Measurement of Trace Elements (Zinc, Copper, Magnesium, and Iron) in the Saliva of Horses: Validation Data and Changes in Equine Gastric Ulcer Syndrome (EGUS)
Alberto Muñoz-PrietoJ. M. Castro CerónFernando TeclesMaría Martín CuervoMaría Dolores Contreras-AguilarIgnacio AyalaAdrián Oudada-GuillénLuis Pardo-MarínSanni Hansen
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The objective of this study was to evaluate the possible use of spectrophotometric assays for the measurement of trace elements, including Zinc (Zn), Copper (Cu), Magnesium (Mg), and iron (Fe) in the saliva of horses and study their possible changes in equine gastric ulcer syndrome (EGUS). EGUS is a highly prevalent disease, with a current high incidence due to the increase in intensive management conditions. There are two EGUS diseases: equine squamous gastric disease (ESGD) and equine glandular gastric disease (EGGD), which can appear individually or together. For this purpose, automated spectrophotometric assays for measuring these analytes in horse saliva were analytically validated. Then, these analytes were measured in the saliva of horses with only ESGD, only EGGD, both ESGD and EGGD and a group of healthy horses. The methods used to measure the analytes were precise and accurate. Horses diagnosed with EGGD presented significantly lower levels of Zn and Mg. Fe concentrations were significantly lower in the saliva of horses with ESGD and EGGD. Overall, these results indicate that there are changes in trace elements in saliva in EGUS that could reflect the physiopathological mechanisms involved in this process and open the possibility of using trace elements as biomarkers of this syndrome.Cite
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Abstract Background Saliva is an attractive sample for detecting SARS-CoV-2. However, contradictory reports exist concerning the sensitivity of saliva versus nasal swabs. Methods We followed close contacts of COVID-19 cases for up to 14 days from last exposure and collected self-reported symptoms, mid-turbinate swabs (MTS), and saliva every two or three days. Ct values, viral load, and frequency of viral detection by MTS and saliva were compared. Results 58 contacts provided 200 saliva-MTS pairs; 14 contacts (13 with symptoms) had one or more positive samples. Saliva and MTS had similar rates of viral detection (p=0.78) and substantial agreement (κ=0.83). However, sensitivity varied significantly with time since symptom onset. Early on (days -3 to 2), saliva had 12 times (95%CI: 1.2, 130) greater likelihood of viral detection and 3.2 times (95% CI: 2.8, 3.8) higher RNA copy numbers compared to MTS. After day 2 post-symptoms, there was a non-significant trend toward greater sensitivity using MTS. Conclusion Saliva and MTS demonstrated high agreement making saliva a suitable alternative to MTS for COVID-19 detection. Saliva was more sensitive early in the infection when transmission is most likely to occur, suggesting that it may be a superior and cost-effective screening tool for COVID-19.
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The findings of this manuscript are increasingly important with new variants that appear to have shorter incubation periods emerging, which may be more prone to detection in saliva before detection in nasal swabs. Therefore, there is an urgent need to provide the science to support the use of a detection method that is highly sensitive and widely acceptable to the public to improve screening rates and early detection.
2019-20 coronavirus outbreak
Betacoronavirus
Sars virus
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During dental erosion, tooth minerals are dissolved, leading to a softening of the surface and consequently to irreversible surface loss. Components from human saliva form a pellicle on the tooth surface, providing some protection against erosion. To assess the effect of different components and compositions of saliva on the protective potential of the pellicle against enamel erosion, we prepared four different kinds of saliva: human whole stimulated saliva (HS), artificial saliva containing only ions (AS), human saliva dialysed against artificial saliva, containing salivary proteins and ions (HS/AS), and human saliva dialysed against deionised water, containing only salivary proteins but no ions (HS/DW). Enamel specimens underwent four cycles of immersion in either HS, AS, HS/AS, HS/DW, or a humid chamber (Ctrl), followed by erosion with citric acid. During the cycling process, the surface hardness and the calcium released from the surface of the specimens were measured. The different kinds of saliva provided different levels of protection, HS/DW exhibiting significantly better protection than all the other groups (p < 0.0001). Different components of saliva, therefore, have different effects on the protective properties of the pellicle and the right proportions of these components in saliva are critical for the ability to form a protective pellicle.
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Human tooth
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A rapid assay for detection of cytomegalovirus (CMV) in saliva was evaluated as a screening method for congenital infection. Samples of saliva were examined by detection of early antigen fluorescent foci (DEAFF) and standard tissue culture (TC). Results were compared with those from urine DEAFF. CMV was detected in saliva from 31 (1.7%) of 1870 newborns, 26 by DEAFF and TC, 1 by DEAFF alone, and 4 by TC alone. Urine DEAFF was positive in 28 of these 31 newborns. The sensitivities of various tests were saliva TC, 96.8%; saliva DEAFF, 87.1 %; and urine DEAFF, 90.3%. A change in transport medium for 825 saliva samples resulted in improved sensitivities: saliva TC and saliva DEAFF, 100%; urine DEAFF, 92.3%. Screening saliva of newborns for CMV appears to be at least as sensitive a method for detecting congenital infection as detection of viruria; saliva can be collected with less difficulty and expense than urine.
