A monitoring apparatus

Abstract

A method of monitoring organism behaviour comprising the steps of exposing at least one organism to a substance and observing a response of the organism to the substance. The response is compared to m

Description

The present disclosure relates to a monitoring apparatus and to a method of using a monitoring apparatus, preferably for monitoring the behaviour of organisms in response to a substance. It is known to have a chamber for housing an organism for behavioural monitoring. According to an aspect of the invention, there is provided a method of monitoring organism behaviour, the method comprising the steps of: exposing at least one organism to a substance; and observing a response of the at least one organism to the substance; comparing the response to multiple objective endpoints, wherein the multiple objective endpoints are or correspond to respective phenotypes of the at least one organism; and identifying at least one mode of action of the substance based on the comparison of the response to the multiple objective endpoints. In embodiments of the invention, the method may include the step of recording a video of the response of the at least one organism to the substance. In other embodiments of the invention, the method may include the step of recording an image of the response of the at least one organism to the substance. In further embodiments of the invention, the method may include the step of recording a plurality of videos or images of the response of the at least one organism to the substance. Each video or image may be recorded at a respective different time. According to another aspect of the invention, there is provided a computer-implemented method of monitoring organism behaviour, the method comprising the step of: using a recording of a response of at least one organism to a substance, comparing the response to multiple objective endpoints, wherein the multiple objective endpoints are or correspond to respective phenotypes of the at least one organism; and identifying at least one mode of action of the substance based on the comparison of the response to the multiple objective endpoints. Various types of phenotypes can be observed using the method according to the invention. Non-limiting examples of phenotypes include: • Mortality; • Motility, mobility, motor function quality and / or paralysis; • Deterrence, attraction, seeking and / or avoidance; • Feeding behaviour and / or feeding inhibition; • Morphological change, such as colour change and / or size change; • Spatial organisation; • Fecundity; • Cleaning; • Agitation; • Growth inhibition and / or developmental halting. The method of the invention may include the step of, using the recording of the response, continuously observing a response of the at least one organism to the substance. Furthermore, the method may include the step of using an artificial intelligence algorithm or model to analyse the observed response of the at least one organism to the substance in order to compare the response to the multiple objective endpoints and identify the at least one mode of action of the substance based on the comparison of the response to the multiple objective endpoints. According to yet another aspect of the invention, there is provided a computer-implemented method of identifying at least one mode of action of a substance, the method comprising the steps of: collecting a set of data by carrying out the method according to any one of the preceding aspects of the invention and their embodiments, wherein the collected set of data includes the identified at least one mode of action of the substance based on the comparison of the response to the multiple objective endpoints; creating a training set including the collected set of data; training a machine learning algorithm or model using the training set; and identifying the at least one mode of action of the substance based on the comparison of the response to the multiple objective endpoints based on an output of the machine learning algorithm or model. According to a further aspect of the invention, there is provided a computer program comprising computer code configured to perform the method of the invention. In the invention, the at least one organism is preferably an insect, a pest or an arthropod. According to another further aspect of the invention, there is provided a monitoring apparatus for monitoring organism behaviour, the monitoring apparatus comprising: at least one chamber configured for housing at least one organism, the or each chamber configurable to selectively expose the at least one organism to a substance; a monitoring device configured to, in use, record a response of the at least one organism to the substance; a processor configured to, using the recording of the response, compare the response to multiple objective endpoints, wherein the multiple objective endpoints are or correspond to respective phenotypes of the at least one organism, and identifying at least one mode of action of the substance based on the comparison of the response to the multiple objective endpoints. The monitoring device may be configured to, in use, record a video or image of the response of the at least one organism to the substance. The monitoring apparatus may include a memory including computer program code. The memory and computer program code may be configured to, with the processor, enable the monitoring apparatus to carry out various processing functions. The processor and memory may form part of one or more of an electronic device, a portable electronic device, a portable telecommunications device, a mobile phone, a personal digital assistant, a tablet, a phablet, a laptop computer, a server, a cloud computing network, a smartphone, a smartwatch, smart eyewear, and a module for one or more of the same. It will be appreciated that references to a memory or a processor may encompass a plurality of memories or processors. According to a still further aspect of the invention, there is provided a monitoring apparatus for monitoring organism behaviour, the monitoring apparatus comprising: at least one chamber configured for housing at least one organism; a barrier configured to retain the or each organism within the or each chamber; an actuator configured to, in use, control the barrier to selectively open and close the or each chamber, the barrier permitting release of the or each organism from the or each chamber when the barrier is configured to open the or each chamber, the barrier retaining the or each organism within the or each chamber when the barrier is configured to close the or each chamber; and a monitoring device configured to, in use, monitor the at least one released organism. Features of the method of the invention apply mutatis mutandis to the monitoring apparatus of the invention. It will be appreciated that the method of the invention is not limited to being performed by the monitoring apparatus of the invention. It is envisaged that different aspects of the monitoring apparatus of the invention may be used in combination. The monitoring apparatus may include a plurality of chambers, each of which is configured for housing at least one respective organism. The barrier may be configured to retain the organisms