PCR Diagnosis of Some Citrus Viroids in Field Samples

Effective application of slag-forming mixtures (SFM), being fed into continuous castingg machine (CCM) moulds, depends on their even distribution on the melt surface. Manual feeding of the SFM which is widely usedd does not provide this condition, resulting in the necessity to actualize the work to elaborate systems of SFM mechanized feedingg into moulds of various types CCM. A concept of the designing of a system of SFM feeding into CCM moulds presented with the ratte strictly correspondent to the casting speed and providing formation of an even layer of fine material of given thickness on the whoole surface of liquid steel. The proposed methods of designing of the SFM mechanized feeding systems based on three-dimensional computer simulation with the subsequent verification of the correctness of the adopted technical solutions on field samples. Informattion is presented on the design features of the adjusted facilities intended for continuous supply of finely granulated and powder mixtuures on metal mirror in moulds at the production of high-quality billets, blooms and slabs. Variants of mechanical and pneumo-mechaanical SFM supply elaborated. At the mechanical supply the fine material from the feeding hopper is moved at a adjusted distance bby a rigid horizontally located screw. At the pneumo-mechanical supply the metered doze of the granular mixture is delivered by a sshort vertical screw, the lower part of which is located in the mixing chamber attached from below to the hopper and equipped with ann ejector serving for pneumatic supply of the SFM in a stream of transporting gas. It was proposed to use flexible spiral screws in the ffuture facilities of mechanical SFM feeding. It will enable to eliminate the restrictions stipulated by the lack of free surface for locatiion of the facility in the working zone of the tundish, as well as to decrease significantly the mass of its movable part and to decreaase the necessary power of the carriage moving mechanism driver. The novelty of the proposed technical solutions is protected by thhree patents. The reduction of 10–15% in the consumption of slag-forming mixtures during the transition from manual to mechanizeed feeding confirmed. The resulting economic effect from the implementation of technical development enables to recoup the costs inncurred within 8–10 months.

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Isolation and identification of Marek’s disease virus (MDV) from feather follicle epithelium and internal organs of diseased chickens in Dakahlia Governorate, Egypt

Mansoura Veterinary Medical Journal ◽

10.35943/mvmj.2019.22.102 ◽

2019 ◽

Vol 20 (2) ◽

pp. 6-11

Author(s):

Aly El-Kenawy ◽

Mohamed El-Tholoth ◽

Emad A

Keyword(s):

Real Time ◽

Disease Virus ◽

Real Time Pcr ◽

Marek's Disease Virus ◽

Marek's Disease ◽

Marek’S Disease Virus ◽

Marek’S Disease ◽

Field Samples ◽

Feather Follicle ◽

Positive Results

In the present study, a total of 16 samples including feather follicle epithelium, ovary, spleen and kidney (4 samples for each organ) were collected from diseased chicken flocks suspected to be infected with Marek’s disease virus (MDV) at Dakahlia Governorate, Egypt during the period from October 2016 to October 2017. Each sample was pooled randomly from three to five birds (90 to 360 days old). The isolation of the suspected virus from the collected samples was carried out via chorioallantoic membranes (CAMs) of 12 days old embryonated chicken eggs (ECEs). Three egg passages were carried out for each sample. Hyperimmune serum was prepared against standard MDV. MDV in both field and egg passaged samples (after 3rd passage) was identified by agar gel precipitation test (AGPT) and indirect fluorescence antibody test (IFAT). Molecular identification of virus was carried out by conventional polymerase chain reaction (PCR) and real- time PCR in four selected samples. The results revealed that 14 samples (87.5%) including 4 (100%) samples from feather follicle epithelium, ovary and kidney and 2 (50%) samples from spleen, showed positive results in virus isolation after 3rd passage. The positive results percentage by AGPT for field samples were 50% (8 out of 16 samples), while after the 3rd passage in ECEs were 37.5% (6 out of 16 samples) and the positive results percentage by IFAT for field samples were 62.5% (10 out of 16 samples), while after the 3rd passage in ECEs were 81.25 % (13 out of 16 samples). Viral nucleic acid was detected in all selected samples by conventional and real- time PCR. The results indicate that feather follicle epithelium is the best organ for MDV detection. IFAT is superior over AGPT in virus detection. Conventional and real - time PCR could be efficiently used for molecular detection of the virus.

