阅读说明：本技术 内容物填充系统中的初始菌确认方法以及无菌性验证方法 (Method for confirming initial bacteria and method for verifying sterility in content filling system ) 是由 早川睦 和田唯子 玉川隆一 时元翼 于 2017-03-07 设计创作，主要内容包括：本发明提供内容物填充系统中的初始菌确认方法以及无菌性验证方法,首先,在不利用容器杀菌装置(13)对瓶(30)进行杀菌的情况下将瓶(30)传送到填充装置(20)中,使用填充装置(20)向瓶(30)内填充培养基。接着,使用盖体安装装置(16)用盖体(33)封闭瓶(30)。其后,验证在瓶(30)内的培养基中是否有菌的存活或繁殖。(The invention provides an initial bacteria confirmation method and a sterility verification method in a content filling system, firstly, a bottle (30) is conveyed to a filling device (20) without sterilizing the bottle (30) by a container sterilizing device (13), and a culture medium is filled into the bottle (30) by using the filling device (20). Then, the bottle (30) is closed with a cap (33) using a cap attachment device (16). Thereafter, it was verified whether bacteria survived or propagated in the medium in the flask (30).)

1. An initial bacteria confirmation method for confirming initial bacteria in a container using a content filling system, the content filling system comprising: a container sterilization apparatus for sterilizing the container, a filling apparatus for filling the container with a content, a gas replacement apparatus for replacing a gas in a head space in the container with an inert gas, and a lid attachment apparatus for closing the container with a lid, the method comprising:

a step of transferring the container into the filling device without sterilizing the container by the container sterilizing device,

a step of filling the culture medium into the container using the filling device,

a step of closing the container with the lid using the lid attachment device, and

verifying the presence or absence of the survival or growth of the bacteria in the culture medium in the vessel,

in the step of filling the culture medium, air is supplied from the gas replacement device to replace the inert gas.

2. The method for confirming initial bacteria according to claim 1, further comprising: and a step of adjusting the sterilization conditions in the container sterilization device based on the result of the verification.

3. An initial bacteria confirmation method for confirming an initial bacteria of a lid body using a content filling system, the content filling system comprising: a filling device for filling a container with contents, a gas replacement device for replacing a gas in a head space in the container with an inert gas, a lid body sterilization device for sterilizing the lid body, and a lid body attachment device for closing the container with the lid body, the method comprising:

a step of filling the culture medium into the vessel using the filling device,

a step of transferring the lid body to the lid body attachment device without sterilizing the lid body by the lid body sterilization device,

a step of closing the container with the lid using the lid attachment device, and

verifying the presence or absence of the survival or growth of the bacteria in the culture medium in the vessel,

in the step of filling the culture medium, air is supplied from the gas replacement device to replace the inert gas.

4. The method of confirming initial bacteria according to claim 3, further comprising: and a step of preparing the sterilization conditions in the cap sterilization device based on the result of the verification.

5. The method of confirming initial bacteria according to claim 1, wherein the contents are acidic, and the pH of the medium is 3.5 or more and 4.6 or less.

6. The method of confirming initial bacteria according to claim 1, wherein the content is neutral, and the pH of the medium is 6 or more and 8 or less.

7. The method of confirming initial bacteria according to claim 1, wherein the content filling system comprises a carbon dioxide dissolving device for adding carbon dioxide to the content,

in the step of filling the culture medium, the supply of carbon dioxide to be added to the content by the carbon dioxide dissolution device is stopped, or air is added to the content by the carbon dioxide dissolution device.

8. The method of confirming initial bacteria according to claim 1, further comprising, before starting the culture of the vessel, the steps of: the culture medium is surely brought into contact with the inner surface of the container by tilting or inverting the container filled with the culture medium.

9. The method of confirming initial bacteria according to claim 1, wherein the content filling system has an aseptic chamber that houses: an air rinsing device for supplying sterile air to the container, the container sterilizing device, the filling device, and the cap attaching device,

the container sterilization apparatus includes: a conduit for supplying a sterilizing agent to the container, the conduit being provided with a 1 st valve for controlling the passage of the sterilizing agent in the conduit; and a bypass conduit communicating with the inside of the sterile chamber, the bypass conduit being provided with a 2 nd valve for controlling the passage of the bactericide in the bypass conduit,

the step of delivering the container to the filling apparatus comprises closing the 1 st valve while opening the 2 nd valve, or opening the 1 st valve while opening the 2 nd valve, thereby bypassing the sterilizing agent from being introduced into the container, and the step of delivering the container to the filling apparatus comprises bypassing the sterile air from being introduced into the container and supplying the sterile air into the sterile chamber.

10. The method for confirming initial bacteria according to claim 1, wherein the content filling system comprises: a sterile water rinsing device for rinsing the container sterilized by the container sterilizing device with sterile water,

in the step of transferring the container to the filling device, the sterile water rinsing device reduces the flow rate of the sterile water to a level where the sterile water does not contact the container while supplying the sterile water to the container.

11. A method of verifying sterility, which is a method of verifying sterility using a content filling system having: a container sterilization apparatus for sterilizing a container and a filling apparatus for filling the container with a content, the method comprising:

a step of individually performing a test for each component of the content filling system to determine whether or not sterility is ensured,

a step of filling the culture medium having a pH adjusted to correspond to the pH of the content in advance into the container using the filling device after the step of performing the detection alone,

a step of closing the vessel filled with the culture medium,

a step of bringing the culture medium into reliable contact with the inner surface of the container by inclining or inverting the container filled with the culture medium before starting the culture of the container, and

and verifying the presence or absence of the survival or growth of the bacteria in the culture medium in the container.

12. The sterility verification method of claim 11, further comprising: and a step of adjusting the sterilization conditions in the container sterilization device based on the result of the verification.

13. A method of verifying sterility, which is a method of verifying sterility using a content filling system having: a filling device for filling a container with contents, a lid body sterilization device for sterilizing the lid body, and a lid body attachment device for closing the container with the lid body, the method comprising:

a step of individually performing a test for each component of the content filling system to determine whether or not sterility is ensured,

a step of filling the culture medium having a pH adjusted to correspond to the pH of the content in advance into the container using the filling device after the step of performing the detection alone,

a step of transferring the cover to the cover attachment device,

a step of closing the container with the lid by using the lid attachment device,

a step of bringing the culture medium into reliable contact with the inner surface of the container by inclining or inverting the container filled with the culture medium before starting the culture of the container, and

and verifying the presence or absence of the survival or growth of the bacteria in the culture medium in the container.

14. The sterility verification method of claim 13, further comprising: and a step of preparing the sterilization conditions in the cap sterilization device based on the result of the verification.

15. The sterility test method according to claim 11, wherein the content is acidic, and the pH of the medium is 3.5 or more and 4.6 or less, or

The content is neutral, and the pH of the medium is set to 6 or more and 8 or less.

Technical Field

The present invention relates to a method for confirming initial bacteria in a content filling system, a method for verifying a content filling system, and a medium used in such a verification method.

Background

An aseptic filling system is widely known in which a sterilized container (PET bottle) is filled with a sterilized content in an aseptic environment, and the container is then closed with a cap. Specifically, in the aseptic filling system, the molded container is supplied to the aseptic filling system, and an aqueous hydrogen peroxide solution as a bactericide is sprayed into the container in the aseptic filling system. The container is thereafter dried to sterilize it, and then the contents are aseptically filled into the container. As another method, there is also the following method: in the case of forming a container, a small amount of a sterilizing agent is dropped onto the inner surface of the container, the mouth portion is sealed, the inner surface of the container is sterilized by vapor of the vaporized sterilizing agent (hydrogen peroxide), the sterilized container is fed into an aseptic filling system, the outer surface of the container is sterilized in the aseptic filling system, and thereafter the mouth portion is opened to aseptically fill the contents.

In addition, for example, in the initial stage of the aseptic filling system, it is necessary to confirm whether or not the sterility of the system can be ensured before actually starting the filling of the container. Various tests were therefore performed to confirm the sterility of the system. After such various tests, an evaluation method using a container filled with a culture medium has been performed in order to comprehensively evaluate the sterility of the aseptic filling system in the final stage.

For example, patent document 1 discloses a method for verifying the sterility level of a container by filling the container with a culture medium after a sterilization treatment.

However, consideration is also given to the fact that in aseptic filling systems the container or cap is contaminated from the outset. In such a case, even if the contents are aseptically filled into the container using an aseptic filling system, there is a possibility that bacteria may multiply inside the finished beverage product, and thus measures such as increasing the amount of the bactericide are required. Therefore, it is important to accurately grasp in advance how much the container or the cap is contaminated with bacteria, or the initial number of bacteria in the container or the cap (the number of bacteria adhering to the container or the cap before filling).

In the detection using such a medium, a neutral medium (pH6 or more and 8 or less) is generally used as the medium for the reason that almost all bacteria can be detected. By using a neutral medium, it is possible to cope with whatever kind of contents is filled by the aseptic filling system. However, in practice, the contents filled by the aseptic filling system may be limited to a specific type. In this case, performing detection for detecting all bacteria imposes a load on the system more than necessary, and increases costs for equipment, chemicals, energy, and the like.

Documents of the prior art

Patent document

Patent document 1: japanese laid-open patent publication No. 2010-36973

Disclosure of Invention

The present invention has been made in view of the above problems, and an object thereof is to provide an initial bacteria confirmation method by which initial bacteria adhering to a container or a lid can be accurately grasped in a content filling system using actual manufacturing equipment. Further, an object of the present invention is to provide a method for verifying a content filling system and a culture medium, which can suppress costs required for equipment, a drug, energy, and the like by using an appropriate culture medium corresponding to the content filled by the content filling system.

