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WO2004014253A1 - Systeme non invasif permettant de controler l'etat physiologique ou sanitaire d'animaux par prelevement d'un echantillon de salive - Google Patents

Systeme non invasif permettant de controler l'etat physiologique ou sanitaire d'animaux par prelevement d'un echantillon de salive Download PDF

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Publication number
WO2004014253A1
WO2004014253A1 PCT/GB2003/003521 GB0303521W WO2004014253A1 WO 2004014253 A1 WO2004014253 A1 WO 2004014253A1 GB 0303521 W GB0303521 W GB 0303521W WO 2004014253 A1 WO2004014253 A1 WO 2004014253A1
Authority
WO
WIPO (PCT)
Prior art keywords
saliva
animal
ovulation
sampling
sample
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/GB2003/003521
Other languages
English (en)
Inventor
Toby Mottram
Paul Berry
Amanda Pickard
John P. Hart
Roy Pemberton
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
SILSOE RESEARCH INSTITUTE
Original Assignee
SILSOE RESEARCH INSTITUTE
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by SILSOE RESEARCH INSTITUTE filed Critical SILSOE RESEARCH INSTITUTE
Priority to AU2003283178A priority Critical patent/AU2003283178A1/en
Publication of WO2004014253A1 publication Critical patent/WO2004014253A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61D—VETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
    • A61D17/00—Devices for indicating trouble during labour of animals ; Methods or instruments for detecting pregnancy-related states of animals
    • A61D17/002—Devices for indicating trouble during labour of animals ; Methods or instruments for detecting pregnancy-related states of animals for detecting period of heat of animals, i.e. for detecting oestrus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B10/00—Instruments for taking body samples for diagnostic purposes; Other methods or instruments for diagnosis, e.g. for vaccination diagnosis, sex determination or ovulation-period determination; Throat striking implements
    • A61B10/0012—Ovulation-period determination
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B10/00—Instruments for taking body samples for diagnostic purposes; Other methods or instruments for diagnosis, e.g. for vaccination diagnosis, sex determination or ovulation-period determination; Throat striking implements
    • A61B10/0045—Devices for taking samples of body liquids
    • A61B10/0051—Devices for taking samples of body liquids for taking saliva or sputum samples

