scholarly journals RPA-based point-of-need detection assay for the invasive Mediterranean fanworm Sabella spallanzanii

2021 ◽  
Vol 4 ◽  
Author(s):  
Martin Zirngibl ◽  
Ulla von Ammon ◽  
Xavier Pochon ◽  
Anastasija Zaiko
Keyword(s):  
New Zealand ◽  
Molecular Detection ◽  
Environmental Dna ◽  
Great Effort ◽  
Detection Techniques ◽  
Context Sensitive ◽  
Detection Assay ◽  
Amplification Assay ◽  
Flow Detection

The Mediterranean fanworm Sabella spallanzanii was first sighted in New Zealand in 2009 (most likely introduced via hull fouling) and has spread across multiple coastal locations. The species presents significant risks to ecological, economic, and societal values, and therefore is subject to targeted surveillance in 11 major ports and marinas, that were identified as high-risk marine biosecurity sites. Great effort and financial resources are put into bi-annual diving surveys that include removal of individuals to contain population growth and spread. In that context, sensitive molecular detection techniques gain great interest and are being increasingly tested for the fanworm detection in New Zealand and Australia. However, conventional molecular detection via PCR assays from environmental DNA (eDNA) samples requires specific laboratory resources and scientific expertise, which restricts the applicability of this approach by biosecurity practitioners or citizen scientists. In order to provide end-users with a fast, easy, and highly specific way to detect S. spallanzanii at the site of interest, a recombinase polymerase amplification assay (RPA) was designed for read-out with lateral flow detection. RPA generates amplification within 20 minutes at 39°C, which makes it possible to run assays hand-held. Because of this, RPA presents an excellent tool for the point-of-need application in targeted biosecurity surveillance. Here we present the results of the successful in silico and in vitro validation of the newly designed RPA assay for S. spallanzanii.

scholarly journals Using a roller pump for establishing extra-corporal membrane oxygenation (ECMO) – technical considerations for times of crisis

Perfusion ◽  
2021 ◽  
pp. 026765912199618
Author(s):  
Mirko Kaluza ◽  
Benjamin May ◽  
Torsten Doenst
Keyword(s):  
Flow Characteristics ◽  
Centrifugal Pumps ◽  
Roller Pump ◽  
Silicone Tubing ◽  
Heart Lung Machine ◽  
Surface Irregularities ◽  
Set Up ◽  
Flow Detection ◽  
Silicon Tubing

Objective: The COVID-19 pandemic requires thinking about alternatives to establish ECMO when often-limited hardware resources are exhausted. Heart-lung-machines may potentially be used for ECMO but contain roller pumps as compared to centrifugal pumps in ECMO-circuits. We here tested roller pumps as rescue pump for ECMO-establishment. Methods: We set up in vitro circuits on roller pumps from C5 heart-lung-machine with 5 l/minutes flow. In two series, we placed either PVC or silicon tubing for an ECMO circuit into the roller pump. We assessed the mechanical stress on the tubing (aiming to run the pump for at least 1 week), measured the temperature increase generated by the friction and assessed flow characteristics and its measurement in simulated situations resembling tube kinking and suction. Results: The roller pumps led to expected and unexpected adverse events. PVC tubing burst between 36 and 78 hours, while silicon tubing lasted for at least 7 days. At 7 days, the silicone tubing showed significant signs of roller pump wear visible on the outside. The inside, however, was free of surface irregularities. Using these tubings in a roller pump led to a remarkable increase in circuit temperature (PVC: +12.0°C, silicone +2.9°C). Kinking or suction on the device caused the expected dramatic flow reduction (as assessed by direct measurement) while the roller pump display continued to show the preset flow. The roller pump is therefore not able to reliably determine the true flow rate. Conclusion: Roller pumps with silicone tubing but not PVC tubing may be used for running ECMO circuits. Silicone tubing may endure the roller pump shear forces for up to 1 week. Thus, repeated tubing repositioning may be a solution. Circuit heating and substantial limitations in flow detection should increase attention if clinical use in situations of crisis is considered.