The present invention relates to an initial bacteria confirmation method for confirming initial bacteria in a container using a content filling system, the content filling system including: a container sterilization apparatus for sterilizing the container, a filling apparatus for filling the container with a content, and a lid attachment apparatus for closing a lid for the container, the method comprising: a step of transferring the container to the filling device without sterilizing the container by the container sterilizing device; filling the culture medium into the container using the filling device; a step of closing the container with the lid by using the lid attachment device; and verifying the presence or absence of the survival or propagation of the bacteria in the culture medium in the container.

The initial bacteria verification method of the present invention is characterized by further comprising a step of preparing sterilization conditions in the container sterilization device based on the verification result.

The present invention relates to an initial bacteria confirmation method for confirming initial bacteria in a lid body using a content filling system, the content filling system including: a filling device for filling a container with contents, a lid body sterilization device for sterilizing the lid body, and a lid body attachment device for closing the container with the lid body, the method comprising the steps of: filling the culture medium in the container by using the filling device; a step of transferring the lid body to the lid body attachment device without sterilizing the lid body by the lid body sterilization device; a step of closing the container with the lid by using the lid attachment device; and verifying the presence or absence of the survival or propagation of the bacteria in the culture medium in the container.

The initial bacteria verification method of the present invention is characterized by further comprising a step of preparing sterilization conditions in the cap sterilization device based on the verification result.

The method for confirming the initial bacteria of the present invention is characterized in that the contents are acidic, and the pH of the medium is 3.5 or more and 4.6 or less.

The method for confirming an initial bacterium of the present invention is characterized in that the content is neutral, and the pH of the medium is set to 6 or more and 8 or less.

In the initial bacteria confirmation method of the present invention, in the verification step, physical movement is applied to the culture medium in the container.

According to the present invention, initial bacteria adhering to the container or the lid can be detected.

The present invention relates to a method for verifying a content filling system using a culture medium, the method comprising: a step of supplying a container to the content filling system; filling the culture medium into the container in the content filling system, and then sealing the container; and verifying whether or not bacteria survive or proliferate in the medium in the container; the characteristics of the culture medium are made equivalent to those of the contents filled in the contents filling system, that is, characteristics affecting the growth of bacteria.

The method of the present invention is characterized in that the characteristic of the content is the pH of the content, and the pH of the medium is adjusted to correspond to the pH of the content.

The verification method of the present invention is characterized in that the pH of the medium is 3.5 or more and 4.6 or less.

The verification method of the present invention is characterized in that the content contains carbon dioxide and carbon dioxide is dissolved in the culture medium.

The verification method of the present invention is characterized in that the content does not contain at least one of a carbon source and a nitrogen source, and the medium does not contain at least one of a carbon source and a nitrogen source.

The verification method of the present invention is characterized in that the content does not contain at least one of a carbon source and a nitrogen source, and catechin is dissolved in the medium.

The verification method of the present invention is characterized in that the characteristic of the content is a total organic carbon amount, and the total organic carbon amount of the culture medium is adjusted to be equivalent to the total organic carbon amount of the content.

The verification method of the present invention is characterized in that the content contains catechin, and catechin is dissolved in the medium.

The present invention relates to a culture medium used in the above-described verification method, wherein the characteristics of the culture medium are equivalent to the characteristics of the content filled by the content filling system, that is, the characteristics affecting the growth of bacteria.

According to the present invention, by matching the characteristics of the culture medium with the characteristics of the contents filled by the contents filling system, it is possible to verify whether or not bacteria have grown in the culture medium in the container while suppressing the costs required for facilities, chemicals, energy, and the like.

Drawings

Fig. 1 is a schematic plan view showing a content filling system used in an initial bacteria verification method according to embodiment 1 of the present invention.

Fig. 2 is a flowchart illustrating an initial bacteria confirmation method according to embodiment 1 of the present invention.

Fig. 3 is a schematic plan view showing a content filling system when the initial bacteria verification method according to embodiment 1 of the present invention is performed.

Fig. 4 is a flowchart showing an initial bacteria confirmation method according to a modification example of embodiment 1 of the present invention.

Fig. 5 is a schematic cross-sectional view showing a sterilization apparatus of a content filling system used in the initial bacteria verification method according to embodiment 1 of the present invention.

Fig. 6 is a schematic plan view showing a content filling system used in the verification method according to embodiment 2 of the present invention.

Fig. 7 is a flowchart showing a verification method according to embodiment 2 of the present invention.

Fig. 8 is a schematic plan view showing a content filling system when the verification method according to embodiment 2 of the present invention is performed.

Detailed Description

(embodiment 1)

Embodiment 1 of the present invention will be described below with reference to the drawings. Fig. 1 to 3 are views showing embodiment 1 of the present invention.

(Contents filling System)

First, a content filling system (aseptic filling system ) according to the present embodiment will be described with reference to fig. 1.

The content filling system 10 shown in fig. 1 is a system for filling a bottle (container) 30 with a content such as a beverage. The bottle 30 can be produced by subjecting a preform, which is produced by injection molding of a synthetic resin material, to biaxial stretching blow molding. As a material of the bottle 30, a thermoplastic resin, particularly PE (polyethylene), PP (polypropylene), PET (polyethylene terephthalate), or PEN (polyethylene naphthalate), is preferably used. In addition, the container may be glass, a can, paper, a bag, or a composite container thereof. In the present embodiment, a case where a bottle is used as a container will be described as an example.

As shown in fig. 1, the content filling system 10 includes a bottle supply unit 21, a sterilization device 11, an air rinsing device 14, a sterile water rinsing device 15, a filling device (filler) 20, a cap attachment device (capping machine, hemming and sealing machine) 16, and a product bottle carrying-out unit 22. These bottle supply unit 21, sterilizing device 11, air rinsing device 14, sterile water rinsing device 15, filling device 20, lid attachment device 16, and product bottle carrying-out unit 22 are arranged in this order from the upstream side to the downstream side in the conveying direction. Further, 2 or more transfer wheels 12 for transferring bottles 30 between the sterilizing apparatus 11 and the air rinsing apparatus 14 and the sterile water rinsing apparatus 15 and between the filling apparatus 20 and the cap attaching apparatus 16 are provided.

The bottle supply unit 21 sequentially receives empty bottles 30 from the outside into the content filling system 10, and transfers the received bottles 30 to the sterilizer 11.

A bottle molding section (not shown) for molding the bottle 30 by biaxially stretch blow molding a preform may be provided upstream of the bottle supply section 21. In this way, the steps from the supply of the preform through the molding of the bottle 30 to the filling and closing of the content in the bottle 30 can be continuously performed. In this case, the preform having a small volume can be transported from the outside to the content filling system 10, not in the form of the bottle 30 having a large volume, and thus the transportation cost can be reduced.

Sterilizing apparatus 11 sprays a sterilizing agent into bottle 30 to sterilize the inside of bottle 30. Thus, the bottle 30 is sterilized with the sterilizing agent before the contents are filled, and spores of bacteria are allowed to survive but vegetative cells, mold, and yeast of bacteria are not allowed to survive. As the bactericide, for example, an aqueous hydrogen peroxide solution is used. Mist or gas of the hydrogen peroxide solution is generated in the sterilizer 11, and the mist or gas is sprayed on the inner and outer surfaces of the bottle 30. Since the inside of the bottle 30 is sterilized with the mist or gas of the hydrogen peroxide solution in this manner, the inner surface of the bottle 30 is uniformly sterilized.

The air rinsing device 14 supplies sterile heated air or normal temperature air to the bottle 30 to activate the hydrogen peroxide and remove foreign substances, hydrogen peroxide, and the like from the bottle 30.

The sterile water rinsing device 15 cleans the bottle 30 sterilized with hydrogen peroxide as a sterilizing agent with sterile water at 15 to 85 ℃. This allows the hydrogen peroxide adhering to the bottle 30 to be washed away and foreign matter to be removed. The sterile water rinsing device 15 may not be provided.

In the present embodiment, the container sterilization device 13 is configured by the above-described sterilization device 11.

The filling device 20 fills the bottle 30 with the content that has been sterilized in advance from the mouth of the bottle 30. In the filling device 20, the empty bottle 30 is filled with the content. In this filling device 20, the contents are filled into the bottles 30 while rotating (revolving) 2 or more bottles 30. The content may be filled into the bottle 30 at normal temperature. The contents are sterilized by heating or the like in advance, cooled to a room temperature of 3 ℃ or higher and 40 ℃ or lower, and then filled in the bottle 30. Spores of the bacteria are allowed to survive within the vial 30, as described above. Therefore, it is not necessary to heat and sterilize product bottle 35 (described later) from the outside, which is filled with the content in a state heated to a high temperature in bottle 30, which is kept for a long time after the content is filled in bottle 30, or which is filled with the content in bottle 30 and closed with lid 33, as in the conventional case.

The contents filled by the filling device 20 have predetermined characteristics that affect the growth of bacteria. In the present embodiment, the predetermined characteristic may be the pH of the content. More specifically, the contents may be constituted by an acidic beverage. The beverage preferably has an acidity of less than ph4.6, more preferably less than ph 4.0. Examples of the beverage having a pH of 4.0 or more and a pH of 4.6 or less include tomato juice, vegetable juice, etc., and examples of the beverage having a pH of less than 4.0 include lemon tea, orange juice, milk carbonated beverage, functional beverage, carbonated lemon juice, grape juice, fruit juice (fruit juice ジュース), etc.

In general, spores of bacteria maintain a sterile state without germination in a liquid having a high acidity to some extent (for example, a pH of less than 4.6, preferably less than 4.0), and therefore, the contents can be stored without causing putrefaction. Therefore, as described above, the beverage can be prevented from being deteriorated or spoiled by filling the bottle 30 with the sterilized contents having an acidity at which spores of bacteria remain in a viable state in the bottle 30 before filling by the filling device 20, but germination of the spores of bacteria can be suppressed (for example, a pH of less than 4.6, preferably less than 4.0).

