Search results for: zoonotic viruses

Authors: Sandisiwe Mangali, Graeme Bradley

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Genome is complete set of the organism's hereditary information encoded as either deoxyribonucleic acid or ribonucleic acid in most viruses. The three different types of genomes are nuclear, mitochondrial and the plastid genome and their sequences which are uncovered by genome sequencing are known as an archive for all genetic information and enable researchers to understand the composition of a genome, regulation of gene expression and also provide information on how the whole genome works. These sequences enable researchers to explore the population structure, genetic variations, and recent demographic events in threatened species. Particularly, genome sequencing refers to a process of figuring out the exact arrangement of the basic nucleotide bases of a genome and the process through which all the afore-mentioned genomes are sequenced is referred to as whole or complete genome sequencing. Gelidium pristoides is South African endemic Rhodophyta species which has been harvested in the Eastern Cape since the 1950s for its high economic value which is one motivation for its sequencing. Its endemism further motivates its sequencing for conservation biology as endemic species are more vulnerable to anthropogenic activities endangering a species. As sequencing, mapping and annotating the Gelidium pristoides genome is the aim of this study. To accomplish this aim, the genomic DNA was extracted and quantified using the Nucleospin Plank Kit, Qubit 2.0 and Nanodrop. Thereafter, the Ion Plus Fragment Library was used for preparation of a 600bp library which was then sequenced through the Ion S5 sequencing platform for two runs. The produced reads were then quality-controlled and assembled through the SPAdes assembler with default parameters and the genome assembly was quality assessed through the QUAST software. From this assembly, the plastid and the mitochondrial genomes were then sampled out using Gelidiales organellar genomes as search queries and ordered according to them using the Geneious software. The Qubit and the Nanodrop instruments revealed an A260/A280 and A230/A260 values of 1.81 and 1.52 respectively. A total of 30792074 reads were obtained and produced a total of 94140 contigs with resulted into a sequence length of 217.06 Mbp with N50 value of 3072 bp and GC content of 41.72%. A total length of 179281bp and 25734 bp was obtained for plastid and mitochondrial respectively. Genomic data allows a clear understanding of the genomic constituent of an organism and is valuable as foundation information for studies of individual genes and resolving the evolutionary relationships between organisms including Rhodophytes and other seaweeds.

Keywords: Gelidium pristoides, genome, genome sequencing and assembly, Ion S5 sequencing platform

Procedia PDF Downloads 123

Authors: Justo Rodriguez, Daming Chen, Amador M. Guzman

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The detection, treatment, and control of rapidly propagating, deadly viruses such as COVID-19, require the development of inexpensive, fast, and accurate devices to address the urgent needs of the population. Microfluidics-based sensors are amongst the different methods and techniques for detection that are easy to use. A micro analyzer is defined as a microfluidics-based sensor, composed of a network of microchannels with varying functions. Given their size, portability, and accuracy, they are proving to be more effective and convenient than other solutions. A micro analyzer based on the concept of “Lab on a Chip” presents advantages concerning other non-micro devices due to its smaller size, and it is having a better ratio between useful area and volume. The integration of multiple processes in a single microdevice reduces both the number of necessary samples and the analysis time, leading the next generation of analyzers for the health-sciences. In some applications, the flow of solution within the microchannels is originated by a pressure gradient, which can produce adverse effects on biological samples. A more efficient and less dangerous way of controlling the flow in a microchannel-based analyzer is applying an electric field to induce the fluid motion and either enhance or suppress the mixing process. Electrokinetic flows are characterized by no less than two non-dimensional parameters: the electric Rayleigh number and its geometrical aspect ratio. In this research, stable and unstable flows have been studied numerically (and when possible, will be experimental) in a T-shaped microchannel. Additionally, unstable electrokinetic flows for Rayleigh numbers higher than critical have been characterized. The flow mixing enhancement was quantified in relation to the stretching and folding that fluid particles undergo when they are subjected to supercritical electrokinetic flows. Computational simulations were carried out using a finite element-based program while working with the flow mixing concepts developed by Gollub and collaborators. Hundreds of seeded massless particles were tracked along the microchannel from the entrance to exit for both stable and unstable flows. After post-processing, their trajectories, the folding and stretching values for the different flows were found. Numerical results show that for supercritical electrokinetic flows, the enhancement effects of the folding and stretching processes become more apparent. Consequently, there is an improvement in the mixing process, ultimately leading to a more homogenous mixture.

Keywords: microchannel, stretching and folding, electro kinetic flow mixing, micro-analyzer

Procedia PDF Downloads 98

Authors: Yunjia Yang, Peng Li, Gordon Xu, Timothy Mahony, Bing Zhang, Neena Mitter, Karishma Mody

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Sheep flystrike is one of the most economically important diseases affecting the Australian sheep and wool industry (>356M/annually). Currently, control of Lucillia cuprina relies almost exclusively on chemicals controls and the parasite has developed resistance to nearly all control chemicals used in the past. It is therefore critical to develop an alternative solution for the sustainable control and management of flystrike. RNA interference (RNAi) technologies have been successfully explored in multiple animal industries for developing parasites controls. This research project aims to develop a RNAi based biological control for sheep blowfly. Double-stranded RNA (dsRNA) has already proven successful against viruses, fungi and insects. However, the environmental instability of dsRNA is a major bottleneck for successful RNAi. Bentonite polymer (BenPol) technology can overcome this problem, as it can be tuned for the controlled release of dsRNA in the gut challenging pH environment of the blowfly larvae, prolonging its exposure time to and uptake by target cells. To investigate the potential of BenPol technology for dsRNA delivery, four different BenPol carriers were tested for their dsRNA loading capabilities, and three of them were found to be capable of affording dsRNA stability under multiple temperatures (4°C, 22°C, 40°C, 55°C) in sheep serum. Based on stability results, dsRNA from potential targeted genes was loaded onto BenPol carriers and tested in larvae feeding assays, three genes resulting in knockdowns. Meanwhile, a primary blowfly embryo cell line (BFEC) derived from L. cuprina embryos was successfully established, aim for an effective insect cell model for testing RNAi efficacy for preliminary assessments and screening. The results of this study establish that the dsRNA is stable when loaded on BenPol particles, unlike naked dsRNA rapidly degraded in sheep serum. The stable nanoparticle delivery system offered by BenPol technology can protect and increase the inherent stability of dsRNA molecules at higher temperatures in a complex biological fluid like serum, providing promise for its future use in enhancing animal protection.

Keywords: flystrike, RNA interference, bentonite polymer technology, Lucillia cuprina

Procedia PDF Downloads 58

Authors: Johannes Mertl

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Our surrounding air contains various particles. Besides typical representatives of inorganic dust, such as soot and ash, also particles originating from animals, microorganisms or plants are floating through the air, so-called bioaerosols. The group of bioaerosols consists of a broad spectrum of particles of different size, including fungi, bacteria, viruses, spores, or tree, flower and grass pollen that are of high relevance for allergy sufferers. In dependence of the environmental climate and the actual season, these allergenic particles can be found in enormous numbers in the air and are inhaled by humans via the respiration tract, with a potential for inflammatory diseases of the airways, such as asthma or allergic rhinitis. As a consequence air filter systems of ventilation and air conditioning devices are required to meet very high standards to prevent, or at least lower the number of allergens and airborne germs entering the indoor air. Still, filter systems are merely classified for their separation rates using well-defined mineral test dust, while no appropriate sufficiently standardized test methods for bioaerosols exist. However, determined separation rates for mineral test particles of a certain size cannot simply be transferred to bioaerosols, as separation efficiency of particularly fine and respirable particles (< 10 microns) is dependent not only on their shape and particle diameter, but also defined by their density and physicochemical properties. For this reason, the OFI developed a test method, which directly enables a testing of filters and filter media for their separation rates on bioaerosols, as well as a classification of filters. Besides allergens from an intact or fractured tree or grass pollen, allergenic proteins bound to particulates, as well as allergenic fungal spores (e.g. Cladosporium cladosporioides), or bacteria can be used to classify filters regarding their separation rates. Allergens passing through the filter can then be detected by highly sensitive immunological assays (ELISA) or in the case of fungal spores by microbiological methods, which allow for the detection of even one single spore passing the filter. The test procedure, which is carried out in laboratory scale, was furthermore validated regarding its sufficiency to cover real life situations by upscaling using air conditioning devices showing great conformity in terms of separation rates. Additionally, a clinical study with allergy sufferers was performed to verify analytical results. Several different air conditioning filters from the car industry have been tested, showing significant differences in their separation rates.

Keywords: airborne germs, allergens, classification of filters, fine dust

Procedia PDF Downloads 224

Authors: Umut Kokbas, Mustafa Nisari

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Immunoglobulins, also known as antibodies, are glycoprotein molecules produced by activated B cells that transform into plasma cells and result in them. Antibodies are critical molecules of the immune response to fight, which help the immune system specifically recognize and destroy antigens such as bacteria, viruses, and toxins. Immunoglobulin classes differ in their biological properties, structures, targets, functions, and distributions. Five major classes of antibodies have been identified in mammals: IgA, IgD, IgE, IgG, and IgM. Evaluation of the immunoglobulin isotype can provide a useful insight into the complex humoral immune response. Evaluation and knowledge of immunoglobulin structure and classes are also important for the selection and preparation of antibodies for immunoassays and other detection applications. The immunoglobulin test measures the level of certain immunoglobulins in the blood. IgA, IgG, and IgM are usually measured together. In this way, they can provide doctors with important information, especially regarding immune deficiency diseases. Hypogammaglobulinemia (HGG) is one of the main groups of primary immunodeficiency disorders. HGG is caused by various defects in B cell lineage or function that result in low levels of immunoglobulins in the bloodstream. This affects the body's immune response, causing a wide range of clinical features, from asymptomatic diseases to severe and recurrent infections, chronic inflammation and autoimmunity Transient infant hypogammaglobulinemia (THGI), IgM deficiency (IgMD), Bruton agammaglobulinemia, IgA deficiency (SIgAD) HGG samples are a few. Most patients can continue their normal lives by taking prophylactic antibiotics. However, patients with severe infections require intravenous immune serum globulin (IVIG) therapy. The IgE level may rise to fight off parasitic infections, as well as a sign that the body is overreacting to allergens. Also, since the immune response can vary with different antigens, measuring specific antibody levels also aids in the interpretation of the immune response after immunization or vaccination. Immune deficiencies usually occur in childhood. In Immunology and Allergy clinics, apart from the classical methods, it will be more useful in terms of diagnosis and follow-up of diseases, if it is fast, reliable and especially in childhood hypogammaglobulinemia, sampling from children with a method that is more convenient and uncomplicated. The antibodies were attached to the electrode surface via the poly hydroxyethyl methacrylamide cysteine nanopolymer. It was used to evaluate the anodic peak results obtained in the electrochemical study. According to the data obtained, immunoglobulin determination can be made with a biosensor. However, in further studies, it will be useful to develop a medical diagnostic kit with biomedical engineering and to increase its sensitivity.

