EQUIPMENT FOR THE CHROMATIC DISCRIMINATION AND COUNTING OF ORGANISMS

Abstract

The present disclosure relates to a device for chromatic discrimination and counting of organisms in a liquid medium with application to organisms belonging to the genus Daphnia, Ceriodaphnia, Artemia, as well Collembola; or similar organisms. The device can be applied in the field of ecotoxicology and involves technologies in the areas of analytical instrumentation, electronics, computer science and biology.

Claims

1. A device for chromatic discrimination and counting of organisms in liquid medium, comprising: a channel for conducting the sample, wherein the channel has an inner section so that the bodies are driven sequentially one-by-one through channel; a collector for conducting the sample to that channel; a light emitter to emit a chromatic signal to that channel; a light receiver to capture the chromatic signal of the organism intercepting that channel; an electronic data processor configured to differentiate the organisms counted according to the chromatic signal received.

2. The device according to claim 1 wherein the channel is tubular, preferably a stylet.

3. The device according to claim 1 wherein the collector is a funnel.

4. The device according to claim 1 wherein the collector is positioned relative to the channel such that the sample is driving by gravity.

5. The device according to claim 1 in which the chromatic signal present in that channel is the chromatic signal reflected, absorbed, refracted and/or self-issued by the organism.

6. The device according to claim 1 wherein the light receiver is placed at 180° from the emitter to capture the chromatic signal transmittance of this organism in that channel.

7. The device according to claim 1 wherein the light receiver is placed at an angle less than 180° from the emitter to capture the chromatic signal reflection, refraction and/or the self-emission organism present in that channel.

8. The device according to claim 1 wherein the light emitter and the receiver are included in a coupled lens array in the cross section of that channel.

9. Device for chromatic discrimination and counting of organisms according to the previous claim 1 further comprising an optical component comprising an optical fiber, lens or filter for colour filtering to highlight the distinction between the color of the chromatic signal received from the organism against non-organism, in particular being the chromatic filtration with a green colour filter.

10. The device according to claim 1 further comprising a second light emitter and a second light receiver for chromatic characterization of the sample medium.

11. The device according to claim 10 wherein the second light emitter and the second light receiver are arranged upstream of the first light emitter and the first light receiver.

12. The device according to claim 1 further comprising a display screen or any other integrated counting display.

13. The device according to claim 1 further comprising a data link to send the count data.

14. A system comprising two or more devices for discrimination and organism counting according to claim 1 wherein the devices are coupled in parallel for simultaneous processing of samples, or coupled in series for sequentially processing samples with different characteristics, or coupled in combination series-parallel.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0047] For an easier understanding of the solution, the attached figures represent preferred embodiments of the invention, however, the figures do not intend to limit the object of the present disclosure.

[0048] FIG. 1—Schematic representation of an embodiment of the device, illustrating an embodiment of claims 1 to 9, followed by a detailed description of the meaning of constituent parts listed.

[0049] FIG. 2—Representation of an accomplishment for multiple chromatic characterization and counting of several replicates, common procedure in ecotoxicology tests.

[0050] FIG. 3—Schematic representation of a diagram in the form of singular achievement of the counting equipment.

[0051] FIG. 4—Schematic representation of an embodiment comprising two or more counting devices connected to a single processing unit.

[0052] In ecotoxicology, conducting assay tests with model organisms, eg. Daphnia sp., requires the counting of the total number of organisms in various replicates for evaluation of several indexes of toxicity. Currently, the counting is carried out by technicians through direct observation of organisms exposed backlit. However, this counting procedure using “naked eye” is slow, tiresome and prone to human error, as well as may have adverse effects on visual acuity of the technical staff involved in the count. This release automates that counting process with gains in runtime and counting accuracy, and also allows the chromatic characterization of organisms.

[0053] The device is intended for counting and chromatic characterization of small-sized bodies in suspension, in particular Daphnia sp., Ceriodaphnia, Artemia or fish eggs, allowing differentiation between specific type of organism to count and any artefacts using different chromatic features. Additionally, this device allows evaluation of ingestion, for example tests on Daphnia or Ceriodaphnia, by chromatic characterization of the medium in which these organisms are.

[0054] In one embodiment, the apparatus has a sampler (1) for organisms in suspension in a liquid. The container in figure is only illustrative of a possible embodiment. The liquid could come from another device, for example, through tubes.

[0055] In one embodiment, the serialization component of all organisms present, consists of: a collector of the sample segment (2.1) in the upper section with a funnel-like functionality; and a cylindrical segment, stylet (2.2), in the lower section, with an internal diameter adapted to the size of the bodies. The serialization component, or part thereof, is inserted into a darkroom to eliminate interference from outside light.

[0056] In one embodiment, the optical component is composed of one or more light emitters (3.1) and one or more light receivers (3.2), radially distributed around an optical window (3.3) through ducts (3.4) for passage of the beam and it may contain optical fibber. The optical component may also include optical lenses or filters (3.5) placed immediately after the light emitting and/or before the light receivers. The light output is adjustable, eventually with a spectrum adapted to the organism features, ensuring minimal contrast between organisms and the surrounding medium.