The cap attaching device 16 closes the bottle 30 by attaching the cap 33 to the mouth of the bottle 30. In the cap mounting device 16, the mouth of the bottle 30 is closed and sealed by the cap 33 so that external air or microorganisms do not intrude into the bottle 30. In the cap attaching device 16, the cap 33 is attached to the mouth portion of 2 or more bottles 30 filled with the content while rotating (revolving). Thus, product bottle 35 is obtained by attaching cap 33 to the mouth of bottle 30.

The lid 33 is sterilized in advance by the lid sterilizer 18. The cap sterilization device 18 is disposed, for example, inside the sterile chamber 70 (described later) and in the vicinity of the cap attachment device 16. In the cap sterilization device 18, a plurality of caps 33 loaded from the outside of the content filling system 10 are collected in advance and conveyed in a row toward the cap attachment device 16. On the way of the lid 33 toward the lid attachment device 16, mist or gas of hydrogen peroxide is blown toward the inner and outer surfaces of the lid 33, and then dried with hot air to perform sterilization treatment.

The product bottle carrying-out section 22 continuously carries out the product bottles 35 to which the caps 33 are attached by the cap attachment device 16, to the outside of the content filling system 10.

It is noted that the content filling system 10 has a sterile chamber 70. The sterilization apparatus 11, the air rinsing apparatus 14, the sterile water rinsing apparatus 15, the filling apparatus 20, the lid sterilization apparatus 18, and the lid attachment apparatus 16 are housed in the sterile chamber 70. Such a content filling system 10 may be constituted by, for example, an aseptic filling system. In this case, the inside of the aseptic chamber 70 is maintained in an aseptic state.

Alternatively, the content filling system 10 may be a high-temperature filling system that fills the content at a high temperature of 85 ℃ or higher and less than 100 ℃. It may be a medium-temperature filling system for filling the content at a medium temperature of 55 ℃ or higher and less than 85 ℃. The technical idea of the present embodiment can also be applied to aseptic packaging using post-sterilization such as autoclaving.

(method of filling content)

Next, a content filling method using the content filling system 10 (fig. 1) will be described. A filling method in a normal case, that is, a method of filling contents such as beverages into the bottle 30 to manufacture the product bottle 35, will be described below.

First, 2 or more empty bottles 30 are sequentially supplied to the bottle supply unit 21 from the outside of the content filling system 10. The bottles 30 are conveyed from the bottle supply unit 21 to the sterilizer 11 by the conveyor wheel 12 (container supply step).

Next, in sterilization apparatus 11 constituting container sterilization apparatus 13, bottle 30 is sterilized with an aqueous hydrogen peroxide solution as a sterilizing agent (sterilization step). At this time, the hydrogen peroxide solution is once vaporized to a gas or mist at a boiling point or higher, and is supplied to the bottle 30. The mist of the hydrogen peroxide solution adheres to the entire inner surface of the bottle 30, and sterilizes vegetative cells of bacteria, mold, and yeast in the bottle 30. The amount of the mist of hydrogen peroxide supplied into the bottle 30 is, for example, 5 μ L/bottle or more and 50 μ L/bottle or less, and 1mg/L or more and 5mg/L or less in the case of hydrogen peroxide gas, and the bactericidal activity is considered to be such that the vegetative cells, mold, and yeast of bacteria can be sterilized but spores of bacteria are not sterilized. Thereby enabling the amount of hydrogen peroxide to be reduced.

Next, the bottles 30 are conveyed to the air rinsing device 14 by the conveying wheel 12, and the air rinsing device 14 activates hydrogen peroxide by supplying sterile heated air or normal temperature air, and removes foreign substances, hydrogen peroxide, and the like from the bottles 30. Next, the bottle 30 is transferred to the sterilized water rinsing device 15 by the transfer wheel 12. In the aseptic water rinsing apparatus 15, washing is performed with aseptic water at 15 to 85 ℃ (rinsing step). Specifically, sterile water at 15 to 85 ℃ is supplied into the bottle 30 at a flow rate of 5L/min to 15L/min. In this case, it is preferable that the bottle 30 is inverted, and sterile water is supplied into the bottle 30 from the mouth portion facing downward, and the sterile water flows out of the bottle 30 from the mouth portion. The hydrogen peroxide adhering to the bottle 30 is washed away by the hot water, and foreign substances are removed.

The bottles 30 are then transferred by the transfer wheel 12 to the filling device 20. In this filling device 20, the contents are filled into the bottle 30 from the mouth while rotating (revolving) the bottle 30 (filling step).

Before the bottle 30 is filled with the filling device 20, the contents are prepared in advance and heat-sterilized. As described above, the content may have a predetermined pH as a characteristic affecting the growth of bacteria. In particular, the contents may consist of an acidic beverage preferably less than pH4.6, more preferably less than pH4. The heating temperature is usually about 60 ℃ or higher and 120 ℃ or lower when the acidity of the contents is less than ph4.0, and about 115 ℃ or higher and 150 ℃ or lower when the acidity is at ph4.0 or higher. In this way, all microorganisms that can grow in the product bottle 35 in the content before filling are sterilized. The contents after the heat sterilization treatment are cooled to a normal temperature of about 3 ℃ to 40 ℃.

In the filling device 20, the sterilized bottle 30 is filled with the contents that have been sterilized and cooled to room temperature at room temperature. The temperature of the contents during filling is, for example, about 3 ℃ to 40 ℃. The acidity of the contents is preferably less than pH4.6, more preferably less than pH4 as described above, and specifically, tomato juice, vegetable juice, lemon tea, orange juice, milk carbonated beverage, functional beverage, carbonated lemon juice, grape juice, fruit juice, and the like can be mentioned. That is, by such a content filling method, the product bottle 35 filled with almost all kinds of beverages except barley tea, mixed tea, and milk-containing beverage having a ph of 4.6 or more can be manufactured. Of course, cola and soda water containing no animal or plant components and having a carbon dioxide pressure of 1.0kg/cm can also be produced²A carbonated beverage product bottle 35 at a temperature of 20 ℃ or higher.

Next, the bottle 30 filled with the content is conveyed to the cap mounting device 16 by the conveying wheel 12.

On the other hand, the lid 33 is sterilized in advance by the lid sterilizer 18 (lid sterilization step). During this time, first, the lid 33 is carried into the lid sterilizer 18 from the outside of the content filling system 10. Next, mist or gas of hydrogen peroxide is blown onto the lid 33 in the lid sterilizer 18 to sterilize the inner and outer surfaces thereof, and then dried with hot air and sent to the lid attachment device 16.

Next, the cap unit 16 attaches the sterilized cap 33 to the mouth of the bottle 30 transferred from the filling unit 20, thereby obtaining a product bottle 35 (cap attaching step).

Thereafter, the product bottle 35 is conveyed from the cap attaching device 16 to the product bottle carrying-out section 22, and carried out to the outside of the content filling system 10.

The steps from the sterilization step to the lid attachment step are performed in an aseptic atmosphere surrounded by the aseptic chamber 70, that is, in an aseptic environment. The aseptic chamber 70 is previously sterilized by spraying hydrogen peroxide, discharging hot water, or the like so as to allow the spores of bacteria to survive but not allow vegetative cells of bacteria, mold, and yeast to survive. After the sterilization process, the sterile air is supplied at a positive pressure into the sterile chamber 70 so that the sterile air is always blown out of the sterile chamber 70.

The production (transfer) speed of the bottle 30 in the content filling system 10 is preferably 100bpm or more and 1500bpm or less. The bpm here means a transport speed of the bottle 30 every 1 minute.

(method of confirming initial bacteria in Contents filling System)

Next, an initial bacteria confirmation method for verifying the sterility of the bottle 30 will be described using the content filling system 10 (fig. 1) described above.

The initial bacteria checking method of the present embodiment is a method of checking whether or not the sterility of the bottle 30 filled with the content is ensured in the content filling system 10. This initial bacteria confirmation method is performed, for example, at an initial stage immediately after the completion of the content filling system 10, that is, before the product bottle 35 is manufactured by actually starting the filling of the bottle 30 using the content filling system 10. Alternatively, the initial bacteria confirmation method according to the present embodiment may be performed when there is a possibility that sterility is affected, for example, when the process or the apparatus in the content filling system 10 is changed at all or when the content filling system 10 is not used for a certain period of time. Alternatively, the initial bacteria confirmation method of the present invention is periodically performed every predetermined filling cycle regardless of whether or not there is a possibility of affecting sterility.

First, before the initial bacteria checking method of the present embodiment is performed, whether or not the sterility is ensured is individually checked for each component of the content filling system 10. Specifically, for example, detection of whether the supply line of the content is properly warmed (SIP warm-up confirmation detection), detection of whether bottle 30 and lid 33 are properly sterilized (bottle sterilization detection, lid sterilization detection), detection of whether sterile chamber 70 is sterilized (chamber sterilization detection), and the like are performed.

After such detection, the method for confirming initial bacteria according to the present embodiment is performed in order to evaluate the sterility of the bottle 30. Specifically, when a plurality of bottles 30 are made to flow through the content filling system 10 and the bottles 30 are not sterilized by the container sterilization apparatus 13 (sterilization apparatus 11), each bottle 30 is filled with a predetermined culture medium instead of the actually filled content and is closed by the lid 33. Thereafter, it was confirmed that the medium filled in each bottle 30 did not decay after a certain period of time had elapsed (method for confirming the initial bacteria in the container).

The method for confirming the initial bacteria of the content filling system 10 (the method for confirming the initial bacteria of the container) according to the present embodiment will be further described below with reference to fig. 2 and 3. Fig. 2 is a flowchart illustrating an initial bacteria confirmation method according to the present embodiment, and fig. 3 is a schematic plan view illustrating a content filling system when the initial bacteria confirmation method according to the present embodiment is performed. In fig. 3, the same components as those of the content filling system 10 shown in fig. 1 are denoted by the same reference numerals.