Keywords: biosensor, immunosensor, immunoglobulin, infection

Procedia PDF Downloads 65

Authors: Nareshkumar Harale, B. B. Meshram

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The continued exponential growth of successful cyber intrusions against today’s businesses has made it abundantly clear that traditional perimeter security measures are no longer adequate and effective. We evolved the network trust architecture from trust-untrust to Zero-Trust, With Zero Trust, essential security capabilities are deployed in a way that provides policy enforcement and protection for all users, devices, applications, data resources, and the communications traffic between them, regardless of their location. Information exchange over the Internet, in spite of inclusion of advanced security controls, is always under innovative, inventive and prone to cyberattacks. TCP/IP protocol stack, the adapted standard for communication over network, suffers from inherent design vulnerabilities such as communication and session management protocols, routing protocols and security protocols are the major cause of major attacks. With the explosion of cyber security threats, such as viruses, worms, rootkits, malwares, Denial of Service attacks, accomplishing efficient and effective intrusion detection and prevention is become crucial and challenging too. In this paper, we propose a design and analysis model for next generation network intrusion detection and protection system as part of layered security strategy. The proposed system design provides intrusion detection for wide range of attacks with layered architecture and framework. The proposed network intrusion classification framework deals with cyberattacks on standard TCP/IP protocol, routing protocols and security protocols. It thereby forms the basis for detection of attack classes and applies signature based matching for known cyberattacks and data mining based machine learning approaches for unknown cyberattacks. Our proposed implemented software can effectively detect attacks even when malicious connections are hidden within normal events. The unsupervised learning algorithm applied to network audit data trails results in unknown intrusion detection. Association rule mining algorithms generate new rules from collected audit trail data resulting in increased intrusion prevention though integrated firewall systems. Intrusion response mechanisms can be initiated in real-time thereby minimizing the impact of network intrusions. Finally, we have shown that our approach can be validated and how the analysis results can be used for detecting and protection from the new network anomalies.

Keywords: network intrusion detection, network intrusion prevention, association rule mining, system analysis and design

Procedia PDF Downloads 200

Authors: Guler D. Uzel, Andrew G. Parker, Robert L. Mach, Adly Abd-Alla

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Several tsetse fly species (Diptera: Glossinidae) in natural or colonized populations can be infected with the salivary gland hypertrophy virus (SGHV), a circular dsDNA virus (Hytrosaviridae). The virus infection is mainly asymptomatic but, in some species under certain conditions, the infection can produce salivary gland hypertrophy (SGH) symptoms. In the laboratory colonized tsetse, flies with SGH have reduced fertility, which negatively affects colony performance. Therefore, a high prevalence of SGH in insect mass rearing represents a major challenge for tsetse control using the sterile insect technique. The main objective of this study is to analyze the impact of Glossina pallidipes SGHV infection in various tsetse species on mortality and productivity and its impact on the symbiotic bacteria. Hypertropied salivary glands (SG) were collected from G. pallidipes into phosphate buffered saline (PBS) to prepare suspension; 2 µl aliquots were injected into adults of several tsetse species (G. pallidipes (Gp), G. p. gambiensis (Gpg), G. brevipalpis (Gb), G. morsitans morsitans (Gmm), G. morsitans centralis (Gmc) and G. fuscipes (Gf)) and the change in virus and symbiont titers were analyzed using qPCR. The development of SGH in the F1 was detected by dissection 10 days after emergence and virus infection was confirmed by PCR. The impact of virus infection on fly mortality and productivity was recorded. 2 µl aliquots were also injected into 3rd instar larvae of the different species and the adult SGs assayed by PCR for virus. Virus positive SGs from each species were homogenized in PBS and pooled within species for injection into larvae of the same species. Flies injected with PBS were used as control. Injecting teneral flies with SGHV caused increasing virus titer over time in all species but no SGH was detected. Dissection of the F1 also showed no development of SGH except in Gp (the homologous host). Injection of SGHV did not have any impact on the prevalence of the tsetse symbionts, but an increase in Sodalis titer was observed correlated with fly age regardless of virus infection. The virus infection had a negative impact on productivity and mortality. SGHV injection into larvae of the different species produced SGHV infected glands in the adults determined by PCR with a rate of 60%, 27%, 16%, 7% and 7% for Gp, Gf, Gpg, Gmm and Gmc, respectively. Virus positive SGs observed in the heterologous species were smaller than SGH found in Gp. No virus positive SG was detected by PCR in Gb and no SGH was observed in any adults except in Gp. Injecting virus suspension from the virus positive SGs into conspecific larvae did not produce any adults with infected SGs (except in Gp). SGHV can infect all tested tsetse species. Although the virus can infect and increase in titer in other tsetse species and affect fly mortality and productivity, no vertical virus transmission was observed in other tsetse species with might indicate a transmission barrier in these species, and virus collected from flies injected as larvae was not infective by injection.

Keywords: DNA viruses, glossina, hytrosaviridae, symbiotic bacteria, tsetse

Procedia PDF Downloads 184

Authors: Lakmini Wijesooriya, Vicki Chalker, Jessica Day, Priyantha Perera, N. P. Sunil-Chandra

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M. pneumoniae has been identified as an etiology for exacerbation of asthma (EQA), although viruses play a major role in EOA. M. pneumoniae infection is treated empirically with macrolides, and its antibiotic sensitivity is not detected routinely. Characterization of the organism by genotyping and determination of macrolide resistance is important epidemiologically as it guides the empiric antibiotic treatment. To date, there is no such characterization of M. pneumoniae performed in Sri Lanka. The present study describes the characterization of M. pneumoniae detected from a child with EOA following a screening of 100 children with EOA. Of the hundred children with EOA, M. pneumoniae was identified only in one child by Real-Time polymerase chain reaction (PCR) test for identifying the community-acquired respiratory distress syndrome (CARDS) toxin nucleotide sequences. The M. pneumoniae identified from this patient underwent detection of macrolide resistance via conventional PCR, amplifying and sequencing the region of the 23S rDNA gene that contains single nucleotide polymorphisms that confer resistance. Genotyping of the isolate was performed via nested Multilocus Sequence Typing (MLST) in which eight (8) housekeeping genes (ppa, pgm, gyrB, gmk, glyA, atpA, arcC, and adk) were amplified via nested PCR followed by gene sequencing and analysis. As per MLST analysis, the M. pneumoniae was identified as sequence type 14 (ST14), and no mutations that confer resistance were detected. Resistance to macrolides in M. pneumoniae is an increasing problem globally. Establishing surveillance systems is the key to informing local prescriptions. In the absence of local surveillance data, antibiotics are started empirically. If the relevant microbiological samples are not obtained before antibiotic therapy, as in most occasions in children, the course of antibiotic is completed without a microbiological diagnosis. This happens more frequently in therapy for M. pneumoniae which is treated with a macrolide in most patients. Hence, it is important to understand the macrolide sensitivity of M. pneumoniae in the setting. The M. pneumoniae detected in the present study was macrolide sensitive. Further studies are needed to examine a larger dataset in Sri Lanka to determine macrolide resistance levels to inform the use of macrolides in children with EOA. The MLST type varies in different geographical settings, and it also provides a clue to the existence of macrolide resistance. The present study enhances the database of the global distribution of different genotypes of M. pneumoniae as this is the first such characterization performed with the increased number of samples to determine macrolide resistance level in Sri Lanka. M. pneumoniae detected from a child with exacerbation of asthma in Sri Lanka was characterized as ST14 by MLST and no mutations that confer resistance were detected.

Keywords: mycoplasma pneumoniae, Sri Lanka, characterization, macrolide resistance

Procedia PDF Downloads 154

Authors: Yunjia Yang, Peng Li, Gordon Xu, Timothy Mahony, Bing Zhang, Neena Mitter, Karishma Mody

Abstract:

Sheep flystrike is one of the most economically important diseases affecting the Australian sheep and wool industry (>356M/annually). Currently, control of Lucillia cuprina relies almost exclusively on chemicals controls, and the parasite has developed resistance to nearly all control chemicals used in the past. It is, therefore, critical to develop an alternative solution for the sustainable control and management of flystrike. RNA interference (RNAi) technologies have been successfully explored in multiple animal industries for developing parasites controls. This research project aims to develop a RNAi based biological control for sheep blowfly. Double-stranded RNA (dsRNA) has already proven successful against viruses, fungi, and insects. However, the environmental instability of dsRNA is a major bottleneck for successful RNAi. Bentonite polymer (BenPol) technology can overcome this problem, as it can be tuned for the controlled release of dsRNA in the gut challenging pH environment of the blowfly larvae, prolonging its exposure time to and uptake by target cells. To investigate the potential of BenPol technology for dsRNA delivery, four different BenPol carriers were tested for their dsRNA loading capabilities, and three of them were found to be capable of affording dsRNA stability under multiple temperatures (4°C, 22°C, 40°C, 55°C) in sheep serum. Based on stability results, dsRNA from potential targeted genes was loaded onto BenPol carriers and tested in larvae feeding assays, three genes resulting in knockdowns. Meanwhile, a primary blowfly embryo cell line (BFEC) derived from L. cuprina embryos was successfully established, aim for an effective insect cell model for testing RNAi efficacy for preliminary assessments and screening. The results of this study establish that the dsRNA is stable when loaded on BenPol particles, unlike naked dsRNA rapidly degraded in sheep serum. The stable nanoparticle delivery system offered by BenPol technology can protect and increase the inherent stability of dsRNA molecules at higher temperatures in a complex biological fluid like serum, providing promise for its future use in enhancing animal protection.

