scholarly journals Comparison of PCR versus Culture for Detection of Mycobacterium bovis after Experimental Inoculation of Various Matrices Held under Environmental Conditions for Extended Periods

2013 ◽  
Vol 79 (20) ◽  
pp. 6501-6506 ◽  
Author(s):  
Angela P. Adams ◽  
Steven R. Bolin ◽  
Amanda E. Fine ◽  
Carole A. Bolin ◽  
John B. Kaneene
Keyword(s):  
Mycobacterium Bovis ◽  
Nested Pcr ◽  
Weather Conditions ◽  
Content Type ◽  
Mycobacterial Culture ◽  
Monthly Sample ◽  
Sample Testing ◽  
Weekly Sample ◽  
Primer Sets ◽  
Initial Contamination

ABSTRACTThe purpose of this study was to compare the performance of a molecular detection technique (nested PCR) with that of mycobacterial culture in the detection ofMycobacterium bovisDNA in a set of 687 samples of experimentally inoculated environmental substrates (hay, soil, corn, water) exposed to natural weather conditions in Michigan. Four replicates of each substrate were used; half were autoclaved for sterilization, all were inoculated with 50,000 CFU ofM. bovisisolated from Michigan livestock, and all were placed in outdoor enclosures, with half under shade and the other half exposed to direct sunlight. Samples were tested for the presence ofM. bovisduring one 12-month period, with monthly sample testing and during three 12-week periods (winter, spring, summer) with weekly sample testing. Samples were subjected to mycobacterial culture for isolation ofM. bovisand a nested PCR with two primer sets targeting IS6110to detectM. bovisDNA. In 128 samples tested during the 12-month period,M. boviswas not detectable by culture after 2 months butM. bovisDNA was detectable by PCR for at least 7 months. Of the 559 samples tested during the 12-week periods, PCR detectedM. bovisDNA for up to 88 days in all of the sample types. There were no significant differences in the detection ofM. bovisbetween shade and sun samples or between sterile and unsterilized samples, regardless of the detection method (PCR or culture). For use in epidemiologic investigations, the PCR assay was more rapid than mycobacterial culture, was not hindered by contaminating organisms, and detectedM. bovisDNA in environment samples much longer after initial contamination than mycobacterial culture did.

scholarly journals Mycobacterium bovis DNA Detection in Colostrum as a Potential Indicator of Vaccination Effectiveness against Bovine Tuberculosis

2013 ◽  
Vol 20 (4) ◽  
pp. 627-633 ◽  
Author(s):  
Sara E. Herrera-Rodríguez ◽  
María Alejandra Gordiano-Hidalgo ◽  
Gonzálo López-Rincón ◽  
Luis Bojorquez-Narváez ◽  
Francisco Javier Padilla-Ramírez ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Nested Pcr ◽  
Bovine Tuberculosis ◽  
Dna Detection ◽  
Dairy Farms ◽  
Primary Vaccination ◽  
Content Type ◽  
Booster Immunization ◽  
Culture Filtrate Proteins

ABSTRACTBovine tuberculosis (bTB) remains a problem on many dairy farms in Mexico, as well as a public health risk. We previously found a high frequency ofMycobacterium bovisDNA in colostrum from dairy cows using a nested PCR to detectmpb70. Since there are no reliablein vivotests to determine the effectiveness of boosterMycobacterium bovisBCG vaccination against bTB, in this work we monitoredM. bovisDNA in colostrum by using this nested PCR. In order to decrease the risk of adverse reactions in animals likely containing viableM. bovis, a single application of BCG and a subunit vaccine (EEP-1) formulated withM. bovisculture filtrate proteins (CFP) and a copolymer as the adjuvant was performed in tuberculin skin test-negative cattle (TST−), while TST reactor animals (TST+) received EEP-1 only. Booster immunization using EEP-1 was applied to both groups, 2 months after primary vaccination to whole herds and 12 months later to lactating cows. Colostrum samples were collected from 6 farms where the cows were vaccinated over a 12-month period postvaccination and, for comparison, from one control farm where the cows were not vaccinated with comparable bTB prevalence. We observed an inverse relationship between the frequency ofM. bovisDNA detection and time postvaccination at the first (P< 0.001) and second (P< 0.0001) 6-month periods. Additionally, the concentration of gamma interferon (IFN-γ) was higher inmpb70PCR-positive colostrum samples (P= 0.0003). These results suggest thatM. bovisDNA frequency in colostrum could be a potentially useful biomarker for bTB vaccine efficacy on commercial dairy farms.