First, as shown in fig. 3, an empty bottle 30 for verification is made to flow in the content filling system 10. In this case, empty bottles 30 are supplied from the outside to the bottle supply unit 21 of the content filling system 10 (container supply step, step S1 in fig. 2). The number of bottles 30 may be predetermined, and may be, for example, a predetermined number of 100 to 300,000 (preferably 1,000 to 30,000).

Subsequently, the bottle 30 is sent to the sterilizer 11 of the container sterilizer 13. The sterilization apparatus 11, the air rinsing apparatus 14, and the sterile water rinsing apparatus 15 of the container sterilization apparatus 13 are stopped in advance, and the bottle 30 is not sterilized. Therefore, bottle 30 is conveyed to filling device 20 directly through sterilizing device 11, air rinsing device 14, and sterile water rinsing device 15 without sterilizing bottle 30 by sterilizing device 11 (non-sterilized container conveying step, step S2 in fig. 2). Note that bottle 30 may be sent to filling device 20 by using another bypass route instead of sterilization device 11, air rinsing device 14, and sterile water rinsing device 15.

A method of transferring only the bottle 30 into the aseptic chamber 70 without sterilization and filling the culture medium with the filling device (filler) 20 as described below will be described here. First, in the case of the hydrogen peroxide system, the hydrogen peroxide gas is bypassed or the supply of the hydrogen peroxide gas is stopped in the sterilization apparatus 11 so as not to be introduced into the bottle 30. Further, if air is supplied into the bottle 30, the initial number of bacteria in the bottle 30 cannot be accurately measured, and therefore, the air needs to be bypassed.

Fig. 5 is a schematic sectional view showing the sterilization device 11. As shown in fig. 5, a disc 51 that rotates by power from a predetermined drive source is horizontally attached to a turning shaft 53 that stands on a base 52. The support column 54 extends upward from the disk surface of the disk 51, and a manifold 55 for flowing hydrogen peroxide gas is fixed to the upper end of the support column 54. A conduit 56 extends upward from the center of the upper portion of the manifold 55 on an extension of the axis of the swing shaft 53, and the conduit 56 is held by a bearing 57 to a frame member of the aseptic chamber 70 connected to the machine table 52. Thereby, the manifold 55 can rotate around the swivel shaft 53 integrally with the disk 51.

Further, another support 58 extends upward from the disk surface of the wheel 51, and a holder 60 for the bottle 30 is attached to an upper portion of the support 58. A plurality of stays 58 and holders 60 are arranged at predetermined intervals around the wheel 51. The plurality of holders 60 are coupled to the disc 51 via struts 58 and rotate together with the rotation of the disc 51.

Hydrogen peroxide gas supply pipes 59 extend from the periphery of the manifold 55 to the holders 60, and nozzles 61 are attached to the tips of the supply pipes 59. The nozzle 61 is fixed to the support post 58, and an opening at the tip thereof faces the mouth of the bottle 30 held by the holder 60. When the wheel 51 is rotated in this way, the nozzle 61 rotates around the turning shaft 53 together with the bottle 30 held by the holder 60, and the hydrogen peroxide gas is blown to the bottle 30. Further, a passage 62 is provided around the wheel 51 so as to surround the travel path of the bottle 30 held by the holder 60. The nozzle 61 may be provided with a guide member (not shown) for discharging the hydrogen peroxide gas introduced into the bottle 30 to the outside of the bottle 30 while contacting the screw mouth portion (see fig. 4 and 5 of japanese patent No. 4526820).

A conduit 63 is connected to an upper end of the conduit 56 of the manifold 55 via a seal member 64. The conduit 56 rotates integrally with the manifold 55 with respect to the conduit 63, and the seal member 64 prevents the hydrogen peroxide gas from leaking out from the connection between the conduits 56 and 63. The conduit 63 is provided with a 1 st valve 41 for controlling the passage of the hydrogen peroxide gas in the conduit 63. A bypass conduit 67 branches from the upstream side of the 1 st valve 41. The bypass conduit 67 communicates with the interior of the sterile chamber 70. The bypass conduit 67 is provided with a 2 nd valve 43 for controlling the passage of the hydrogen peroxide gas in the bypass conduit 67. The bypass duct 67 may extend from between the bearing 57 and the nozzle 61.

A gas supply device including a blower 65, a HEPA (High Efficiency Air Filter) Filter 66, and an electric heater 69 is provided on the upstream side of the duct 63. The hydrogen peroxide adding device 68 is inserted into one or both of the front and rear of the electric heater 69. When the hydrogen peroxide adding device 68 is provided downstream of the electric heater 69, the hydrogen peroxide gas may be mixed in the piping. When hydrogen peroxide is not in a gaseous state, the residual value of hydrogen peroxide tends to increase. On the other hand, when the hydrogen peroxide adding device 68 is provided upstream of the electric heater 69, hydrogen peroxide may be added to the piping in a liquid state such as a mist. In this case, the set temperature of the electric heater 69 is preferably not less than the boiling point of the supplied bactericide, but may be not less than 100 ℃ (preferably not less than 130 ℃) in accordance with the sterilization intensity of the bottle 30. Alternatively, a separate electric heater may be provided further upstream of the spray to spray the mist into hot sterile air (80 ℃ C. or higher). Alternatively, the hydrogen peroxide adding device 68 may be inserted into both front and rear sides of the electric heater 69. When the bottle 30 is made of PET (polyethylene terephthalate), hydrogen peroxide is easily adsorbed and the residual value is easily increased; when the material is HDPE (high density polyethylene), the amount of hydrogen peroxide adsorbed is extremely small, and is 1/5 to 1/20. Therefore, not only a method of adding hydrogen peroxide into sterile air by gasifying hydrogen peroxide, but also a method of spraying hydrogen peroxide and mixing the gas may be adopted. The hydrogen peroxide gas is blown out from the nozzles 61 to the bottle 30 through the supply pipes 59, and the bottle 30 is sterilized. A pressure gauge 71 for measuring the pressure in the sterile chamber 70 is attached to the sterile chamber 70. The bactericide may contain hydrogen peroxide at a concentration of 1% or more. A reagent obtained by diluting 35% hydrogen peroxide with ethanol may also be used.

In fig. 5, in normal production, the 1 st valve 41 of the sterilization apparatus 11 is opened while the 2 nd valve 43 is closed, and hydrogen peroxide gas is introduced into the bottle 30 through a normally used conduit 63. On the other hand, when the initial bacteria of the container is checked, the 1 st valve 41 is closed and the 2 nd valve 43 on the bypass side is opened. Thus, the hydrogen peroxide gas passes through the bypass conduit 67 and is not introduced into the bottle 30. When the initial bacteria of the container is checked, the 1 st valve 41 may be opened and the 2 nd valve 43 on the bypass side may be opened at the same time.

In the subsequent air rinsing process using the air rinsing device 14, it is also necessary to bypass the sterile air or stop the supply of the sterile air so that the inside of the bottle 30 is not replaced with the sterile air. However, it is preferable to supply sterile air into the sterile chamber 70 in the same manner as in the normal manufacturing process so that the sterile chamber 70 does not contain any unwanted bacteria in a positive pressure state. The configuration of the air rinsing device 14 may be substantially the same as that of the sterilization device 11 shown in fig. 5.

In the case of a device equipped with the sterile water rinsing device 15 as in the present embodiment, since it is impossible to grasp the accurate initial number of bacteria when the bottle 30 is washed with sterile water, it is necessary to reduce the flow rate of the sterile water to such an extent that the sterile water does not contact the bottle 30. When the drying operation is performed with the aseptic water rinsing stopped, the water guide of the aseptic water rinsing device 15 may be worn or damaged, and therefore, it is preferable to supply the aseptic water while minimizing the flow rate (for example, the flow rate per 1 nozzle is 3L/min or less). The same idea can be applied to the chemical rinsing method using a peracetic acid preparation.

Next, in the filling device 20, a predetermined amount of the culture medium is filled into the bottle 30 from the mouth of the bottle 30 (culture medium filling step, step S3 in fig. 2).

Before the flask 30 is filled with the medium by the filling device 20, the medium is prepared in advance and heat-sterilized. The characteristics of the medium are equivalent to those of the content filled by the content filling system 10, that is, characteristics affecting the growth of bacteria. In the present embodiment, the pH of the medium is adjusted to be acidic to the pH of the contents, for example, to a pH of 4.0 to 4.6. More specifically, in the case where the pH of the contents is less than pH4.0, the pH of the medium is preferably adjusted to pH4.0 as the upper limit thereof. When the pH of the content is not less than 4 and less than 4.6, the pH of the medium is preferably adjusted to pH4.6 as the upper limit. When the product having the highest pH in the product bottle 35 to be produced is, for example, pH 3.5. + -. 0.2, the test can be carried out by adjusting the pH of the culture medium to pH3.5, pH3.7 as the upper limit, or pH3.8 or pH3.9 slightly higher than the upper limit.

In this way, by adjusting the pH of the medium to 3.5 or more and 4.6 or less, preferably 4.0 or more and 4.6 or less, corresponding to the characteristics of the contents, the medium becomes an environment in which spores of bacteria are allowed to survive but vegetative cells of bacteria, molds, and yeasts are not allowed to survive. Therefore, the growth environment of the bacteria in the culture medium can be made similar to the content actually filled.

Such a medium is usually prepared by dissolving in water 0.2 to 3% by weight of monosaccharides such as glucose and dextrose, disaccharides, polysaccharides or sodium carbonate and sodium bicarbonate as inorganic carbon sources as carbon sources, 0.5 to 3% by weight of casein peptone, chicken peptone, myocardial peptone, gelatin peptone, soybean peptone, polypeptone, yeast extract, meat extract, ammonium sulfate, magnesium sulfate, nitrate as nitrogen sources (including coenzyme), and 0.05 to 1% by weight of sodium chloride, monopotassium phosphate (リン acid monobasic カリウム), dipotassium hydrogen phosphate, potassium dihydrogen phosphate (リン acid dihydrate カリウム) as trace minerals or buffers. The pH of the medium is adjusted by dissolving hydrochloric acid, tartaric acid, citric acid, sodium hydroxide, potassium hydroxide, etc. in the medium.