within the plurality of chambers. The actuator may be configured to, in use, control the barrier to selectively open and close the plurality of chambers, the barrier permitting release of at least one of the organisms from the plurality of chambers when the barrier is configured to open the plurality of chambers, the barrier retaining the organisms within the plurality of chambers when the barrier is configured to close the plurality of chambers. The barrier may be formed of a single unitary barrier member configured to retain the organisms within the plurality of chambers. The barrier may include a plurality of discrete barrier members. Each barrier member may be configured to separately retain the organisms within a respective one or more of the plurality of chambers. The barrier may include a plurality of apertures spaced apart from each other by blocking sections. The apertures may be aligned with the plurality of chambers when the barrier is configured to open the plurality of chambers. The blocking sections may be aligned with the plurality of chambers when the barrier is configured to close the plurality of chambers. The actuator may be configured to, in use, control the barrier to synchronously open and close the plurality of chambers. It will be understood that the actuator may be replaced by a plurality of actuators in this embodiment and throughout the specification. The barrier may be removably coupled to the actuator. The barrier may be made of a metallic material. The barrier may be configured to be slidable by the actuator to selectively open and close the or each chamber. The actuator may be a linear actuator. The actuator may be remotely controllable. The monitoring apparatus may further include a programmable controller configured to control the actuator. The programmable controller may be or may include, but is not limited to, a microprocessor or a computer. The monitoring apparatus may further include a sample retention device for holding a sample. The barrier may be configured to, in use, separate the or each organism from the sample retention device when the barrier is configured to close the or each chamber. The barrier may be configured to, in use, provide the or each organism with access to the sample retention device when the barrier is configured to open the or each chamber. The barrier and the sample retention device may be configured to be in sealing cooperation to provide at least one sealed environment for the or each organism and the sample when the barrier is configured to open the or each chamber. In embodiments employing the use of a plurality of chambers, the barrier and the sample retention device may be configured to be in sealing cooperation to provide sealed environments for the organisms and the sample when the barrier is configured to open the plurality of chambers. The sample retention device may be a sample clamping device for clamping the sample. The sample clamping device may include at least one clamping member corresponding to the or each chamber. The or each clamping member may be configured to, in use, position the sample against or towards the or each corresponding chamber. The sample clamping device may include a first clamping member support structure. The or each clamping member may be attached to, or form part of, the first clamping member support structure. The sample clamping device may include a second clamping member support structure. The or each clamping member and the second clamping member support structure may be configured to, in use, clamp the sample between the or each clamping member and the second clamping member support structure. The barrier, the or each clamping member and the second clamping member support structure may be configured to be in sealing cooperation to provide at least one sealed environment for the or each organism and the sample when the barrier is configured to open the or each chamber. In embodiments employing the use of a plurality of chambers, the barrier, the clamping members and the second clamping member support structure may be configured to be in sealing cooperation to provide sealed environments for the organisms and the sample when the barrier is configured to open the plurality of chambers. The monitoring apparatus may include at least one removable well assembly. The or each well assembly may be configured for holding at least one sample and at least one organism. The monitoring device may be configured to, in use, monitor the at least one organism in the or each well assembly. The or each well assembly may be made of a glass material, a metallic material, an aluminium material or a stainless steel material. The monitoring apparatus may include at least one removable tubular conduit. The or each tubular conduit may be configured for holding at least one sample and at least one organism. The monitoring device may be configured to, in use, monitor the at least one organism in the or each tubular conduit. The or each tubular conduit may include at least one sample holder arranged at one end of the tubular conduit. The at least one sample holder may be configured for containing a liquid sample. The sample holder may include two membrane layers separated by a spacing member. The or each tubular conduit may be made of a glass material, a metallic material, an aluminium material or a stainless steel material. The or each removable well assembly and / or the or each tubular conduit may be configured for use after temporarily removing the at least one chamber, the barrier and the actuator from the monitoring apparatus. This is to provide the monitoring apparatus with one or more alternative configurations for monitoring the or each organism. The sample retention device may be made of a metallic material. The monitoring apparatus may further include a housing in which the or each chamber, the barrier and the actuator are located. The monitoring apparatus may further include a light source configured to, in use, illuminate the or each chamber. The monitoring apparatus may further include one or more light blocking masks arranged to, in use, block part of an illumination light provided by the light source. The or each light blocking mask may be used in combination with the or each well assembly and / or the or each tubular conduit. The or each chamber may be made of a glass material, a metallic material, an aluminium material or a stainless steel material. The monitoring device may be or may include an imaging device. The monitoring apparatus may be an insect, pest or arthropod monitoring apparatus. According to another aspect of the invention, there is provided a method of using a monitoring apparatus in accordance with any one of the preceding aspects of the invention and its embodiments, the method comprising the steps of: loading at least one organism into the or each chamber; by the barrier, retaining the or each organism within the or each chamber; providing a substance; by the actuator, controlling the barrier to open the or each chamber to release the or each organism from the or each chamber and thereby expose the or each organism to the substance; and by the monitoring device, monitoring a behaviour of the or each organism in response to the substance. The substance may be a