AU2020100026A4 - Portable device and method for field collection of euglena - Google Patents
Portable device and method for field collection of euglena Download PDFInfo
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- AU2020100026A4 AU2020100026A4 AU2020100026A AU2020100026A AU2020100026A4 AU 2020100026 A4 AU2020100026 A4 AU 2020100026A4 AU 2020100026 A AU2020100026 A AU 2020100026A AU 2020100026 A AU2020100026 A AU 2020100026A AU 2020100026 A4 AU2020100026 A4 AU 2020100026A4
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/08—Flask, bottle or test tube
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/02—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by impregnation, e.g. using swabs or loops
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/24—Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms
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Abstract
The present invention belongs to the field of alga collection, and particularly relates to a device and method for field collection of euglena. The present invention provides a portable device for field collection of euglena. The device includes a hollow tube body, a tube cap matched with the hollow tube body, and a pick-and-place rod. One end of the pick-and-place rod is fixed with a water absorbent collecting body and the pick-and-place rod is disposed in the hollow tube body. The present invention further provides a method for field collection of euglena. The collected euglena is adsorbed on the water absorbent collecting body for facilitating sampling; and the water absorbent collection body can prevent the euglena from swimming in the transportation process (bad environment) and can make the euglena fixed in the materials. After transported to the laboratory, an appropriate amount of culture medium is added to a centrifuge tube, illumination is applied to restore a favorable environment, at this time, the motility of the euglena is enhanced, and the euglena swims from the water absorbent collecting body into the culture medium and grows in a large amount. 23 24 -271
Description
2020100026 08 Jan 2020
PORTABLE DEVICE AND METHOD FOR FIELD COLLECTION OF EUGLENA TECHNICAL FIELD
The present invention belongs to the field of alga collection, and particularly relates to a device and method for field collection of euglena.
BACKGROUND
Euglena is a creature that has been bred on the earth 500 million years ago. It has no cell wall, can swim, and is a kind of protozoa. At the same time, the green euglena cells have green chloroplasts to carry out photosynthesis. Therefore, euglena has the characteristics of both animal and plant. Euglena is an optimal experimental material for studying endosymbiosis, chloroplast development, and photosynthesis. In addition, euglena has a high commercial value. The National Health and Family Planning Commission Announcement on the Approval of Eight New Food Ingredients Such as Euglena” (No. 10, 2013) approved euglena as a new food ingredient on October 30, 2013. Euglena is rich in nutrients: 59 kinds of essential nutrients required for human bodies such as vitamins, mineral nutrients, amino acids, unsaturated fatty acids, chlorophyll, lutein, zeaxanthin, and GABA, which are essential nutrients for staying health. The most important euglena polysaccharide is a unique component of euglena.
At the end of spring and the beginning of summer, euglena may grow explosively in the water surface of fish ponds, thereby easily forming green or rust red blooms. In order to obtain more euglena species to conduct research, it is necessary to collect a large number of samples of euglena blooms in the water samples from different regions for collection and transportation, so as to carry out scientific research such as separation, purification, and cultivation in the next step. However, field sampling research of laboratory researchers not only is time-consuming and laborious, also consumes a large amount of scientific research travel expenses, thus taking up a lot of time for researchers and greatly increasing the scientific research cost. The traditional method for collecting euglena is that local fishermen take samples when the euglena blooms and send the samples to the laboratory. Due to the lack of conditions and concepts of aseptic operation, the ordinary fishermen can only use water bottles (plastic water bottles, or the like) to collect water samples; and at the same time, due to the regulations of the general courier services companies, the unknown water sample will not be mailed, even if it can be mailed, there are at least 3 days of transportation, and the alga conditions such as temperature and illumination cannot be controlled during the express transportation, causing different degrees of damage to the water samples (water smelliness, bacteria multiplication, other dominant species blooms, or the i
2020100026 08 Jan 2020 like), thus losing the research value.
In recent years, only a few people have published articles and patents on alga collection, for example, a patent (publication number: CN208594537U) titled with “Handheld green alga collection netting gear that floats” (see FIG. 1). The device is suitable for green alga that floats less, can be used for concentrated collection of floating green alga, and can also be used for collecting when the euglena is less; but the operation is not convenient, the cost is high, and further operations are required to perform mailing. An another patent (publication number: CN206260301U) is titled with “Shallow water area benthic diatom collector” (see FIG. 2), where this patent focuses on the collection of benthic diatoms, but this patent is not applicable to the collection of euglena because the euglena is unicellular alga and can swim. How to collect euglena in the field and make them survive as much as possible during transportation for subsequent research in the laboratory is an urgent problem to be solved.