The culture medium is heat-sterilized (UHT) or filter-sterilized by a predetermined sterilization method in the liquid treatment facility 23, and filled in the filling apparatus 20. When a carbonated beverage is also produced by the liquid treatment facility 23, a carbon dioxide dissolving device (carbonator) 24 for adding carbon dioxide after filtration sterilization to the product liquid needs to be provided before the filling device 20. When carbon dioxide is added during the filling of the culture medium, the culture medium may have a bacteriostatic action, and thus the supply of carbon dioxide may be stopped or carbon dioxide may be replaced with air. By changing carbon dioxide to air, the sterility of the carbon dioxide sterilization apparatus, not shown, including the filter, can be confirmed.

Then, the bottle 30 filled with the culture medium is sent from the filling device 20 to the lid attachment device 16 through the rear of the gas replacement device 25 for replacing the gas in the head space in the bottle 30. The gas replacement device 25 may blow the inert gas (nitrogen, carbon dioxide) subjected to the filtration sterilization to the mouth of the bottle 30 at the time of normal production, stop the supply of the inert gas (nitrogen, carbon dioxide) at the time of filling the culture medium, or replace the inert gas with air. By changing to air, the sterility of the headspace including the filter of the gas replacement device 25 can also be confirmed.

On the other hand, the lid 33 is sterilized in advance by the lid sterilizer 18 (lid sterilization step, step S8 in fig. 2). The lid 33 is carried from the outside of the content filling system 10 to the lid sterilization apparatus 18, sprayed with mist or gas of hydrogen peroxide, sterilized on the inner and outer surfaces thereof, activated and removed by hot air, washed with sterile water, and sent to the lid attachment apparatus 16. This cap sterilization step is performed in the same manner as the cap sterilization step in the above-described normal content filling method.

Next, in the cap attaching device 16, the cap 33 after sterilization by the cap sterilization device 18 is attached to the mouth of the bottle 30 (cap attaching step, step S4 in fig. 2). The lid attachment step is performed in the same manner as the lid attachment step in the above-described normal content filling method. Thus, the inside of the bottle 30 is filled with the culture medium, and the mouth portion cover 33 is closed, thereby obtaining the bottle 36 for verification.

Subsequently, the verification bottle 36 filled with the culture medium is carried out of the product bottle carrying-out section 22 and packaged in a packaging process. The packed box is tilted (or inverted) manually or automatically on the conveyor so that the culture medium is reliably brought into contact with the inner surface of the bottle 30 (culture medium contact step, step S5 in fig. 4). Thereafter, 2 or more verification bottles 36 are transferred to the constant temperature storage 37 maintained at a predetermined temperature of 25 ℃ to 40 ℃ and are subjected to static culture in the constant temperature storage 37 (culture step, step S6 in fig. 2). When the product bottle 35 is sold by heating using a heating vending machine or the like, it is preferable to check the sterility of the high-temperature bacteria, and the bottle 36 for verification may be cultured at a temperature of 40 ℃ to 65 ℃ in addition to the culture conditions described above.

After a predetermined period of time (for example, 3 days or longer, preferably 7 days or longer), all the verification bottles 36 are taken out from the incubator 37, and it is verified whether or not bacteria survive or propagate in the medium in the verification bottles 36 (verification step, step S7 in fig. 2). If the number of verification bottles 36 for which bacteria survive or proliferate is equal to or less than a predetermined number (for example, zero) as a result of the verification, it is determined that the initial bacteria are not present in the bottle 30, and the sterility is ensured. On the other hand, if the number of verification bottles 36 for verifying the survival or propagation of bacteria is equal to or greater than a predetermined number (for example, equal to or greater than 1) as a result of the verification, it is determined that the initial bacteria are present in the bottle 30, and a countermeasure is taken. For example, sterilization of the transfer and carrying-in paths of the bottles 30 or preparation (intensification) of the sterilization conditions in the container sterilization apparatus 13 (sterilization apparatus 11) may be performed. Further, in some cases, bottle 30 can be sufficiently sterilized by operating container sterilization device 13 (sterilization device 11) according to the type of bacteria, and in such a case, it can be determined that sterilization is sufficient when container sterilization device 13 is operated when the contents such as beverages are actually filled in bottle 30.

In order to shorten the period of time during which the medium in the verification bottle 36 is cultured in the thermostatic chamber 37, the verification bottle 36 may be stored in a state in which the medium moves by applying physical movement to the medium in the verification bottle 36. Examples of such motions include rotation, inversion, reciprocation, vibration, and stirring. By applying physical movement to the culture medium, dissolution of oxygen into the culture medium can be promoted, so that aerobic bacteria can be cultured, thereby enabling the culture speed of the bacteria to be increased. As a result, the predetermined period required for the culture (for example, 3 days or more, preferably 7 days or more) can be shortened, and the presence or absence of the initial bacteria in the bottle 30 can be determined quickly.

For example, according to the experiment of the present inventors, it was found that by applying vibration to the bottle 30 filled with the culture medium in the constant temperature storage 37 (example a), the culture of the bacteria can be promoted as compared with the case where no vibration is applied to the bottle 30 (comparative example B). Specifically, 2 or more bottles 30 each filled with a liquid neutral medium were sealed in a state where the headspace thereof contained therein a colony floating in the laboratory, and stored at 22 ℃ to 27 ℃. Meanwhile, vibration was applied to one set of bottles 30 (example a), and the other set of bottles 30 (comparative example B) was left standing without applying vibration. As a result, as shown in the following table, the positive rate was increased particularly in the case of the number of days of culture from 2 to 3 days. This indicates that the application of vibration enables the cultivation of bacteria in a shorter number of days than the case where vibration is not applied. Here, the positive rate is a ratio of the number of bottles 30 that are positive at a predetermined time to the number of bottles 30 (total number of positive) in which bacteria finally survive or proliferate (positive). Specifically, the positive rate was calculated based on the expression "positive rate (%) ═ number of positive observed by eye/total number of positive observed) × 100". All positive numbers were numbers that could be visually confirmed as positive after 13 days of culture.

[ Table 1]

As described above, according to the present embodiment, the vessel sterilization apparatus 13 fills the non-sterilized bottle 30 with the culture medium, closes the bottle with the lid 33, and verifies whether or not the bacteria survive or grow in the culture medium in the bottle 30. This makes it possible to accurately determine whether or not the bio-burden (bio-burden) of the initial bacteria is present in the bottle 30, and to take a measure against sterilization of the bottle 30 before the bottle 30 is actually filled with the contents such as a beverage.

In addition, according to the present embodiment, since the pH of the medium used in the verification is 4.0 or more and 4.6 or less, in contrast to the acidic content actually filled, the medium allows the spores of bacteria to survive but does not allow vegetative cells of bacteria, molds, and yeasts to survive. Thus, when the sterility of the content filling system 10 is comprehensively evaluated using a culture medium, it is possible to verify that the growth environment of bacteria is close to the actual content. This eliminates the need to use a large amount of equipment for sterilization, reduces the amount of chemicals and heat energy required for sterilization, and reduces the manufacturing cost of product bottle 35. For example, the temperature of steam, hot water, or the like used when performing an SIP (in-line sterilization) treatment in the beverage supply system piping of the content filling system 10 can be reduced, or the time for circulating steam, hot water, or the like can be shortened. In addition, it is possible to shorten the time required for COP (offline Cleaning out of Place) processing or SOP (offline sterilization out of Place) processing in the aseptic chamber 70.

In the case where the content to be actually filled is a low-acidic or neutral beverage, the pH of the medium may be in a low-acidic or neutral range of 7.0(6.0 to 8.0) as in the case of a general medium. In this case, substantially all of the bacteria can be detected.

The case where the sterilization apparatus for performing hydrogen peroxide sterilization and hot water sterilization is used as the sterilization apparatus for the container has been described above, but the present invention is not limited thereto. The present invention can be applied to all sterilization apparatuses, including a peracetic acid sterilization method in which the inside and outside surfaces of a bottle are sterilized by rinsing with peracetic acid and then the inside and outside surfaces are rinsed with sterile water, an electron beam sterilization method in which the inside and outside surfaces of a bottle are sterilized by irradiating an electron beam from the inside or outside surfaces of the bottle and then the bottle is rinsed with sterile air, and UV sterilization. It can be used not only for sterilizing bottles, but also for sterilizing preforms, cups, bags and paper containers. In addition, since the case where a PET bottle is used as the container has been described above as an example, the culture medium for aerobic bacteria is used as the culture medium, but the present invention is not limited thereto. When a retort container such as a tank is used, a culture medium for anaerobic bacteria may be used as the culture medium.

(modification example)

Next, a modified example of the present embodiment will be explained.

In the above-described embodiment, the case where the presence or absence of the initial bacteria in the bottle 30 is verified is described as an example. However, the present invention is not limited to this, and it is also possible to verify the presence or absence of the initial bacteria in the lid 33. That is, after a plurality of bottles 30 are circulated through the content filling system 10 and each bottle 30 is sterilized, a predetermined culture medium is filled into each bottle 30 without adding the content actually filled. The bottle 30 is then closed with the cap 33 that has not been sterilized by the cap sterilizer 18. After that, it was confirmed that the medium filled in each bottle 30 did not decay after a certain period of time (initial bacteria confirmation method of the lid).

An initial bacteria confirmation method (initial bacteria confirmation method of the lid) of the content filling system 10 according to the present modification will be described below with reference to fig. 3 and 4. Fig. 4 is a flowchart showing an initial bacteria confirmation method according to a modification.

First, as described above, the empty bottle 30 for verification is circulated through the content filling system 10. In this case, empty bottles 30 are supplied from the outside to the bottle supply unit 21 of the content filling system 10 (container supply step, step S11 in fig. 4). The number of bottles 30 may be predetermined, and may be, for example, 1,000 to 300,000 (preferably 3,000 to 30,000).