stimulus-generating substance that produces a stimulus to which the at least one organism may respond. The substance may be an odourgenerating substance. The substance may be a leaf. The substance may be an uncut leaf. The substance may be or may include at least one crop material, at least one insect target, at least one microbe, at least one microbial extract, at least one natural product, at least one metabolite, at least one metabolite mixture, at least one chemical active, at last one chemical compound and / or at least one biological material. The step of providing a substance may include adding a substance as a sample to the sample retention device prior to the barrier being controlled to open the or each chamber. The method may further include the step of controlling the barrier to synchronously open the plurality of chambers to release the organisms from the plurality of chambers and thereby expose the organisms to the substance at the same time. According to yet another aspect of the invention, there is provided a monitoring apparatus comprising: at least one tubular conduit configured for holding at least one sample and at least one organism, wherein the or each tubular conduit includes at least one sample holder for holding the at least one sample, the sample holder including two membrane layers separated by a spacing member, a monitoring device configured to, in use, monitor the at least one organism in the or each tubular conduit. The or each tubular conduit may include at least one sample holder arranged at one end of the tubular conduit. The at least one sample holder may be configured for containing a liquid sample. The tubular conduit may be made of a glass material, a metallic material, an aluminium material or a stainless steel material. It will be appreciated that the use of the terms "first" and "second", and the like, in this patent specification may be used to help distinguish between similar features (e.g., the first and second clamping structures) and is not intended to indicate the relative importance of one feature over another feature, unless otherwise specified. Within the scope of this application it is expressly intended that the various aspects, embodiments, examples and alternatives set out in the preceding paragraphs, and the claims and / or the following description and drawings, and in particular the individual features thereof, may be taken independently or in any combination. That is, all embodiments and all features of any embodiment can be combined in any way and / or combination, unless such features are incompatible. The applicant reserves the right to change any originally filed claim or file any new claim accordingly, including the right to amend any originally filed claim to depend from and / or incorporate any feature of any other claim although not originally claimed in that manner. Preferred embodiments of the invention will now be described by way of non-limiting examples with reference to the following drawings, in which: Figures 1 and 2 respectively show perspective and exploded views of a plurality of chambers, a barrier, an actuator and a sample retention device of a monitoring apparatus according to an embodiment of the invention; Figures 3 and 4 illustrate a method of using a monitoring apparatus according to an embodiment of the invention; Figures 5 and 6 respectively show interior and exterior views of a monitoring apparatus according to an embodiment of the invention; Figures 7A to 7F illustrate steps of reconfiguring a monitoring apparatus according to the invention; Figures 8A to 8C show an alternative screening configuration of a monitoring apparatus according to the invention; Figures 9A to 9D show another alternative screening configuration of a monitoring apparatus according to the invention; and Figure 10 illustrates a degree of response of microbes with respect to various phenotypes. The figures are not necessarily to scale, and certain features and certain views of the figures may be shown exaggerated in scale or in schematic form in the interests of clarity and conciseness. A plurality of chambers 20, a barrier 30, an actuator 40 and a sample retention device 50 of a monitoring apparatus according to an embodiment of the invention is shown in Figures 1 and 2. The sample retention device 50 is optional, and it is envisaged that the sample retention device 50 may not be included in all embodiments of the invention. Each of the plurality of chambers 20 may house an organism. In further embodiments, each chamber 20 may house more than one organism. In some embodiments, the organism may be an insect or an agricultural pest or an arthropod. The plurality of chambers 20 may be in the form of a well assembly (e.g. a 256-well assembly) in which each chamber 20 is a well. In other embodiments, the well assembly may be removable for replacement by another well assembly. In further other embodiments, the plurality of chambers 20 may be in the form of multiple well assemblies in which each well assembly includes some of the chambers 20. The plurality of chambers 20 may be made of a metallic material. In alternative embodiments, the plurality of chambers 20 may be made of another material which does not retain or can be cleansed of stimulus-generating substances, odourgenerating substances or odours. For example, and not limitation, a chamber 20 may be made of glass, a non-porous material, or a specialised synthetic material or polymer. The barrier 30 is configured, in its closed configuration, to retain the organisms within each of the plurality of chambers 20. In some embodiments, the barrier 30 may be formed of a plurality of discrete barrier members 32. In other embodiments, the barrier 30 may be formed of a single unitary barrier member. The barrier 30 may be at least partially comprised of a metallic material, glass, a non-porous material, or a specialised synthetic material or polymer. The material of the barrier 30 preferably does not retain odours or must be completely sanitisable to remove any substances, stimuli or odours, e.g. by UV light, boiling water, solvents or heating (e.g. using autoclaves). The barrier 30 may be formed of a plurality of discrete barrier members 32 which are configured to separately retain organisms within a subset of the plurality of chambers 20. In some embodiments, the subset may be a single "row" of chambers 20, or any number of rows of chambers 20. In the pictured embodiment, each row of chambers 20 is closed by the barrier 30 which is formed of a plurality of long, single-row barrier members 32 (e.g. sliding keys). Each single-row barrier member 32 takes the form of a strip having a plurality of apertures 34 along its length so that the apertures 34 are spaced apart from each other by blocking sections. The individual barrier members 32 are controlled by an actuator 40. The barrier members 32 may be permanently or detachably coupled to the actuator 40. The actuator 40 controls the barrier 30 to selectively retain and selectively release the organisms in the plurality of chambers 20. The actuator 40 controls the movement of the barrier 30 from a closed configuration to an open configuration, and from an open configuration to a closed configuration. The barrier 30 permits release of the