Evaluation of Modification of the 3M™ Molecular Detection Assay (MDA) Salmonella Method (2013.09) for the Detection of Salmonella in Selected Foods: Collaborative Study

2014 ◽  
Vol 97 (5) ◽  
pp. 1329-1342 ◽  
Author(s):  
Patrick Bird ◽  
Kiel Fisher ◽  
Megan Boyle ◽  
Travis Huffman ◽  
M Joseph Benzinger ◽  
...  
Keyword(s):  
Confidence Intervals ◽  
Molecular Detection ◽  
Collaborative Study ◽  
Ground Beef ◽  
Inoculum Level ◽  
Test Portion ◽  
Low Level ◽  
Dog Food ◽  
Detection Assay ◽  
The U.S

Abstract The 3M™ Molecular Detection Assay (MDA) Salmonella utilizes isothermal amplification of nucleic acid sequences with high specificity, efficiency, rapidity and bioluminescence to detect amplification of Salmonella spp. in food, food-related, and environmental samples after enrichment. A method modification and matrix extension study of the previously approved AOAC Official MethodSM 2013.09 was conducted, and approval of the modification was received on March 20, 2014. Using an unpaired study design in a multilaboratory collaborative study, the 3M MDA Salmonella method was compared to the U.S. Department of Agriculture/Food Safety and Inspection Service (USDA/FSIS) Microbiology Laboratory Guidebook (MLG) 4.05 (2011), Isolation and Identification of Salmonella from Meat, Poultry, Pasteurized Egg, and Catfish Products for raw ground beef and the U.S. Food and Drug Administration (FDA)/Bacteriological Analytical Manual (BAM) Chapter 5, Salmonella reference method for wet dog food following the current AOAC guidelines. A total of 20 laboratories participated. For the 3M MDA Salmonella method, raw ground beef was analyzed using 25 g test portions, and wet dog food was analyzed using 375 g test portions. For the reference methods, 25 g test portions of each matrix were analyzed. Each matrix was artificially contaminated with Salmonella at three inoculation levels: an uninoculated control level (0 CFU/test portion), a low inoculum level (0.2–2 CFU/test portion), and a high inoculum level (2–5 CFU/test portion). In this study, 1512 unpaired replicate samples were analyzed. Statistical analysis was conducted according to the probability of detection (POD). For the low-level raw ground beef test portions, the following dLPOD (difference between the LPODs of the reference and candidate method) values with 95% confidence intervals were obtained: –0.01 (–0.14, +0.12). For the low-level wet dog food test portions, the following dLPOD with 95% confidence intervals were obtained: –0.04 (–0.16, +0.09). No significant differences were observed in the number of positive samples detected by the 3M MDA Salmonella method versus either the USDA/FSIS-MLG or FDA/BAM methods.

scholarly journals Experimental Evidence for Therapeutic Potentials of Propolis

Nutrients ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 2528
Author(s):  
Priyanshu Bhargava ◽  
Debajit Mahanta ◽  
Ashish Kaul ◽  
Yoshiyuki Ishida ◽  
Keiji Terao ◽  
...  
Keyword(s):  
New Zealand ◽  
Experimental Evidence ◽  
Antioxidant Properties ◽  
Geographic Location ◽  
Caffeic Acid Phenethyl Ester ◽  
Candidate Drug ◽  
Brazilian Propolis ◽  
Brazilian Green Propolis

Propolis is produced by honeybees from materials collected from plants they visit. It is a resinous material having mixtures of wax and bee enzymes. Propolis is also known as bee glue and used by bees as a building material in their hives, for blocking holes and cracks, repairing the combs and strengthening their thin borders. It has been extensively used since ancient times for different purposes in traditional human healthcare practices. The quality and composition of propolis depend on its geographic location, climatic zone and local flora. The New Zealand and Brazilian green propolis are the two main kinds that have been extensively studied in recent years. Their bioactive components have been found to possess a variety of therapeutic potentials. It was found that Brazilian green propolis improves the cognitive functions of mild cognitive impairments in patients living at high altitude and protects them from neurodegenerative damage through its antioxidant properties. It possesses artepillin C (ARC) as the key component, also known to possess anticancer potential. The New Zealand propolis contains caffeic acid phenethyl ester (CAPE) as the main bioactive with multiple therapeutic potentials. Our lab performed in vitro and in vivo assays on the extracts prepared from New Zealand and Brazilian propolis and their active ingredients. We provided experimental evidence that these extracts possess anticancer, antistress and hypoxia-modulating activities. Furthermore, their conjugation with γCD proved to be more effective. In the present review, we portray the experimental evidence showing that propolis has the potential to be a candidate drug for different ailments and improve the quality of life.