The importance of public support in the implementation of green transportation in smart cities using smart vehicle bicycle communication transport

2020 ◽  
Vol 38 (5/6) ◽  
pp. 997-1011
Author(s):  
Ning Li ◽  
Parthasarathy R. ◽  
Harshila H. Padwal
Keyword(s):  
Smart Cities ◽  
Optimization Technique ◽  
Public Support ◽  
Weather Conditions ◽  
Transport Systems ◽  
Content Type ◽  
Signaling Systems ◽  
Traffic Conditions ◽  
Green Transportation ◽  
Smart Vehicle

Purpose Smart mobility is a major guideline in the development of Smart Cities’ transport systems and management. The issue of transition into green, secure and sustainable transport modes, such as using bicycles, should be implemented in this case, along with the subjectivism of management. Design/methodology/approach The proposed technology reflects the Smart Bicycle vehicle model, which tracks cyclists and weather conditions and turns to electric motors in critical circumstances. Findings This reduces the physical load and battery consumption of cyclists which affects the Smart Cities’ ecology positively. Originality/value In Smart Vehicle Bicycle Communication Transport, the vehicle movement optimization technique is used for traffic scenarios to analyze traffic signaling systems that give better results in variable and dense traffic conditions.

scholarly journals New Recombinant Mycobacterium bovis BCG Expression Vectors: Improving Genetic Control over Mycobacterial Promoters

2016 ◽  
Vol 82 (8) ◽  
pp. 2240-2246 ◽  
Author(s):  
Alex I. Kanno ◽  
Cibelly Goulart ◽  
Henrique K. Rofatto ◽  
Sergio C. Oliveira ◽  
Luciana C. C. Leite ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Fluorescent Protein ◽  
Vaccine Development ◽  
Expression Vectors ◽  
Content Type ◽  
Expression Levels ◽  
Wide Range ◽  
Mycobacteriophage L5 ◽  
Green Fluorescent ◽  
Selection Of

ABSTRACTThe expression of many antigens, stimulatory molecules, or even metabolic pathways in mycobacteria such asMycobacterium bovisBCG orM. smegmatiswas made possible through the development of shuttle vectors, and several recombinant vaccines have been constructed. However, gene expression in any of these systems relied mostly on the selection of natural promoters expected to provide the required level of expression by trial and error. To establish a systematic selection of promoters with a range of strengths, we generated a library of mutagenized promoters through error-prone PCR of the strong PL5promoter, originally from mycobacteriophage L5. These promoters were cloned upstream of the enhanced green fluorescent protein reporter gene, and recombinantM. smegmatisbacteria exhibiting a wide range of fluorescence levels were identified. A set of promoters was selected and identified as having high (pJK-F8), intermediate (pJK-B7, pJK-E6, pJK-D6), or low (pJK-C1) promoter strengths in bothM. smegmatisandM. bovisBCG. The sequencing of the promoter region demonstrated that it was extensively modified (6 to 11%) in all of the plasmids selected. To test the functionality of the system, two different expression vectors were demonstrated to allow corresponding expression levels of theSchistosoma mansoniantigen Sm29 in BCG. The approach used here can be used to adjust expression levels for synthetic and/or systems biology studies or for vaccine development to maximize the immune response.

A new IR and sprinkler based embedded controller directed robotic arm for automatic cleaning of solar panel

2019 ◽  
Vol 18 (4) ◽  
pp. 905-921
Author(s):  
Manish Kumar Ghodki ◽  
Akhilesh Swarup ◽  
Yash Pal
Keyword(s):  
Pulse Width Modulation ◽  
Weather Conditions ◽  
Solar Panel ◽  
Performance Ratio ◽  
Robotic Arm ◽  
Pv System ◽  
Content Type ◽  
Average Value ◽  
Ir Sensor ◽  
Embedded Controller

Purpose The purpose of this paper is to design and develop an IR and sprinkler based embedded controller operated robotic arm for automatic dust removal system to mitigate the dust effect on the solar panel surface, since dust accumulation normally affected by real weather conditions is one of the serious concern for the deterioration of photovoltaic (PV) system output. Design/methodology/approach The system is a wet cleaning device which provides a cheap silicon rubber-based wiping operation controlled by the pulse width modulation-operated motors of robotic arm. The IEEE 1149.1-compliant mixed signal-embedded platform of C8051F226DK is involved to command the complete system. Findings A prototype of 30 WP system is capable of producing an inspiring average value of 11.26 per cent in energy increase, 13.63 per cent in PV module efficiency and 85.20 per cent in performance ratio of the system after 73 days of cleaning in summer season. In addition, a total of 1,617.93 W power; 1,0516.55 Wh energy; and 350.55 KWh/KWP final yield was found during the entire cleaning period. Originality/value A novel technique of the implementation of IR sensor and sprinkler in dust mitigation is proposed in this paper. The IR sensor is used as a versatile object which can manage the robotic arm setting and control the automatic switching between cleaning and charging, as well as identify the thermal condition of solar panel for overheating.