Keywords: lucilia cuprina, primary cell line establishment, RNA interference, insect cell transfection

Procedia PDF Downloads 47

Authors: Miguel García-Ferrús, Yolanda Domínguez, M Angeles Castillo, M Antonia Ferrús, Ana Jiménez-Belenguer

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Antibiotic resistance is a growing public health concern worldwide, and it is now regarded as a critical issue within the "One Health" approach that affects human and animal health, agriculture, and environmental waste management. This concept focuses on the interconnected nature of human, animal and environmental health, and WHO highlights zoonotic diseases, food safety, and antimicrobial resistance as three particularly relevant areas for this framework. Fresh vegetables are garnering attention in the food chain due to the presence of pathogens and because they can act as a reservoir for Antibiotic Resistance Bacteria (ARB) and Antibiotic Resistance Genes (ARG). These fresh products are frequently consumed raw, thereby contributing to the spread and transmission of antibiotic resistance. Therefore, the aim of this research was to study the microbiological quality, the prevalence of ARB, and their role in the dissemination of ARG in fresh vegetables intended for human consumption. For this purpose, 102 samples of fresh vegetables (30 lettuce, 30 cabbage, 18 strawberries and 24 spinach) from different retail establishments in Valencia (Spain) have been analyzed to determine their microbiological quality and their role in spreading ARB and ARG. The samples were collected and examined according to standardized methods for total viable bacteria, coliforms, Shiga toxin-producing Escherichia coli (STEC), Listeria monocytogenes and Salmonella spp. Isolation was made in culture media supplemented with antibiotics (cefotaxime and meropenem). A total of 239 strains resistant to beta-lactam antibiotics (Third-Generation Cephalosporins and Carbapenems) were isolated. Thirty Gram-negative isolates were selected and biochemically identified or partial sequencing of 16S rDNA. Their sensitivity to 12 antibiotic discs was determined using the Kirby-Bauer disc diffusion technique to different therapeutic groups. To determine the presence of ARG, PCR assays for the direct sample and selected isolate DNA were performed for main expanded spectrum beta-lactamase (ESBL)-, carbapenemase-encoding genes and plasmid-mediated quinolone resistance genes. From the total samples, 68% (24/24 spinach, 28/30 lettuce and 17/30 cabbage) showed total viable bacteria levels over the accepted standard 10(2)-10(5) cfu/g range; and 48% (24/24 spinach, 19/30 lettuce and 6/30) showed coliforms levels over the accepted standard 10(2)-10(4) cfu/g range. In 9 samples (3/24 spinach, 3/30 lettuce, 3/30 cabbage; 9/102 (9%)) E. coli levels were higher than the standard 10(3) cfu/g limit. Listeria monocytogenes, Salmonella and STEC have not been detected. Six different bacteria species were isolated from samples. Stenotrophomonas maltophilia (64%) was the prevalent species, followed by Acinetobacter pitii (14%) and Burkholderia cepacia (7%). All the isolates were resistant to at least one tested antibiotic, including meropenem (85%) and ceftazidime (46%). Of the total isolates, 86% were multidrug-resistant and 68% were ESBL productors. Results of PCR showed the presence of resistance genes to beta-lactams blaTEM (4%) and blaCMY-2 (4%), to carbapenemes blaOXA-48 (25%), blaVIM (7%), blaIMP (21%) and blaKPC (32%), and to quinolones QnrA (7%), QnrB (11%) and QnrS (18%). Thus, fresh vegetables harboring ARB and ARG constitute a potential risk to consumers. Further studies must be done to detect ARG and how they propagate in non-medical environments.

Keywords: ESBL, β-lactams, resistances, fresh vegetables.

Procedia PDF Downloads 38

Authors: Hosein I. Hosein, Ragab Azzam, Ahmed M. S. Menshawy, Sherin Rouby, Khaled Hendy, Ayman Mahrous, Hany Hussien

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Brucellosis is a highly contagious bacterial zoonotic disease of a worldwide spread and has different names; Infectious or enzootic abortion and Bang's disease in animals; and Mediterranean or Malta fever, Undulant Fever and Rock fever in humans. It is caused by the different species of genus Brucella which is a Gram-negative, aerobic, non-spore forming, facultative intracellular bacterium. Brucella affects a wide range of mammals including bovines, small ruminants, pigs, equines, rodents, marine mammals as well as human resulting in serious economic losses in animal populations. In human, Brucella causes a severe illness representing a great public health problem. The disease was reported in Egypt for the first time in 1939; since then the disease remained endemic at high levels among cattle, buffalo, sheep and goat and is still representing a public health hazard. The annual economic losses due to brucellosis were estimated to be about 60 million Egyptian pounds yearly, but actual estimates are still missing despite almost 30 years of implementation of the Egyptian control programme. Despite being the gold standard, bacterial isolation has been reported to show poor sensitivity for samples with low-level of Brucella and is impractical for regular screening of large populations. Thus, serological tests still remain the corner stone for routine diagnosis of brucellosis, especially in developing countries. In the present study, a total of 1533 cows (256 from Beni-Suef Governorate, 445 from Al-Fayoum Governorate and 832 from Damietta Governorate), were employed for estimation of relative sensitivity, relative specificity, positive predictive value and negative predictive value of buffered acidified plate antigen test (BPAT), rose bengal test (RBT) and complement fixation test (CFT). The overall seroprevalence of brucellosis revealed (19.63%). Relative sensitivity, relative specificity, positive predictive value and negative predictive value of BPAT,RBT and CFT were estimated as, (96.27 %, 96.76 %, 87.65 % and 99.10 %), (93.42 %, 96.27 %, 90.16 % and 98.35%) and (89.30 %, 98.60 %, 94.35 %and 97.24 %) respectively. BPAT showed the highest sensitivity among the three employed serological tests. RBT was less specific than BPAT. CFT showed the least sensitivity 89.30 % among the three employed serological tests but showed the highest specificity. Different tissues specimens of 22 seropositive cows (spleen, retropharyngeal udder, and supra-mammary lymph nodes) were subjected for bacteriological studies for isolation and identification of Brucella organisms. Brucella melitensis biovar 3 could be recovered from 12 (54.55%) cows. Bacteriological examinations failed to classify 10 cases (45.45%) and were culture negative. Bruce-ladder PCR was carried out for molecular identification of the 12 Brucella isolates at the species level. Three fragments of 587 bp, 1071 bp and 1682 bp sizes were amplified indicating Brucella melitensis. The results indicated the importance of using several procedures to overcome the problem of escaping of some infected animals from diagnosis.Bruce-ladder PCR is an important tool for diagnosis and epidemiologic studies, providing relevant information for identification of Brucella spp.

Keywords: brucellosis, relative sensitivity, relative specificity, Bruce-ladder, Egypt

Procedia PDF Downloads 314

Authors: Damiano Omari, Topirian Kerempe, Dibo Sama, Walter Wafula, Sharon Chepkoech, Chrispine Juma, Gilbert Kirui, Simon Mburu, Susan Keino

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The One Health concept was officially adopted by the international organizations and scholarly bodies in 1984. It aims at combining human, animal and environmental components to address global health challenges. Using collaborative efforts optimal health to people, animals, and the environment can be achieved. One health approach plays a significant approach role in prevention and control of zoonosis diseases. It has also been noted that 75% of new emerging human infectious diseases are zoonotic. In Kenya, one health has been embraced and strongly advocated for by One Health East and Central Africa (OHCEA). It was inaugurated on 17th of October 2010 at a historic meeting facilitated by USAID with participants from 7 public health schools, seven faculties of veterinary medicine in Eastern Africa and 2 American universities (Tufts and University of Minnesota) in addition to respond project staff. The study was conducted in Loitoktok Sub County, specifically in the Amboseli Ecosystem. The Amboseli ecosystem covers an area of 5,700 square kilometers and stretches between Mt. Kilimanjaro, Chyulu Hills, Tsavo West National park and the Kenya/Tanzania border. The area is arid to semi-arid and is more suitable for pastoralism with a high potential for conservation of wildlife and tourism enterprises. The ecosystem consists of the Amboseli National Park, which is surrounded by six group ranches which include Kimana, Olgulului, Selengei, Mbirikani, Kuku and Rombo in Loitoktok District. The Manyatta of study was Osiram Cultural Manyatta in Mbirikani group ranch. Apart from visiting the Manyatta, we also visited the sub-county hospital, slaughter slab, forest service, Kimana market, and the Amboseli National Park. The aim of the study was to identify the one health issues facing the community. This was done by a conducting a community needs assessment and prioritization. Different methods were used in data collection for the qualitative and numerical data. They include among others; key informant interviews and focus group discussions. We also guided the community members in drawing their Resource Map this helped identify the major resources in their land and also help them identify some of the issues they were facing. Matrix piling, root cause analysis, and force field analysis tools were used to establish the one health related priority issues facing community members. Skits were also used to present to the community interventions to the major one health issues. Some of the prioritized needs among the community were water scarcity and inadequate markets for their beadwork. The group intervened on the various needs of the Manyatta. For water scarcity, we educated the community on water harvesting methods using gutters as well as proper storage by the use of tanks and earth dams. The community was also encouraged to recycle and conserve water. To improve markets; we educated the community to upload their products online, a page was opened for them and uploading the photos was demonstrated to them. They were also encouraged to be innovative to attract more clients.