Next, the bottle 30 is sent to the sterilization apparatus 11 of the container sterilization apparatus 13, and the bottle 30 is sterilized by the sterilization apparatus 11 using the hydrogen peroxide solution as a sterilizing agent (sterilization step, step S12 in fig. 4). The sterilization step is performed in the same manner as the sterilization step in the above-described normal content filling method.

Next, the bottle 30 is sequentially sent to the air rinsing device 14 and the sterile water rinsing device 15, and the bottle 30 is cleaned with air and sterile water in the air rinsing device 14 and the sterile water rinsing device 15 (rinsing step, step S13 in fig. 4). The rinsing step is similar to the rinsing step in the above-described normal content filling method.

Next, the bottle 30 is transferred to the filling device 20. In the filling apparatus 20, a predetermined amount of the sterilized culture medium is filled into the bottle 30 from the mouth of the bottle 30 (culture medium filling step, step S14 in fig. 4). The medium filling step is the same as the medium filling step in the method for confirming the initial bacteria in the container.

On the other hand, the lid sterilization device 18 is stopped in advance, and the sterilization process for the lid 33 is not performed. Therefore, the lid 33 passes through the lid sterilization apparatus 18 without being sterilized by the lid sterilization apparatus 18 and is conveyed to the lid attachment apparatus 16 (non-sterilization lid conveyance step, step S18 in fig. 4). The lid 33 may be sent to the lid attachment device 16 by using another bypass passage instead of the lid sterilization device 18.

Then, the bottle 30 filled with the culture medium is sent to the lid unit 16. In the cap attaching device 16, the cap 33 that has not been sterilized is attached to the mouth of the bottle 30 (cap attaching step, step S15 in fig. 4). Thus, the culture medium was filled in the bottle 30, and the mouth was closed with the non-sterilized lid 33 to obtain the verification bottle 36.

Subsequently, the verification bottle 36 filled with the culture medium is carried out of the product bottle carrying-out section 22 and packaged in a packaging process. The packed box is tilted (or reversed) manually or automatically on the conveyor so that the culture medium is reliably brought into contact with the inner surface of the lid 33 (culture medium contact step, step S16 in fig. 4). Thereafter, 2 or more verification bottles 36 are transferred to the incubator 37, and are subjected to static culture in the incubator 37 (culture step, step S17 in fig. 4). The culture step is the same as the culture step in the method for confirming the initial bacteria in the container.

After a predetermined period of time (for example, 3 days or longer, preferably 7 days or longer), all the verification bottles 36 are taken out from the incubator 37, and it is verified whether or not bacteria survive or propagate in the medium in the verification bottles 36 (verification step, step S18 in fig. 4). From the verification result, the actual biological load of the lid body can be accurately grasped. On the other hand, if the number of verification bottles 36 for verifying the survival or propagation of bacteria is equal to or greater than a predetermined number (for example, equal to or greater than 1) as a result of the verification, it is determined that the initial bacteria are present in the lid 33, and a countermeasure is taken. For example, sterilization of the conveyance and carrying-in passages of the lid 33, or preparation (strengthening) of sterilization conditions in the lid sterilization apparatus 18 may be performed. In addition, the cap sterilization device 18 may be operated according to the type of bacteria to sufficiently sterilize the cap 33, and in this case, it can be determined that sterilization is sufficient when the cap sterilization device 18 is operated when the bottle 30 is actually filled with contents such as beverages.

As described above, according to this modification, after filling the sterilized bottle 30 with the culture medium and closing the bottle with the cap 33 that has not been sterilized by the cap sterilizer 18, it is verified whether or not bacteria survive or proliferate in the culture medium in the bottle 30. This makes it possible to accurately determine whether or not the initial bacteria are present in the lid 33, and to take a measure for sterilizing the lid 33 before the bottle 30 is actually filled with the contents such as a beverage.

It should be noted that embodiment 1 and the modification example may be combined to verify the presence or absence of the initial bacteria in either of the bottle 30 and the lid 33 at a time. That is, bottle 30 is transferred to filling apparatus 20 without sterilizing bottle 30 by container sterilization apparatus 13, and the sterilized medium is filled into unsterilized bottle 30 by using filling apparatus 20. Next, the cap 33 is transferred to the cap attaching device 16 without sterilizing the cap 33 by the cap sterilizing device 18, and the bottle 30 is closed by the cap 33 that is not sterilized by the cap attaching device 16. Thereafter, it can be verified whether bacteria survive or multiply in the medium in the flask 30.

(examples)

Next, a specific example of embodiment 1 will be explained.

(example 1-1)

A beverage filling system of 600bpm (bottle per minute) was used for filling a sterilized beverage at room temperature into a PET bottle of a capacity of 500mL sterilized in a sterile atmosphere, and sealing the bottle with a sterilized cap. Using this beverage filling system, 3000 unsterilized PET bottles were filled with the sterilized acidic medium having a pH of 4.0 at room temperature and closed with a sterilized lid. These were then incubated at 27 ℃ for 1 week, and all of the PET bottles were tested, and as a result, only 1 PET bottle in which the medium was deteriorated was present. The identification of the bacteria contained in the putrefactive medium resulted in bacteria with low drug resistance (Cladosporium cladosporioides). Therefore, it was determined that sufficient sterilization could be performed by operating the container sterilization apparatus when the PET bottle was actually filled with the beverage.

(examples 1 to 2)

A beverage filling system of 600bpm (bottle per minute) was used for filling a sterilized beverage at room temperature into a PET bottle of a capacity of 500mL sterilized in a sterile atmosphere, and sealing the bottle with a sterilized cap. Using this beverage filling system, 3000 sterilized PET bottles were filled with the sterilized acidic medium of ph4.0 at room temperature, and closed with a cap that was not sterilized. These were then incubated at 27 ℃ for 1 week, and all of the PET bottles were tested, and as a result, only 1 PET bottle in which the medium was deteriorated was present. Aspergillus niger (a. niger) was presumed to be a result of identification of bacteria contained in the rotten medium. Therefore, it was determined that sufficient sterilization could be performed by operating the container sterilization apparatus when the PET bottle was actually filled with the beverage.

(embodiment 2)

Next, embodiment 2 of the present invention will be explained with reference to the drawings. Fig. 6 to 8 are views showing embodiment 2 of the present invention. In fig. 6 to 8, the same components as those in embodiment 1 shown in fig. 1 to 5 are denoted by the same reference numerals, and detailed description thereof is omitted.

(Contents filling System)

First, a content filling system (aseptic filling system ) according to the present embodiment will be described with reference to fig. 6.

The content filling system 10 shown in fig. 6 includes a bottle supply unit 21, a sterilization device 11, a hot water rinsing device 15A, a filling device (filler) 20, a lid attachment device (capping machine, hemming and sealing machine) 16, and a product bottle carrying-out unit 22. These bottle supply unit 21, sterilizing device 11, hot water rinsing device 15A, filling device 20, lid attachment device 16, and product bottle carrying-out unit 22 are arranged in order from the upstream side to the downstream side in the conveying direction. Further, 2 or more transfer wheels 12 for transferring bottles 30 between the sterilization apparatus 11, the hot water rinsing apparatus 15A, the filling apparatus 20, and the cap attaching apparatus 16 are provided.

The hot water rinsing device 15A sterilizes the bottle 30 sterilized with the hydrogen peroxide solution as the bactericide by using hot water. Specifically, hot water at a temperature of, for example, 65 ℃ or higher and 80 ℃ or lower is supplied into the bottle 30.

The contents filled by the filling device 20 have predetermined characteristics that affect the growth of bacteria. In the present embodiment, the predetermined characteristic is the pH of the contents. More specifically, the contents consist of an acidic beverage. The beverage preferably has an acidity of less than ph4.6, more preferably less than ph 4.0.

The sterilization apparatus 11, the hot water rinsing apparatus 15A, the filling apparatus 20, and the lid attachment apparatus 16 are housed inside the sterile chamber 70. Such a content filling system 10 may be constituted by, for example, an aseptic filling system. In this case, the inside of the aseptic chamber 70 is maintained in an aseptic state.

In the present embodiment, the configurations of the bottle supply unit 21, the sterilization apparatus 11, the filling apparatus 20, the lid attachment apparatus 16, and the product bottle carrying-out unit 22 are substantially the same as those in embodiment 1, and therefore, a detailed description thereof is omitted.

(method of filling content)

Next, a content filling method using the content filling system 10 (fig. 6) will be described. A filling method in a normal case, that is, a method of filling contents such as beverages into the bottle 30 to manufacture the product bottle 35, will be described below.

First, 2 or more empty bottles 30 are sequentially supplied to the bottle supply unit 21 from the outside of the content filling system 10. The bottles 30 are conveyed from the bottle supply unit 21 to the sterilizer 11 by the conveyor wheel 12 (container supply step).

Next, the bottle 30 is sterilized by using the hydrogen peroxide solution as a sterilizing agent in the sterilizing apparatus 11 (sterilizing step). This sterilization step is substantially the same as the sterilization step in embodiment 1.

Next, the bottle 30 is conveyed to the hot water rinsing device 15A by the conveying wheel 12. In the hot water rinsing apparatus 15A, hot water sterilization is performed on the bottle 30 sterilized with hydrogen peroxide as a sterilizing agent (hot water rinsing step). Specifically, hot water at a temperature of 65 ℃ to 75 ℃ is supplied into the bottle 30 at a flow rate of 5L/min to 15L/min. At this time, it is preferable that the bottle 30 is inverted, hot water is supplied into the bottle 30 from the mouth portion facing downward, and the hot water flows out of the bottle 30 from the mouth portion. The hot water sterilizes vegetative cells of mold, yeast, bacteria, and the like damaged by the hydrogen peroxide. The hot water washes off the excess hydrogen peroxide solution remaining in the bottle 30 and discharges the same to the outside of the bottle 30. In some cases, the outer surface of the bottle 30 may be rinsed with hot water in the same manner as the inner surface.