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Identification of a novel CBFB-MYH11 transcript: implications for RT–PCR diagnosis

The Hematology Journal ◽

10.1038/sj.thj.6200103 ◽

2001 ◽

Vol 2 (3) ◽

pp. 206-209 ◽

Cited By ~ 17

Author(s):

Bert A Van der Reijden ◽

Linda de Wit ◽

Sonja van der Poel ◽

Erna B Luiten ◽

Marina Lafage-Pochitaloff ◽

...

Keyword(s):

Rt Pcr ◽

Pcr Diagnosis

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Performance of a Handheld Chlorophyll-a Fluorometer: Potential Use for Rapid Algae Monitoring

Water ◽

10.3390/w13101409 ◽

2021 ◽

Vol 13 (10) ◽

pp. 1409

Author(s):

Hamdhani Hamdhani ◽

Drew E. Eppehimer ◽

David Walker ◽

Michael T. Bogan

Keyword(s):

Chlorophyll A ◽

Surface Waters ◽

Harmful Algal Blooms ◽

Algal Blooms ◽

Water Quality Monitoring ◽

Ambient Light ◽

Calibration Equation ◽

Field Samples ◽

Potential Use

Chlorophyll-a measurements are an important factor in the water quality monitoring of surface waters, especially for determining the trophic status and ecosystem management. However, a collection of field samples for extractive analysis in a laboratory may not fully represent the field conditions. Handheld fluorometers that can measure chlorophyll-a in situ are available, but their performance in waters with a variety of potential light-interfering substances has not yet been tested. We tested a handheld fluorometer for sensitivity to ambient light and turbidity and compared these findings with EPA Method 445.0 using water samples obtained from two urban lakes in Tucson, Arizona, USA. Our results suggested that the probe was not sensitive to ambient light and performed well at low chlorophyll-a concentrations (<25 µg/L) across a range of turbidity levels (50–70 NTU). However, the performance was lower when the chlorophyll-a concentrations were >25 µg/L and turbidity levels were <50 NTU. To account for this discrepancy, we developed a calibration equation to use for this handheld fluorometer when field monitoring for potential harmful algal blooms in water bodies.

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Genomic Analyses of Globodera pallida, A Quarantine Agricultural Pathogen in Idaho

Pathogens ◽

10.3390/pathogens10030363 ◽

2021 ◽

Vol 10 (3) ◽

pp. 363

Author(s):

Sulochana K. Wasala ◽

Dana K. Howe ◽

Louise-Marie Dandurand ◽

Inga A. Zasada ◽

Dee R. Denver

Keyword(s):

Genetic Variation ◽

Potato Production ◽

Globodera Pallida ◽

Fixation Index ◽

Parasitic Nematodes ◽

Nucleotide Polymorphisms ◽

Single Nucleotide ◽

Genome Wide ◽

Field Samples ◽

Multiple Introduction

Globodera pallida is among the most significant plant-parasitic nematodes worldwide, causing major damage to potato production. Since it was discovered in Idaho in 2006, eradication efforts have aimed to contain and eradicate G. pallida through phytosanitary action and soil fumigation. In this study, we investigated genome-wide patterns of G. pallida genetic variation across Idaho fields to evaluate whether the infestation resulted from a single or multiple introduction(s) and to investigate potential evolutionary responses since the time of infestation. A total of 53 G. pallida samples (~1,042,000 individuals) were collected and analyzed, representing five different fields in Idaho, a greenhouse population, and a field in Scotland that was used for external comparison. According to genome-wide allele frequency and fixation index (Fst) analyses, most of the genetic variation was shared among the G. pallida populations in Idaho fields pre-fumigation, indicating that the infestation likely resulted from a single introduction. Temporal patterns of genome-wide polymorphisms involving (1) pre-fumigation field samples collected in 2007 and 2014 and (2) pre- and post-fumigation samples revealed nucleotide variants (SNPs, single-nucleotide polymorphisms) with significantly differentiated allele frequencies indicating genetic differentiation. This study provides insights into the genetic origins and adaptive potential of G. pallida invading new environments.