Definitions

  • the present invention concerns a non-invasive system for monitoring one or more aspects of the physiological or health status of animals.
  • a physiological aspect especially addressed relates to determination of ovulation and notably provides a method and apparatus suitable for determining the timing of ovulation in pigs.
  • the method and apparatus is particularly useful to enable, for example, insemination of 10 sows to be carried out at the right time to optimise fertilisation to improve both farrowing rate and litter size.
  • the apparatus may also be configured to enable the health of the animal to be monitored.
  • a more accurate system for predicting the timing of ovulation would overcome these problems, enabling fertilisation to be achieved more reliably from a single insemination event.
  • biomarkers for health status monitoring may include known pathogen molecules or other molecules or metabolites associated with ill health such as, for example, IgF, antibodies, corticosteroids or acute phase proteins such as NAGase, haptoglobin or serum amyloid A.
  • the present invention provides a non-invasive system for monitoring one or more aspects of the physiological or health status of animals which comprises:
  • a biosensor or conductivity probe operatively associated with the saliva sampling device to sense the level of a hormone or other biomarker in the saliva or the ionic balance of the saliva to provide corresponding signals to a storage means or display.
  • Any changes in level of the biomarker or in ionic balance/ ionic composition of the saliva over time may be monitored and used as a diagnostic tool.
  • the system may be programmed to carry out analysis automatically and suitably to provide a diagnosis or prediction automatically.
  • the automated sampling device suitably has a saliva sampling tube and a suction device, eg pump, to draw a sample of saliva therein for analysis. This preferably is activated automatically when an animal presents itself to the sampling device.
  • the present invention provides a method of determining the timing of ovulation in a pig or other animal which entails taking samples of saliva from the animal at intervals and analysing the samples for level of key hormones, suitably 17 ⁇ -estradiol (E 2 ) and / or LH, to monitor for changes indicative of the timing of ovulation.
  • level of key hormones suitably 17 ⁇ -estradiol (E 2 ) and / or LH
  • the present invention also provides an automated system for determining the timing of ovulation in a mammal and which comprises an automatic saliva sampling apparatus to take samples of saliva from individual animals at intervals, in combination with a biosensor to sense the concentrations of the one or more key hormones in the saliva.
  • the present invention provides a novel solution to the problem of reliably and accurately detecting oestrus in dry sows.
  • the system identifies the pattern of diurnal concentrations of key hormones expressed in saliva, to provide a non-invasive automatic method of , sampling saliva using a low cost biosensor capable of detecting concentrations of the hormones down to levels of picogrammes per ml of saliva.
  • saliva provides a good source for non-invasive sampling to monitor steroid hormone levels of the ovarian cycle.
  • the steroid hormone 17 ⁇ -estradiol (E 2 ) in saliva has been established to be a good analyte for ovarian cycle analysis and ovulation prediction in sows.
  • Saliva provides a good source for non-invasive sampling to monitor other biomarkers than ovulation cycle stage biomarkers and can be used to good effect for monitoring for preclinical disease in pigs.
  • Figure 1 is a schematic diagram ot one preferred embodiment of the ovulation detection system
  • Figure 2 is a perspective view of the detection system mounted in a single animal occupancy stall for sampling one farm animal, e.g. a single sow, at a time;
  • Figure 3 is a schematic diagram of a prototype of the ovulation detection system where the saliva sampler is integrated with a drinking water supply unit;
  • Figure 4 is a schematic diagram of a suitable ELISA for assaying the saliva samples for the key hormone 17 ⁇ -estradiol (E 2 );
  • Figure 5 is a schematic diagram corresponding to the Figure 5 ELISA as adapted to operation on a screen-printed carbon electrode (SPCE); and
  • Figure 6 is a schematic diagram of a further preferred embodiment of the ovulation detection system with multiple sample pots and an associated manifold to deliver the sampled saliva filling a first pot and then each adjacent pot in turn.
  • the system comprises a station 1 that is adapted for automated taking of samples of sow saliva.
  • the station was actually adapted to supply water when the sampling line 5 inlet was mouthed.
  • a switch was provided that the pig had to nuzzle (eg. in the drinker 6) that would trigger the dispensing of water from a supply 12 .
  • nuzzle eg. in the drinker 6
  • a small suction pump 7 was arranged to take a saliva sample at sample chamber 13. This, however, proved not to be the best way of ensuring salivation of the pig and taking of samples.
  • the best solution to ensuring effective regular saliva sampling was the use of a chewable and suitably digestible (and even nutritious/ tasty) and continuously replenishable sampling line 5 as shown in Figure 1. This is preferably pliable to enhance chewability but robust enough to perform its role. It may be reinforced suitably in zones/ bands with nylon or other tough plastics to ensure that it remains sufficiently intact for sampling to be carried out.
  • the saliva sampling line 5 is reeled around a motorised reel 21 to provide the desired replenishing of the exposed line 5 following each sample-taking session.
  • a cutting head 22 is provided to cut off the previously chewed free end of the line 5 and at the same time perforates the free end suitably providing multiple lateral apertures 25 to optimise saliva uptake and avoid any occlusion of the end that may occur in the cutting step.
  • a flushing water line 23 also suitably feeds into the apparatus at this stage.
  • the sampling line 5 selectively couples with one of a plurality of sample pots 24 in a rotating carousel 25 so that the sample from one sampling session is isolated from samples from other sessions and may thus be assayed independently and with regard to the identified animal involved and time and date of the sample.
  • Suction is applied by pump 7 once the transponder and linked processor has identified the animal presenting itself to the sampling station 1.
  • a pressure sensor 26 monitors the vacuum pressure to maintain it within appropriate levels.
  • a liquid level sensor 27 at the pot 24 determines when the pot is full and will trigger termination of suction.
  • the carousel 25 will then automatically switch the pot 24, moving it away for the assay to be carried out and replacing it with an empty pot 24 for the next session.
  • the flushing line 23 will be opened to flush the sample line 5 and the line 5 will be cut and perforated ready for the next animal.
  • the station 1 is housed in an individual stall 10 with baffles 11 to prevent access of neighbouring animals to the sampler 5 from the sides. An individual animal must approach from the front entrance to the stall 10.