Keywords: Amboseli ecosystem, community interventions, community needs assessment and prioritization, one health issues

Procedia PDF Downloads 139

Authors: Serap Gedikli, Hakan Çakmak, M. Buğra Güldiken, Duygu Yalnızoğlu

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Air, which is necessary for living things to survive; while it carries some useful substances in it, it can also carry foreign particles of different sizes that may be harmful to the health. All airborne organic substances of biological origin, including bacteria, fungi, fungal spores, viruses, pollen, and their components, are called "bioaerosols". Nowadays, everyone spends most of their time in closed areas such as home, workplace, school, etc. Although it is known that outdoor air pollution affects health, it is not known that indoor air pollution has harmful effects in terms of health. In this study, indoor air microbial load and SARS-CoV-2 virus cleaning efficiency of Froumann brand air cleaners were studied. This work in 300 m³, 600 m³, and 1000 m³ completely closed areas without any air circulation with Froumann N80, N90, and N100 air-cleaning devices. Analyzes were performed for both areas at 60 minutes before and after the device was operated using a particle measuring device (Particles Plus 7302) and an air sampler (Mas-100 ECO). The measurements were taken by placing the test equipment 1.5-2 m away from the air cleaner. At the same time, the efficiency of the HEPA filter was evaluated by taking samples from the air outlet point of the HEPA filter using the air sampling device (Mas-100 ECO) after the device was started. Nutrient agar and malt agar are used as total mesophilic bacteria and total fungi. The number of colony-forming units per m³ (cfu/m³) was calculated by counting colonies in Petri dishes after incubation for 48 hours at 37°C for bacteria and 72 hours at 30°C for fungi. The change in the number of colonies and the decrease in the microbial load was calculated as a percentage value. SARS-CoV-2 activity analysis studies were carried out by İnönü University Microbiology Department in accordance with the World Health Organization regulations. Finally, the HEPA filter in the devices used was taken and kept under a certain temperature and humidity, and the change in the microbial load on it was monitored over a 6-month period. At the end of the studies, a 91%-94% reduction was determined in the total mesophilic bacteria count of Frouman brand N80, N90, and N100 model air cleaners. A decrease of 94%-96% was detected in the total number of yeast/molds. HEPA filter efficiency was evaluated, and at the end of the analysis, 98% of the bacterial load and approximately 100% of yeast/mold load at the HEPA filter air outlet point were decreased. According to the SARS- CoV-2 analysis results, when the device is operating at the medium airflow level 3, it can filter virus-carrying aerosols by 99%. As a result, it was determined that the Froumann model air cleaner was effective in controlling and reducing the microbial load in the indoor air.

Keywords: HEPA filter, indoor air quality, microbial load, SARS-CoV-2

Procedia PDF Downloads 165

Authors: Viviana Manzulli, Luigina Serrecchia, Adelia Donatiello, Valeria Rondinone, Sabine Zange, Alina Tscherne, Antonio Parisi, Antonio Fasanella

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Anthrax is a zoonotic disease that affects a wide range of animal species (primarily ruminant herbivores), and can be transmitted to humans through consumption or handling of contaminated animal products. The etiological agent B.anthracis is able to survive in unfavorable environmental conditions by forming endospore which remain viable in the soil for many decades. Furthermore, B.anthracis is considered as one of the most feared agents to be potentially misused as a biological weapon and the importance of the disease and its treatment in humans has been underscored before the bioterrorism events in the United States in 2001. Due to the often fatal outcome of human cases, antimicrobial susceptibility testing plays especially in the management of anthrax infections an important role. In Italy, animal anthrax is endemic (predominantly found in the southern regions and on islands) and is characterized by sporadic outbreaks occurring mainly during summer. Between 2012 and 2017 single human cases of cutaneous anthrax occurred. In this study, 90 diverse strains of B.anthracis, isolated in Italy from 2001 to 2017, were screened to their susceptibility to sixteen clinically relevant antimicrobial agents by using the broth microdilution method. B.anthracis strains selected for this study belong to the strain collection stored at the Anthrax Reference Institute of Italy located inside the Istituto Zooprofilattico Sperimentale of Puglia and Basilicata. The strains were isolated at different time points and places from various matrices (human, animal and environmental). All strains are a representative of over fifty distinct MLVA 31 genotypes. The following antibiotics were used for testing: gentamicin, ceftriaxone, streptomycin, penicillin G, clindamycin, chloramphenicol, vancomycin, linezolid, cefotaxime, tetracycline, erythromycin, rifampin, amoxicillin, ciprofloxacin, doxycycline and trimethoprim. A standard concentration of each antibiotic was prepared in a specific diluent, which were then twofold serial diluted. Therefore, each wells contained: bacterial suspension of 1–5x104 CFU/mL in Mueller-Hinton Broth (MHB), the antibiotic to be tested at known concentration and resazurin, an indicator of cell growth. After incubation overnight at 37°C, the wells were screened for color changes caused by the resazurin: a change from purple to pink/colorless indicated cell growth. The lowest concentration of antibiotic that prevented growth represented the minimal inhibitory concentration (MIC). This study suggests that B.anthracis remains susceptible in vitro to many antibiotics, in addition to doxycycline (MICs ≤ 0,03 µg/ml), ciprofloxacin (MICs ≤ 0,03 µg/ml) and penicillin G (MICs ≤ 0,06 µg/ml), recommend by CDC for the treatment of human cases and for prophylactic use after exposure to the spores. In fact, the good activity of gentamicin (MICs ≤ 0,25 µg/ml), streptomycin (MICs ≤ 1 µg/ml), clindamycin (MICs ≤ 0,125 µg/ml), chloramphenicol(MICs ≤ 4 µg/ml), vancomycin (MICs ≤ 2 µg/ml), linezolid (MICs ≤ 2 µg/ml), tetracycline (MICs ≤ 0,125 µg/ml), erythromycin (MICs ≤ 0,25 µg/ml), rifampin (MICs ≤ 0,25 µg/ml), amoxicillin (MICs ≤ 0,06 µg/ml), towards all tested B.anthracis strains demonstrates an appropriate alternative choice for prophylaxis and/or treatment. All tested B.anthracis strains showed intermediate susceptibility to the cephalosporins (MICs ≥ 16 µg/ml) and resistance to trimethoprim (MICs ≥ 128 µg/ml).

Keywords: Bacillus anthracis, antibiotic susceptibility, treatment, minimum inhibitory concentration

Procedia PDF Downloads 186

Authors: Marcin Debowski, Marcin Zielinski, Magdalena Zielinska, Paulina Rusanowska

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The problem with digestate management is one of the most important factors influencing on the development and operation of biogas plant. Turbidity and bacterial contamination negatively affect the growth of algae, which can limit the use of the effluent in the production of algae biomass on a large scale. These problems can be overcome by cultivating of algae species resistant to environmental factors, such as Chlorella sp., Scenedesmus sp., or reducing load of organic compounds to prevent bacterial contamination. The effluent requires dilution and/or purification. One of the methods of effluent treatment is the use of a membrane technology such as microfiltration (MF), ultrafiltration (UF), nanofiltration (NF) and reverse osmosis (RO), depending on the membrane pore size and the cut off point. Membranes are a physical barrier to solids and particles larger than the size of the pores. MF membranes have the largest pores and are used to remove turbidity, suspensions, bacteria and some viruses. UF membranes remove also color, odor and organic compounds with high molecular weight. In treatment of wastewater or other waste streams, MF and UF can provide a sufficient degree of purification. NF membranes are used to remove natural organic matter from waters, water disinfection products and sulfates. RO membranes are applied to remove monovalent ions such as Na⁺ or K⁺. The effluent was used in UF for medium to cultivation of two microalgae: Chlorella sp. and Phaeodactylum tricornutum. Growth rates of Chlorella sp. and P. tricornutum were similar: 0.216 d⁻¹ and 0.200 d⁻¹ (Chlorella sp.); 0.128 d⁻¹ and 0.126 d⁻¹ (P. tricornutum), on synthetic medium and permeate from UF, respectively. The final biomass composition was also similar, regardless of the medium. Removal of nitrogen was 92% and 71% by Chlorella sp. and P. tricornutum, respectively. The fermentation effluents after UF and dilution were also used for cultivation of algae Scenedesmus sp. that is resistant to environmental conditions. The authors recommended the development of biorafinery based on the production of algae for the biogas production. There are examples of using a multi-stage membrane system to purify the liquid fraction from digestate. After the initial UF, RO is used to remove ammonium nitrogen and COD. To obtain a permeate with a concentration of ammonium nitrogen allowing to discharge it into the environment, it was necessary to apply three-stage RO. The composition of the permeate after two-stage RO was: COD 50–60 mg/dm³, dry solids 0 mg/dm³, ammonium nitrogen 300–320 mg/dm³, total nitrogen 320–340 mg/dm³, total phosphorus 53 mg/dm³. However compostion of permeate after three-stage RO was: COD < 5 mg/dm³, dry solids 0 mg/dm³, ammonium nitrogen 0 mg/dm³, total nitrogen 3.5 mg/dm³, total phosphorus < 0,05 mg/dm³. Last stage of RO might be replaced by ion exchange process. The negative aspect of membrane filtration systems is the fact that the permeate is about 50% of the introduced volume, the remainder is the retentate. The management of a retentate might involve recirculation to a biogas plant.

Keywords: digestate, membrane filtration, microalgae cultivation, Chlorella sp.

