scholarly journals Lactoferrin quantification in cattle faeces by ELISA

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e8631
Author(s):  
Andrew S. Cooke ◽  
Kathryn A. Watt ◽  
Greg F. Albery ◽  
Eric R. Morgan ◽  
Jennifer A.J. Dungait

Background Promoting and maintaining health is critical to ruminant welfare and productivity. Within human medicine, faecal lactoferrin is quantified for routine assessment of various gastrointestinal illnesses avoiding the need for blood sampling. This approach might also be adapted and applied for non-invasive health assessments in animals. Methods In this proof-of-concept study, a bovine lactoferrin enzyme-linked immunosorbent assays (ELISA), designed for serum and milk, was applied to a faecal supernatant to assess its potential for quantifying lactoferrin in the faeces of cattle. Faecal lactoferrin concentrations were compared to background levels to assess the viability of the technique. A comparison was then made against serum lactoferrin levels to determine if they were or were not reflective of one another. Results The optical densities of faecal samples were significantly greater than background readings, supporting the hypothesis that the assay was effective in quantifying faecal lactoferrin (T13, 115 = 11.99, p < 0.0005). The mean faecal lactoferrin concentration was 0.269 µg mL−1 (S.E. 0.031) and the mean serum concentration 0.074 µg mL−1 (S.E. 0.005). Lactoferrin concentrations of faecal and serum samples, taken from the same animals on the same day, were significantly different (T21 = 2.20, p = 0.039) and did not correlate (r = 0.2699, p = 0.238). Conclusion Results support the hypothesis that lactoferrin can be quantified in cattle faeces by ELISA. Whilst further research is required to determine the physiological source of the lactoferrin, this highlights the potential of the method for non-invasive assessment of cattle immunology and pathology.

2019 ◽  
Author(s):  
Andrew S. Cooke ◽  
Kathryn Watt ◽  
Greg F. Albery ◽  
Eric R. Morgan ◽  
Jennifer A. J. Dungait

Background: Promoting and maintaining health is critical to ruminant welfare and productivity. Within human medicine, faecal lactoferrin is quantified for routine assessment of various gastrointestinal illnesses avoiding the need for blood sampling. This approach might also be adapted and applied for non-invasive health assessments in animals. Methods: In this proof-of-concept study a bovine lactoferrin enzyme-linked immunosorbent assays (ELISA), designed for serum and milk, was applied to a faecal supernatant to assess its potential for quantifying lactoferrin in the faeces of cattle. Faecal lactoferrin concentrations were compared to background levels to assess the viability of the technique. A comparison was then made against serum lactoferrin levels to determine if they were or were not reflective of one another. Results: The optical densities of faecal samples were significantly greater than background readings, supporting the hypothesis that the assay was effective in quantifying faecal lactoferrin (T13, 115 = 11.99, p < 0.0005, n = 115). Lactoferrin concentrations of faecal and serum samples, taken from the same animals on the same day, were significantly different (T21 = 2.49, p = 0.022) and did not correlate (r = 0.069, p = 0.767).


2019 ◽  
Author(s):  
Andrew S. Cooke ◽  
Kathryn Watt ◽  
Greg F. Albery ◽  
Eric R. Morgan ◽  
Jennifer A. J. Dungait

Background: Promoting and maintaining health is critical to ruminant welfare and productivity. Within human medicine, faecal lactoferrin is quantified for routine assessment of various gastrointestinal illnesses avoiding the need for blood sampling. This approach might also be adapted and applied for non-invasive health assessments in animals. Methods: In this proof-of-concept study a bovine lactoferrin enzyme-linked immunosorbent assays (ELISA), designed for serum and milk, was applied to a faecal supernatant to assess its potential for quantifying lactoferrin in the faeces of cattle. Faecal lactoferrin concentrations were compared to background levels to assess the viability of the technique. A comparison was then made against serum lactoferrin levels to determine if they were or were not reflective of one another. Results: The optical densities of faecal samples were significantly greater than background readings, supporting the hypothesis that the assay was effective in quantifying faecal lactoferrin (T13, 115 = 11.99, p < 0.0005, n = 115). Lactoferrin concentrations of faecal and serum samples, taken from the same animals on the same day, were significantly different (T21 = 2.49, p = 0.022) and did not correlate (r = 0.069, p = 0.767).