  • a transponder antenna 2 built into the station is responsive to signals from a tag 3 worn by each animal in order to identify each individual pig and ensure that the samples from each are handled and logged solely in relation to that particular animal.
  • Each saliva sample may then be analysed suitably in situ within the station 1 for oestrogens and other hormones by a biosensor.
  • a preferred biosensor is as illustrated in Figure 5, comprising a screen printed carbon electrode 20.
  • a herd management database records the oestrogen concentrations. When a preprogrammed scenario is achieved (e.g. oestrogen declines to 50% of its maximum value or to within mean +/- 2SD of the preceding baseline value), the stockman is suitably alerted of the impending ovulation.
  • the operating principle of the biosensor that we have adopted is an immunoassay with an electrochemical signal transduced using the screen-printed electrodes 5.
  • the screen-printed carbon electrode electrochemical biosensor 5 for the determination of 17 ⁇ -estradiol (E 2 17 ⁇ -estradiol (E 2 ) or LH.
  • This biosensor is capable of monitoring oestradiol/LH levels in pig saliva at pg/ml concentrations.
  • 96-well plates were coated with rabbit anti-mouse IgG (Sigma) in carbonate buffer, pH9.6, followed by mouse anti-E 2 mAb in PBS. E 2 dilutions were prepared in ethanol. Protocol was as described in Fig.4, using a competitive format with alkaline-phosphatase (AP)-labelled E 2 .
  • AP alkaline-phosphatase
  • SPCEs Screen-printed carbon electrodes
  • a working electrode was fabricated using D14 carbon ink, printed onto a polyester substrate. This was overlaid with a dielectric layer to leave an exposed circular working area of approx. 9 mm 2 . The working area was then coated with rlgG and mAb, as used for ELISA plates.
  • Electrochemical Biosensor Assay The competitive electrochemical assay for E 2 was performed using the protocol described in Fig.5 The final measurement step as performed by placing the biosensors in a prototype thin-layer electrochemical cell and performing a chronoamperometric measurement, recording the current response at 10s post- application of the optimised potential. In the results obtained a highly sensitive assay was established. This was made even more sensitive through use of increasing amounts of r IgG .
  • the present invention provides a radical new approach to improve fertility management in the pig sector.
  • the present invention predicts ovulation in sows. It can also be used to predict ovulation in gilts as well as for detecting pregnancy automatically.
  • Oestrous behaviour is initiated by the secretion of oestrogen by the developing ovarian follicles.
  • Oestrogen injections into ovariectomised gilts initiates oestrous behaviour and higher oestrogen doses result in an increased duration of oestrus.
  • Oestrogen secretion also stimulates a surge in the release of luteinizing hormone (LH) from the pituitary gland, which is essential for the ovulation of the matured oocytes.
  • LH luteinizing hormone
  • Plasma oestrogen concentrations relate predictably to the preovulatory LH surge and the time of ovulation.
  • Our automatic system based on the measurement of salivary oestrogen hormone excretion using biosensor technology provides an objective method for predicting ovulation in a given individual. Hence the optimal time for insemination can be indicated to the stockman and maximum fertility achieved.
  • a major advantage of using a biosensor is that the biological elements of the immunoassays are combined with an electrode transducer into a single device capable of generating an electrical signal in response to a given concentration of the analyte of interest.
  • screen-printing technology it is possible to mass-produce the transducer element and to fabricate individual sensors at low cost, enabling them to be single-use, disposable devices.
  • Devices employing electrochemical measurements offer an advantage over optical devices since they are independent of interference from contaminants which absorb UV-visible radiation.
  • the ELISA assays operate effectively using the saliva samples and the immunology and signal transduction performed well using the electrochemical approach.
  • a row of ten sampling pots is provided beneath a selectively gated manifold 30 to enable saliva drawn up the sampling tube 5 to be deposited firstly in the first sample pot 24a and then, when a sensor detects that the first pot is filled to a chosen level, the next 24b and so on until the required sample volume has been taken and distributed substantially evenly across the pots 24.
  • the system has a biosensor 20 mounted to each sample pot 24 or automatically deliverable to one or more pots. Indeed it is preferred that several different biosensors are provided so that a profile of a range of different biomarkers may be monitored. For example when" monitoring for sow ovulation both biomarkers, 17 ⁇ - estradiol (E 2 ) and LH, may be monitored for and suitably the automated system is programmed to deploy/ activate the biosensor for 17 ⁇ -estradiol (E 2 ) first to observe its rise and then to deploy/ activate the LH biosensor. Indeed programming of the system for selective deployment and/ or activation of different biomarker biosensors in a manner that takes into account expected and/or actual changes in the levels of the biomarkers over time provides a very valuable tool for analysis and prediction.
  • biosensors are suitably co-mounted on a single substrate, eg silicon wafer, but if to be deployed selectively/ independently are suitably each mounted on their own respective support.
  • the biosensors may be reusable if thoroughly cleaned but generally, are s'uitably designed to be disposable.
  • the pig recognition sensor operates to start the sampling sequence as in the earlier embodiment but in a refinement to the preliminary steps the drive rollers/ feed mechanism 31 for advancing the sampling pipe 5 suitably is programmed to do so as an overture, alternately shortly advancing and retracting the pipe 5 to wave it at passing pigs to attract their attention.
  • a movement or contact sensor 32 suitably a vibration sensor, at or near the end of the pipe 5 will be activated to sense and signal to the system when the pig nuzzles/ mouths the pipe 5.
  • the vacuum pump 33 is automatically switched on, the sample level detector of the first sample pot 24a is activated and the saliva drawn into it through the open gate of the gated manifold 30.
  • the vacuum pump is deactivated automatically and the pipe 5 retracted, the chewed end of the pipe 5 is cut off by the tube cutter 34 and the pipe 5 then fed out again to the required length prior to the next session.
  • Washing of the pipe 5 is automatically carried out at the end of the cycle of operation , with opening of the water valves 35 , 36 to allow the water to pass through and rinse the manifold 30 throughout and rinse through the suction pipe 5.
  • the water valves are than shut/ switched over to allow passage of air from the pump 33, the air being to blown through the rinsed pipework to clear it of water and dry it out.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Biophysics (AREA)
  • Pregnancy & Childbirth (AREA)
  • Engineering & Computer Science (AREA)
  • Animal Husbandry (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