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P052 Leflunomide and severe COVID-19 outcome: a cautionary observation from the COVID-19 Scottish Registry of Autoimmune Rheumatic Diseases (SCAR-19)

Rheumatology ◽

10.1093/rheumatology/keab247.049 ◽

2021 ◽

Vol 60 (Supplement_1) ◽

Author(s):

Maira Karabayas ◽

James Brock ◽

Gillian Fordyce ◽

Neil Basu

Keyword(s):

Rheumatic Diseases ◽

Clinical Characteristics ◽

National Registry ◽

Favourable Outcome ◽

Non Invasive ◽

Autoimmune Rheumatic Diseases ◽

Non Invasive Ventilation ◽

Pcr Diagnosis ◽

Related Symptom ◽

Disease Modifying

Abstract Background/Aims Leflunomide, a conventional disease modifying drug (csDMARD), is used in a variety of autoimmune rheumatic diseases (ARD) due to its immunomodulating, immunosuppressive and antiproliferative properties. This agent does however confer a greater infection risk and, due to its long half-life, drug washout procedures are often advised in the context of serious infections. Interestingly, Leflunomide is currently being tested as a potential therapy for COVID-19 in the general population. It is unknown whether leflunomide therapy is associated with a poor or favourable outcome among ARD patients infected with COVID-19. Methods A Scottish-wide registry was rapidly developed in March 2020. Clinical characteristics and outcomes of infected cases were collated across all Scottish health boards. Eligible patients included any adult leflunomide treated ARD patients with a confirmed (clinically or PCR) diagnosis of COVID-19. Results Of the 69 cases included in the registry, n = 4 were treated with leflunomide (75% female; mean age 61, SD 4.2). N = 2 were treated with combination baricitinib or hydroxychloroquine respectively, whilst n = 1 received recent corticosteroid therapy (intramuscular Kenalog). Comorbidities observed in this sub-cohort include diabetes mellitus n = 3, hypertension n = 2, cardiovascular disease n = 1, lung disease n = 1 and latent TB n = 1. At presentation, all patients (n = 4) experienced the established COVID-19 related symptom triad of dyspnoea, cough and fever and promptly developed acute respiratory syndrome. Diarrhoea was also recorded in n = 2 and constitutional upset n = 3. All patients suffered a serious COVID-19 disease outcome (defined as a requirement of invasive or non-invasive ventilation (n = 4) and/ or death (n = 2). P052 Table 1:Patient demographics, clinical characteristics and outcomesPatient 1Patient 2Patient 3Patient 4Age58635766SexFemaleFemaleMaleFemaleRheumatic diagnosisRheumatoid arthritisPsoriatic arthritisPsoriatic arthritisRheumatoid ArthritisComorbiditiesDiabetesHypertension Diabetes COPDNilIschaemic heart disease Hypertension Diabetes Latent TBClinical presentationDyspnoea Cough Fever Confusion Constitutional upsetDyspnoea Cough Fever Diarrhoea Constitutional upsetDyspnoea Cough Fever Constitutional upsetDyspnoea Cough Fever Diarrhoea Constitutional upsetAdditional csDMARD*NilNilNilHydroxychloroquinebDMARD**/ tsDMARD***BaricitinibNilNilNilSteroid therapyNilNilNilIM KenalogInvasive or non-invasive ventilationYesYesYesYesDeathNoNoYesYes* conventional disease modifying drug,**biologic disease modifying drug,***targeted synthetic disease modifying drug. Conclusion Preliminary data from this Scotland-wide registry has identified only a small number of leflunomide treated ARD patients infected with COVID-19. However, it is concerning that all cases experienced a serious outcome. Given the relatively infrequent prescription of this drug, combining similar national registry data is necessary to ensure this observation is not spurious. If confirmed, leflunomide washout procedures should be encouraged among such patients when they first present with COVID-19. Disclosure M. Karabayas: None. J. Brock: None. G. Fordyce: None. N. Basu: None.

