An Enzyme-Linked Immunosorbent Assay for Cortisol in the Saliva of Man and Domestic Farm Animals

Cooper, T.R.; Trunkfield, H.R.; Zanella, Adroaldo José; Booth, W. D.

doi:10.1677/joe.0.123r013

Cited by 76 publications

(41 citation statements)

References 10 publications

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“…avoiding the variations due to blood sampling or restraint stress) some authors have proposed detecting it in different biological materials, such as saliva (Cooper et al, 1989), milk (Shutt and Fell, 1985;Verkerk et al, 1998), faeces and urine (Hay and Mormede, 1998). The level of the cortisol is in some equilibrium with the different compartments and the sampling of these materials is, generally, considered less invasive.…”

Section: Cortisolmentioning

confidence: 99%

Some physiological and biochemical methods for acute and chronic stress evaluationin dairy cows

Trevisi

Bertoni

2009

Italian Journal of Animal Science

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Stress factors are so numerous and so diverse in their strength and duration that the consequences on animal welfare can be quite varied. The first important distinction concerns the characterization of acute and chronic stress conditions. Acute stress is a short-lived negative situation that allows a quick and quite complete recovery of the physiological balance (adaptation), while chronic stress is a long lasting condition from which the subject cannot fully recover (maladaptation). In the latter case, the direct effects of the stress factors (heat, low energy, anxiety, suffering etc.), as well as the indirect ones (changes occurring at endocrinological, immune system or function level) can be responsible for pre-pathological or pathological consequences which reduce animal welfare. To evaluate the possible chronic stress conditions in single animals or on a farm (in particular a farm of dairy cows), some parameters of the direct or indirect effects can be utilised. They are physiological (mainly hormone changes: cortisol, β-endorphin), behavioural (depression), biochemical (metabolites, acute phase proteins, glycated proteins etc.), as well as performance parameters (growing rate, milk yield, fertility, etc.). Special attention has been paid to the interpretation of cortisol levels and to its changes after an ACTH challenge. Despite fervent efforts, well established and accepted indices of chronic stress (distress) are currently lacking; but without this objective evaluation, the assessment of animal welfare and, therefore, the optimization of the livestock production, could prove more difficult.

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Section: Cortisolmentioning

confidence: 99%

Some physiological and biochemical methods for acute and chronic stress evaluationin dairy cows

Trevisi

Bertoni

2009

Italian Journal of Animal Science

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“…Although traditionally cortisol concentrations have been determined in plasma or serum, their determination in other fluids or organic tissues (saliva, milk, muscle, urine, faeces and hair) can be of interest in animal welfare studies of different species (Cooper et al, 1989;Fritsche and Steinhart, 1998;Verkerk et al, 1998;Cirimele et al, 1999;Fritsche et al, 1999;Antignac et al, 2000;Morrow, 2000;Palme et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

Validation of an EIA [enzyme immunoassay] technique for the determination of salivary cortisol in cattle

Pérez

Laita

Portal

et al. 2004

Span. j. agric. res.