Studies of Virulence Factors of Avian Pathogenic Escherichia coli in Avian Colibacillosis by In vitro Method and PCR

2021 ◽  
Author(s):  
Ayushi Singh ◽  
Daljeet Chhabra ◽  
Rakhi Gangil ◽  
Rakesh Sharda ◽  
Ravi Sikrodia ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Molecular Detection ◽  
Bacterial Disease ◽  
Avian Pathogenic Escherichia Coli ◽  
E Coli ◽  
Pathological Conditions ◽  
Biofilm Production ◽  
Pathogenic Escherichia Coli ◽  
Stx1 Gene

Background: Avian colibacillosis is considered as major cause of morbidity and mortality in poultry. It is a common bacterial disease of poultry and many virulence factors of E. coli are associated with the disease. The current study was aimed to investigate the presence of some virulence factors of E. coli isolated from the cases of colibacillosis.Methods: In present study, total 150 samples (liver, heart, lungs, air sacs and feaces) of chicken exhibiting pathological conditions of colibacillosis were collected from various poultry farms (organized and backyard) situated in and around Mhow and Indore cities. E.coli was isolated and identified from the samples on the basis of cultural characteristics and biochemical test. All E. coli isolates were further subjected to evaluate the presence of virulence factors such as biofilm production, haemolysis, invasiveness and molecular detection of fimH and stx1 gene.Result: Out of these 51.33% of incidence of E. coli was recorded. E. coli O84 and O149 serotypes were found most prevalent. Out of 77 isolates, 46 (59.7%) and 45 (58.4%) were positive for biofilm formation by tube method and modified CRA method, respectively. All E. coli isolates were showing invasiveness in congo red binding assay while none of the isolates was found haemolytic. Molecular detection revealed the presence of fimH (508bp) gene in 33.3% of tested samples while stx1 gene could not be detected in any isolates.

scholarly journals Comparison of the Echinocandin Caspofungin with Amphotericin B for Treatment of Histoplasmosis following Pulmonary Challenge in a Murine Model

2000 ◽  
Vol 44 (7) ◽  
pp. 1850-1854 ◽  
Author(s):  
Steve Kohler ◽  
L. Joseph Wheat ◽  
Patricia Connolly ◽  
Carol Schnizlein-Bick ◽  
Michelle Durkin ◽  
...  
Keyword(s):  
Amphotericin B ◽  
Histoplasma Capsulatum ◽  
Clinical Laboratory ◽  
Antigen Detection ◽  
National Committee ◽  
Slight Effect ◽  
Fungal Burden ◽  
Detection Techniques ◽  
The Mean

ABSTRACT Twenty clinical isolates of Histoplasma capsulatum were tested for their in vitro susceptibilities to caspofungin in comparison to those to amphotericin B by following National Committee for Clinical Laboratory Standards guidelines for yeasts. The mean MICs were 16.6 μg/ml (range, 8 to 32 μg/ml) for caspofungin and 0.56 μg/ml (range, 0.5 to 1.0 μg/ml) for amphotericin B. Survival experiments used a 105 dose in a pulmonary challenge model with B6C3F1 mice. All mice that received amphotericin B at 2 mg/kg of body weight every other day (q.o.d.), 30% of mice that received caspofungin at 8 mg/kg/day, and 20% of mice that received caspofungin at 4 mg/kg/day survived to day 15, while mice that received caspofungin at 2 mg/kg/day and all control mice that received the vehicle died by day 14. Amphotericin B at 2 mg/kg q.o.d. markedly reduced the fungal burden in the lungs and spleens, as measured byHistoplasma antigen detection techniques and quantitative cultures, for each comparison. Caspofungin at 10 mg/kg twice a day (b.i.d.) did not reduce the fungal burden, as measured by antigen detection techniques, but slightly reduced the levels of fungi in both the lungs and spleens, as determined by quantitative cultures. Caspofungin at 5 mg/kg b.i.d. did not affect fungal burden. Overall, caspofungin had only a slight effect on survival or fungal burden.

scholarly journals Unexplored diversity of microscopic myxomycetes: evidence from environmental DNA

2019 ◽  
Vol 152 (3) ◽  
pp. 499-506 ◽  
Author(s):  
Oleg N. Shchepin ◽  
Martin Schnittler ◽  
Nikki H.A. Dagamac ◽  
Dmitry V. Leontyev ◽  
Yuri K. Novozhilov
Keyword(s):  
New Species ◽  
18S Rdna ◽  
Sequence Similarity ◽  
Phylogenetic Analyses ◽  
Environmental Dna ◽  
Full Length ◽  
Detection Techniques ◽  
Rdna Sequences ◽  
18S Rdna Sequences ◽  
Environmental Sequences

