Using a Microscale Approach To Rapidly Separate and Characterize Three Photosynthetic Pigment Species from Fern

Ayudhya, Theppawut Israsena Na; Posey, F.; Tyus, Jessica C.; Dingra, N. N.

doi:10.1021/ed500344c

Cited by 29 publications

(19 citation statements)

References 19 publications

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“…All faecal samples collected in this study showed similar pro le and trends. Two distinct bands were observed around 576 nm and 670 nm, which were previously found to be associated with Hb and chlorophyll, respectively (41,49,50). Subsequently, for the prediction of Hb in sheep faeces in the present study, all analyses were carried out within the wavelength range of 387-609 nm to limit the effects of the 670 nm chlorophyll peak.…”

Section: Resultsmentioning

confidence: 83%

Influence of environmental factors on the detection of blood in sheep faeces using visible-near infrared spectroscopy as a measure of Haemonchus contortus infection

Kho

Fernandes

Kotze

et al. 2020

Preprint

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Background: Existing diagnostic methods for the parasitic gastrointestinal nematode, Haemonchus contortus, are time consuming and require specialised expertise, limiting their utility in the field. A practical, on-farm diagnostic tool could facilitate timely treatment decisions, preventing production and welfare loss in the flock. We previously demonstrated the ability of visible-near infrared (vis-NIR) spectroscopy to detect and quantify blood in sheep faeces with high accuracy. Here we investigate whether variation in sheep type and environment affect the prediction accuracy of vis-NIR spectroscopy in quantifying blood in faeces.Methods: Vis-NIR spectra were obtained from worm-free sheep faeces from different environments in South Australia (SA) and New South Wales (NSW), Australia and spiked with various sheep blood concentrations collected. Spectra were analysed using principal component analysis (PCA), and calibration models were built around the haemoglobin (Hb) wavelength region (387 – 609 nm) using partial least squares (PLS) regression. Models were used to predict Hb concentrations in spiked faeces from SA and naturally infected Queensland (QLD) faeces. Naturally occurring blood in QLD samples was quantified using Hemastix® and FAMACHA© scores.Results: PCA showed that location, class of sheep and pooled/individual samples were factors affecting the Hb predictions in sheep faeces. The calibration models successfully differentiated ‘healthy’ SA samples from those requiring anthelmintic treatment with moderate to good prediction accuracy (sensitivity: 57 – 94%, specificity: 44 – 79%). The models were not predictive for naturally infected QLD samples, which may be due in part to variability of faecal background and blood chemistry between samples, or the difference in validation methods used for blood quantification. PCA of QLD samples, however, identified a difference between samples containing high and low quantities of blood.Conclusion: This study demonstrates the potential of vis-NIR spectroscopy for estimating blood concentration in faeces from various types of sheep and environmental backgrounds. However, the calibration models developed here did not capture enough environmental variation to accurately predict Hb in faeces collected from environments different to those used in the calibration model. Consequently, it will be necessary to establish models that incorporate samples that are more representative of areas where H. contortus is endemic for the accurate prediction of H. contortus infections in these regions.

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Section: Resultsmentioning

confidence: 83%

Influence of environmental factors on the detection of blood in sheep faeces using visible-near infrared spectroscopy as a measure of Haemonchus contortus infection

Kho

Fernandes

Kotze

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…All faecal samples collected in this study showed similar pro le and trends. Two distinct bands were observed around 576 nm and 670 nm, which were previously found to be associated with Hb and chlorophyll, respectively [42,51,52]. Subsequently, for the prediction of Hb in sheep faeces in the present study, all analyses were carried out within the wavelength range of 387 -609 nm to limit the effects of the 670 nm chlorophyll peak.…”

Section: Resultsmentioning

confidence: 83%

Influence of environmental factors on the detection of blood in sheep faeces using visible-near infrared spectroscopy as a measure of Haemonchus contortus infection