Un aspect de cette invention concerne un système non invasif permettant de contrôler un ou plusieurs aspects de l'état physiologique ou sanitaire d'animaux. Ce système comprend: une station (1) conçue pour recevoir un animal devant être examiné, laquelle station est doté d'un dispositif de prélèvement d'échantillon conçu pour prélever un échantillon de salive sur l'animal. Le système comprend également un biocapteur (20) associé de manière opérationnelle au dispositif de prélèvement d'échantillon de salive de manière à détecter le niveau d'une hormone ou d'un autre marqueur biologique dans la salive afin de fournir des signaux correspondants à un moyen de stockage ou à un dispositif d'affichage. Le dispositif de prélèvement d'échantillon est automatisé et il est doté d'un tube de prélèvement de salive (5) et d'un dispositif d'aspiration, par exemple, d'une pompe (7), permettant l'acheminement d'un échantillon de salive à l'intérieur du tube pour y être analysé. Le dispositif de prélèvement d'échantillon est programmé pour déclencher le processus susmentionné lorsqu'un animal se présente tout seul sur la station de prélèvement d'échantillon (1). Cette invention est particulièrement utile pour contrôler des maladies précliniques et pour prévoir la période d'ovulation chez des porcs d'élevage.
PCT/GB2003/003521 2002-08-13 2003-08-13 Systeme non invasif permettant de controler l'etat physiologique ou sanitaire d'animaux par prelevement d'un echantillon de salive Ceased WO2004014253A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2003283178A AU2003283178A1 (en) 2002-08-13 2003-08-13 Non-invasive system for monitoring physiological or health status of animals by sampling saliva

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB0218798A GB0218798D0 (en) 2002-08-13 2002-08-13 Improvements to determination of ovulation
GB0218798.7 2002-08-13