SUMMARY
The existing euglena field collection equipment is rough and requires professional operation, which is difficult to meet the requirements of fast mailing, and even if it can be mailed, it cannot be mailed by air transportation and can only be mailed by land transportation, which takes a long time, thereby leading to the problems that alga growth conditions such as temperature and illumination cannot be controlled, causing different degrees of damage to the water samples (water smelliness, bacteria multiplication, other dominant species blooms, or the like), and losing the research value. The present invention provides a portable device for field collection of euglena. The device includes a hollow tube body, a tube cap matched with the hollow tube body, and a pick-and-place rod. One end of the pick-and-place rod is fixed with a water absorbent collecting body and the pick-and-place rod is disposed in the hollow tube body.
Specifically, in the portable device for field collection of euglena provided in the present invention, the water absorbent collecting body is a cotton or sponge or super absorbent resin.
Preferably, in the portable device for field collection of euglena provided in the present invention, the hollow tube body is made of opaque materials.
Preferably, in the portable device for field collection of euglena provided in the present invention, the bottom of the hollow tube body is conical, so that the pressing force, applied by the tube wall, of the water absorbent collecting body is becoming small.
In addition, the present invention provides a method for field collection of euglena, which uses the portable device for field collection of euglena provided in the present invention, and
2020100026 08 Jan 2020 includes the following steps:
(1) before collection, subjecting the portable device for field collection of euglena to biocidal treatment;
(2) adding an euglena culture medium containing antibiotics to the hollow tube body under aseptic conditions, wherein the amount of the culture medium is 0.2-0.4 times the water absorption amount of the water absorbent collecting body, and tightly closing the cap;
(3) taking the portable device for field collection of euglena into a collection place, holding the pick-and-place rod in hand and using the water absorbent collecting body to slightly sweep the surface of the euglena bloom until the water absorbent collecting body is green or rust red, immediately putting it back into the hollow tube body, and covering the tube cap; and collecting water samples below the surface of the euglena bloom;
(4) after the alga sample is sent to the demanding place, adding collected water samples filtered and sterilized by a filtration membrane to the hollow tube body to restore the activity of the euglena, and then adding euglena culture medium; and (5) detecting the collected euglena samples: coating a plate and observing the bacteria contamination of each sample after the euglena samples are cultured.
Specifically, in the method for filed collection of euglena provided in the present invention, the culture medium used is PolyA culture medium.
Specifically, in the method for filed collection of euglena provided in the present invention, the antibiotic contained in the used culture medium is ampicillin.
Specifically, in the method for filed collection of euglena provided in the present invention, the content of the ampicillin is 50-100 mg/mL.
The portable device and method for field collection of euglena of the present invention have the following advantages:
The water absorbent collecting body is added with a small amount of liquid culture medium for transporting quickly; the collected euglena is adsorbed on the water absorbent collecting body for facilitating sampling; and the water absorbent collection body can prevent the euglena from swimming in the transportation process (bad environment) and can make the euglena fixed in the materials. After transported to the laboratory, an appropriate amount of culture medium is added to a centrifuge tube, light is applied to restore a favorable environment, at this time, the motility
2020100026 08 Jan 2020 of the euglena is enhanced, and the euglena swims from the water absorbent collecting body into the culture medium and grows in a large amount. In this process, some other species introduced by sampling are greatly reduced due to poor environment and poor motility.
Comparison of the portable device for field collection of euglena of the present invention and a conventional sampling device:
| Conventional sampling device | Device of the present invention |
| Easy to operate, sampling requires to reach the bottle into water | Easy to operate, sampling only requires to slightly sweep the water surface by using a cotton ball with a rod |
| Massive bacterial reproduction | Antibiotics inhibit the bacterial growth |
| Undemutrition, darkness or high temperature, and alga death | Semi-wet environment, other alga and floating small animals death; euglena is under dormancy to avoid the harsh environment |
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is a schematic diagram of a handheld green alga collection netting gear that floats described in the background;
FIG. 2 is a schematic diagram of a shallow water area benthic diatom collector described in the background;
FIG. 3 is a schematic diagram of a portable device for field collection of euglena according to an embodiment of the present invention;
FIG. 4 shows a culture result of field collection of euglena under different operating conditions by using the portable device for field collection of euglena according to the present invention; and
FIG. 5 shows a plate culture result of a water sample of field collection of euglena under different operating conditions by using the portable device for field collection of euglena according to the present invention.
DETAILED DESCRIPTION
For better presentation of the conception of the present invention, the following describes the technical solutions of the present invention in detail with reference to specific embodiments and attached drawings.