organisms from the chambers 20 when the barrier 30 is configured to align the apertures 34 with the plurality of chambers 20 and thereby open the chambers 20 (i.e. the open configuration). The barrier 30 retains the organisms within the chambers 20 when the barrier 30 is configured to align the blocking sections with the plurality of chambers 20 and thereby close the chambers 20 (i.e. the closed configuration). In the pictured embodiment, the actuator 40 is a linear actuator which is configured to synchronously control all barrier members 32 in the barrier 30 to open all chambers 20 simultaneously. A guide rail may be employed to guide the linear movement of the actuator 40. Guide rails 36 may be employed to aid the linear sliding movement of the barrier members 32 as controlled by the actuator 40. In other embodiments, the actuator 40 may control the barrier 30 to open some chambers 20 non-synchronously. For example, the actuator 40 may be configured to control a number of barrier members 32 to open the chambers 20 "row by row", or in specified regions. In yet other embodiments, there may be a further actuator or actuators to control different barrier members 32. In some embodiments, the actuator 40 may be controlled remotely by a programmable controller and / or a human operator. The remote control prevents the human operator from directly affecting the organisms inside the chambers 20, e.g. by odour contamination. The programmable controller may be programmed or configured to control the actuator 40 to selectively open or close the barrier 30 after a predetermined time or in response to a predetermined signal or command. In other embodiments a human operator may use their judgment to control the actuator 40 to selectively open or close the barrier 30. A sample retention device 50 may be included in some embodiments. The sample retention device 50 may be comprised of a single unitary piece or multiple pieces. An embodiment of the sample retention device 50 is shown in Figures 3 and 4. In the pictured embodiment, the sample retention device 50 is a sample clamping device comprising first and second clamping member support structures 52, 54. Both clamping member support structures 52, 54 are in the form of plates respectively. The second clamping member support structure 54 further includes a receptacle surrounded by a frame wall, whereby the receptacle is shaped to receive the first clamping member support structure 52. In some aspects, the sample retention device 50 (or specifically the second clamping member support structure 54) may be configured to be removably attached to a casing 56 storing the plurality of chambers 20. The attachment may be performed by an external clamping device or by a clamping device which is part of the sample retention device 50. In some embodiments the attachment may be performed by a plurality of removable screws or a different form of fastener or securing mechanism. In the pictured embodiment, the second clamping member support structure 54 is attached to the casing 56 by way of a plurality of screws 56. In use, a sample 58 is placed on the second clamping member support structure 54, as shown in Figure 3. The first clamping member support structure 52 is lowered onto the second clamping member support structure 54 so as to clamp the sample 58 between the first and second clamping member support structures 52, 54, as shown in Figure 4. The first clamping member support structure 52 may be held in place simply by the weight of the first clamping member support structure 52, or may be held in place either by complementary fit with the second clamping member support structure 54 or by another fastening or securing mechanism. The sample retention device 50 may include a plurality of clamping members, each corresponding to a respective one of the plurality of chambers 20 and configured to position the sample 58 against or towards the corresponding chamber 20. The clamping members may be in the form of projections. The clamping members may form part of or be attached to the first clamping member support structure 52 and may be configured to position and retain the stimulus-generating substance against openings in the second clamping member support structure 54 corresponding to the chambers 20. The barrier 30 in its closed configuration separates the organisms within the chambers 20 from the first and second clamping member support structures 52, 54 and the clamped sample 58. The barrier 30 in its open configuration provides the organisms within the chambers 20 with access to the first and second clamping member support structures 52, 54 and the clamped sample 58. In this way, when the apertures 34 of the barrier members 32 are aligned with the plurality of chambers 20, the organisms in the chambers 20 are exposed to the clamped sample 58 via the openings in the second clamping member support structure 54. Preferably, in the open configuration of the barrier 30, the barrier 30 and the first and second clamping member support structures 52, 54 are engaged with each other so as to be in sealing cooperation with each other and thereby create sealed environments that include the plurality of chambers 20. This beneficially creates environments for at least part of the sample 58 and the organisms that is sealed against external stimulus such as unwanted odours. In other embodiments, in the open configuration of the barrier 30, the barrier 30 and the first and second clamping member support structures 52, 54 may be engaged with each other to physically contain at least part of the sample 58 and organisms within an enclosed environment, without necessarily being in sealing cooperation with each other. The clamping members may be transparent. When the barrier 30 is in the open configuration, the transparent clamping members and the openings in the second clamping member support structure 54 permit light from a light source to pass through the chambers 20, the openings in the second clamping member support structure 54, the sample 58, and the transparent clamping members in that order. A monitoring device 70 may be arranged to receive the light that has passed through the chambers 20, the openings and the transparent clamping members. Further details of the monitoring device 70 are described elsewhere in this specification. Components of the sample retention device 50 may be wholly or partially comprised of a metallic material, glass, a non-porous material, or a specialised synthetic material or polymer. The material of the components of the sample retention device 50 preferably does not retain odours or must be completely sanitisable to remove any substances, stimuli or odours, e.g. by UV light, boiling water, solvents or heating (e.g. using autoclaves). The light source 60 may be located beneath the chambers 20 to provide backlighting through a glass panel 90. Alternatively the light source 60 may be located to the side or in another location and be configured to illuminate the chambers 20 from below, for example and not limitation by directing light beneath the chambers 20. The monitoring apparatus may further include one or more mirrors positioned to redirect the light to illuminate the chambers 20, or the light may be redirected in another manner. The light source 