[0057] In one embodiment, the electronic component performs conditioning of the electrical signals received in the receptors and subsequent analogue-to-digital conversion.

[0058] Computer component, portable or otherwise, for counting and/or chromatic characterization of the organisms, may also register other information deemed relevant as the date, location, temperature, etc.

[0059] Collector of organisms and solution, may be a container, as illustrated in the figure, or other device

[0060] This embodiment has the following constituent parts: [0061] Replication of the serialization components; [0062] Electronic component shared; [0063] Computational component shared.

[0064] One application for this device can be in ecotoxicology studies with Daphnia organisms, particularly in neonates counting as stipulated in the OECD standard number 221, but can be applied in the counting and chromatic characterization of other organisms.

[0065] The process is conducted with the serialization of all organisms present, using a device, simultaneously lightweight, space-saving, and with high portability. The device allows the replacement of the current “naked eye” process of organisms counting, namely Daphnia in ecotoxicology tests. The device uses an automatic system of high reliability; minimizing duration of counting, human error and possible harmful consequences for the health of technicians. Furthermore, a possible embodiment of this device allows the adaptation to the chromatic features of the medium or the individual organism, minimizing possible counting errors. In addition, the chromatic characterization of the medium can be used as a parameter for evaluating the uptake by organisms, particularly useful in ingestion tests. Regardless of this possibility, with this device, the counting procedure and the counting repetition is a simple and fast process.

[0066] Due to the simplicity of the device, it can also be used for automatic counting procedures of other organisms such as Ceriodaphnia and Artemia, and also fish eggs, differentiating fertilized eggs from unfertilized eggs. In addition to the aquatic organisms counting, this device can be used to count other entities provided they are placed in suspension, such as organisms of the genus Collembola.

[0067] In addition, chromatic characterization can be used to evaluate the colour of the organisms, an indicator of its health. With the chromatic characterization, this indicator can be translated into a digital quantitative parameter which quantitatively describes the change in the health status resulting from exposure to certain chemicals or environmental factors.

[0068] This device also allows the evaluation of ingestion rates, for example in tests on Daphnia magna and Ceriodaphnia through the chromatic characterization of the medium in which these daphnids are in two different time moments.

Description of Daphnia Counting in a Sample Containing Daphnia Organisms and Daphnia Shells Using the Device for Chromatic Discrimination and Counting of Organisms in the Present Disclosure

[0069] For Daphnia neonates counting, the solution with the organisms is placed in the collector of the serialization unit, being the organisms serialized and directed to intercept the beams in the optical component, thereby generating a proper signal to be conditioned and sent to computer unit for processing and result achievement. The total counting, typically a number between 0 and 60 individuals per 100 ml solution, is obtained in about 60 seconds; however, the number of countings per test to be carried out can amount to hundreds. At the beginning of each session, a calibration with a set of the organisms should be made in order to calibrate the equipment according to the organisms in test—Daphnia neonates and shells of Daphnia neonates. The individual chromatic signal produced by each element of the neonates group shows a similar intra-group pattern but different and distinct from the pattern shown by the elements belonging to the shells of neonates group, and it is based on this chromatic differentiation that the computation unit will discern the group to which belongs each element or item analysed in the optical component, summing up—one by one—the discerned elements to one or to the other group in study, according to the chromatic pattern produced by each element. In routine work, it can be used a “factory” calibration provided with the equipment.

Description of Fertilized Fish Eggs and Unfertilized Fish Eggs Countings Using the Device for Chromatic Discrimination and Counting of the Present Disclosure Organisms

[0070] In one embodiment for fish egg counting, fertilized and unfertilized eggs are placed in approximately 100 ml of an aqueous solution, or other, and subsequently placed on the collector serialization unit, following the procedure described in the previous paragraph. At the beginning of each session, a calibration should be done with 3 sets of eggs; for example, proceeding with the preparation of a standard consisting in 50 fertilized eggs, another standard with 50 unfertilized eggs, and a mixture, for example, 50 fertilized eggs+50 unfertilized eggs, as an evaluation measure to the equipment. The counting indicates the total number of eggs, the total number of fertilized eggs and the total number of unfertilized eggs, and it may also indicate a number of eggs which classification was inconclusive.

[0071] The word “comprises” or “comprising” when used in this document is intended to indicate the presence of features, elements, integers, steps and components mentioned, but does not preclude the presence or addition of one or more other features, elements, integers, steps, components, or groups thereof.

[0072] The embodiments described above are combinable with one another.

[0073] The invention is of course not in any way restricted to the embodiments described herein and a person of ordinary skill in the area can provide many modification possibilities thereof and replacements of technical features by other equivalent, depending on the requirements of each situation, as defined in the appended claims.

[0074] The following claims further define preferred embodiments.