Pathogens ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 243
Author(s):  
Julieta Rousseau ◽  
Mónia Nakamura ◽  
Helena Rio-Maior ◽  
Francisco Álvares ◽  
Rémi Choquet ◽  
...  

Sarcoptic mange is globally enzootic, and non-invasive methods with high diagnostic specificity for its surveillance in wildlife are lacking. We describe the molecular detection of Sarcoptes scabiei in non-invasively collected faecal samples, targeting the 16S rDNA gene. We applied this method to 843 Iberian wolf Canis lupus signatus faecal samples collected in north-western Portugal (2006–2018). We further integrated this with serological data (61 samples from wolf and 20 from red fox Vulpes vulpes, 1997–2019) in multi-event capture–recapture models. The mean predicted prevalence by the molecular analysis of wolf faecal samples from 2006–2018 was 7.2% (CI95 5.0–9.4%; range: 2.6–11.7%), highest in 2009. The mean predicted seroprevalence in wolves was 24.5% (CI95 18.5–30.6%; range: 13.0–55.0%), peaking in 2006–2009. Multi-event capture–recapture models estimated 100% diagnostic specificity and moderate diagnostic sensitivity (30.0%, CI95 14.0–53.0%) for the molecular method. Mange-infected individually identified wolves showed a tendency for higher mortality versus uninfected wolves (ΔMortality 0.150, CI95 −0.165–0.458). Long-term serology data highlights the endemicity of sarcoptic mange in wild canids but uncovers multi-year epidemics. This study developed and evaluated a novel method for surveying sarcoptic mange in wildlife populations by the molecular detection of S. scabiei in faecal samples, which stands out for its high specificity and non-invasive character.


2021 ◽  
Author(s):  
Shiny Lizia Manohar ◽  
Hemamalini Arambakkam Janardhanam ◽  
Latha Ravichandran

Abstract IntroductionObesity is a major threat to the health and development of children. While hormones in serum samples have been extensively investigated in childhood obesity research, our study attempts to explore salivary metabolic hormones as biomarkers of childhood obesity.MethodsObese (n=83) and non-obese (n=83) school children aged 6-11 years were recruited as cases and controls, respectively, for this case-control study. Salivary concentrations of the hormones, Ghrelin, Leptin, Adiponectin and Insulin were assessed and compared between obese and non-obese children to evaluate their role in childhood obesity. Results The mean ghrelin (1.69 ± 0.38 pg/ml) and adiponectin (95.53 ± 64.92 ng/ml) levels of the non-obese children were higher than the mean values of the obese, while the mean values of the hormones leptin (265.95 ± 83.16 pg/ml) and insulin (224.56 ± 184.86 pg/ml) were found to be higher among the obese. Higher levels of the hormones insulin (OR: 1.09; 95% CI: 1.05-1.12) and leptin (OR: 1.03; 95% CI: 1.02-1.04) posed higher risks for childhood obesity, followed by adiponectin, which was found to be lower among the cases (OR: 0.99; 95% CI: 0.99995-0.99998) which increased the risk of childhood obesity.ConclusionThis study demonstrates the efficacy of saliva as a promising non-invasive tool in childhood obesity research and highlights the significance of appetite regulatory hormones as biomarkers of childhood obesity.