60 may be configured to illuminate all or some of the chambers 20. The organism to be monitored by the monitoring apparatus may be an insect, an agricultural pest or an arthropod. The monitoring apparatus may be suitable for use with other insects or organisms and especially for monitoring the response of agricultural pests to a stimulus. The stimulus may be, for example and not limitation, a sample in the form of a leaf (which may be untreated), a leaf coated in or treated with a stimulus-generating substance, or a stimulus-generating substance. The stimulus-generating substance may be an odour-generating substance. The stimulusgenerating substance may be a powder or liquid and may be coated on or otherwise applied to a leaf. In some aspects, a whole uncut leaf by itself, or the whole leaf coated in or treated with the stimulus-generating substance, may be retained by the sample retention device 50 for exposure to the organisms. It will be appreciated that the reference to the leaf is to help illustrate the working of the invention, and that the invention is applicable to other types of stimulus-generating substances, such as or including at least one crop material, at least one insect target, at least one microbe, at least one microbial extract, at least one natural product, at least one metabolite, at least one metabolite mixture, at least one chemical active, at last one chemical compound and / or at least one biological material. In the pictured embodiment in Figure 5, the monitoring apparatus 70 is located above the sample retention device 50 and arranged to face downwards towards the first clamping member support structure 50 so as to enable the monitoring apparatus 70 to receive the light that has passed through the chambers 20, the openings in the second clamping member support structure 54, the sample 58, and the transparent clamping members in that order. In other embodiments, the monitoring apparatus may further include one or more mirrors positioned to redirect the light to the monitoring apparatus, or the light may be redirected in another manner. The monitoring device 70 may be or include a camera, an imaging device, or another light-sensitive sensor. The monitoring device 70 may further include a programmable controller which may be pre-programmed by a user or manufacturer to control the monitoring device 70. The monitoring device 70 may be configured to begin monitoring the behaviour or movement of the organisms upon detection of a predetermined cue such as, for example and not limitation, a user pressing a key or button, the light source 60 switching to an "on" state, light from the light source 60 being received by the monitoring device 70, a threshold amount of light being received by the monitoring device 70, a predetermined time limit elapsing, the activation of the actuator 40 or the opening of the barriers 30. The predetermined cue may be a combination of the aforementioned examples or another cue. The monitoring device 70 may be configured to detect movement of the organisms, for example by way of a detected change in the light (e.g. change in cast shadow) detected by the monitoring device 70. In this way the monitoring device 70 may detect whether the organisms are repelled by the stimulus, attracted to the stimulus, are consuming the sample or have another behavioural reaction to the stimulus. In some embodiments the monitoring device 70 may be configured to transmit data for analysis (using wired or wireless connection) and / or store the data using an onboard memory. In further embodiments the monitoring device 70 may transmit the data in a "live feed" or to be viewed in real time by the user. The data may include still image data and / or video data. The data may include time stamps and / or date stamps. The monitoring apparatus may further comprise a housing 100 inside which the other components including the plurality of chambers 20, the barrier 30, the actuator 40, the sample retention device 50, the light source 60 and the monitoring device 70 are housed. In further embodiments, the light source 60 and / or the monitoring device 70 may be located outside the case or enclosure. The housing 100 may be sealable to be, for example, light-tight, odour-tight and / or stimulus-tight. The housing 100 may insulate the organisms and sample from the external environment to ensure that the observed behaviour by the organisms is a result of exposure to the target sample, stimulus or stimulus-generating substance, and not any other sample, stimulus or stimulus-generating substance. An exemplary embodiment of a monitoring apparatus inside a sealable housing 100 is shown in Figures 5 and 6. A cutout of an embodiment of a monitoring apparatus including a sealable housing 100 is shown in Figure 5 to illustrate exemplary locations of the plurality of chambers 20, the barrier 30, the actuator 40, the sample retention device 50, the light source 60 and the monitoring device 70. Figure 6 shows the exterior of the housing 100, which may include a display screen 102 for showing still images or videos transmitted from the monitoring device 70. An exemplary method of using the monitoring apparatus comprises the following steps: a) Using the actuator 40 or manually, slide the barrier members 32 to open the chambers 20, and load organisms into the opened chambers 20. The chamber opening and the organism loading can be carried out simultaneously or row-by-row. b) Using the actuator 40 or manually, slide the barrier members 32 to close the chambers 20 so as to retain the organisms within the chambers 20. c) Place the second clamping member support structure 54 over the casing 56 and use the screws to attach the second clamping member support structure 54 to the casing 56. d) Place the sample 58 over a selection of the openings of the second clamping member support structure 54. e) Use the first clamping member support structure 52 to clamp the sample 58 between the first and second clamping member support structures 52, 54. f) Turning on the light source 60 to apply backlighting to the chambers 20. g) Using the actuator 40, slide the barrier members to synchronously open some or all of the chambers 20, thereby releasing the organisms from the opened chambers 20 and exposing the organisms to the sample 58. h) Using the monitoring device 70, monitoring a behaviour of the organisms exposed to the sample 50. Each chamber 20 will be seen as a lit circle, with each organism being observed as a shadow. The movement of the shadow is recorded by the monitoring device 70. i) After completing the monitoring step, removing the organisms from the chambers 20 by performing steps a) to e) in reverse. The foregoing features of the monitoring apparatus results in a standalone platform with a high throughput screening configuration for monitoring and tracking the behaviour of live insects exposed to a stimulus in a controlled environment. The features of the monitoring apparatus, particularly the controlled actuation of the barrier members 32 to synchronously release the insects, enable high levels of repeatability which is useful for producing reliable behavioural and efficacy data collection for high numbers of insects exposed to same or different types of stimuli. This is in contrast to