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Comparison of the ColiPlate™ Kit with Two Common E. coli Enumeration Methods for Water

Water ◽

10.3390/w13131804 ◽

2021 ◽

Vol 13 (13) ◽

pp. 1804

Author(s):

Cassi J. Gibson ◽

Abraham K. Maritim ◽

Jason W. Marion

Keyword(s):

High Risk ◽

Natural Water ◽

Membrane Filtration ◽

Fecal Contamination ◽

Reference Method ◽

World Health ◽

Risk Levels ◽

E Coli ◽

Field Samples ◽

Very High

Quantitatively assessing fecal indicator bacteria in drinking water from limited resource settings (e.g., disasters, remote areas) can inform public health strategies for reducing waterborne illnesses. This study aimed to compare two common approaches for quantifying Escherichia coli (E. coli) density in natural water versus the ColiPlate™ kit approach. For comparing methods, 41 field samples from natural water sources in Kentucky (USA) were collected. E. coli densities were then determined by (1) membrane filtration in conjunction with modified membrane-thermotolerant E. coli (mTEC) agar, (2) Idexx Quanti-Tray® 2000 with the Colilert® substrate, and (3) the Bluewater Biosciences ColiPlate kit. Significant correlations were observed between E. coli density data for all three methods (p < 0.001). Paired t-test results showed no difference in E. coli densities determined by all the methods (p > 0.05). Upon assigning modified mTEC as the reference method for determining the World Health Organization-assigned “very high-risk” levels of fecal contamination (> 100 E. coli CFU/100 mL), both ColiPlate and Colilert exhibited excellent discrimination for screening very high-risk levels according to the area under the receiver operating characteristic curve (~89%). These data suggest ColiPlate continues to be an effective monitoring tool for quantifying E. coli density and characterizing fecal contamination risks from water.

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Detection of Antimicrobial Residues in Poultry Litter: Monitoring a Risk through a Selective and Sensitive HPLC–MS/MS Method

Animals ◽

10.3390/ani11051399 ◽

2021 ◽

Vol 11 (5) ◽

pp. 1399

Author(s):

Karina Yévenes ◽

Ekaterina Pokrant ◽

Lina Trincado ◽

Lisette Lapierre ◽

Nicolás Galarce ◽

...

Keyword(s):

High Performance ◽

Limit Of Detection ◽

Poultry Litter ◽

Chromatography Tandem Mass Spectrometry ◽

Antimicrobial Residues ◽

Field Samples ◽

Liquid Chromatography Tandem Mass ◽

Commercial Poultry ◽

Agriculture Fertilizer ◽

And Control

Tetracyclines, sulphonamides, and quinolones are families of antimicrobials (AMs) widely used in the poultry industry and can excrete up to 90% of AMs administrated, which accumulate in poultry litter. Worryingly, poultry litter is widely used as an agriculture fertilizer, contributing to the spread AMs residues in the environment. The aim of this research was to develop a method that could simultaneously identify and quantify three AMs families in poultry litter by high-performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS). Samples of AMs free poultry litter were used to validate the method according to 657/2002/EC and VICH GL49. Results indicate that limit of detection (LOD) ranged from 8.95 to 20.86 μg kg−1, while limits of quantitation (LOQ) values were between 26.85 and 62.58 µg kg−1 of tetracycline, 4-epi-tetracycline, oxytetracycline, 4-epi-oxytetracycline, enrofloxacin, ciprofloxacin, flumequine, sulfachloropyridazine, and sulfadiazine. Recoveries obtained ranged from 93 to 108%. The analysis of field samples obtained from seven commercial poultry flocks confirmed the adequacy of the method since it detected means concentrations ranging from 20 to 10,364 μg kg−1. This provides us an accurate and reliable tool to monitor AMs residues in poultry litter and control its use as agricultural fertilizer.