The bottles 30 are then transferred to the filling device 20 using the transfer wheel 12. In this filling device 20, the contents are filled into the bottle 30 from the mouth while rotating (revolving) the bottle 30 (filling step).

Before the bottle 30 is filled with the filling device 20, the contents are mixed in advance and subjected to a heat sterilization treatment. As described above, the content has a predetermined pH as a characteristic affecting the growth of bacteria. Specifically, the contents consist of an acidic beverage preferably less than pH4.6, more preferably less than pH4. The filling step is substantially the same as the filling step in embodiment 1.

Next, the bottle 30 filled with the content is transferred to the cap mounting device 16 by the transfer wheel 12. Next, in cap attaching device 16, a sterilized cap (not shown) is attached to the mouth of bottle 30, thereby obtaining product bottle 35 (cap attaching step).

Thereafter, the product bottle 35 is conveyed from the cap attaching device 16 to the product bottle carrying-out section 22, and carried out to the outside of the content filling system 10.

(method of verifying content filling System)

Next, a method of verifying the sterility of the content filling system 10 (fig. 6) will be described.

The verification method of the present embodiment is a method of confirming whether or not the sterility of the content filling system 10 is ensured. This verification method is performed, for example, at an initial stage immediately after completion of the content filling system 10, that is, before the product bottle 35 is manufactured by actually starting filling the bottle 30 with the content filling system 10. Alternatively, the verification method according to the present embodiment may be performed when sterility may be affected, for example, when the process or the apparatus in the content filling system 10 is changed at all times or when the content filling system 10 is not used for a certain period of time. Alternatively, the verification method of the present invention may be performed periodically with respect to a predetermined filling cycle, regardless of whether there is a possibility of affecting sterility.

First, before the verification method of the present embodiment is performed, whether or not the sterility is ensured is individually checked for each component of the content filling system 10. Specifically, for example, detection of whether the supply line of the content is properly warmed (SIP warm-up confirmation detection), detection of whether the bottle 30 or the cap is properly sterilized (bottle sterilization detection, cap sterilization detection), detection of whether the sterile chamber 70 is sterilized (chamber sterilization detection), and the like are performed. These tests can be carried out by methods known in the art.

After such detection, in order to comprehensively evaluate the sterility of the content filling system 10 at the final stage, the verification method according to the present embodiment is performed using the bottle 30 filled with the culture medium. Specifically, a plurality of bottles 30 are circulated through the content filling system 10, and each bottle 30 is filled with a predetermined culture medium instead of the content actually filled, and is sealed. Thereafter, it was confirmed that the medium filled in each bottle 30 was not putrefied after a certain period of time.

The verification method of the content filling system 10 based on the present embodiment is further described below with reference to fig. 7 and 8. Fig. 7 is a flowchart illustrating the verification method according to the present embodiment, and fig. 8 is a schematic plan view illustrating the content filling system when the verification method according to the present embodiment is performed. In fig. 8, the same components as those of the content filling system 10 shown in fig. 6 are denoted by the same reference numerals.

First, as shown in fig. 8, an empty bottle 30 for verification is circulated through the content filling system 10. In this case, empty bottles 30 are supplied from the outside to the bottle supply unit 21 of the content filling system 10 (container supply step, step S11 in fig. 7). The number of bottles 30 may be predetermined, and may be, for example, 1,000 to 300,000 (preferably 3,000 to 30,000).

Next, the bottle 30 is sent to the sterilizer 11, and the sterilizer 11 sterilizes the bottle 30 with an aqueous hydrogen peroxide solution as a sterilizer (sterilization step, step S12 in fig. 7). The sterilization step is performed in the same manner as the sterilization step in the above-described normal content filling method.

Next, the bottle 30 is sent to the hot water rinsing device 15A, and the bottle 30 is sterilized by hot water in the hot water rinsing device 15A (hot water rinsing step, step S13 of fig. 7). The hot water rinsing step is similar to the hot water rinsing step in the above-described normal content filling method.

Next, the bottle 30 is transferred to the filling device 20. In the filling apparatus 20, a predetermined amount of the culture medium is filled into the bottle 30 from the mouth of the bottle 30 (culture medium filling step, step S14 in fig. 7).

Before the flask 30 is filled with the medium by the filling device 20, the medium is prepared in advance and heat-sterilized. The characteristics of the medium are equivalent to those of the content filled by the content filling system 10, that is, characteristics affecting the growth of bacteria. In the present embodiment, the pH of the medium is adjusted to be acidic, for example, 4.0 to 4.6, corresponding to the pH of the contents. More specifically, in the case where the pH of the contents is less than pH4.0, the pH of the medium is preferably adjusted to pH4.0 as the upper limit thereof. When the pH of the content is not less than 4 and less than 4.6, the pH of the medium is preferably adjusted to pH4.6 as the upper limit. When the product having the highest pH in the product bottle 35 to be produced is, for example, pH3.5 ± 0.2, the sterility test can be performed by adjusting the pH of the medium to pH3.5, pH3.7 as the upper limit, or pH3.8 or pH3.9 slightly higher than the upper limit.

In this way, by adjusting the pH of the medium to 3.5 or more and 4.6 or less, preferably 4.0 or more and 4.6 or less, corresponding to the characteristics of the contents, the medium becomes an environment in which spores of bacteria are allowed to survive but vegetative cells of bacteria, molds, and yeasts are not allowed to survive. Therefore, the growth environment of the bacteria in the culture medium can be made similar to the content actually filled.

Such a medium is usually prepared by dissolving in water 0.2 to 3% by weight of monosaccharides such as glucose and dextrose, disaccharides, polysaccharides or sodium carbonate and sodium bicarbonate as inorganic carbon sources as carbon sources, 0.5 to 3% by weight of casein peptone, chicken peptone, myocardial peptone, gelatin peptone, soybean peptone, polypeptone, yeast extract, meat extract, ammonium sulfate, magnesium sulfate, nitrate as nitrogen sources (including coenzyme), and 0.05 to 1% by weight of sodium chloride, monopotassium phosphate (リン acid monobasic カリウム), dipotassium hydrogen phosphate, potassium dihydrogen phosphate (リン acid dihydrate カリウム) as trace minerals or buffers. The pH of the medium is adjusted by dissolving hydrochloric acid, tartaric acid, citric acid, sodium hydroxide, potassium hydroxide, etc. in the medium.

In the present embodiment, a medium is also provided, which is a medium used in the verification method and has characteristics equivalent to those of the content filled by the content filling system 10.

Then, the bottle 30 filled with the culture medium is sent to the lid unit 16. In the cap attaching apparatus 16, the cap after sterilization is attached to the mouth of the bottle 30 (cap attaching step, step S15 in fig. 7). The lid attachment step is performed in the same manner as the lid attachment step in the above-described normal content filling method. Thus, the inside of the bottle 30 is filled with the culture medium, and the mouth portion is closed with the lid, thereby obtaining the bottle 36 for verification.

Subsequently, the verification bottle 36 filled with the culture medium is carried out from the product bottle carrying-out section 22 to the outside. Thereafter, 2 or more verification bottles 36 are transferred to the constant temperature storage 37 maintained at a predetermined temperature of 25 ℃ to 40 ℃ and are subjected to static culture in the constant temperature storage 37 (culture step, step S16 in fig. 7). When the product bottle 35 is sold by heating using a heating vending machine or the like, it is necessary to confirm the sterility of the thermophilic bacteria and to culture the verification bottle 36 at a temperature of 40 ℃ to 65 ℃.

After a predetermined period of time (for example, 3 days or longer, preferably 7 days or longer), all the verification bottles 36 are taken out from the incubator 37, and it is verified whether or not bacteria survive or propagate in the medium in the verification bottles 36 (verification step, step S17 in fig. 7). If the number of verification bottles 36 for the survival or propagation of bacteria is equal to or less than a predetermined number (for example, zero), it is determined that the sterility of the content filling system 10 is ensured, and the production of product bottles 35 actually filled with the contents such as beverages is started. In order to shorten the period of time during which the medium in the verification bottle 36 is cultured in the incubator 37, the verification bottle 36 may be stored in a state in which the medium is moved by applying physical movement to the medium in the verification bottle 36, as in the case of embodiment 1.

As described above, according to the present embodiment, since the pH of the medium used at the time of verification is 4.0 or more and 4.6 or less as compared with the characteristics of the content actually filled, the medium allows the spores of bacteria to survive but does not allow vegetative cells of bacteria, molds, and yeasts to survive. Thus, when the sterility of the content filling system 10 is comprehensively evaluated using a culture medium, it is possible to verify that the growth environment of bacteria is close to the actual content. This eliminates the need to use a large amount of equipment for sterilization, reduces the amount of chemicals and heat energy required for sterilization, and reduces the manufacturing cost of product bottle 35. For example, the temperature of steam, hot water, or the like used when performing an SIP (in-line sterilization) treatment in the beverage supply system piping of the content filling system 10 can be reduced, or the time for circulating steam, hot water, or the like can be shortened. In addition, it is possible to shorten the time required for COP (offline Cleaning out of Place) processing or SOP (offline sterilization out of Place) processing in the aseptic chamber 70.