manually exposing the insects to a stimulus, which is prohibitively slow and lacks controllability over exposure timing and duration. In addition to the high throughput screening configuration described above, the monitoring apparatus of the invention may be reconfigured into one or more alternative screening configurations. The or each alternative screening configuration allows for lower throughput screening, which may be suitable for rapid early screening (especially if high numbers of insects are not required) or for running different monitoring experiments (e.g. using larger or smaller organisms, different types of organisms, different types of stimulus, different environments). This is particularly advantageous due to the lower setup time and the shorter loading times. As the throughput is lower (i.e. the number of insects tested per run is lower), synchronised exposure to the stimulus in the same rune is less critical because a time difference between the first insect and last insect being loaded will be more acceptable. Thereafter, the high throughput screening configuration may be used to confirm the results of the rapid early screening by obtaining reliable statistical data using a high number of insects. Non-limiting examples of alternative screening configurations are described as follows. The barrier 30, the actuator 40 and the sample retention device 50 are not essential for the alternative screening configurations. Figures 7A to 7F show a deconstruction sequence of a monitoring apparatus according to an exemplary embodiment of the invention, as follows: 1) [Figure 7A] The first clamping member support structure 52 is removed from the second clamping member support structure 54. 2) [Figure 7A] The screws are removed, and the second clamping member support structure 54 is removed from the casing 56. 3) [Figure 7C] The barrier members 32 are decoupled from the actuator 40. 4) [Figure 7D] The barrier members 32 are removed from the casing 56. 5) [Figure 7E] The guide rails 36 for the barrier members 32 are removed. 6) [Figure 7F] The well assembly defining the chambers 20 are removed from the casing 56, leaving the backlighting glass panel 90 exposed. Following the deconstruction sequence, the monitoring apparatus is then reconfigured to an alternative screening configuration. A first alternative screening configuration based on removable well assemblies 110 is shown in Figures 8A to 8C. In the pictured embodiments, each well assembly 110 includes 6 wells, but it is envisaged that other numbers of wells may be used. In Figure 8A, a light blocking mask 112 is placed over the glass panel 90. The light blocking mask 112 has an opaque region that acts to block part of an illumination light provided by the light source 60, and has openings that permit the transmission of light therethrough. In Figure 8B, well assemblies 110 (such as well plates) are placed over the openings in the light blocking mask 112. Figure 8C shows four well assemblies 110 in situ, completing the alternative screening configuration. Each well 110 contains a sample and an organism. The well assemblies 110 may be pre-loaded with the organisms, or the organisms may be loaded into the well assemblies 110 after the alternative screening configuration is set up. Preferably each well 110 is larger than the wells 20 used in the high throughput screening configuration. The monitoring device 70 detects the transmitted light and thereby detects the shadows cast by the organisms within the wells 110. The first alternative screening configuration permits the use of standardised well assemblies. Furthermore, different well assemblies can be used to allow for a range of different insect retention environments. The different well assemblies may have, for example, different types of wells, different numbers of wells, different sizes of wells and / or different footprints of well assemblies. A second alternative screening configuration based on tubular conduits is shown in Figures 9A to 9D. Each tubular conduit is in the form of a specimen tube 120. In Figure 9A, the specimen tubes 120 are placed over the glass panel 90. The light blocking mask 112 act to block part of an illumination light provided by the light source 60, as shown in Figures 9A and 9B. The tubes 120 are placed over the openings in the light blocking mask 112. In Figure 9C, a sample holder is arranged at the top of each tube 120. The sample holder comprises two parafilm membrane layers 122 separated by a spacing member 124 in the form of an O-ring. A space between the two parafilm membrane layers 122 contains a liquid active, or another type of solid or liquid stimulus-generating substance. An organism is loaded into the tube 120 from the bottom of the tube. In Figure 9D, the organism has reached the top of the tube 120 and feeds on the liquid active through the lower parafilm membrane layer 122. The monitoring device 70 detects the transmitted light through the membrane layers 122 and thereby detects the shadows cast by the organisms within the tubes 120. Furthermore, two sample holders may be arranged at respective top and bottom ends of each tube 120, thus enabling the testing of two stimulus-generating substance in the same run in a controlled way. The two stimulus-generating substances may be the same or different. Conventionally insect screening is carried out manually, which is time-consuming and has low throughput. Such screening is based on the monitoring of a single end point, involves the use of subjective scoring, and has poor temporal resolution. As a result, the conventional insect screening provides limited information on a mode of action of a given substance, which is defined as a functional or anatomical change to an organism . In a preferred method of monitoring organism behaviour, one or more organisms (preferably insects, pests or arthropods) is exposed to a stimulus-generating substance, for example, by using any of the aforementioned screening configurations. A video (or a plurality of videos recorded at different times) of a response of the or each organism to a stimulus provided by the stimulus-generating substance is recorded using a video monitoring device. Using the recorded video, it becomes possible to compare the response to multiple objective endpoints, wherein the multiple objective endpoints are or correspond to respective phenotypes of the at least one organism; and identify one or more mode of actions of the stimulus-generating substance based on the comparison of the response tp the multiple objective endpoints. Such comparison and identification are performed with the assistance of a processor and, optionally, an artificial intelligence algorithm or model. It is envisaged that the video monitoring device may be replaced by an image monitoring device. Phenotypes observable by the method of the invention include: • Mortality (including mortality rates); • Motility, mobility, motor function quality and / or paralysis (including paralysis progression); • Deterrence, attraction, seeking and / or avoidance; • Feeding behaviour and / or feeding inhibition; • Morphological change, such as colour change (e.g. pigmentation change, melanisation pattern) and / or size change; • Spatial organisation; • Fecundity; • Cleaning; • Agitation; • Growth