Procedia PDF Downloads 325

Authors: Sofia Papadimitriou

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INTRODUCTION: The differential diagnosis between acute viral and bacterial infections is an important cost-effectiveness parameter at the stage of the treatment process in order to achieve the maximum benefits in therapeutic intervention by combining the minimum cost to ensure the proper use of antibiotics.The discovery of sensitive and robust molecular diagnostic tests in response to the role of the host in infections has enhanced the accurate diagnosis and differentiation of infections. METHOD: The study used a sample of six independent blood samples (total=756) which are associated with human proteins-proteins, each of which at the transcription stage expresses a different response in the host network between viral and bacterial infections.Τhe individual blood samples are subjected to a sequence of computer filters that identify a gene panel corresponding to an autonomous diagnostic score. The data set and the correspondence of the gene panel to the diagnostic patents a new Bangalore -Viral Bacterial (BL-VB). FINDING: We use a biomarker based on the blood of 10 genes(Panel-VB) that are an important prognostic value for the detection of viruses from bacterial infections with a weighted average AUROC of 0.97(95% CL:0.96-0.99) in eleven independent samples (sets n=898). We discovered a base with a patient score (VB 10 ) according to the table, which is a significant diagnostic value with a weighted average of AUROC 0.94(95% CL: 0.91-0.98) in 2996 patient samples from 56 public sets of data from 19 different countries. We also studied VB 10 in a new cohort of South India (BL-VB,n=56) and found 97% accuracy in confirmed cases of viral and bacterial infections. We found that VB 10 (a)accurately identifies the type of infection even in unspecified cases negative to the culture (b) shows its clinical condition recovery and (c) applies to all age groups, covering a wide range of acute bacterial and viral infectious, including non-specific pathogens. We applied our VB 10 rating to publicly available COVID 19 data and found that our rating diagnosed viral infection in patient samples. RESULTS: Τhe results of the study showed the diagnostic power of the biomarker VB 10 as a diagnostic test for the accurate diagnosis of acute infections in recovery conditions. We look forward to helping you make clinical decisions about prescribing antibiotics and integrating them into your policies management of antibiotic stewardship efforts. CONCLUSIONS: Overall, we are developing a new property of the RNA-based biomarker and a new blood test to differentiate between viral and bacterial infections to assist a physician in designing the optimal treatment regimen to contribute to the proper use of antibiotics and reduce the burden on antimicrobial resistance, AMR.

Keywords: acute infections, antimicrobial resistance, biomarker, blood transcriptome, systems biology, classifier diagnostic score

Procedia PDF Downloads 128

Authors: Halkiopoulos Constantinos, Koutsopoulou Ioanna, Gkintoni Evgenia, Antonopoulou Hera

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Immunization appears to be an essential part of health care service in times of pandemics such as covid-19 and aims not only to protect the health of the population but also the health and sustainability of the economies of the countries affected. It is reported that more than 3.44 billion doses have been administered so far, which accounts for 45 doses for 100 people. Vaccination programs in various countries have been promoted and accepted by people differently and therefore they proceeded in different ways and speed; most countries directing them towards people with vulnerable chronic or recent health statuses. Large scale restriction measures or lockdown, personal protection measures such as masks and gloves and a decrease in leisure and sports activities were also implemented around the world as part of the protection health strategies against the covid-19 pandemic. This research aims to present an analysis based on variations on people’s attitudes towards vaccination based on demographic, social and epidemiological characteristics, and health status on the one hand and perception of health, health satisfaction, pain, and quality of life on the other hand. 1500 Greek e-consumers participated in the research, mainly through social media who took part in an online-based survey voluntarily. The questionnaires included demographic, social and medical characteristics of the participants, and questions asking people’s willingness to be vaccinated and their opinion on whether there should be a vaccine against covid-19. Other stressor factors were also reported in the questionnaires and participants’ loss of someone close due to covid-19, or staying at home quarantine due to being infected from covid-19. WHOQUOL-BREF and GLOBAL PSYCHOTRAUMA SCREEN- GPS were used with kind permission from WHO and from the International Society for Traumatic Stress Studies in this study. Attitudes towards vaccination varied significantly related to aging, level of education, health status and consumer behavior. Health professionals’ attitudes also varied in relation to age, level of education, profession, health status and consumer needs. Vaccines have been the most common technological aid of human civilization so far in the fight against viruses. The results of this study can be used for health managers and digital marketers of pharmaceutical companies and also other staff involved in vaccination programs and for designing health policy immunization strategies during pandemics in order to achieve positive attitudes towards vaccination and larger populations being vaccinated in shorter periods of time after the break out of pandemic. Health staff needs to be trained, aided and supervised to go through with vaccination programs and to be protected through vaccination programs themselves. Feedback in each country’s vaccination program, short backs, deficiencies and delays should be addressed and worked out.

Keywords: consumer behavior, cognitive models, vaccination policy, pandemic, Covid-19, Greece

Procedia PDF Downloads 156

Authors: Mattias Bergstrom

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Significance of the Study: This research will provide new insights into the risks with digital embedded infrastructure. Through this research, we will analyze each risk and its potential negation strategies, especially for AI and autonomous automation. Moreover, the analysis that is presented in this paper will convey valuable information for future research that can create more stable, secure, and efficient autonomous systems. To learn and understand the risks, a large IoT system was envisioned, and risks with hardware, tampering, and cyberattacks were collected, researched, and evaluated to create a comprehensive understanding of the potential risks. Potential solutions have then been evaluated on an open source IoT hardware setup. This list shows the identified passive and active risks evaluated in the research. Passive Risks: (1) Hardware failures- Critical Systems relying on high rate data and data quality are growing; SCADA systems for infrastructure are good examples of such systems. (2) Hardware delivers erroneous data- Sensors break, and when they do so, they don’t always go silent; they can keep going, just that the data they deliver is garbage, and if that data is not filtered out, it becomes disruptive noise in the system. (3) Bad Hardware injection- Erroneous generated sensor data can be pumped into a system by malicious actors with the intent to create disruptive noise in critical systems. (4) Data gravity- The weight of the data collected will affect Data-Mobility. (5) Cost inhibitors- Running services that need huge centralized computing is cost inhibiting. Large complex AI can be extremely expensive to run. Active Risks: Denial of Service- It is one of the most simple attacks, where an attacker just overloads the system with bogus requests so that valid requests disappear in the noise. Malware- Malware can be anything from simple viruses to complex botnets created with specific goals, where the creator is stealing computer power and bandwidth from you to attack someone else. Ransomware- It is a kind of malware, but it is so different in its implementation that it is worth its own mention. The goal with these pieces of software is to encrypt your system so that it can only be unlocked with a key that is held for ransom. DNS spoofing- By spoofing DNS calls, valid requests and data dumps can be sent to bad destinations, where the data can be extracted for extortion or to corrupt and re-inject into a running system creating a data echo noise loop. After testing multiple potential solutions. We found that the most prominent solution to these risks was to use a Peer 2 Peer consensus algorithm over a blockchain to validate the data and behavior of the devices (sensors, storage, and computing) in the system. By the devices autonomously policing themselves for deviant behavior, all risks listed above can be negated. In conclusion, an Internet middleware that provides these features would be an easy and secure solution to any future autonomous IoT deployments. As it provides separation from the open Internet, at the same time, it is accessible over the blockchain keys.

Keywords: IoT, security, infrastructure, SCADA, blockchain, AI

Procedia PDF Downloads 69

Authors: Pakize Ozlem Kurt Polat

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GMO (genetically modified organism) is the result of a laboratory process where genes from the DNA of one species are extracted and artificially forced into the genes of an unrelated plant or animal. This technology includes; nucleic acid hybridization, recombinant DNA, RNA, PCR, cell culture and gene cloning techniques. The studies are divided into three groups of properties transferred to the transgenic plant. Up to 59% herbicide resistance characteristic of the transfer, 28% resistance to insects and the virus seems to be related to quality characteristics of 13%. Transgenic crops are not included in the commercial production of each product; mostly commercial plant is soybean, maize, canola, and cotton. Day by day increasing GMO interest can be listed as follows; Use in the health area (Organ transplantation, gene therapy, vaccines and drug), Use in the industrial area (vitamins, monoclonal antibodies, vaccines, anti-cancer compounds, anti -oxidants, plastics, fibers, polyethers, human blood proteins, and are used to produce carotenoids, emulsifiers, sweeteners, enzymes , food preservatives structure is used as a flavor enhancer or color changer),Use in agriculture (Herbicide resistance, Resistance to insects, Viruses, bacteria, fungi resistance to disease, Extend shelf life, Improving quality, Drought , salinity, resistance to extreme conditions such as frost, Improve the nutritional value and quality), we explain all this methods step by step in this research. GMO has advantages and disadvantages, which we explain all of them clearly in full text, because of this topic, worldwide researchers have divided into two. Some researchers thought that the GMO has lots of disadvantages and not to be in use, some of the researchers has opposite thought. If we look the countries law about GMO, we should know Biosafety law for each country and union. For this Biosecurity reasons, the problems caused by the transgenic plants, including Turkey, to minimize 130 countries on 24 May 2000, ‘the United Nations Biosafety Protocol’ signed nudes. This protocol has been prepared in addition to Cartagena Biosafety Protocol entered into force on September 11, 2003. This protocol GMOs in general use by addressing the risks to human health, biodiversity and sustainable transboundary movement of all GMOs that may affect the prevention, transit covers were dealt and used. Under this protocol we have to know the, ‘US Regulations GMO’, ‘European Union Regulations GMO’, ‘Turkey Regulations GMO’. These three different protocols have different applications and rules. World population increasing day by day and agricultural fields getting smaller for this reason feeding human and animal we should improve agricultural product yield and quality. Scientists trying to solve this problem and one solution way is molecular biotechnology which is including the methods of GMO too. Before decide to support or against the GMO, should know the GMO protocols and it effects.

