine, heart and spleen. Upon immune challenges, it showed an up-regulated pattern of expression with Edwardsiella tarda, rock bream irido virus and poly I:C up to 6 h post injection and up to 24 h with LPS. However, In the presence of Streptococcus iniae rbGal like B showed an up and down pattern of expression with the peak at 6 - 12 h. Results from the present study revealed the phylogenetic position and role of rbGal like B in response to microbial infection in rock bream.

Keywords: galectin like protein B, immune response, Oplegnathus fasciatus, molecular characterization

Procedia PDF Downloads 354

Authors: Sonia Amariei, Ionut Avramia, Florin Ursachi, Ancuta Chetrariu, Ancuta Petraru

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The food industry is one of the major generators of plastic waste derived from conventional synthetic petroleum-based polymers, which are non-biodegradable, used especially for packaging. These packaging materials, after the food is consumed, accumulate serious environmental concerns due to the materials but also to the organic residues that adhere to them. It is the concern of specialists, researchers to eliminate problems related to conventional materials that are not biodegradable or unnecessary plastic and replace them with biodegradable and edible materials, supporting the common effort to protect the environment. Even though environmental and health concerns will cause more consumers to switch to a plant-based diet, most people will continue to add more meat to their diet. The paper presents the possibility of replacing the polyethylene packaging from the surface of the trays for meat preparations with biodegradable packaging obtained from biopolymers. During the storage of meat products may occur deterioration by lipids oxidation and microbial spoilage, as well as the modification of the organoleptic characteristics. For this reason, different compositions of polymer mixtures and film conditions for obtaining must be studied to choose the best packaging material to achieve food safety. The compositions proposed for packaging are obtained from alginate, agar, starch, and glycerol as plasticizers. The tensile strength, elasticity, modulus of elasticity, thickness, density, microscopic images of the samples, roughness, opacity, humidity, water activity, the amount of water transferred as well as the speed of water transfer through these packaging materials were analyzed. A number of 28 samples with various compositions were analyzed, and the results showed that the sample with the highest values for hardness, density, and opacity, as well as the smallest water vapor permeability, of 1.2903E-4 ± 4.79E-6, has the ratio of components as alginate: agar: glycerol (3:1.25:0.75). The water activity of the analyzed films varied between 0.2886 and 0.3428 (aw< 0.6), demonstrating that all the compositions ensure the preservation of the products in the absence of microorganisms. All the determined parameters allow the appreciation of the quality of the packaging films in terms of mechanical resistance, its protection against the influence of light, the transfer of water through the packaging. Acknowledgments: This work was supported by a grant of the Ministry of Research, Innovation, and Digitization, CNCS/CCCDI – UEFISCDI, project number PN-III-P2-2.1-PED-2019-3863, within PNCDI III.

Keywords: meat products, alginate, agar, starch, glycerol

Procedia PDF Downloads 167

Authors: Ghaneshree Moonsamy, Nodumo N. Zulu, Rajesh Lalloo, Suren Singh, Santosh O. Ramchuran

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The abalone industry of South Africa is under severe pressure due to illegal harvesting and poaching of this seafood delicacy. These abalones are harvested excessively; as a result, these animals do not have a chance to replace themselves in their habitats, ensuing in a drastic decrease in natural stocks of abalone. Abalone has an extremely slow growth rate and takes approximately four years to reach a size that is market acceptable; therefore, it was imperative to investigate methods to boost the overall growth rate and immunity of the animal. The University of Cape Town (UCT) began to research, which resulted in the isolation of two microorganisms, a yeast isolate Debaryomyces hansenii and a bacterial isolate Vibrio midae, from the gut of the abalone and characterised them for their probiotic abilities. This work resulted in an internationally competitive concept technology that was patented. The next stage of research was to develop a suitable bioprocess to enable commercial production. Numerous steps were taken to develop an efficient production process for V. midae, one of the isolates found by UCT. The initial stages of research involved the development of a stable and robust inoculum and the optimization of physiological growth parameters such as temperature and pH. A range of temperature and pH conditions were evaluated, and data obtained revealed an optimum growth temperature of 30ᵒC and a pH of 6.5. Once these critical growth parameters were established further media optimization studies were performed. Corn steep liquor (CSL) and high test molasses (HTM) were selected as suitable alternatives to more expensive, conventionally used growth medium additives. The optimization of CSL (6.4 g.l⁻¹) and HTM (24 g.l⁻¹) concentrations in the growth medium resulted in a 180% increase in cell concentration, a 5716-fold increase in cell productivity and a 97.2% decrease in the material cost of production in comparison to conventional growth conditions and parameters used at the onset of the study. In addition, a stable market-ready liquid probiotic product, encompassing the viable but not culturable (VBNC) state of Vibrio midae cells, was developed during the downstream processing aspect of the study. The demonstration of this technology at a full manufacturing scale has further enhanced the attractiveness and commercial feasibility of this production process.

