scholarly journals Lactoferrin quantification in cattle faeces by ELISA

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).


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.


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.


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.


Diagnostics ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1345
Author(s):  
Mahathir Humaidi ◽  
Wei Ping Tien ◽  
Grace Yap ◽  
Choon Rong Chua ◽  
Lee Ching Ng

Dengue diagnosis is largely dependent on clinical symptoms and routinely confirmed with laboratory detection of dengue virus in patient serum samples collected via phlebotomy. This presents a challenge to patients not amenable to venipuncture. Non-invasive methods of dengue diagnosis have the potential to enhance the current dengue detection algorithm. In this study, samples from dengue infected patients were collected between January 2012 until September 2012 and September 2013 until December 2013 in two different setups. Panel A samples (blood, urine, and saliva) were collected daily when the 39 patients were hospitalised and during their follow-up visits while Panel B samples (saliva) were collected from 23 patients during the acute stage of dengue. Using DENV PCR on Panel A, from day 2 to day 4 post fever onset, serum showed the best overall positivity followed by saliva and urine (100%/82.1%/67.9%). From day 5 until day 10 post fever onset, serum and urine had similar positivity (67.4%/61.2%), followed by saliva (51.3%). Beyond day 10 post fever onset, DENV was undetectable in sera, but urine and saliva showed 56.8% and 28.6% positivity, respectively. DENV in urine was detectable up until 32 days post fever. Panel B results showed overall sensitivity of 32.4%/36% (RNA/NS1) for DENV detection in saliva. Our results suggest that the urine-based detection method is useful especially for late dengue detection, where DENV is undetected in sera but still detectable in urine. This provides a potential tool for the physician to pick up new cases in an area where there is ongoing dengue transmission and subsequently prompt for intensified vector control activities.


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.


2017 ◽  
Vol 57 (1) ◽  
pp. 8 ◽  
Author(s):  
Andra Kurnianto ◽  
Herman Bermawi ◽  
Afifa Darmawanti ◽  
Erial Bahar

Background The gold standard for diagnosis of neonatal jaundice is total serum bilirubin (TSB) measurement. This method, however, is invasive, painful, and costly in terms of workload, time, and money. Moreover, repeated blood sampling may lead to significant blood loss, which is of particular concern in preterm infants. To overcome these drawbacks, non-invasive methods of bilirubin measurement have been proposed. Transcutaneous bilirubinometry (TcB) determines the yellowness of the subcutaneous tissue of a newborn infant by measuring the difference between optical densities for light in the blue and green wavelength regions.Objective To evaluate the accuracy of transcutaneous bilirubinometry for estimating TSB levels in neonatal jaundice.Methods Subjects were infants aged < 28 days with jaundice who had never been treated with phototherapy or exchange transfusion. The study was done from February to July 2016 in Mohammad Hoesin Hospital. Subjects underwent transcutaneous bilirubin (TcB) and TSB assays, with a maximum interval of 15 minutes between tests.Results One hundred fifty patients were included in this study. The TcB values > 5 mg/dL were correlated to TSB > 5 mg/dL, with 100% sensitivity and 83.3% specificity. This cut-off point was obtained from a receiver-operator characteristic (ROC) curve with AUC 99.3% (95%CI 97.9 to 100%; P< 0.001).The correlation coefficients (r) for TSB and TcB measurements on the forehead were 0.897 (P<0.001).Conclusion Transcutaneous bilirubinometry can be used to accurately estimate TSB levels in neonatal jaundice, and may be useful in clinical practice as a non-invasive method to reduce blood sampling.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Imteyaz Ahmad Khan ◽  
Safoora Rashid ◽  
Nidhi Singh ◽  
Sumaira Rashid ◽  
Vishwajeet Singh ◽  
...  

AbstractEarly-stage diagnosis of pancreatic ductal adenocarcinoma (PDAC) is difficult due to non-specific symptoms. Circulating miRNAs in body fluids have been emerging as potential non-invasive biomarkers for diagnosis of many cancers. Thus, this study aimed to assess a panel of miRNAs for their ability to differentiate PDAC from chronic pancreatitis (CP), a benign inflammatory condition of the pancreas. Next-generation sequencing was performed to identify miRNAs present in 60 FFPE tissue samples (27 PDAC, 23 CP and 10 normal pancreatic tissues). Four up-regulated miRNAs (miR-215-5p, miR-122-5p, miR-192-5p, and miR-181a-2-3p) and four down-regulated miRNAs (miR-30b-5p, miR-216b-5p, miR-320b, and miR-214-5p) in PDAC compared to CP were selected based on next-generation sequencing results. The levels of these 8 differentially expressed miRNAs were measured by qRT-PCR in 125 serum samples (50 PDAC, 50 CP, and 25 healthy controls (HC)). The results showed significant upregulation of miR-215-5p, miR-122-5p, and miR-192-5p in PDAC serum samples. In contrast, levels of miR-30b-5p and miR-320b were significantly lower in PDAC as compared to CP and HC. ROC analysis showed that these 5 miRNAs can distinguish PDAC from both CP and HC. Hence, this panel can serve as a non-invasive biomarker for the early detection of PDAC.


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