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This work involves the development and validation of an enzyme immunoassay technique (EIA) for the measurement of the cortisol concentration in cattle saliva. Saliva samples present several advantages over plasma samples in animal welfare studies. Saliva collection avoids venipuncture as a stress factor. Also, saliva components do not affect EIA as plasma components do. At present, there is no validated commercial method for saliva cortisol determination in cattle. Commercially available radioimmunoassay kits for human plasma (detection range: 10-100 ng ml -1 ) are not sensitive enough for animals with low concentrations of salivary cortisol (< 4 ng ml -1 ). Thus, EIA is the method of choice in cattle. Sensitivity, specificity, precision and accuracy EIA tests showed this method to be suitable and reliable. The detection limit was found to be 0.024 ng ml -1 , representing an improvement on previously described techniques. Intra-assay and inter-assay variation coefficients were 1.47-7.30% and 2.40-9.78%, respectively. The recovery rates for cortisol added to saliva samples were 91.36-126.5%. Parallelism tests showed that saliva cortisol levels can be determined in cattle samples without extraction. The correlation between saliva and plasma cortisol was positive (r = 0.75) and the saliva/plasma cortisol ratio was around 10%. Therefore, saliva samples are a suitable alternative to plasma samples in bovine HPA (hypothalamic-pituitary-adrenal) axis evaluation.Key words: immunoassay, HPA axis, cow, stress. ResumenValidación de una técnica de EIA para la determinación de cortisol en saliva de ganado bovino En este trabajo se desarrolla y valida una técnica de enzimoinmunoensayo (EIA) para la determinación directa de cortisol en muestras de saliva de ganado vacuno. La saliva como muestra tiene gran importancia en los estudios de bienestar animal, ya que presenta ventajas respecto al plasma, como son el menor estrés al que se somete a los animales para su obtención y la menor interferencia que producen los componentes de la saliva en la determinación directa en el EIA. Hoy en día no existe en el mercado ningún método validado para la determinación de cortisol en saliva en la especie bovina. Las técnicas de radioinmunoensayo comercializadas para plasma de la especie humana (rango de detección de 10 a 100 ng ml -1 ) no son suficientemente sensibles para el ganado bovino, que presenta concentraciones de cortisol salival muy reducidas (< 4 ng ml -1 ), por ello no es posible su adaptación para esta especie, siendo los métodos de EIA los de elección. El límite de detección de la técnica EIA fue de 0,024 ng ml -1 , mejorando la sensibilidad de técnicas descritas previamente; los coeficientes de variación intra e interensayo fueron de 1,47-7,30% y 2,40-9,78%, respectivamente. La recuperación de cortisol añadido a muestras de saliva fue de 91,36-126,5%. Las pruebas de paralelismo demostraron que es posible la determinación directa de muestras de saliva sin necesidad de extracción previa. La correlación existente entre las...

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“…or salivary cortisol has been used as a biomarker of stress (Cooper et al 1989, Beerda et al 1996, Whitten 1998, Dreschel & Granger 2005, Hellhammer et al 2009, Montanha et al 2009. It has been shown that salivary and plasma levels of cortisol are correlated (Beerda et al 1996) and the measure of the basal cortisol levels in domestic cats using saliva samples was previously studied and validated (Siegford et al 2003).…”

Section: Introductionmentioning

confidence: 99%

Effect of a synthetic analogue of the feline facial pheromone on salivary cortisol levels in the domestic cat

et al. 2017

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This study aimed to evaluate the ability of a saliva collection device (Salivette®) to measure cortisol levels in saliva samples of domestic cats and to assess the effect of a synthetic analogue of the feline facial pheromone fraction F3 (Feliway®) on cortisol levels. A total of 28 domestic cats from a private high-quality sanctuary were sampled before exposure to the facial pheromone and after 35 days of exposure. Two pheromone devices were placed in the area where the animals ate to guarantee the exposure of all cats. The collecting device yielded a sufficient volume of saliva (≥0.20mL) to allow cortisol measurement. Cortisol measurements ranged from 0.02g/dL to 0.16μg/dL, with a difference between before (42.1%) and after (62.6%) exposure to the pheromone (F=3.2351; p≤0.0002). No difference in cortisol levels was observed between before (x =0.078μg/dL) and after (x =0.066μg/dL) (t=1.79; p=0.08) exposure. However, salivary cortisol levels decreased in 75% (21/28) of the cats after exposure (x 2=12.07; p=0.0005), suggesting that the animals have different susceptibilities to the pheromone or that they spent different lengths of time in the area where the pheromone devices were installed.

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An Enzyme-Linked Immunosorbent Assay for Cortisol in the Saliva of Man and Domestic Farm Animals

Cited by 76 publications

References 10 publications

Some physiological and biochemical methods for acute and chronic stress evaluationin dairy cows

Some physiological and biochemical methods for acute and chronic stress evaluationin dairy cows

Validation of an EIA [enzyme immunoassay] technique for the determination of salivary cortisol in cattle

Effect of a synthetic analogue of the feline facial pheromone on salivary cortisol levels in the domestic cat

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