inhibition and / or developmental halting. Morphological change is defined as the physical alteration in organism appearance or structure. This can provide an indication of, for example, metabolic disruption, molting issues, or physiological stress. This provides an early indicator of sublethal effects or insect pest modifying behaviour before mortality. Paralysis is defined as loss of voluntary movement ability, either partial or complete. This can provide an indicator of, for example, neurotoxic effects or muscle function disruption. This is a critical safety measure as paralysis often precedes mortality. Mortality is defined as the death of the organism. This provides, for example, a measure of acute toxicity. Developmental halting is defined as interruption of normal life cycle progression. This can provide an indication of, for example, interference with molting, metamorphosis, or growth. This provides long-term population control without acute toxicity. Feeding behaviour is defined as changes in food consumption patterns or feeding cessation. This can provide an indication of, for example, appetite suppression or inability to feed. This is critical to crop protection without the involvement of direct mortality. Spatial organization relates to how insects position themselves relative to each other and environment. This can indicate, for example, changes in social behavior or environmental response. This can reveal repellent or attractant effects. Fecundity is defined as reproductive capacity and egg-laying behavior. This can indicate, for example, the impact on population growth potential. This can provide long-term pest control through reproductive suppression. Seeking is defined as active search behavior for food, mates, or habitat. This can show, for example, changes in motivation or ability to locate resources. This can reveal disruption of essential behaviors. Avoidance is defined as active movement away from specific stimuli or areas. This can indicate, for example, recognition and rejection of treated areas. This is key for repellent-based crop protection strategies. Motor function quality is defined as quality and coordination of movement. This can indicate, for example, neuromuscular system impacts. This can reveal subtle effects missed by mortality screening. Cleaning is defined as grooming and maintenance behaviors. This can indicate, for example, changes in basic behavioural patterns. This provides an early indicator of neurological or physiological stress. Agitation is defined as increased movement or erratic behavior. This can indicate, for example, nervous system disruption or stress response. This provides an early warning of insecticide effects. Other phenotypes include developmental transitions, behavioural patterns, time-to-effect and dose response curves. Non-limiting examples of modes of action identifiable by the invention include: 1) Nervous system disruption Action: Interference with the insect's nervous system, leading to paralysis or hyperactivity, followed by death. Examples of substances related to this mode of action include: • Acetylcholinesterase inhibitors: Organophosphates (e.g., malathion) and carbamates (e.g., carbaryl); • Sodium channel modulators: Pyrethroids (e.g., permethrin); • Nicotinic acetylcholine receptor (nAChR) agonists / antagonists: Neonicotinoids (e.g., imidacloprid). 2) Muscle Function Disruption Action: Targets the muscles, preventing contraction or causing excessive contraction. Examples of substances related to this mode of action include: • Ryanodine receptor modulators: Diamides (e.g., chlorantraniliprole). 3) Growth and Development Interference Action: Disruption of normal insect growth or molting processes. Examples of substances related to this mode of action include: • Chitin synthesis inhibitors: Benzoylureas (e.g., diflubenzuron); • Juvenile hormone analogs: Pyriproxyfen (used in mosquito control). 4) Energy Metabolism Inhibition Action: Disruption of energy production, affecting cellular respiration. Examples of substances related to this mode of action include: • Mitochondrial electron transport inhibitors: Rotenone; • ATP synthase inhibitors: Spirotetramat. 5) Feeding Inhibition Action: Stops the insect from feeding by affecting behavior or physiology. Examples of substances related to this mode of action include: • Antifeedants: Azadirachtin (from neem). 6) Gut Disruption Action: Damage to the insect's gut lining, leading to starvation or septicaemia. Examples of substances related to this mode of action include: • Bacterial toxins: Bacillus thuringiensis (Bt) toxins. • Toxins from spinosyns: Spinosad. 7) Physical Disruption Action: Physically harms or suffocates the insect. Examples of substances related to this mode of action include: • Oils and soaps: Disrupt the waxy cuticle. • Desiccants: Diatomaceous earth. The invention is applicable to a wide range of organisms, including insects, pests and arthropods. The comparison of the observed response to multiple objective endpoints facilities the generation of more data that enables not only accurate identification of the mode(s) of action of the stimulus-generating substance but also new modes of action that cannot be identified through traditional screening, and thereby provides more information on subtle changes in insect behaviour that cannot be detected by the conventional insect screening method. For example, a combination of observed behaviours would be indicative of a particular type of mode of action, while another combination of observed behaviours would be indicative of another type of mode of action. Such data can be used to map out a response of a given organism in terms of different phenotypes and / or different types of substances. An exemplary map is illustrated in Figure 10 that shows the degree of response of microbes with respect to various phenotypes. This level of information is not available by simply monitoring a single objective endpoint and identifying a single mode of action which conventionally is mortality. A machine learning algorithm or model may be trained to improve the identification of at least one mode of action of a stimulus-generating substance. The training may involve the following steps: • collecting a set of data that includes the identified at least one mode of action of the stimulus-generating substance; • creating a training set including the collected set of data; • training a machine learning algorithm or model using the training set; and identifying the at least one mode of action of the stimulus-generating substance based on the comparison of the response to the multiple objective endpoints based on an output of the machine learning algorithm or model. 5 Preferences and options for a given aspect, feature or parameter of the invention should, unless the context indicates otherwise, be regarded as having been disclosed in combination with any and all preferences and options for all other aspects, features and parameters of the invention. 10 The listing or discussion of an apparently prior published document or apparently published information in this specification should not necessarily be taken as an acknowledgement that the document is part of the state of the art or is common general knowledge.