Keywords: biotechnology, GMO (genetically modified organism), molecular marker

Procedia PDF Downloads 209

Authors: N. Manjunatha, K. T. Rangswamy, N. Nagaraju, H. A. Prameela, P. Rudraswamy, M. Krishnareddy

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The identification of virus in cowpea especially potyviruses is confusing. Even though there are several studies on viruses causing diseases in cowpea, difficult to distinguish based on symptoms and serological detection. The differentiation of potyviruses considering as a constraint, the present study is initiated for molecular characterization, host plant resistance and epidemiology of the BCMV infecting cowpea. The etiological agent causing cowpea mosaic was identified as Bean Common Mosaic Virus (BCMV) on the basis of RT-PCR and electron microscopy. An approximately 750bp PCR product corresponding to coat protein (CP) region of the virus and the presence of long flexuous filamentous particles measuring about 952 nm in size typical to genus potyvirus were observed under electron microscope. The characterized virus isolate genome had 10054 nucleotides, excluding the 3’ terminal poly (A) tail. Comparison of polyprotein of the virus with other potyviruses showed similar genome organization with 9 cleavage sites resulted in 10 functional proteins. The pairwise sequence comparison of individual genes, P1 showed most divergent, but CP gene was less divergent at nucleotide and amino acid level. A phylogenetic tree constructed based on multiple sequence alignments of the polyprotein nucleotide and amino acid sequences of cowpea BCMV and potyviruses showed virus is closely related to BCMV-HB. Whereas, Soybean variant of china (KJ807806) and NL1 isolate (AY112735) showed 93.8 % (5’UTR) and 94.9 % (3’UTR) homology respectively with other BCMV isolates. This virus transmitted to different leguminous plant species and produced systemic symptoms under greenhouse conditions. Out of 100 cowpea genotypes screened, three genotypes viz., IC 8966, V 5 and IC 202806 showed immune reaction in both field and greenhouse conditions. Single marker analysis (SMA) was revealed out of 4 SSR markers linked to BCMV resistance, M135 marker explains 28.2 % of phenotypic variation (R2) and Polymorphic information content (PIC) value of these markers was ranged from 0.23 to 0.37. The correlation and regression analysis showed rainfall, and minimum temperature had significant negative impact and strong relationship with aphid population, whereas weak correlation was observed with disease incidence. Path coefficient analysis revealed most of the weather parameters exerted their indirect contributions to the aphid population and disease incidence except minimum temperature. This study helps to identify specific gaps in knowledge for researchers who may wish to further analyse the science behind complex interactions between vector-virus and host in relation to the environment. The resistant genotypes identified are could be effectively used in resistance breeding programme.

Keywords: cowpea, epidemiology, genotypes, virus

Procedia PDF Downloads 203

Authors: Zahra Mohajer, Afsaneh Soltani

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Doping is a major issue in competitive sports and is favored by vast groups of athletes. The feeling of being higher-ranking than others and gaining fame has caused many athletes to misuse drugs. The definition of doping is to use prohibited substances and/or methods that help physical or mental performances or both. Doping counts as the illegal use of chemical substances or drugs, excessive amounts of physiological substances to increase the performance at or out of competition or even the use of inappropriate medications to treat an injury to gain the ability to participate in a competition. The International Olympic Committee (IOC) and World Anti-Doping Agency (WADA) have forbidden these substances to ensure fair and equal competition and also the health of the competitors. As of 2004 WADA has published an international list of illegal substances used for doping, which is updated annually. In the process of the Genome Project scientists have gained the ability to treat numerous diseases by gene therapy, which may result in bodily performance increase and therefore a potential opportunity to misuse by some athletes. Gene doping is defined as the non-therapeutic direct and indirect genetic modifications using genetic materials that can improve the performances in sports events. Biosynthetic drugs are a form of indirect genetic engineering. The method can be performed in three ways such as injecting the DNA directly into the muscle, inserting the genetically engineered cells, or transferring the DNA using a virus as a vector. Erythropoietin is a hormone majorly released by the kidney and in small amounts by the liver. Its function is to stimulate the erythropoiesis and therefore the more production of red blood cells (RBC) which causes an increase in Hemoglobin (Hb). During this process, the oxygen delivery to muscles will increase, which will improve athletic performance and postpone exhaustion. There are ways to increase the oxygen transferred to muscles such as blood transfusion, stimulating the production of red blood cells by using Erythropoietin (EPO), and also using allosteric effectors of Hemoglobin. EPO can either be injected as a protein or can be inserted into the cells as the gene which encodes EPO. Adeno-associated viruses have been employed to deliver the EPO gene to the cells. Employing the genes that naturally exist in the human body such as the EPO gene can reduce the risk of detecting gene doping. The first research about blood doping was conducted in 1947. The study has shown that an increase in hematocrit (HCT) up to 55% following homologous transfusion makes it more unchallenging for the body to perform the exercise at the altitude. Thereafter athletes’ attraction to blood infusion escalated. Also, a study has demonstrated that by reinfusing their own blood 4 weeks after being drawn, three men have shown a rise in Hb level which improved the oxygen uptake, and a delay in exhaustion. The list of performance-enhancing drugs is published by WADA annually and includes the following drugs: anabolic agents, hormones, Beta-2 agonists, Beta-blockers, Diuretics, Stimulants, narcotics, cannabinoids, and corticosteroids.

Keywords: doping, PEDs, sports, WADA

Procedia PDF Downloads 81

Authors: Niamh Higgins, Dawn Howard

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The Irish agri-food sector is of major importance to Ireland’s manufacturing sector and to the Irish economy through employment and the exporting of animal products worldwide. Infectious diseases and parasites have an impact on farm animal health causing profitability and productivity to be affected. For the sustainability of Irish dairy farming, there must be the highest standard of animal health. There can be a lack of information in accounting for > 1% of complete microbial diversity in an environment. There is the tendency of culture-based methods of microbial identification to overestimate the prevalence of species which grow easily on an agar surface. There is a need for new technologies to address these issues to assist with animal health. Metagenomic approaches provide information on both the whole genome and transcriptome present through DNA sequencing of total DNA from environmental samples producing high determination of functional and taxonomic information. Nanopore Next Generation Technologies have the ability to be powerful sequencing technologies. They provide high throughput, low material requirements and produce ultra-long reads, simplifying the experimental process. The aim of this study is to use a metagenomics approach to analyze dairy cattle faecal samples using the Oxford Nanopore MinION Next Generation Sequencer and to establish an in-house pipeline for metagenomic characterization of complex samples. Faecal samples will be obtained from Irish dairy farms, DNA extracted and the MinION will be used for sequencing, followed by bioinformatics analysis. Of particular interest, will be the parasite Buxtonella sulcata, which there has been little research on and which there is no research on its presence on Irish dairy farms. Preliminary results have shown the ability of the MinION to produce hundreds of reads in a relatively short time frame of eight hours. The faecal samples were obtained from 90 dairy cows on a Galway farm. The results from Oxford Nanopore ‘What’s in my pot’ (WIMP) using the Epi2me workflow, show that from a total of 926 classified reads, 87% were from the Kingdom Bacteria, 10% were from the Kingdom Eukaryota, 3% were from the Kingdom Archaea and < 1% were from the Kingdom Viruses. The most prevalent bacteria were those from the Genus Acholeplasma (71 reads), Bacteroides (35 reads), Clostridium (33 reads), Acinetobacter (20 reads). The most prevalent species present were those from the Genus Acholeplasma and included Acholeplasma laidlawii (39 reads) and Acholeplasma brassicae (26 reads). The preliminary results show the ability of the MinION for the identification of microorganisms to species level coming from a complex sample. With ongoing optimization of the pipe-line, the number of classified reads are likely to increase. Metagenomics has the potential in animal health for diagnostics of microorganisms present on farms. This would support wprevention rather than a cure approach as is outlined in the DAFMs National Farmed Animal Health Strategy 2017-2022.

Keywords: animal health, buxtonella sulcata, infectious disease, irish dairy cattle, metagenomics, minION, next generation sequencing

Procedia PDF Downloads 125

Authors: Maryam Nazari

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COVID-19 is a respiratory disease triggered by the novel coronavirus, SARS-CoV-2, that has reached pandemic status today. Acute inflammation and immune cells infiltration into lung injuries result in multi-organ failure. The presence of other non-communicable diseases (NCDs) with systemic inflammation derived from COVID-19 may exacerbate the patient's situation and increase the risk for adverse effects and mortality. This pandemic is a novel situation and the scientific community at this time is looking for vaccines or drugs to treat the pathology. One of the biggest challenges is focused on reducing inflammation without compromising the correct immune response of the patient. In this regard, addressing the nutritional factors should not be overlooked not only as a matter of avoiding the presence of NCDs with severe infections but also as an adjunctive way to modulate the inflammatory status of the patients. Despite the pivotal role of nutrition in modifying immune response, due to the novelty of the COVID-19 disease, information about the effects of specific dietary agents is limited in this area. From the macronutrients point of view, protein deficiency (quantity or quality) has negative effects on the number of functional immunoglobulins and gut-associated lymphoid tissue (GALT). High biological value proteins or some amino acids like arginine and glutamine are well known for their ability to augment the immune system. Among lipids, fish oil has the ability to inactivate enveloped viruses, suppress pro-inflammatory prostaglandin production and block platelet-activating factors and their receptors. In addition, protectin D1, which is an Omega-3 PUFAs derivation, is a novel antiviral drug. So it seems that these fatty acids can reduce the severity and/or improve recovery of patients with COVID-19. Carbohydrates with lower glycemic index and fibers are associated with lower levels of inflammatory cytokines (CRP, TNF-α, and IL-6). Short-Chain Fatty acids not only exert a direct anti-inflammatory effect but also provide appropriate gut microbial, which is important in gastrointestinal issues related to COVID-19. From the micronutrients point of view, Vitamins A, C, D, E, iron, magnesium, zinc, selenium and copper play a vital role in the maintenance of immune function. Inadequate status in these nutrients may result in decreased resistance against COVID-19 infection. There are specific bioactive compounds in the diet that interact with the ACE2 receptor, which is the gateway for SARS and SARS-CoV-2, and thus controls the viral infection. Regarding this, the potential benefits of probiotics, resveratrol (a polyphenol found in grape), oleoylethanolamide (derived from oleic acid), and natural peroxisome proliferator-activated receptor γ agonists in foodstuffs (like curcumin, pomegranate, hot pepper) are suggested. Yet, it should be pointed out that most of these results have been reported in animal models and further human studies are needed to be verified.