scholarly journals Predicting the effect of decompression sickness on survival following submarine tower escape

2021 ◽  
Vol ahead-of-print (ahead-of-print) ◽  
Author(s):  
Geoff A.M. Loveman ◽  
Joel J.E. Edney
Keyword(s):  
Subject Matter ◽  
Decompression Sickness ◽  
Survival Rates ◽  
Weather Conditions ◽  
Subject Matter Expert ◽  
Content Type ◽  
Probability Of Survival ◽  
Predicted Probability ◽  
Standard Probability ◽  
Current Advice

Purpose The purpose of the present study was the development of a methodology for translating predicted rates of decompression sickness (DCS), following tower escape from a sunken submarine, into predicted probability of survival, a more useful statistic for making operational decisions. Design/methodology/approach Predictions were made, using existing models, for the probabilities of a range of DCS symptoms following submarine tower escape. Subject matter expert estimates of the effect of these symptoms on a submariner’s ability to survive in benign weather conditions on the sea surface until rescued were combined with the likelihoods of the different symptoms occurring using standard probability theory. Plots were generated showing the dependence of predicted probability of survival following escape on the escape depth and the pressure within the stricken submarine. Findings Current advice on whether to attempt tower escape is based on avoiding rates of DCS above approximately 5%–10%. Consideration of predicted survival rates, based on subject matter expert opinion, suggests that the current advice might be considered as conservative in the distressed submarine scenario, as DCS rates of 10% are not anticipated to markedly affect survival rates. Originality/value According to the authors’ knowledge, this study represents the first attempt to quantify the effect of different DCS symptoms on the probability of survival in submarine tower escape.

scholarly journals Assessing Performance of Spore Samplers in Monitoring Aeromycobiota and Fungal Plant Pathogen Diversity in Canada

2018 ◽  
Vol 84 (9) ◽  
Author(s):  
Wen Chen ◽  
Sarah Hambleton ◽  
Keith A. Seifert ◽  
Odile Carisse ◽  
Moussa S. Diarra ◽  
...  
Keyword(s):  
Prince Edward Island ◽  
Fungal Pathogens ◽  
Plant Pathogens ◽  
High Throughput Sequencing ◽  
Climatic Factors ◽  
Weather Conditions ◽  
Pilot Project ◽  
Sampling Location ◽  
Fungal Community Composition ◽  
Content Type

ABSTRACTSpore samplers are widely used in pathogen surveillance but not so much for monitoring the composition of aeromycobiota. In Canada, a nationwide spore-sampling network (AeroNet) was established as a pilot project to assess fungal community composition in air and rain samples collected using three different spore samplers in the summers of 2010 and 2011. Metabarcodes of the internal transcribed spacer (ITS) were exhaustively characterized for three of the network sites, in British Columbia (BC), Québec (QC), and Prince Edward Island (PEI), to compare performance of the samplers. Sampler type accounted for ca. 20% of the total explainable variance in aeromycobiota compositional heterogeneity, with air samplers recovering more Ascomycota and rain samplers recovering more Basidiomycota. Spore samplers showed different abilities to collect 27 fungal genera that are plant pathogens. For instance,Cladosporiumspp.,Drechsleraspp., andEntylomaspp. were collected mainly by air samplers, whileFusariumspp.,Microdochiumspp., andUstilagospp. were recovered more frequently with rain samplers. The diversity and abundance of some fungi were significantly affected by sampling location and time (e.g.,AlternariaandBipolaris) and weather conditions (e.g.,MycocentrosporaandLeptosphaeria), and depended on using ITS1 or ITS2 as the barcoding region (e.g.,EpicoccumandBotrytis). The observation that Canada's aeromycobiota diversity correlates with cooler, wetter conditions and northward wind requires support from more long-term data sets. Our vision of the AeroNet network, combined with high-throughput sequencing (HTS) and well-designed sampling strategies, may contribute significantly to a national biovigilance network for protecting plants of agricultural and economic importance in Canada.IMPORTANCEThe current study compared the performance of spore samplers for collecting broad-spectrum air- and rain-borne fungal pathogens using a metabarcoding approach. The results provided a thorough characterization of the aeromycobiota in the coastal regions of Canada in relation to the influence of climatic factors. This study lays the methodological basis to eventually develop knowledge-based guidance on pest surveillance by assisting in the selection of appropriate spore samplers.