Author(s):  
R.M. Sarode ◽  
A. Das ◽  
Y. Bhardwaj ◽  
A.K. Sharma

Background: Captive Indian leopards are exposed to different kind of stresses part of which can be alleviated through supplementation of carotenoids. However, invasive monitoring of antioxidants is often pro-act as a stressor itself. Hence, use of non-invasive monitoring of antioxidants and stress would be desirable. However, before recommendation, such non-invasive procedures must be compared with conventional ones. Hence, this experiment was designed to compare the cortisol and total antioxidants (TAA) status measured either in serum or faecal samples. Methods: Three diets were tested on twelve adult leopards (7 M and 5 F, BW ranging from 45-63 kg) in an experiment based on replicated Latin square design comprising of three treatments, three periods and three sequences and four animals in each sequence. The ratio of buffalo meat: chicken carcass was 100:0, 90:10 and 80:10 in groups CON, GI and GII, respectively. Result: Intake and absolute quantity of carotenoids absorbed increased (P less than 0.01) with increased level of chicken carcass; however, efficiency of absorption was lower (P less than 0.05) in GII as compared to other two groups. Faecal concentrations of cortisol decreased (P less than 0.0001) and TAA increased (P less than 0.0001) with increased level of carotenoids in the diet. Irrespective of the dietary treatments, concentration of cortisol was lower (P less than 0.0001) and TAA was higher in faecal as compared to serum samples. Regression analysis revealed positive relationship between serum and faecal sample assay with respect to both TAA and cortisol. Thus, faecal samples could be used to monitor cortisol and antioxidant status in Indian leopards. Assay of faecal samples indicated that replacement of buffalo meat with chicken at 20% in the diet improved the antioxidants and alleviated stress in captive leopards.


1996 ◽  
Vol 76 (06) ◽  
pp. 0925-0931 ◽  
Author(s):  
John F Carroll ◽  
Keith A Moskowitz ◽  
Niloo M Edwards ◽  
Thomas J Hickey ◽  
Eric A Rose ◽  
...  

SummaryTwenty-one cardiothoracic surgical patients have been treated with fibrin as a topical hemostatic/sealing agent, prepared from bovine fibrinogen clotted with bovine thrombin. Serum samples have been collected before treatment with fibrin and postoperatively between 1 and 9 days, 3 and 12 weeks, and 6 and 8 months. The titers of anti-bovine fibrinogen antibodies, measured by ELISA specific for immunoglobulins IgG or IgM, increased to maximal values after about 8 or 6 weeks, respectively. After 8 months, IgG titers were on average 20-fold lower than the mean maximal value, while IgM titers returned to the normal range. IgG was the predominant anti-bovine fibrinogen immunoglobulin as documented by ELISA, affinity chromatography and electrophoresis. Anti-bovine fibrinogen antibodies present in patients reacted readily with bovine fibrinogen, but did not cross-react with human fibrinogen as measured by ELISA or by immunoelectrophoresis. A significant amount of antibodies against bovine thrombin and factor V has been found, many cross-reacting with the human counterparts. No hemorrhagic or thrombotic complications, or clinically significant allergic reactions, occurred in any patient, in spite of antibody presence against some bovine and human coagulation factors. The treatment of patients with bovine fibrin, without induction of immunologic response against human fibrinogen, appeared to be an effective topical hemostatic/sealing measure.


1992 ◽  
Vol 67 (05) ◽  
pp. 507-509 ◽  
Author(s):  
John Gibson ◽  
Margaret Nelson ◽  
Ross Brown ◽  
Hatem Salem ◽  
Harry Kronenberg

SummaryIn order to investigate the possibility that autoantibodies to thrombomodulin (TM) may exist in patients with the lupus anticoagulant (LA) and perhaps be implicated in the pathogenesis of recurrent thrombosis seen in such patients, we developed an enzyme-immunoassay to screen serum samples for anti-human TM activity. The major technical problem encountered in developing this assay was to reduce the non-specific binding of serum components from both the LA positive and the negative population. Considerable reduction of non-specific binding was achieved by use of a phosphate/citrate buffer at pH 8.0 and the use of an optimal sample dilution of 1/40. In addition, samples were always tested in parallel in blank wells and results are expressed as an OD ratio. Samples from 113 patients with the LA were assayed and compared to 78 patients referred for LA testing but found to be negative. The mean OD values for the LA positive patients (± SD) was 1.36 (0.44) with a range of 0.78-2.57. This was virtually identical to the values for the LA negative population (1.38 ± 0.40, range 0.76-2.77). The results of this study indicate that there is no evidence for the presence of a significant autoantibody activity to TM in patients with the LA when compared to LA negative patients. If such autoantibodies do exist their frequency must be quite low.