Keywords: Covid-19, inflammation, nutrition, dietary agents

Procedia PDF Downloads 146

Authors: Zatovska T. V., Nesterova N. V., Baranova G. V., Zagorodnya S. D.

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In recent years, biosensor technologies based on the phenomenon of surface plasmon resonance (SPR) are becoming increasingly used in biology and medicine. Their application facilitates exploration in real time progress of binding of biomolecules and identification of agents that specifically interact with biologically active substances immobilized on the biosensor surface (biochips). Special attention is paid to the use of Biosensor analysis in determining the antibody-antigen interaction in the diagnostics of diseases caused by viruses and bacteria. According to WHO, the diseases that are caused by the herpes simplex virus (HSV), take second place (15.8%) after influenza as a cause of death from viral infections. Current diagnostics of HSV infection include PCR and ELISA assays. The latter allows determination the degree of immune response to viral infection and respective stages of its progress. In this regard, the searches for new and available diagnostic methods are very important. This work was aimed to develop Biosensor chip for detection of specific antibodies to HSV-1 in the human blood serum. The proteins of HSV1 (strain US) were used as antigens. The viral particles were accumulated in cell culture MDBK and purified by differential centrifugation in cesium chloride density gradient. Analysis of the HSV1 proteins was performed by polyacrylamide gel electrophoresis and ELISA. The protein concentration was measured using De Novix DS-11 spectrophotometer. The device for detection of antigen-antibody interactions was an optoelectronic two-channel spectrometer ‘Plasmon-6’, using the SPR phenomenon in the Krechman optical configuration. It was developed at the Lashkarev Institute of Semiconductor Physics of NASU. The used carrier was a glass plate covered with 45 nm gold film. Screening of human blood serums was performed using the test system ‘HSV-1 IgG ELISA’ (GenWay, USA). Development of Biosensor chip included optimization of conditions of viral antigen sorption and analysis steps. For immobilization of viral proteins 0.2% solution of Dextran 17, 200 (Sigma, USA) was used. Sorption of antigen took place at 4-8°C within 18-24 hours. After washing of chip, three times with citrate buffer (pH 5,0) 1% solution of BSA was applied to block the sites not occupied by viral antigen. It was found direct dependence between the amount of immobilized HSV1 antigen and SPR response. Using obtained biochips, panels of 25 positive and 10 negative for the content of antibodies to HSV-1 human sera were analyzed. The average value of SPR response was 185 a.s. for negative sera and from 312 to. 1264 a.s. for positive sera. It was shown that SPR data were agreed with ELISA results in 96% of samples proving the great potential of SPR in such researches. It was investigated the possibility of biochip regeneration and it was shown that application of 10 mM NaOH solution leads to rupture of intermolecular bonds. This allows reuse the chip several times. Thus, in this study biosensor chip for detection of specific antibodies to HSV1 was successfully developed expanding a range of diagnostic methods for this pathogen.

Keywords: biochip, herpes virus, SPR

Procedia PDF Downloads 395

Authors: Kaomud Tyagi, Rajasree Chakraborty, Shantanu Kundu, Devkant Singha, Kailash Chandra, Vikas Kumar

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The melon thrips, Thrips palmi is a serious pest of a wide range of agriculture crops and also act as vectors for plant viruses (genus Tospovirus, family Bunyaviridae). More molecular data on this species is required to understand the cryptic speciation and evolutionary affiliations. Mitochondrial genomes have been widely used in phylogenetic and evolutionary studies in insect. So far, mitogenomes of five thrips species (Anaphothrips obscurus, Frankliniella intonsa, Frankliniella occidentalis, Scirtothrips dorsalis and Thrips imaginis) is available in the GenBank database. In this study, we sequenced the first complete mitogenome T. palmi and compared it with available thrips mitogenomes. We assembled the mitogenome from the whole genome sequencing data generated using Illumina Hiseq2500. Annotation was performed using MITOS web-server to estimate the location of protein coding genes (PCGs), transfer RNA (tRNAs), ribosomal RNAs (rRNAs) and their secondary structures. The boundaries of PCGs and rRNAs was confirmed manually in NCBI. Phylogenetic analyses were performed using the 13 PCGs data using maximum likelihood (ML) in PAUP, and Bayesian inference (BI) in MrBayes 3.2. The complete mitogenome of T. palmi was 15,333 base pairs (bp), which was greater than the genomes of A. obscurus (14,890bp), F. intonsa (15,215 bp), F. occidentalis (14,889 bp) and S. dorsalis South Asia strain (SA1) (14,283 bp), but smaller than the genomes of T. imaginis (15,407 bp) and S. dorsalis East Asia strain (EA1) (15,343bp). Like in other thrips species, the mitochondrial genome of T. palmi was represented by 37 genes, including 13 PCGs, large and small ribosomal RNA (rrnL and rrnS) genes, 22 transfer RNA (tRNAs) genes (with one extra gene for trn-Serine) and two A+T-rich control regions (CR1 and CR2). Thirty one genes were observed on heavy (H) strand and six genes on the light (L) strand. The six tRNA genes (trnG,trnK, trnY, trnW, trnF, and trnH) were found to be conserved in all thrips species mitogenomes in their locations relative to a protein-coding or rRNA gene upstream or downstream. The gene arrangements of T. palmi is very close to T. imaginis except the rearrangements in tRNAs genes: trnR (arginine), and trnE (glutamic acid) were found to be located between cox3 and CR2 in T. imaginis which were translocated between atp6 and CR1 in T. palmi; trnL1 (Leucine) and trnS1(Serine) were located between atp6 and CR1 in T. imaginis which were translocated between cox3 and CR2 in T. palmi. The location of CR1 upstream of nad5 gene was suggested to be ancestral condition of the thrips species in subfamily Thripinae, was also observed in T. palmi. Both the Maximum likelihood (ML) and Bayesian Inference (BI) phylogenetic trees generated resulted in similar topologies. The T. palmi was clustered with T. imaginis. We concluded that more molecular data on the diverse thrips species from different hierarchical level is needed, to understand the phylogenetic and evolutionary relationships among them.

Keywords: thrips, comparative mitogenomics, gene rearrangements, phylogenetic analysis

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Authors: Andreas Latz, Daniela Heinz, Fatima Hashemi, Melek Baygül

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Introduction: Feline leukemia virus (FeLV), feline immunodeficiency virus (FIV), and feline coronavirus (FCoV) are serious infectious diseases affecting cats worldwide. Transmission of these viruses occurs primarily through close contact with infected cats (via saliva, nasal secretions, faeces, etc.). FeLV, FIV, and FCoV infections can occur in combination and are expressed in similar clinical symptoms. Diagnosis can therefore be challenging: Symptoms are variable and often non-specific. Sick cats show very similar clinical symptoms: apathy, anorexia, fever, immunodeficiency syndrome, anemia, etc. Sample volume for small companion animals for diagnostic purposes can be challenging to collect. In addition, multiplex diagnosis of diseases can contribute to an easier, cheaper, and faster workflow in the lab as well as to the better differential diagnosis of diseases. For this reason, we wanted to develop a new diagnostic tool that utilizes less sample volume, reagents, and consumables than multiplesingleplex ELISA assays Methods: The Multiplier from Dynextechonogies (USA) has been used as platform to develop a Multiplex diagnostic tool for the detection of antibodies against FIV and FCoV/FIP and antigens for FeLV. Multiplex diagnostics. The Dynex®Multiplier®is a fully automated chemiluminescence immunoassay analyzer that significantly simplifies laboratory workflow. The Multiplier®ease-of-use reduces pre-analytical steps by combining the power of efficiently multiplexing multiple assays with the simplicity of automated microplate processing. Plastic beads have been coated with antigens for FIV and FCoV/FIP, as well as antibodies for FeLV. Feline blood samples are incubated with the beads. Read out of results is performed via chemiluminescence Results: Bead coating was optimized for each individual antigen or capture antibody and then combined in the multiplex diagnostic tool. HRP: Antibody conjugates for FIV and FCoV antibodies, as well as detection antibodies for FeLV antigen, have been adjusted and mixed. 3 individual prototyple batches of the assay have been produced. We analyzed for each disease 50 well defined positive and negative samples. Results show an excellent diagnostic performance of the simultaneous detection of antibodies or antigens against these feline diseases in a fully automated system. A 100% concordance with singleplex methods like ELISA or IFA can be observed. Intra- and Inter-Assays showed a high precision of the test with CV values below 10% for each individual bead. Accelerated stability testing indicate a shelf life of at least 1 year. Conclusion: The new tool can be used for multiplex diagnostics of the most important feline infectious diseases. Only a very small sample volume is required. Fully automation results in a very convenient and fast method for diagnosing animal diseases.With its large specimen capacity to process over 576 samples per 8-hours shift and provide up to 3,456 results, very high laboratory productivity and reagent savings can be achieved.