Livestock online marketplace “Pashushala.com”: standardizing unstandardized operating procedures

2021 ◽  
Vol 11 (3) ◽  
pp. 1-18
Author(s):  
Aasha Jayant Sharma ◽  
Shashank Bhat
Keyword(s):  
Business Model ◽  
Standard Operating Procedure ◽  
Chief Executive ◽  
Weather Conditions ◽  
Supply Chain Risk Management ◽  
Content Type ◽  
Link Type ◽  
Online Marketplace ◽  
Academic Level ◽  
Business Model Canvas

Learning outcomes It enables students to understand and design a business model canvas, design standard operating procedure (SOP) for very unorganized business operations and also devise base pricing for vendor negotiation. Case overview/synopsis Mr Gaurav Chaudhary, chief executive officer and Founder of Pashushala.com, established a first-ever Livestock online marketplace in India, leveraging the penetration of internet users in 2019. Pashsuhala.com evolved as an all-inclusive ecosystem that offered an innovative business model by bundling financial aid, logistics, veterinary and insurance solutions to its buyers and sellers. While every other aspect seemed to have had fallen in place, Gaurav was not convinced with the everyday handling of the cattle especially during transportation. Transporting cattle was the most challenging task tempered with issues such as changing weather conditions, stock density, lack of training on handling cattle while loading and unloading, long journey hours, feeding and watering procedures and many more for which Gaurav had to depend on the logistics partners. Gaurav was in a dilemma whether to have his own fleet armed with trained personnel for transporting the cattle or to streamline the existing operating procedures into SOP to be followed by logistics partners. If he continued with logistics partners he also had to work on standard costs i.e. fixed and variable costs incurred during the transportation of livestock. The case deals with business concepts such as supply chain risk management in the livestock sector, SOPs for a very unstructured and unpredictable ecosystem, pricing strategies and business model canvas. Complexity academic level Masters in business administration (MBA) and Executive MBA level. Supplementary materials Teaching Notes are available for educators only. Please contact your library to gain login details or email [email protected] to request teaching notes. Subject code CSS 9: Operations and Logistics.

scholarly journals Stable Expression of Lentiviral Antigens by Quality-Controlled Recombinant Mycobacterium bovis BCG Vectors

2015 ◽  
Vol 22 (7) ◽  
pp. 726-741 ◽  
Author(s):  
Bryan E. Hart ◽  
Rose Asrican ◽  
So-Yon Lim ◽  
Jaimie D. Sixsmith ◽  
Regy Lukose ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Antigen Expression ◽  
Full Length ◽  
Content Type ◽  
Immunodeficiency Virus ◽  
Vaccine Stability ◽  
Hiv Gp120 ◽  
Recombinant Bcg

ABSTRACTThe well-established safety profile of the tuberculosis vaccine strain,Mycobacterium bovisbacille Calmette-Guérin (BCG), makes it an attractive vehicle for heterologous expression of antigens from clinically relevant pathogens. However, successful generation of recombinant BCG strains possessing consistent insert expression has encountered challenges in stability. Here, we describe a method for the development of large recombinant BCG accession lots which stably express the lentiviral antigens, human immunodeficiency virus (HIV) gp120 and simian immunodeficiency virus (SIV) Gag, using selectable leucine auxotrophic complementation. Successful establishment of vaccine stability stems from stringent quality control criteria which not only screen for highly stable complemented BCG ΔleuCDtransformants but also thoroughly characterize postproduction quality. These parameters include consistent production of correctly sized antigen, retention of sequence-pure plasmid DNA, freeze-thaw recovery, enumeration of CFU, and assessment of cellular aggregates. Importantly, these quality assurance procedures were indicative of overall vaccine stability, were predictive for successful antigen expression in subsequent passaging bothin vitroandin vivo, and correlated with induction of immune responses in murine models. This study has yielded a quality-controlled BCG ΔleuCDvaccine expressing HIV gp120 that retained stable full-length expression after 1024-fold amplificationin vitroand following 60 days of growth in mice. A second vaccine lot expressed full-length SIV Gag for >1068-fold amplificationin vitroand induced potent antigen-specific T cell populations in vaccinated mice. Production of large, well-defined recombinant BCG ΔleuCDlots can allow confidence that vaccine materials for immunogenicity and protection studies are not negatively affected by instability or differences between freshly grown production batches.