Keywords: probiotics, abalone aquaculture, bioprocess technology, manufacturing scale technology development

Procedia PDF Downloads 152

Authors: Zh. Karaulova, D. Baizhigitov

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At the Aktogay deposit, the oxidized ore section has been developed since 2015; by now, the reserves of easily enriched ore are decreasing, and a large number of copper-poor, difficult-to-enrich ores has been accumulated in the dumps of the KAZ Minerals Aktogay deposit, which is unprofitable to mine using the traditional mining methods. Hence, another technology needs to be implemented, which will significantly expand the raw material base of copper production in Kazakhstan and ensure the efficient use of natural resources. Heap and dump bacterial recovery are the most acceptable technologies for processing low-grade secondary copper sulfide ores. Test objects were the copper ores of Aktogay deposit and chemolithotrophic bacteria Leptospirillum ferrooxidans (L.f.), Acidithiobacillus caldus (A.c.), Sulfobacillus Acidophilus (S.a.), which are mixed cultures were both used in bacterial oxidation systems. They can stay active in the 20-400C temperature range. These bacteria were the most extensively studied and widely used in sulfide mineral recovery technology. Biocatalytic acceleration was achieved as a result of bacteria oxidizing iron sulfides to form iron sulfate, which subsequently underwent chemical oxidation to become sulfate oxide. The following results have been achieved at the initial stage: the goal was to grow and maintain the life activity of bacterial cultures under laboratory conditions. These bacteria grew the best within the pH 1,2-1,8 range with light stirring and in an aerated environment. The optimal growth temperature was 30-33оC. The growth rate decreased by one-half for each 4-5°C fall in temperature from 30°C. At best, the number of bacteria doubled every 24 hours. Typically, the maximum concentration of cells that can be grown in ferrous solution is about 107/ml. A further step researched in this case was the adaptation of microorganisms to the environment of certain metals. This was followed by mass production of inoculum and maintenance for their further cultivation on a factory scale. This was done by adding sulfide concentrate, allowing the bacteria to convert the ferrous sulfate as indicated by the Eh (>600 mV), then diluting to double the volume and adding concentrate to achieve the same metal level. This process was repeated until the desired metal level and volumes were achieved. The final stage of bacterial recovery was the transportation and irrigation of secondary sulfide copper ores of the oxidized ore section. In conclusion, the project was implemented at the Aktogay mine since the bioleaching process was prolonged. Besides, the method of bacterial recovery might compete well with existing non-biological methods of extraction of metals from ores.

Keywords: bacterial recovery, copper ore, bioleaching, bacterial inoculum

Procedia PDF Downloads 74

Authors: Marie Davin, Marie-Laure Fauconnier, Gilles Colinet

Abstract:

In Wallonia, there are 6,000 estimated brownfields (rising to over 3.5 million in Europe) that require remediation. Polycyclic Aromatic Hydrocarbons (PAHs) are a class of recalcitrant carcinogenic/mutagenic organic compounds of major concern as they accumulate in the environment and represent 17% of all encountered pollutants. As an alternative to environmentally aggressive, expensive and often disruptive soil remediation strategies, a lot of research has been directed to developing techniques targeting organic pollutants. The following experiment, based on the observation that PAHs soil content decreases in the presence of plants, aimed at improving our understanding of the underlying mechanisms involved in phytoremediation. It focusses on plant root exudates and whether they improve PAHs solubilization, which would make them more available for bioremediation by soil microorganisms. The effect of saponin, a natural surfactant found in some plant roots such as members of the Fabaceae family, on PAHs solubilization was also investigated as part of the implementation of the experimental protocol. The experiments were conducted on soil collected from a brownfield in Saint-Ghislain (Belgium) and presenting weathered PAHs contamination. Samples of soil were extracted with different solutions containing either plant root exudates or commercial saponin. Extracted PAHs were determined in the different aqueous solutions using High-Performance Liquid Chromatography and Fluorimetric Detection (HPLC-FLD). Both root exudates of alfalfa (Medicago sativa L.) or red clover (Trifolium pratense L.) and commercial saponin were tested in different concentrations. Distilled water was used as a control. First of all, results show that PAHs are more extracted using saponin solutions than distilled water and that the amounts generally rise with the saponin concentration. However, the amount of each extracted compound diminishes as its molecular weight rises. Also, it appears that passed a certain surfactant concentration, PAHs are less extracted. This suggests that saponin might be investigated as a washing agent in polluted soil remediation techniques, either for ex-situ or in-situ treatments, as an alternative to synthetic surfactants. On the other hand, preliminary results on experiments using plant root exudates also show differences in PAHs solubilization compared to the control solution. Further results will allow discussion as to whether or not there are differences according to the exudates provenance and concentrations.

Keywords: brownfield, Medicago sativa, phytoremediation, polycyclic aromatic hydrocarbons, root exudates, saponin, solubilization, Trifolium pratense

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Authors: Paula I. Buonfiglio, Carlos David Bruque, Lucia Salatino, Vanesa Lotersztein, Sebastián Menazzi, Paola Plazas, Ana Belén Elgoyhen, Viviana Dalamón

Abstract:

Hearing loss (HL) is the most prevalent sensorineural disorder affecting about 10% of the global population, with more than half due to genetic causes. About 1 in 500-1000 newborns present congenital HL. Most of the patients are non-syndromic with an autosomal recessive mode of inheritance. To date, more than 100 genes are related to HL. Therefore, the Whole-exome sequencing (WES) technique has become a cost-effective alternative approach for molecular diagnosis. Nevertheless, new challenges arise from the detection of novel variants, in particular missense changes, which can lead to a spectrum of genotype-to-phenotype correlations, which is not always straightforward. In this work, we aimed to identify the genetic causes of HL in isolated and familial cases by designing a multistep approach to analyze target genes related to hearing impairment. Moreover, we performed in silico and in vivo analyses in order to further study the effect of some of the novel variants identified in the hair cell function using the zebrafish model. A total of 650 patients were studied by Sanger Sequencing and Gap-PCR in GJB2 and GJB6 genes, respectively, diagnosing 15.5% of sporadic cases and 36% of familial ones. Overall, 50 different sequence variants were detected. Fifty of the undiagnosed patients with moderate HL were tested for deletions in STRC gene by Multiplex ligation-dependent probe amplification technique (MLPA), leading to 6% of diagnosis. After this initial screening, 50 families were selected to be analyzed by WES, achieving diagnosis in 44% of them. Half of the identified variants were novel. A missense variant in MYO6 gene detected in a family with postlingual HL was selected to be further analyzed. A protein modeling with AlphaFold2 software was performed, proving its pathogenic effect. In order to functionally validate this novel variant, a knockdown phenotype rescue assay in zebrafish was carried out. Injection of wild-type MYO6 mRNA in embryos rescued the phenotype, whereas using the mutant MYO6 mRNA (carrying c.2782C>A variant) had no effect. These results strongly suggest the deleterious effect of this variant on the mobility of stereocilia in zebrafish neuromasts, and hence on the auditory system. In the present work, we demonstrated that our algorithm is suitable for the sequential multigenic approach to HL in our cohort. These results highlight the importance of a combined strategy in order to identify candidate variants as well as the in silico and in vivo studies to analyze and prove their pathogenicity and accomplish a better understanding of the mechanisms underlying the physiopathology of the hearing impairment.

Keywords: diagnosis, genetics, hearing loss, in silico analysis, in vivo analysis, WES, zebrafish

Procedia PDF Downloads 94