Cytomegalovirus
Betaherpesvirinae
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In order to clarify how we collect saliva for analyzing salivary protein in aged subjects who can not eat well, we compared the effects of suction, spitting and the swab saliva collection method on the yield of protein components in saliva samples from normal volunteers. The saliva collected by suction, spitting and the swab method were designated as, Saliva I, II and III, respectively.The saliva volume collected by Saliva I was about 2-fold greater than that by of Saliva II and III. This is mainly due to the fact that saliva secretion was stimulated by the suction itself. The content of total protein, S-IgA, trypsin-like activity and human airway trypsin-like protease (HAT) were almost the same in Saliva I and II, and significantly lower in Saliva III than in Saliva I and II. Kallikrein activity was almost the same in Saliva I, II and III. The concentration of each total protein, S-IgA, kallikrein activity, trypsin activity and HAT in Saliva I were significantly positively correlated with that in Saliva II.These results indicate that we can obtain information of change of salivary protein by analyzing saliva collected by suction method, although this method caused the stimulation of saliva to some extent. J. Med. Invest. 53: 140-146, February, 2006
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Here, we develop a simple molecular test for SARS-CoV-2 in saliva based on reverse transcription loop-mediated isothermal amplification. The test has two steps: (1) heat saliva with a stabilization solution and (2) detect virus by incubating with a primer/enzyme mix. After incubation, saliva samples containing the SARS-CoV-2 genome turn bright yellow. Because this test is pH dependent, it can react falsely to some naturally acidic saliva samples. We report unique saliva stabilization protocols that rendered 295 healthy saliva samples compatible with the test, producing zero false positives. We also evaluated the test on 278 saliva samples from individuals who were infected with SARS-CoV-2 but had no symptoms at the time of saliva collection, and from 54 matched pairs of saliva and anterior nasal samples from infected individuals. The Saliva TwoStep test described herein identified infections with 94% sensitivity and >99% specificity in individuals with sub-clinical (asymptomatic or pre-symptomatic) infections.
Asymptomatic carrier
2019-20 coronavirus outbreak
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Saliva diagnostics have become increasingly popular due to their non-invasive nature and patient-friendly collection process. Various collection methods are available, yet these are not always well standardized for either quantitative or qualitative analysis. In line, the objective of this study was to evaluate if measured levels of various biomarkers in the saliva of healthy individuals were affected by three distinct saliva collection methods: 1) unstimulated saliva, 2) chew stimulated saliva, and 3) oral rinse. Saliva samples from 30 healthy individuals were obtained by the three collection methods. Then, the levels of various salivary biomarkers such as proteins and ions were determined. It was found that levels of various biomarkers obtained from unstimulated saliva were comparable to those in chew stimulated saliva. The levels of potassium, sodium, and amylase activity differed significantly among the three collection methods. Levels of all biomarkers measured using the oral rinse method significantly differed from those obtained from unstimulated and chew-stimulated saliva. In conclusion, both unstimulated and chew-stimulated saliva provided comparable levels for a diverse group of biomarkers. However, the results obtained from the oral rinse method significantly differed from those of unstimulated and chew-stimulated saliva, due to the diluted nature of the saliva extract.
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Abstract A CONSIDERABLE number of recent investigations have concerned the zinc requirement of chickens and turkeys. This work was stimulated by the results of studies conducted by O’Dell and Savage (1957) and Supplee et al. (1958). Compounds which have been used as zinc sources in the various investigations include zinc carbonate, zinc chloride, zinc oxide, zinc sulfate and certain zinc proteinates. A few comparisons have been made, but only one detailed study has been conducted to determine the biological availability of zinc from various sources to chicks and poults. Roberson and Schaible (1958) reported that the zinc in zinc sulfate and zinc chloride was equally available to starting chicks. One-hundred parts per million (ppm.) of zinc from each compound was added to a zinc-deficient, purified diet. This level of supplementary zinc, however, was more than three times the amount required by chicks. No definite conclusions regarding availability can be drawn from …
Zinc compounds
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Objective. Tissue inhibitor of metalloproteinases 1 (TIMP‐1) has been identified as a potential biomarker in diseases such as cancer, cardiovascular diseases and diabetes. Since TIMP‐1 resides in most tissues and bodily fluids, we evaluated the potential of using saliva to obtain reproducible TIMP‐1 measurements in a non‐invasive manner. Material and methods. Samples of unstimulated and stimulated whole saliva and saliva collected from individual glands were analysed for TIMP‐1 content. A TIMP‐1 ELISA was validated for use in saliva testing and the most optimal sampling and handling procedures for reproducible measurements identified. Western blotting and MALDI‐TOF mass spectrometry were used for confirmatory analyses. Results. The TIMP‐1 ELISA was found suitable for saliva measurements. All saliva secretions contained TIMP‐1, but in different concentrations ranging from 2.81 ng/mL in submandibular/sublingual saliva to 173.88 ng/mL in parotid saliva. TIMP‐1 concentrations were influenced to a varying degree by fluctuations in flow. We found the lowest output in submandibular/sublingual saliva stimulated with 0.5 % citric acid (3.56 ng/min) and highest output in chewing‐stimulated whole saliva (267.01 ng/min). Conclusion. This study shows that saliva contains authentic TIMP‐1, the concentration of which was found to depend on gland type and salivary flow. Stimulated whole saliva is suggested as a reliable and easily accessible source for TIMP‐1 determinations in bodily fluids.
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