Keywords: Multiplex, FIV, FeLV, FCoV, FIP

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Authors: Sivananth Murugesan, I. Regupathi, B. Vishwas Prabhu, Ankit Devatwal, Vishnu Sivan Pillai

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Pectinase is an important enzyme which has a wide range of applications including textile processing and bioscouring of cotton fibers, coffee and tea fermentation, purification of plant viruses, oil extraction etc. Selective separation and purification of pectinase from fermentation broth and recover the enzyme form process stream for reuse are cost consuming process in most of the enzyme based industries. It is difficult to identify a suitable medium to enhance enzyme activity and retain its enzyme characteristics during such processes. The cost effective, selective separation of enzymes through the modified Liquid-liquid extraction is of current research interest worldwide. Reverse micellar extraction, globally acclaimed Liquid-liquid extraction technique is well known for its separation and purification of solutes from the feed which offers higher solute specificity and partitioning, ease of operation and recycling of extractants used. Surfactant concentrations above critical micelle concentration to an apolar solvent form micelles and addition of micellar phase to water in turn forms reverse micelles or water-in-oil emulsions. Since, electrostatic interaction plays a major role in the separation/purification of solutes using reverse micelles. These interaction parameters can be altered with the change in pH, addition of cosolvent, surfactant and electrolyte and non-electrolyte. Even though many chemical based commercial surfactant had been utilized for this purpose, the biosurfactants are more suitable for the purification of enzymes which are used in food application. The present work focused on the partitioning of pectinase from the synthetic aqueous solution within the reverse micelle phase formed by a biosurfactant, Soybean Lecithin dissolved in chloroform. The critical micelle concentration of soybean lecithin/chloroform solution was identified through refractive index and density measurements. Effect of surfactant concentrations above and below the critical micelle concentration was considered to study its effect on enzyme activity, enzyme partitioning within the reverse micelle phase. The effect of pH and electrolyte salts on the partitioning behavior was studied by varying the system pH and concentration of different salts during forward and back extraction steps. It was observed that lower concentrations of soybean lecithin enhanced the enzyme activity within the water core of the reverse micelle with maximizing extraction efficiency. The maximum yield of pectinase of 85% with a partitioning coefficient of 5.7 was achieved at 4.8 pH during forward extraction and 88% yield with a partitioning coefficient of 7.1 was observed during backward extraction at a pH value of 5.0. However, addition of salt decreased the enzyme activity and especially at higher salt concentrations enzyme activity declined drastically during both forward and back extraction steps. The results proved that reverse micelles formed by Soybean Lecithin and chloroform may be used for the extraction of pectinase from aqueous solution. Further, the reverse micelles can be considered as nanoreactors to enhance enzyme activity and maximum utilization of substrate at optimized conditions, which are paving a way to process intensification and scale-down.

Keywords: pectinase, reverse micelles, soybean lecithin, selective partitioning

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Authors: Miriam I. Jimenez-Perez, Mariana C. Orellana-Haro, Carolina Guzman-Brambila

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As part of microbiology and dietetics courses, students of medicine and nutrition analyze the main pathogenic microorganisms and perform dietary analyzes. The course of microbiology describes in a general way the main pathogens including bacteria, viruses, fungi, and parasites, as well as their interaction with the human species. We hypothesize that lack of practical application of the course causes the students not to find the value and the clinical application of it when in reality it is a matter of great importance for healthcare in our country. The courses of the medical microbiology and dietetics are mostly theoretical and only a few hours of laboratory practices. Therefore, it is necessary the incorporation of new innovative techniques that involve more practices and community fieldwork, real cases analysis and real-life situations. The purpose of this intervention was to incorporate real-world learning experiences in the instruction of medical microbiology and dietetics courses, in order to improve the learning process, understanding and the application in the field. During a period of 6 months, medicine and nutrition students worked in a community of urban poverty. We worked with 90 children between 4 and 6 years of age from low-income families with no access to medical services, to give an infectious diagnosis related to nutritional status in these children. We expect that this intervention would give a different kind of context to medical microbiology and dietetics students improving their learning process, applying their knowledge and laboratory practices to help a needed community. First, students learned basic skills in microbiology diagnosis test during laboratory sessions. Once, students acquired abilities to make biochemical probes and handle biological samples, they went to the community and took stool samples from children (with the corresponding informed consent). Students processed the samples in the laboratory, searching for enteropathogenic microorganism with RapID™ ONE system (Thermo Scientific™) and parasites using Willis and Malloy modified technique. Finally, they compared the results with the nutritional status of the children, previously measured by anthropometric indicators. The anthropometric results were interpreted by the OMS Anthro software (WHO, 2011). The microbiological result was interpreted by ERIC® Electronic RapID™ Code Compendium software and validated by a physician. The results were analyses of infectious outcomes and nutritional status. Related to fieldwork community learning experiences, our students improved their knowledge in microbiology and were capable of applying this knowledge in a real-life situation. They found this kind of learning useful when they translate theory to a real-life situation. For most of our students, this is their first contact as health caregivers with real population, and this contact is very important to help them understand the reality of many people in Mexico. In conclusion, real-world or fieldwork learning experiences empower our students to have a real and better understanding of how they can apply their knowledge in microbiology and dietetics and help a much- needed population, this is the kind of reality that many people live in our country.

Keywords: real-world learning experiences, medical microbiology, dietetics, nutritional status, infectious status.

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Authors: Mohanad Elemary, Imose Itua, Rajeswari B. Matam

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Virologists and public health experts have been predicting potential pandemics from coronaviruses for decades. The viruses which caused the SARS and MERS pandemics and the Nipah virus led to many lost lives, but still, the COVID-19 pandemic caused by the SARS-CoV2 virus surprised many scientific communities, experts, and governments with its ease of transmission and its pathogenicity. Governments of various countries reacted by locking down entire populations to their homes to combat the devastation caused by the virus, which led to a loss of livelihood and economic hardship to many individuals and organizations. To revive national economies and support their citizens in resuming their lives, governments focused on the development and use of contact tracing apps as a digital way to track and trace exposure. Google and Apple introduced the Exposure Notification Systems (ENS) framework. Independent organizations and countries also developed different frameworks for contact tracing apps. The efficiency, popularity, and adoption rate of these various apps have been different across countries. In this paper, we present a critical analysis of the different contact tracing apps with respect to their efficiency, adoption rate and general perception, and the governmental strategies and policies, which led to the development of the applications. When it comes to the European countries, each of them followed an individualistic approach to the same problem resulting in different realizations of a similarly functioning application with differing results of use and acceptance. The study conducted an extensive review of existing literature, policies, and reports across multiple disciplines, from which a framework was developed and then validated through interviews with six key stakeholders in the field, including founders and executives in digital health startups and corporates as well as experts from international organizations like The World Health Organization. A framework of best practices and tactics is the result of this research. The framework looks at three main questions regarding the contact tracing apps; how to develop them, how to deploy them, and how to regulate them. The findings are based on the best practices applied by governments across multiple countries, the mistakes they made, and the best practices applied in similar situations in the business world. The findings include multiple strategies when it comes to the development milestone regarding establishing frameworks for cooperation with the private sector and how to design the features and user experience of the app for a transparent, effective, and rapidly adaptable app. For the deployment section, several tactics were discussed regarding communication messages, marketing campaigns, persuasive psychology, and the initial deployment scale strategies. The paper also discusses the data privacy dilemma and how to build for a more sustainable system of health-related data processing and utilization. This is done through principles-based regulations specific for health data to allow for its avail for the public good. This framework offers insights into strategies and tactics that could be implemented as protocols for future public health crises and emergencies whether global or regional.

Keywords: contact tracing apps, COVID-19, digital health applications, exposure notification system

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Authors: Sonika Sharma, Mohan G. Vairale, Sibnarayan Datta, Soumya Chatterjee, Dharmendra Dubey, Rajesh Prasad, Raghvendra Budhauliya, Bidisha Das, Vijay Veer

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Bacteriophages are viruses that parasitize specific bacteria and multiply in metabolising host bacteria. Bacteriophages hunt for a single or a subset of bacterial species, making them potential antibacterial agents. Utilizing the ability of phages to control bacterial populations has several applications from medical to the fields of agriculture, aquaculture and the food industry. However, harnessing phage based techniques in wastewater treatments to improve quality of effluent and sludge release into the environment is a potential area for R&D application. Phage mediated bactericidal effect in any wastewater treatment process has many controlling factors that lead to treatment performance. In laboratory conditions, titer of bacteriophages (coliphages) isolated from effluent water of a specially designed anaerobic digester of human night soil (DRDO Biotoilet) was successfully increased with a modified protocol of the classical double layer agar technique. Enrichment of the same was carried out and efficacy of the phage enriched medium was evaluated at different conditions (specific media, temperature, storage conditions). Growth optimization study was carried out on different media like soybean casein digest medium (Tryptone soya medium), Luria-Bertani medium, phage deca broth medium and MNA medium (Modified nutrient medium). Further, temperature-phage yield relationship was also observed at three different temperatures 27˚C, 37˚C and 44˚C at laboratory condition. Results showed the higher activity of coliphage 27˚C and at 37˚C. Further, addition of divalent ions (10mM MgCl2, 5mM CaCl2) and 5% glycerol resulted in a significant increase in phage titer. Besides this, effect of antibiotics addition like ampicillin and kanamycin at different concentration on plaque formation was analysed and reported that ampicillin at a concentration of 1mg/ml ampicillin stimulates phage infection and results in more number of plaques. Experiments to test viability of phage showed that it can remain active for 6 months at 4˚C in fresh tryptone soya broth supplemented with fresh culture of coliforms (early log phase). The application of bacteriophages (especially coliphages) for treatment of effluent of human faecal matter contaminated effluent water is unique. This environment-friendly treatment system not only reduces the pathogenic coliforms, but also decreases the competition between nuisance bacteria and functionally important microbial populations. Therefore, the phage based cocktail to treat fecal pathogenic bacteria present in black water has many implication in wastewater treatment processes including ‘DRDO Biotoilet’, which is an ecofriendly appropriate and affordable human faecal matter treatment technology for different climates and situations.

Keywords: wastewater, microbes, virus, biotoilet, phage viability

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