In contrast, as a comparative example, almost all bacteria were detected in the case where the pH of the culture medium was in a low acidic to neutral range of 7.0(6.0 to 8.0) as in the case of a general culture medium. However, when the sterility of the content filling system 10 is verified using a culture medium of ph7.0(ph6.0 to 8.0), the content filling system 10 is required to have a sterilizing capability more than necessary in order to reduce the number of verification bottles 36 for the survival or propagation of bacteria to a predetermined number (e.g., zero). In this case, an excessive load may be applied to the sterilization in the process of filling the system 10 with contents, the sterilization of the bottle 30 or the cap, or the sterilization of the aseptic chamber 70. For example, it may be necessary to increase the temperature of steam, hot water, or the like used for sterilization, to prolong the time during which steam, hot water, or the like flows, or to increase the amount of a chemical to be used. In contrast, in the present embodiment, since the content filled in the content filling system 10 is acidic (less than ph4.6), the content is not likely to be putrefied by the spores of the bacteria, and the content filling system 10 does not need to be sterilized to such an extent that the spores of the bacteria are sterilized. Therefore, by using an acidic medium (pH of 4.0 or more and 4.6 or less), it can be confirmed that vegetative cells of bacteria other than spores of bacteria, molds, and yeasts have been killed.

(modification example)

Next, each modified example of the present embodiment will be described.

In the present embodiment, the characteristic of the medium used for verifying the content filling system 10 is the pH of the content, and the case where the pH of the medium is adjusted to 3.5 or more and 4.6 or less, preferably 4.0 or more and 4.6 or less has been described as an example. However, the present invention is not limited thereto.

For example, in the case where the content filled by the content filling system 10 is constituted by a substance containing carbon dioxide, such as a carbonated beverage, carbon dioxide may be dissolved in the culture medium. In this case, the solubility of carbon dioxide dissolved in the medium is preferably a lower limit value of the solubility of carbon dioxide dissolved in the content. In general, when the content filled by the content filling system 10 contains carbon dioxide, the growth of bacteria in the content is suppressed. Therefore, by dissolving carbon dioxide in the culture medium, the sterility can be verified in a state where the growth environment of the bacteria is close to the actual contents. This can suppress the equipment, chemicals, energy, and the like required for sterilization in the content filling system 10.

In addition, when the content to be filled by the content filling system 10 is composed of a substance such as mineral water that does not contain at least one of a carbon source and a nitrogen source (organic matter), the culture medium may not contain at least one of them (or at least one of them may be adjusted to 0.1 wt% or less). In general, when the content does not contain at least one of a carbon source and a nitrogen source, the growth of bacteria that propagate using at least one of a carbon source and a nitrogen source can be suppressed. Accordingly, by appropriately excluding at least one of a carbon source and a nitrogen source from the medium, sterility can be verified in a state where the growth environment of the bacteria is close to the content actually filled. This can suppress the equipment, chemicals, energy, and the like required for sterilization in the content filling system 10.

As a property affecting the growth of bacteria, a value of Total Organic Carbon (TOC) contained in the content may be used. The sterility can also be verified in a state where the value of the total organic carbon content in the culture medium is adjusted to a value close to the total organic carbon content of the content actually filled. For example, the TOC value contained in commercially available mineral water is about 0.1 to 0.3 mg/L. Therefore, when the content is mineral water, the sterility can be evaluated by adding a carbon source and/or a nitrogen source so that the TOC value of the medium to be filled is, for example, 5mg/L (preferably 0.5 mg/L).

In addition, when the content filled by the content filling system 10 does not include at least one of a carbon source and a nitrogen source, or is constituted of, for example, a green tea beverage containing catechin, the sterility can be verified by adding catechin to the medium. Generally, when the total amount of catechins (the total amount is a content of 8 kinds of Epigallocatechin (EGC), epigallocatechin gallate (EGCg), Epicatechin (EC), epicatechin gallate (ECg), Gallocatechin (GC), gallocatechin gallate (GCg), catechin (C), and catechin gallate (Cg)) in the content is 30 mg% or more, the growth of bacteria is suppressed. Therefore, by adding 30 mg% of total catechin to the culture medium, the sterility can be verified in a state where the growth environment of the bacteria is close to the content actually filled. This can suppress the equipment, chemicals, energy, and the like required for sterilization in the content filling system 10.

In embodiment 2 and the modification, the following medium is exemplified as a medium having a property (bacterial growth inhibitory factor) that affects the growth of bacteria: (i) a medium having a pH of 3.5 or more and 4.6 or less (preferably 4.0 or more and 4.6 or less), (ii) a medium in which carbon dioxide is dissolved, (iii) a medium containing no carbon source or nitrogen source, or (iv) a medium in which the amount of total organic carbon is adjusted, or (v) a medium in which catechin is dissolved. The medium is not limited to any of the above characteristics (i) to (v), and a medium having 2 or more of the characteristics (i) to (v) may be used. For example, (i) a medium having a pH of 3.5 or more and 4.6 or less and (ii) a carbon dioxide-dissolved medium can be used.

The case where a sterilization apparatus for performing hydrogen peroxide sterilization and hot water sterilization is used as the sterilization apparatus for the container has been described above, but the present invention is not limited thereto. All sterilization apparatuses such as peracetic acid sterilization, electron beam sterilization, and UV sterilization may be used. In addition, the above description has been given by taking a case where a PET bottle is used as a container as an example, and a culture medium for aerobic bacteria is used as the culture medium, but the present invention is not limited thereto. When a retort container such as a tank is used, a culture medium for anaerobic bacteria may be used as the culture medium.

(examples)

Next, specific examples of the present embodiment will be described.

(example 2-1)

A beverage filling system of 600bpm (number of bottles per minute) for filling a sterilized beverage at normal temperature into a PET bottle of a capacity of 500mL sterilized with a sterile atmosphere and sealing with a sterilized cap was used. In this beverage filling system, a container (bottle, lid), a filling chamber, and a product liquid line are subjected to a process in which vegetative cells of mold, yeast, and bacteria can be sterilized, although bacterial spores can survive. Next, using the above beverage filling system, 10,000 PET bottles were filled with an acidic medium having a pH of 4.0 at room temperature, and these were cultured at 30 ℃ for 1 week. After the culture, all the PET bottles were examined, and as a result, it was confirmed that no PET bottle in which the medium was rotten was present. Then, using the beverage filling system, product bottles were produced by filling an acidic beverage having a ph of less than 4.0 at room temperature, and then these product bottles were examined, and as a result, neither of the acidic beverages in any of the product bottles was spoiled.

(example 2-2)

A beverage filling system of 600bpm (number of bottles per minute) for filling a sterilized beverage at normal temperature into a PET bottle of a capacity of 500mL sterilized with a sterile atmosphere and sealing with a sterilized cap was used. Using this beverage filling system, 10,000 PET bottles were filled with a medium containing carbon dioxide dissolved therein at room temperature, and were cultured at 30 ℃ for 1 week. The amount (volume) of carbon dioxide added was set to 2.0 as the product lower limit gas volume GV. After the culture, all the PET bottles were examined, and as a result, it was confirmed that no PET bottle in which the medium was rotten was present. Then, using the beverage filling system, carbonated beverages having a gas volume of 2.0 or more were filled at room temperature to produce product bottles, and then these product bottles were examined, and as a result, neither of the carbonated beverages in any of the product bottles was spoiled.

(examples 2 to 3)

A beverage filling system of 600bpm (number of bottles per minute) for filling a sterilized beverage at normal temperature into a PET bottle of a capacity of 500mL sterilized with a sterile atmosphere and sealing with a sterilized cap was used. Using this beverage filling system, 10,000 PET bottles were filled with a medium in which the carbon source and the nitrogen source were reduced to 0.05 wt% respectively at room temperature, and they were cultured at 30 ℃ for 3 weeks. After the culture, all the PET bottles were examined, and as a result, it was confirmed that no PET bottle in which the medium was rotten was present. After that, mineral water was filled at normal temperature to produce product bottles, and these product bottles were examined, and as a result, none of the product bottles was spoiled.

(examples 2 to 4)

A beverage filling system of 600bpm (number of bottles per minute) for filling a sterilized beverage at normal temperature into a PET bottle of a capacity of 500mL sterilized with a sterile atmosphere and sealing with a sterilized cap was used. In this beverage filling system, a container (bottle, lid), a filling chamber, and a product liquid line are subjected to a process in which vegetative cells of mold, yeast, and bacteria can be sterilized, although bacterial spores can survive. Then, using the above beverage filling system, a medium containing 30 mg% total catechins was filled into 10,000 PET bottles at room temperature, and these were cultured at 30 ℃ for 1 week. After the culture, all the PET bottles were examined, and as a result, it was confirmed that no PET bottle in which the medium was rotten was present. Then, using the beverage filling system, product bottles were produced by filling tea beverages having a total amount of catechins of 30 mg% or more at normal temperature, and then these product bottles were examined, and as a result, none of the tea beverages in any of the product bottles was spoiled.

(examples 2 to 5)

A beverage filling system of 600bpm (number of bottles per minute) for filling the sterilized beverage into PET bottles of 500mL capacity at a high temperature and sealing with a cap was used. In this beverage filling system, 10,000 PET bottles were filled with an acidic medium having a pH of 4.0 at a high temperature of 85. + -. 5 ℃ and they were cultured at 30 ℃ for 1 week. After the culture, all the PET bottles were examined, and as a result, it was confirmed that no PET bottle in which the medium was rotten was present. Then, product bottles were produced by using the beverage filling system described above and filling acidic beverages having a ph of less than 4.0 at high temperature, and these product bottles were examined, and as a result, neither of the acidic beverages in any of the product bottles was spoiled.

(examples 2 to 6)

A beverage filling system of 600bpm (number of bottles per minute) for filling the sterilized beverage at a medium temperature into a PET bottle of a capacity of 500mL sterilized with a sterile atmosphere and sealing with a sterilized cap was used. In this beverage filling system, 10,000 PET bottles were filled with an acidic medium having a pH of 4.0 at a medium temperature of 65. + -. 5 ℃ and they were cultured at 30 ℃ for 1 week. After the culture, all the PET bottles were examined, and as a result, it was confirmed that no PET bottle in which the medium was rotten was present. After that, product bottles were produced by filling acidic beverages having a ph of less than 4.0 at medium temperature, and then these product bottles were examined, and as a result, neither of the acidic beverages in any of the product bottles was spoiled.