2020100026 08 Jan 2020
As shown in FIG. 3, the portable device for field collection of euglena in an embodiment of the present invention includes a hollow tube body 1, a tube cap 2 matched with the hollow tube body 1, and a pick-and-place rod 3. One end of the pick-and-place rod is fixed with a water absorbent collecting body 31 and the pick-and-place rod 3 is disposed in the hollow tube body 1.
In other embodiments of the present invention, the pick-and-place rod can fix sponge or other polymeric water absorbing materials as the water absorbent collecting body 31, which is not shown in the figure.
The hollow tube body 1 is made of opaque materials.
Collection method
The operation of collecting euglena by using the portable device for field collection of euglena in this embodiment is as follows:
(1) before collection, subjecting the portable device for field collection of euglena to biocidal treatment;
(2) adding an euglena culture medium of PolyA culture medium containing antibiotics (where the content of ampicillin is 50-100 mg/mL) to the hollow tube body under aseptic conditions, where the amount of the PolyA culture medium is 0.2-0.4 times the water absorption amount of the water absorbent collecting body (1-2 ml in this embodiment), and tightly closing the cap;
(3) taking the portable device for field collection of euglena into a collection place, holding the pick-and-place rod in hand and using a cotton ball to slightly sweep the surface of the euglena bloom until the water absorbent collecting body is green or rust red, immediately putting it back into the hollow tube body, and covering the tube cap; and collecting water samples below the surface of the euglena bloom;
(4) after the alga sample is sent to the demanding place, adding 5 mL of collected water samples filtered and sterilized by a filtration membrane to the hollow tube body to restore the activity of the euglena, and then adding the PolyA culture medium; and (5) detecting the collected euglena samples: coating an LB plate and observing the bacteria contamination of each sample after the euglena samples are cultured for three days.
The formula of the PolyA culture medium used in this embodiment is shown in Table 1:
Table 1 PolyA culture medium used for culture in the present invention
2020100026 08 Jan 2020
| Component | g/L |
| Sodium acetate | 2 |
| Yeast extract | 1 |
| Tryptone | 1 |
The water absorbent collecting body used in this embodiment is a 0.2 g cotton ball with the water absorption amount about 5 mL. In the specific embodiment, the condition setting for collecting euglena by using the portable device for field collection of euglena is shown in Table 2 below:
Table 2 Condition setting for collecting euglena
| Conditions | 1 | 2 | 3 | 4 | 5 |
| Water absorbent collecting body | Cotton ball | Cotton ball | Cotton ball | Cotton ball | Cotton ball |
| Sterilization of water absorbent collection body | Non-sterile | Non-sterile | Sterile | Sterile | Sterile |
| Culture medium addition | 1 ml tap water (0.2 times) | 3 ml tap water (0.6 times) | 1 ml sterile polyA culture medium (0.2 times) | 1 ml sterile polyA culture medium (0.2 times) | 1.5 ml sterile polyA culture medium (0.3 times) |
| Content of antibiotics in the added culture medium | 0 mg/mL | 0 mg/mL | 50 mg/mL | 100 mg/mL | 50 mg/mL |
| Aseptic operation | No | No | Yes | Yes | Yes |
| Conditions | 6 | 7 | 8 | 9 | 10 |
| Water absorbent collecting body | Cotton ball | Two cotton swabs | Two cotton swabs | Three cotton swabs | Three cotton swabs |
| Sterilization of water absorbent collection body | Sterile | Sterile | Sterile | Sterile | Sterile |
2020100026 08 Jan 2020
| Culture medium addition | 2 ml sterile polyA culture medium (0.4 times) | 250 uL sterile polyA culture medium | 500 uL sterile polyA culture medium | 500 uL sterile polyA culture medium | 750 uL sterile polyA culture medium |
| Content of antibiotics in the added culture medium | 100 mg/mL | 50 mg/mL | 50 mg/mL | 100 mg/mL | 100 mg/mL |
| Aseptic operation | Yes | Yes | Yes | Yes | Yes |
Euglena collection effect verification
Generally, the indicators for verifying the euglena collection effect include whether the water sample is contaminated with a large amount of bacteria, the survival amount and activity of the euglena in the water body (which can be determined from the color of the water body). As shown in FIG. 4, both No.l and No.2 tubes are added with unsterilized water and cotton balls, while the remaining tubes are added with sterilized cotton balls and culture medium containing antibiotics. It can be learned from the results that, compared with other tubes, the water bodies in the No.l and No.2 tubes are contaminated with a large amount of bacteria, and the water samples are turned yellow. Compared with the results of No. 3, No. 4, No. 5, and No. 6 tubes, we can learn that for 0.2 g cotton balls, the addition of 2 mL (0.4 times the water absorption amount of the cotton ball of the water absorbent collecting body) of PolyA culture medium with the content of ampicillin being 100 mg/mL can make the sampling results better. The No. 3, No. 4, No. 5, and No. 6 tubes are compared with the No. 7, No. 8, No. 9, and No. 10 tubes; the former four tubes use cotton balls, while the latter four tubes use cotton swabs; and it can be learned from the results that the water absorption amount of the cotton swab is not as good as that of the cotton ball, and therefore, the amount of euglena collected by the cotton swab is not as good as that of euglena collected by the cotton ball.