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Virus Adaptation Following Experimental Infection of Chickens with a Domestic Duck Low Pathogenic Avian Influenza Isolate from the 2017 USA H7N9 Outbreak Identifies Polymorphic Mutations in Multiple Gene Segments

Viruses ◽

10.3390/v13061166 ◽

2021 ◽

Vol 13 (6) ◽

pp. 1166

Author(s):

Klaudia Chrzastek ◽

Karen Segovia ◽

Mia Torchetti ◽

Mary Lee Killian ◽

Mary Pantin-Jackwood ◽

...

Keyword(s):

Avian Influenza ◽

Experimental Infection ◽

Clinical Signs ◽

Interspecies Transmission ◽

Clinical Samples ◽

Domestic Duck ◽

Receptor Binding Site ◽

Synonymous Mutations ◽

Field Samples ◽

Virus Adaptation

In March 2017, highly pathogenic (HP) and low pathogenic (LP) avian influenza virus (AIV) subtype H7N9 were detected from poultry farms and backyard birds in several states in the southeast United States. Because interspecies transmission is a known mechanism for evolution of AIVs, we sought to characterize infection and transmission of a domestic duck-origin H7N9 LPAIV in chickens and genetically compare the viruses replicating in the chickens to the original H7N9 clinical field samples used as inoculum. The results of the experimental infection demonstrated virus replication and transmission in chickens, with overt clinical signs of disease and shedding through both oral and cloacal routes. Unexpectedly, higher levels of virus shedding were observed in some cloacal swabs. Next generation sequencing (NGS) analysis identified numerous non-synonymous mutations at the consensus level in the polymerase genes (i.e., PA, PB1, and PB2) and the hemagglutinin (HA) receptor binding site in viruses recovered from chickens, indicating possible virus adaptation in the new host. For comparison, NGS analysis of clinical samples obtained from duck specimen collected during the outbreak indicated three polymorphic sides in the M1 segment and a minor population of viruses carrying the D139N (21.4%) substitution in the NS1 segment. Interestingly, at consensus level, A/duck/Alabama (H7N9) had isoleucine at position 105 in NP protein, similar to HPAIV (H7N9) but not to LPAIV (H7N9) isolated from the same 2017 influenza outbreak in the US. Taken together, this work demonstrates that the H7N9 viruses could readily jump between avian species, which may have contributed to the evolution of the virus and its spread in the region.

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A Comparison of Approaches to Regional Land-Use Capability Analysis for Agricultural Land-Planning

Land ◽

10.3390/land10050458 ◽

2021 ◽

Vol 10 (5) ◽

pp. 458

Author(s):

Tara A. Ippolito ◽

Jeffrey E. Herrick ◽

Ekwe L. Dossa ◽

Maman Garba ◽

Mamadou Ouattara ◽

...

Keyword(s):

Land Use ◽

Land Use Planning ◽

Crop Production ◽

Agricultural Land ◽

Arable Land ◽

Agricultural Activity ◽

Field Samples ◽

Agricultural Potential ◽

Fertility Data ◽

Using Data

Smallholder agriculture is a major source of income and food for developing nations. With more frequent drought and increasing scarcity of arable land, more accurate land-use planning tools are needed to allocate land resources to support regional agricultural activity. To address this need, we created Land Capability Classification (LCC) system maps using data from two digital soil maps, which were compared with measurements from 1305 field sites in the Dosso region of Niger. Based on these, we developed 250 m gridded maps of LCC values across the region. Across the region, land is severely limited for agricultural use because of low available water-holding capacity (AWC) that limits dry season agricultural potential, especially without irrigation, and requires more frequent irrigation where supplemental water is available. If the AWC limitation is removed in the LCC algorithm (i.e., simulating the use of sufficient irrigation or a much higher and more evenly distributed rainfall), the dominant limitations become less severe and more spatially varied. Finally, we used additional soil fertility data from the field samples to illustrate the value of collecting contemporary data for dynamic soil properties that are critical for crop production, including soil organic carbon, phosphorus and nitrogen.

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