scholarly journals Rapid Detection of Serum Antibody by Dual-Path Platform VetTB Assay in White-Tailed Deer Infected with Mycobacterium bovis

2013 ◽  
Vol 20 (6) ◽  
pp. 907-911 ◽  
Author(s):  
Konstantin P. Lyashchenko ◽  
Rena Greenwald ◽  
Javan Esfandiari ◽  
Daniel J. O'Brien ◽  
Stephen M. Schmitt ◽  
...  
Keyword(s):  
Diagnostic Accuracy ◽  
Mycobacterium Bovis ◽  
Skin Testing ◽  
Serum Antibody ◽  
Antibody Responses ◽  
Antibody Detection ◽  
Serum Samples ◽  
Content Type ◽  
In The Wild ◽  
Free Ranging

ABSTRACTBovine tuberculosis (TB) in cervids remains a significant problem affecting farmed herds and wild populations. Traditional skin testing has serious limitations in certain species, whereas emerging serological assays showed promising diagnostic performance. The recently developed immunochromatographic dual-path platform (DPP) VetTB assay has two antigen bands, T1 (MPB83 protein) and T2 (CFP10/ESAT-6 fusion protein), for antibody detection. We evaluated the diagnostic accuracy of this test by using serum samples collected from groups of white-tailed deer experimentally inoculated withMycobacterium bovis,M. aviumsubsp.paratuberculosis, orM. bovisBCG Pasteur. In addition, we used serum samples from farmed white-tailed deer in herds with no history of TB, as well as from free-ranging white-tailed deer culled during field surveillance studies performed in Michigan known to have bovine TB in the wild deer population. The DPP VetTB assay detected antibody responses in 58.1% of experimentally infected animals within 8 to 16 weeks postinoculation and in 71.9% of naturally infected deer, resulting in an estimated test sensitivity of 65.1% and a specificity of 97.8%. The higher seroreactivity found in deer with naturally acquiredM. bovisinfection was associated with an increased frequency of antibody responses to the ESAT-6 and CFP10 proteins, resulting in a greater contribution of these antigens, in addition to MPB83, to the detection of seropositive animals, compared with experimentalM. bovisinfection. Deer experimentally inoculated with eitherM. aviumsubsp.paratuberculosisorM. bovisBCG Pasteur did not produce cross-reactive antibodies that could be detected by the DPP VetTB assay. The present findings demonstrate the relatively high diagnostic accuracy of the DPP VetTB test for white-tailed deer, especially in the detection of naturally infected animals.

scholarly journals Design and Validation of Four New Primers for Next-Generation Sequencing To Target the 18S rRNA Genes of Gastrointestinal Ciliate Protozoa

2014 ◽  
Vol 80 (17) ◽  
pp. 5515-5521 ◽  
Author(s):  
Suzanne L. Ishaq ◽  
André-Denis G. Wright
Keyword(s):  
18S Rrna ◽  
18S Rrna Gene ◽  
Rrna Genes ◽  
Rrna Gene ◽  
Content Type ◽  
Hypervariable Regions ◽  
Blast Database ◽  
Primer Sets ◽  
18S Rrna Genes ◽  
Ciliate Protozoa

ABSTRACTFour new primers and one published primer were used to PCR amplify hypervariable regions within the protozoal 18S rRNA gene to determine which primer pair provided the best identification and statistical analysis. PCR amplicons of 394 to 498 bases were generated from three primer sets, sequenced using Roche 454 pyrosequencing with Titanium, and analyzed using the BLAST database (NCBI) and MOTHUR version 1.29. The protozoal diversity of rumen contents from moose in Alaska was assessed. In the present study, primer set 1, P-SSU-316F and GIC758R (amplicon of 482 bases), gave the best representation of diversity using BLAST classification, and the set amplifiedEntodinium simplexandOstracodiniumspp., which were not amplified by the other two primer sets. Primer set 2, GIC1080F and GIC1578R (amplicon of 498 bases), had similar BLAST results and a slightly higher percentage of sequences that were identified with a higher sequence identity. Primer sets 1 and 2 are recommended for use in ruminants. However, primer set 1 may be inadequate to determine protozoal diversity in nonruminants. The amplicons created by primer set 1 were indistinguishable for certain species within the generaBandia,Blepharocorys,Polycosta, andTetratoxumand betweenHemiprorodon gymnoprosthiumandProrodonopsiscoli, none of which are normally found in the rumen.

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