Kho

Fernandes

Kotze

et al. 2020

Preprint

View full text Add to dashboard Cite

Background: Existing diagnostic methods for the parasitic gastrointestinal nematode, Haemonchus contortus, are time consuming and require specialised expertise, limiting their utility in the field. A practical, on-farm diagnostic tool could facilitate timely treatment decisions, preventing production and welfare loss in the flock. We previously demonstrated the ability of visible-near infrared (vis-NIR) spectroscopy to detect and quantify blood in sheep faeces with high accuracy. Here we investigate whether variation in sheep type and environment affect the prediction accuracy of vis-NIR spectroscopy in quantifying blood in faeces. Methods: Vis-NIR spectra were obtained from worm-free sheep faeces collected from different environments and sheep types in South Australia (SA) and New South Wales (NSW), Australia and spiked with various sheep blood concentrations. Spectra were analysed using principal component analysis (PCA), and calibration models were built around the haemoglobin (Hb) wavelength region (387 – 609 nm) using partial least squares (PLS) regression. Models were used to predict Hb concentrations in spiked faeces from SA and naturally infected sheep faeces from Queensland (QLD). QLD samples were quantified using Hemastix® and FAMACHA © scores. Results: PCA showed that location, class of sheep and pooled/individual samples were factors affecting the Hb predictions. The models successfully differentiated ‘healthy’ SA samples from those requiring anthelmintic treatment with moderate to good prediction accuracy (sensitivity: 57 – 94%, specificity: 44 – 79%). The models were not predictive for blood in naturally infected QLD samples, which may be due in part to variability of faecal background and blood chemistry between samples, or the difference in validation methods used for blood quantification. PCA of QLD samples, however, identified a difference between samples containing high and low quantities of blood. Conclusion: This study demonstrates the potential of vis-NIR spectroscopy for estimating blood concentration in faeces from various types of sheep and environmental backgrounds. However, the calibration models developed here did not capture enough environmental variation to accurately predict Hb in faeces collected from environments different to those used in the calibration model. Consequently, it will be necessary to establish models that incorporate samples that are more representative of areas where H. contortus is endemic.

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“…The selection of a suitable analytical method for the identification and quantification of pigments from the plant matrix depends on the characteristics of required data (Dudek, Strzelewicz, Krasowska, Rybak & Turczyn, 2014). The most frequently used instrumental methodologies are the UV-VIS spectrophotometry, fluorescence spectroscopy and chromatography (Garber, Odendaal & Carlson, 2013;West & Rauch, 1994;Ayudhya, Posey, Tyus & Dingra, 2015;Johnston, Scaggs, Mallory, Haskett, Warner, Brown, et al, 2013;Galloway, Bretz & Novak, 2015;Rollins, 1963;Lalitha, 1994;Kimbrough, 1992;Valverde, This & Vignolle, 2007). Each of these techniques has several advantages; however, the UV-Vis spectrophotometry is the most suitable for the routine determination of the pigment concentration in the extract from the mixed plant samples, as it generally does not require complicated separation and purification processes.…”

Section: Natural Pigmentsmentioning

confidence: 99%

Scientific experiment focused at pigment degradation by polyvinyl chloride combustion in Science education

Jenisová

Braniša

2019

J. Technol. Sci. Educ.

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The Environmental education navigates students towards environmentally friendly life style and securing a quality of life. The implementation of cross-section topics, including the Environmental education, into science classes, is possible through a variety of techniques. One of the least used methods is a real school experiment. The following paper presents the experiment, by which we simulate the combustion of PVC and observe the impact of combustion products on plants via UV-Vis spectrophotometry optical method. This method is suitable for qualification and analysis of vegetable pigments, i.e. chlorophylls, carotenoids, and anthocyanins. The experiment is integrated into the Techniques and Didactics of School Chemistry Experiments as a part of master degree course, which prepares students for the Chemistry teaching profession. The introduced experiment enables pedagogues to integrate the environmental education into teaching process and develop mathematical and science literacy of students in the Chemistry education.

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Using a Microscale Approach To Rapidly Separate and Characterize Three Photosynthetic Pigment Species from Fern

Cited by 29 publications

References 19 publications

Influence of environmental factors on the detection of blood in sheep faeces using visible-near infrared spectroscopy as a measure of Haemonchus contortus infection

Influence of environmental factors on the detection of blood in sheep faeces using visible-near infrared spectroscopy as a measure of Haemonchus contortus infection

Influence of environmental factors on the detection of blood in sheep faeces using visible-near infrared spectroscopy as a measure of Haemonchus contortus infection

Scientific experiment focused at pigment degradation by polyvinyl chloride combustion in Science education

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