FIG. 5 shows the results of collected 10 samples that are cultured on a plate. The plate culture is carried out by a routine experiment method, where 100 uL samples are taken from each tube for performing plate streaking, and then placed in a 37°C incubator for culturing two days. From the plate culture results, a large number of bacteria are grown in NO. 1 and NO. 2 plates after two days of plate culture, while other samples do not grow bacterial colonies after two days of plate culture, which shows that the addition of 50 mg/mL - 100 mg/mL ampicillin to the culture medium can effectively inhibit the growth of bacteria.
Claims (4)
- (1) before collection, subjecting the portable device for field collection of euglena to biocidal treatment;1. A portable device for field collection of euglena, comprising a hollow tube body, a tube cap matched with the hollow tube body, and a pick-and-place rod, wherein one end of the pick-andplace rod is fixed with a water absorbent collecting body and the pick-and-place rod is disposed in the hollow tube body.
- (2) adding an euglena culture medium containing antibiotics to the hollow tube body under aseptic conditions, wherein the amount of the culture medium is 0.2-0.4 times the water absorption amount of the water absorbent collecting body, and tightly closing the cap;2. The portable device for field collection of euglena according to claim 1, wherein the water absorbent collecting body is a cotton or sponge or super absorbent resin.
- (3) taking the portable device for field collection of euglena into a collection place, holding the pick-and-place rod in hand and using the water absorbent collecting body to slightly sweep the surface of the euglena bloom until the water absorbent collecting body is green or rust red, immediately putting it back into the hollow tube body, and covering the tube cap tightly; and collecting water samples below the surface of the euglena bloom;3. The portable device for field collection of euglena according to claim 1, wherein the hollow tube body is made of opaque materials, preferably, wherein the bottom of the hollow tube body is conical; wherein the water absorbent collecting body is a cotton or sponge or super absorbent resin.4. The portable device for field collection of euglena according to claim 1, wherein the bottom of the hollow tube body is conical.5. A method for field collection of euglena, using any one of the portable device for field collection of euglena according to claims 1 to 4, comprising the following steps:
- (4) after the alga sample is sent to the demanding place, adding collected water samples filtered and sterilized by a filtration membrane to the hollow tube body to restore the activity of the euglena, and then adding the euglena culture medium; and (5) detecting the collected euglena samples: coating a plate and observing the bacteria contamination of each sample after the euglena samples are cultured;preferably, wherein the culture medium used is PolyA culture medium; wherein the antibiotic2020100026 08 Jan 2020 contained in the used culture medium is ampicillin; wherein the content of the ampicillin is 50-100 mg/mL.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910749251.2A CN110387319A (en) | 2019-08-14 | 2019-08-14 | A kind of device and method of Portable field acquisition Euglena |
| CN201910749251.2 | 2019-08-14 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| AU2020100026A4 true AU2020100026A4 (en) | 2020-02-13 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| AU2020100026A Ceased AU2020100026A4 (en) | 2019-08-14 | 2020-01-08 | Portable device and method for field collection of euglena |
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| Country | Link |
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| CN (1) | CN110387319A (en) |
| AU (1) | AU2020100026A4 (en) |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113684134B (en) * | 2021-07-13 | 2023-08-29 | 上海师范大学 | Preservation method and device suitable for living algae during field sampling |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN203502260U (en) * | 2013-09-30 | 2014-03-26 | 华北电力大学（保定） | Saliva sampler |
| CN204594720U (en) * | 2015-05-22 | 2015-08-26 | 中华人民共和国宁德出入境检验检疫局 | Aseptic packet of samples |
| AU2016383699A1 (en) * | 2016-02-12 | 2017-08-31 | John A. Hommeyer | Systems, methods, and compositions related to using non-live-bacteria preparations to promote food safety and preservation |
| CN207001179U (en) * | 2017-05-27 | 2018-02-13 | 广西壮族自治区农业科学院花卉研究所 | A kind of plant living body sampler bag |
| CN208752081U (en) * | 2018-07-03 | 2019-04-16 | 陶心江 | Feces collection detection device |
-
2019
- 2019-08-14 CN CN201910749251.2A patent/CN110387319A/en active Pending
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| CN110387319A (en) | 2019-10-29 |
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