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Evaluation of Respiratory Inductive Plethysmography Using the EMKAbelt System in the Conscious Dog and Primate

K Meecham, E Spridgen, S Jordan, S Purbrick, S Moore, D Butler, A French, D Jones, E Peake, V Milner, P Davies and C Auletta

Huntingdon Life Sciences, Huntingdon, Cambridgeshire, UK

Presented at theSociety of Toxicology Annual Meeting11th–15th March 2012San Francisco, California, USA

Conclusion• The study showed a high degree of correlation between the ventilatory parameters

recorded from the EMKABelt RIP system and the conventional pneumotach/mask system.

• Continuous recording of respiratory rate and volume was obtained from both dog and primate throughout 20 hours recording period.

• The EMKABelt RIP system for respiration monitoring has the potential to be used on stand-alone safety pharmacology studies or as a functional end point on toxicology studies using conscious freely moving dogs and primates.

• Phase angle assessment using the two band RIP system provides a non-invasive method for qualitative assessment of airway mechanics in the conscious large animal.

AbstractTo date, respiratory measurements are typically obtained from conscious large animals using a pneumotachograph attached to a face mask. However, this method requires restraint of the animals, restricts measurements to a relatively short time and recordings cannot be performed during inhalation administration of test substances. Respiratory Inductive Plethysmography (RIP) is reported to allow accurate measurement of ventilatory parameters in freely moving animals. The purpose of this study was to evaluate the data obtained from the non-invasive telemetry EMKABelt (jacket) RIP system to that obtained using the face mask and pneumotachograph system. Four dogs and four primates implanted with DSITM telemetry transmitters were habituated to EMKABelt jackets and face masks over 14 days. Preliminary respiratory recordings were obtained in order to evaluate the most appropriate restrained posture (standing or sitting) for calibration of the EMKABelt system with the pneumotach/mask system. Respiratory recordings from both mask and RIP were obtained prior to and following CO2 (via a re-breath manoeuvre), doxapram (i.v.) or inhaled methacholine (5 min). Cardiovascular parameters were monitored by telemetry during all procedures. The data obtained showed a high degree of correlation between the EMKABelt (jacket) RIP system and the pneumotach/mask system. The EMKABelt system provides viable respiratory data and has the potential to be used on stand-alone safety pharmacology studies or as a functional end point on toxicology studies.

IntroductionThe EMKABelt (jacket) Respiratory Inductive Plethysmography (RIP) system is a non-invasive alternative for accurate measurement of ventilatory parameters (tidal volume and respiration rate) in freely moving animals. The system utilises a two band method, allowing comparison of thoracic and abdominal breathing. The purpose of this study was to evaluate the respiratory data obtained from the EMKABelt (jacket) RIP system to that obtained using the conventional face mask and pneumotachograph system and to establish responses to known positive controls in both dog and primate.

MethodsFour beagle dogs (11.6 - 15.0 kg) were habituated to the RIP jackets and face mask/pneumotachograph systems over a 14 day period. Respiratory recordings were obtained from the EMKA IOX (v 2.5.6) telemetry EMKABelt system and via face masks using a Hans Rudolf pneumotach and Ponemah system (v 4.9). The pneumotach was calibrated to a fixed volume of air before approximately 5 minutes of simultaneous recordings were obtained from the pneumotach and EMKABelt chest bands, with the dogs in a predetermined standing position in order to calibrate the EMKABelt system.

Following a pre-treatment recording period, respiration rate and tidal volume were monitored from both systems (sitting posture) following a CO2 re-breath procedure, doxapram (5 mg/kg i.v.) and inhaled methacholine (19.3 µg/kg over 5 min). Additionally, utilising the thoracic and abdominal EMKABelt chest bands, the time (or angle) between the bands movements was recorded in order to evaluate phase angle measurements as a possible surrogate marker for airway resistance.

During all procedures, cardiovascular parameters were monitored by telemetry in order to monitor the dog’s acceptance to the jacket system and for welfare reasons following administration of the reference compounds (data not shown).

For the primate, animals were habituated to RIP jackets and bands over a period of 14 days. Following pre-dose recording, animals were dosed iv with morphine (1.0 mg/kg) (under mild restraint) with continuous post-dose respiratory recordings made in home cages. To avoid stress, animals were trained to brief application of masks for the purpose of calibration only.

ResultsFigure 1: Restrained and unrestrained EMKABelt jacket monitoring of respiration in both dog and primate in pens and cages and in combination with with face mask/pneumotachograph.

Evaluation of Respiratory Inductive Plethysmography Using the EMKAbelt System in the Conscious Dog and PrimateK Meecham, E Spridgen, S Jordan, S Purbrick, S Moore, D Butler, A French, D Jones, E Peake, V Milner, P Davies and C Auletta Huntingdon Life Sciences, Huntingdon, Cambridgeshire, UK

Figure 2: Comparison of respiratory waveforms from the face mask / pneumotach system and EMKABelt RIP system prior to and following administration of doxapram (5 mg/kg i.v.) in the dog. Respiration traces showed a similar pattern of response to the doxapram.

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Figure 3 (dog): Respiratory rate and tidal volume from the EMKABelt (jacket) RIP system and face mask/pneumotachograph system prior to and following CO2 re-breath procedure, doxapram (5 mg/kg i.v.) and inhaled methacholine (19.3 µg/kg over 5 min).

Figure 5 (primate): Time course of tidal volume and respiration rate recorded from EMKABelt RIP system in the primate prior to and following administration of morphine (1.0 mg/kg i.v.).

Figure 4 (dog): An increase in RIP phase angle following inhaled methacholine exposure, consistent with an expected transient increase in airway resistance. The data were somewhat susceptible to changes in rate and movement artefact but the measure does provide a simple non-invasive method to assess for changes in lung compliance or resistance.

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