2020 ◽  
Vol 2 (Supplement_3) ◽  
pp. ii21-ii21
Author(s):  
Shumpei Onishi ◽  
Fumiyuki Yamasaki ◽  
Motoki Takano ◽  
Ushio Yonezawa ◽  
Kazuhiko Sugiyama ◽  
...  

Abstract Objective: Glioblastoma (GBM) and Primary Central Nervous System Lymphoma (PCNSL) are common intracranial malignant tumors. They sometimes present similar radiological findings and diagnoses could be difficult without surgical biopsy. For improving the current management, development of non-invasive biomarkers are desired. In this study, we explored the differently expressed circulating small noncoding RNA (sncRNA) in serum for specific diagnostic tool of GBM and PCNSL. Material & Methods: Serum samples were obtained from three groups: 1) GBM patients (N=26), 2) PCNSL patients (N=14) 3) healthy control (N=114). The total small RNAs were extracted from serum. The whole expression profiles of serum sncRNAs were measured using Next-Generation Sequencing System. We analyzed serum levels of sncRNAs (15–55 nt) in each serum samples. The difference of sncRNAs expression profile among three groups were compared. Data analysis was performed by logistic regression analysis followed by leave-one-out cross-validation (LOOCV). The accuracy of diagnostic models of sncRNAs combination were evaluated by receiver operating characteristic (ROC) analysis. Results: We created the combination models using three sncRNA in each models based on the logistic regression analysis. The model 1 (based on sncRNA-X1, X2 and X3) enabled to differentiate GBM patients form healthy control with a sensitivity of 92.3% and a specificity of 99.2% (AUC: 0.993). The model 2 (based on sncRNA-Y1, Y2 and Y3) enabled to differentiate PCNSL patients form healthy control with a sensitivity of 100% and a specificity of 93.9% (AUC: 0.984). The model 3 (based on sncRNA-Z1, Z2 and Z3) enabled to differentiate GBM patients form PCNSL patients with a sensitivity of 92.3% and a specificity of 78.6% (AUC: 0.920). Conclusion: We found three diagnostic models of serum sncRNAs as non-invasive biomarkers potentially useful for detection of GBM and PCNSL from healthy control, and for differentiation GBM from PCNSL.


Author(s):  
Patrick Veit-Haibach ◽  
Martin W. Huellner ◽  
Martin Banyai ◽  
Sebastian Mafeld ◽  
Johannes Heverhagen ◽  
...  

Abstract Objectives The purpose of this study was the assessment of volumetric CT perfusion (CTP) of the lower leg musculature in patients with symptomatic peripheral arterial disease (PAD) before and after interventional revascularisation. Methods Twenty-nine consecutive patients with symptomatic PAD of the lower extremities requiring interventional revascularisation were assessed prospectively. All patients underwent a CTP scan of the lower leg, and hemodynamic and angiographic assessment, before and after intervention. Ankle-brachial pressure index (ABI) was determined. CTP parameters were calculated with a perfusion software, acting on a no outflow assumption. A sequential two-compartment model was used. Differences in CTP parameters were assessed with non-parametric tests. Results The cohort consisted of 24 subjects with an occlusion, and five with a high-grade stenosis. The mean blood flow before/after (BFpre and BFpost, respectively) was 7.42 ± 2.66 and 10.95 ± 6.64 ml/100 ml*min−1. The mean blood volume before/after (BVpre and BVpost, respectively) was 0.71 ± 0.35 and 1.25 ± 1.07 ml/100 ml. BFpost and BVpost were significantly higher than BFpre and BVpre in the treated limb (p = 0.003 and 0.02, respectively), but not in the untreated limb (p = 0.641 and 0.719, respectively). Conclusions CTP seems feasible for assessing hemodynamic differences in calf muscles before and after revascularisation in patients with symptomatic PAD. We could show that CTP parameters BF and BV are significantly increased after revascularisation of the symptomatic limb. In the future, this quantitative method might serve as a non-invasive method for surveillance and therapy control of patients with peripheral arterial disease. Key Points • CTP imaging of the lower limb in patients with symptomatic PAD seems feasible for assessing hemodynamic differences before and after revascularisation in PAD patients. • This quantitative method might serve as a non-invasive method, for surveillance and therapy control of patients with PAD.


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