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GB (2) GB0218798D0 (fr)
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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103543036A (zh) * 2013-11-05 2014-01-29 山东省动物疫病预防与控制中心 便捷式猪唾液采样器
CN103558058A (zh) * 2013-11-05 2014-02-05 山东省动物疫病预防与控制中心 猪唾液采样装置
CN104546017A (zh) * 2014-12-26 2015-04-29 华南农业大学 一种动物唾液采集装置
EP2950720A4 (fr) * 2013-01-30 2016-10-26 Salimetrics Llc Procédés de détection d'estradiol salivaire et utilisations pour contrôler l'ovulation et la fertilité, et la sélection ou la synchronisation d'extraction et/ou de fertilisation d'ovule
WO2018062666A1 (fr) * 2016-09-27 2018-04-05 주식회사 토브스 Dispositif et procédé de test salivaire, et système et procédé de surveillance d'animal utilisant ceux-ci
WO2019156756A3 (fr) * 2018-02-07 2020-04-09 Idexx Laboratories, Inc. Appareil de collecte d'échantillon de cage d'animal
CN117426802A (zh) * 2023-12-20 2024-01-23 四川省电子产品监督检验所 一种智能化传染病自助检测装置

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009153813A1 (fr) * 2008-06-17 2009-12-23 Pieraldo Inaudi Appareil, système et procédé pour l'extraction de liquides organiques
DE102015007098A1 (de) 2015-06-08 2016-12-08 Boehringer Ingelheim Vetmedica Gmbh Aufnahmekörper und Verfahren zur Gewinnung einer Speichelprobe

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FR2784023A1 (fr) * 1998-10-02 2000-04-07 Michel Gleizes Dispositif de recipient pour la recuperation en vue d'analyse de crachats
CA2290898A1 (fr) * 1999-09-23 2001-03-23 Cleveland Blythe Sonde vaginale munie d'un detecteur ameliore, ainsi que sa methode d'utilisation

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DE3632303A1 (de) * 1985-09-23 1987-04-02 Sarstedt Kunststoff Verfahren zum gewinnen von menschlichem speichel
US5467778A (en) * 1992-08-21 1995-11-21 Unilever Patent Holdings B.V. Monitoring method
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CA2290898A1 (fr) * 1999-09-23 2001-03-23 Cleveland Blythe Sonde vaginale munie d'un detecteur ameliore, ainsi que sa methode d'utilisation

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Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2950720A4 (fr) * 2013-01-30 2016-10-26 Salimetrics Llc Procédés de détection d'estradiol salivaire et utilisations pour contrôler l'ovulation et la fertilité, et la sélection ou la synchronisation d'extraction et/ou de fertilisation d'ovule
CN103543036A (zh) * 2013-11-05 2014-01-29 山东省动物疫病预防与控制中心 便捷式猪唾液采样器
CN103558058A (zh) * 2013-11-05 2014-02-05 山东省动物疫病预防与控制中心 猪唾液采样装置
CN104546017A (zh) * 2014-12-26 2015-04-29 华南农业大学 一种动物唾液采集装置
WO2018062666A1 (fr) * 2016-09-27 2018-04-05 주식회사 토브스 Dispositif et procédé de test salivaire, et système et procédé de surveillance d'animal utilisant ceux-ci
JP2019534987A (ja) * 2016-09-27 2019-12-05 トゥブス 唾液検査装置及び方法とこれを用いた動物モニタリングシステム及び方法
US11439371B2 (en) 2016-09-27 2022-09-13 Wellness Farmer Co., Ltd. Saliva test device and method, and animal monitoring system and method using same
WO2019156756A3 (fr) * 2018-02-07 2020-04-09 Idexx Laboratories, Inc. Appareil de collecte d'échantillon de cage d'animal
US11974546B2 (en) 2018-02-07 2024-05-07 Idexx Laboratories, Inc. Animal cage-sample collection apparatus
CN117426802A (zh) * 2023-12-20 2024-01-23 四川省电子产品监督检验所 一种智能化传染病自助检测装置
CN117426802B (zh) * 2023-12-20 2024-03-19 四川省电子产品监督检验所 一种智能化传染病自助检测装置

Also Published As

Publication number Publication date
AU2003283178A1 (en) 2004-02-25
GB0319017D0 (en) 2003-09-17
GB2392244A (en) 2004-02-25
GB0218798D0 (en) 2002-09-18

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