Background and aims – Recent studies showed the position of two slime mould species with microscopic sporocarps, Echinosteliopsis oligospora and Echinostelium bisporum, within the class Myxomycetes. These minute species are seldom seen in studies based on detection of sporocarps and can easily be confused with protosteloid amoebozoans.Methods – We searched all published ePCR data sets that targeted myxomycete 18S rDNA for the presence of environmental sequences similar to E. oligospora and Echinosteliales in traditional circumscription, and performed phylogenetic analyses that included short environmental sequences and full-length 18S rDNA sequences representing all the major groups of myxomycetes.Key results – We report 19 unique sequences which are closely related to E. bisporum or E. oligospora based on sequence similarity (73.1–95.2% similarity) and which form well-supported monophyletic clades with these species in phylogenetic analyses. They may represent new species that are not yet described. Our phylogeny based on full-length 18S rDNA sequences further confirms the position of E. bisporum and E. oligospora within myxomycetes and the paraphyly of the order Echinosteliales in its traditional circumscription.Conclusions – Our results show that ePCR-based studies can reveal myxomycete taxa that often escape detection by traditional approaches, including potentially new species, and thus provide valuable new data on diversity and ecology of myxomycetes. As such, strategies for studying myxomycetes biodiversity should be revised, focusing also on molecular detection techniques in addition to the sporocarp-based ones.

scholarly journals Thinopyrum intermedium (Host) Barkw. & Dewey - a review, and evaluation of intermediate and pubescent wheatgrass for dryland agriculture in New Zealand

1998 ◽  
pp. 233-241
Author(s):  
B.J. Wills ◽  
G.B. Douglas ◽  
J. Mckenzie ◽  
K.D. Trainor ◽  
A.G. Foote
Keyword(s):  
New Zealand ◽  
Ground Cover ◽  
Climatic Conditions ◽  
Individual Species ◽  
Thinopyrum Intermedium ◽  
Dryland Agriculture ◽  
Alternative Species ◽  
The Individual ◽  
Local Use

Intermediate and pubescent wheatgrasses (Thinopyrum intermedium) are potentially useful as dryland forage grasses in New Zealand. The recent literature on their international and local use in pasture and cropping situations is reviewed. Under semi-arid conditions at Bendigo Station over a 13-year period, cv. Luna wheatgrass has spread several metres from the original drill lines and it provides very high ground cover. In terms of dryland soil resource conservation this contrasts with adjacent cocksfoot and, to a lesser extent, lucerne plots which have wide-spaced plants interspersed with litter and much bare ground. In a mixed drilling containing wheatgrass, hairy dorycnium, birdsfoot trefoil and sheep's burnet, the first two species now dominate. At Takapau, central Hawke's Bay, annual yields from intermediate and pubescent wheatgrasses produced over 80% of the yield of 10 New Zealand cultivars in spring, summer and autumn, and 65% in the winter. Of the wheatgrasses, cv. Topar intermediate wheatgrass and a pubescent wheatgrass (unnamed line) were the lowest yielding, whereas cv. Oahe and cv. Mandan produced the highest yields. Cv. Greenleaf pubescent wheatgrass had higher foliar nitrogen content than all other species, but in vitro organic matter digestibility was less than 700 g OM/kg DM. Although the wheatgrasses had satisfactory herbage yield and nutritional value, better alternative species are available for pastoral farming in central Hawke's Bay. On Belfield in the Hakataramea Valley, cv. Rush has been the most vigorous wheatgrass for establishment and it seems to handle cold winds and frost well. Hay made from a wheatgrass/ lucerne mix is considered better than that from the individual species as this can increase production by 40-50%, depending on soil and climatic conditions. Usually only one cut per year is taken from the mixed pasture. Crop maintenance includes the application of 200 kg/ha of sulphur super extra every 3-4 years and the pH is maintained at about 5.9-6.4 by liming. A significant advantage is that no stock health, pest or plant disease problems have been noted as yet with any of the wheatgrass varieties available in New Zealand. Keywords: dryland agriculture, New Zealand, Thinopyrum, wheatgrass

Sign in / Sign up

Export Citation Format

Share Document