Claims

1. A method of monitoring organism behaviour, the method comprising the steps of:exposing at least one organism to a substance; andobserving a response of the at least one organism to the substance;comparing the response to multiple objective endpoints, wherein the multiple objective endpoints are or correspond to respective phenotypes of the at least one organism; andidentifying at least one mode of action of the substance based on the comparison of the response to the multiple objective endpoints.

2. A method according to any one of the preceding claims including the step of recording a video or image of the response of the at least one organism to the substance.

3. A method according to any one of the preceding claims including the step of recording a plurality of videos or images of the response of the at least one organism to the substance, wherein each video or image is recorded at a respective different time.

4. A computer-implemented method of monitoring organism behaviour, the method comprising the step of:using a recording of a response of at least one organism to a substance, comparing the response to multiple objective endpoints, wherein the multiple objective endpoints are or correspond to respective phenotypes of the at least one organism; andidentifying at least one mode of action of the substance based on the comparison of the response to the multiple objective endpoints.

5. A method according to any one of the preceding claims wherein the at least one phenotype includes mortality.

6. A method according to any one of the preceding claims wherein the at least one phenotype includes motility, mobility, motor function quality and / or paralysis.

7. A method according to any one of the preceding claims wherein the at least one phenotype includes deterrence, attraction, seeking and / or avoidance.

8. A method according to any one of the preceding claims wherein the at least one phenotype includes feeding behaviour and / or feeding inhibition.

9. A method according to any one of the preceding claims wherein the at least one phenotype includes morphological change.

10. A method according to Claim 9 wherein the morphological change includes colour change and / or size change.

11. A method according to any one of the preceding claims wherein the at least one phenotype includes spatial organisation.

12. A method according to any one of the preceding claims wherein the at least one phenotype includes fecundity.

13. A method according to any one of the preceding claims wherein the at least one phenotype includes cleaning.

14. A method according to any one of the preceding claims wherein the at least one phenotype includes agitation.

15. A method according to any one of the preceding claims wherein the at least one phenotype includes growth inhibition and / or developmental halting.

16. A method according to any one of the preceding claims including the step of continuously observing a response of the at least one organism to the stimulus provided by the substance.

17. A method according to any one of the preceding claims including the step of using an artificial intelligence algorithm or model to analyse the observed response of the at least one organism to the stimulus provided by the substance in order to compare the response to the multiple objective endpoints and identify the at least one mode of action of the substance based on the comparison of the response to the multiple objective endpoints.

18. A computer-implemented method of identifying at least one mode of action of a substance, the method comprising the steps of:collecting a set of data by carrying out the method according to any one of the preceding claims, wherein the collected set of data includes the identified at least one mode of action of the substance associated with the multiple objective endpoints;creating a training set including the collected set of data;training a machine learning algorithm or model using the training set; and identifying the at least one mode of action of the at least one organism based on the comparison of the response to the multiple objective endpoints based on an output of the machine learning algorithm or model.

19. A computer program comprising computer code configured to perform the method of any one of Claims 4 to 18.

20. A method according to any one of the preceding claims wherein the at least one organism is an insect, a pest or an arthropod.

21. A monitoring apparatus for monitoring organism behaviour, the monitoring apparatus comprising:at least one chamber configured for housing at least one organism, the or each chamber configurable to selectively expose the at least one organism to a substance;a monitoring device configured to, in use, record a response of the at least one organism to a stimulus provided by the substance;a processor configured to, using the recording of the response, compare the response to multiple objective endpoints, wherein the multiple objective endpoints are or correspond to respective phenotypes of the at least one organism, and identifying at least one mode of action of the substance based on the comparison of the response to one or more of the multiple objective endpoints.

Images