Search: Biological agents regulation

Premises

The premises where work with biological agents is carried out should be carefully designedin order to

Avoid endangering the people working there

Avoid endangering the environment

Work cleanly, efficiently and purposefully

The following sections briefly list the most important standards and guidelines that are relevant to the handling of biological agents and provide information on the layout anddesign of microbiological workspaces.

Biological Agents Ordinance (BioStoffV)

The Biological Agents Ordinance [1] forms the legal basis for working with biological agents, in particular microorganisms. It serves to protect employees against hazards caused by biological agents (also referred to as "biological substances") and to protect other persons insofar as they may beat risk due to the use of biological agentsby employees or business ownerswithout employees.

Important information inthe BioStoffV:

The term biological agentis explained in more detail (§ 2)

The terms "specific" and "non-specific" activities in connection with biological agentsare explained in more detail (§ 2). This distinction is important for choosing the appropriate level of protection.

Biological agents are categorised into risk groups based on the risk of infection they pose (§ 3)

The BioStoffV stipulates that a risk assessment must be carried out in accordance with § 5 of the German Occupational Health and Safety Act (Arbeitsschutzgesetz)before starting work (§ 4)

An explanation of activities with and without protection level classification as well as the employer's documentation and record-keeping obligations(§ 6 and § 7)

Basic obligations of the employer with regard to occupational health and safety and general hygiene measures

Further content and obligations can be taken directly from the BioStoffV [1].

Technical rules for biological agents (TRBA)

While the Biological Agents Ordinance provides the legal framework, the TRBAs substantiatethe current status of the respective requirements in the individual areas.

The most important TRBAs for brewery laboratories are:

TRBA 100: Protective measures for activities involving biological agents in laboratories

TRBA 400: Guidelinefor risk assessment and for the instruction ofemployees regardingactivities involving biological agents

A complete list of TRBAs is provided bythe German Federal Institute for Occupational Safety and Health [2].

Classification of microorganisms into risk groups

Section 3 of the German Ordinance on Biological Agents (BioStoffV) defines four risk groups for biological agents [3]:

Risk group 1: Biological agents that are unlikely to cause disease in humans

Risk group 2: Biological agents that may cause disease in humans and could pose a risk to employees; unlikely to spread to the community; effective prevention or treatment is usually possible

Risk group 3: Biological agents that may cause serious disease in humans and pose a serious risk to employees; theymay presenta risk of spreading to the community, but effective prevention or treatment is normally possible

Risk group 4: Biological agents that cause serious disease in humans and pose a serious risk to employees; they may present a high risk of spreading to the community; there is usually no effective prevention or treatment available

Examples of substances fromthe respective groups can be found in the literature [3].

Microbiologylaboratories in the brewing and beverage industry generally work with organisms in risk class 1. Exceptions include, for example, microbiological drinking water analysis (see assignmentof laboratories toprotection levels and safety levels).

Classification of laboratories into protection levels and safety levels [4, 5, 6]

According to the BioStoffV, four protection levels are assigned to the four risk groups. The protection level and risk group are identical for specific activities.

Activities are considered specific according to § 2 BioStoffVif:

1. The activities are directly focused onone or more biological agents

2. The biological agent or agents is/are known at least by species, and

3. The exposure of employees during normal operation is sufficiently known or can be estimated

If one of these conditions does not apply, the activities are non-specific. In the case of a non-specificactivity, the further procedure is regulated by § 5 BioStoffV. Annex II BioStoffV lists the protective measures for protection levels 2, 3 and 4.

Biological safety levels are not to be confused with protection levels. Biosafety levels are used to categorise the hazards of genetic engineering work in genetic engineering facilities. The safety levels are also arranged in a 4-level model, whereby biosafety level 1 is often referred to as "BSL-1".

The terms "BSL-1 laboratory" and "protection level 1 laboratory" are often used interchangeably in everyday life. However, as brewery laboratories do not work with GMOs, the term "protection level 1 laboratory" would be more correct.

Further details on the categorisation of brewery laboratories can be found in TRBA 100, Section4.4.3. (1). This states: Laboratories in which sterility tests, colony count determinations and other biological quality-assurance work iscarried out that doesnot serve to specifically detectbiological agents of risk group 2 and highercan be carried out under the conditions of protection level 1. This includes, for example, samples from the production of foodstuffs, medical devices, pharmaceuticals or cosmetics.

If, during the course of the activities,biological agents of risk group 2 or 3 are selectively propagated or enriched, the activitiesmust be carried out under the conditions of at least protection level 2 in accordance with the BioStoffV.

Biological protection levels (biosafety level or BSL laboratory) are used to categorise the hazards of biological agents (biological substances). This classification is standardised by EU Directive 200/54/EC on the protection of workers from risks related to exposure to biological agents at work. It is implemented in Germany by the Biological Agents Ordinance (BioStoffV). It categorises laboratories into four protection levels according to the risk groups. These build on each other, i.e. the rules of the lower protection levels also apply to the higher protection levels. A basic distinction must be made between specificand non-specificactivities. If it is a specificactivity, the protection level corresponds to the risk group. For non-specificactivities, the further procedure is also regulated by the BioStoffV (§ 5).

Protection level 1: General hygiene measures must be observed; this applies to structural, technical and organisational requirements; special hygiene measures in accordance with the technical rules for biological agents (TRBA) continue to apply to activities with protection level assignment; for work in the laboratory, the protective measures for activities involving biological agents in the laboratory (TRBA 100) must also be observed.

Protection level 2: The protection level area must be marked as such, including the protection level designation and the biohazard symbol; only named employees are permitted in the laboratory area; activities involving aerosol formation only in the area of a safety cabinet (clean bench) or in technical facilities with an equivalent level of containment; prior to disposal, mandatory inactivation via proven physical or chemical methods (usually autoclave). Suitable equipment for viewing the laboratory from the outside; TRBA 100: in addition to lab coats, protective gloves, face protection if necessary; mandatory eyewash facility; windows and doors must be closed; disinfection after completion of work according to hygiene plan.

Protection level 3: Access restricted todesignated employees; only with access control.

Protection level 4: Laboratory structurally separated; can be sealed for possible fumigation: filters for supply and exhaust air; access via three-chamber airlock.

If work is carried out with genetically modified organisms, the German Genetic Engineering Safety Ordinance (GenTSV) applies in accordance with the German Genetic Engineering Act (GenTG). This defines additional biosafety levels (hazard classification for genetic engineering work in genetic engineering facilities). (biosafety levels 1–4).

General definition: Genetically modified organisms (GMOs) are produced by genetic engineering. Not officiallyrelevant for brewing laboratories.

General information on the layoutof a brewery laboratory [2, 3]

Some general principles apply to the basic design of microbiologylaboratories. These general instructions are specified in TRBA 100 in sections5.2–5.5 for protection levels 1–4.

As an example, only the rules of TRBA 100 for protection level 1 should be mentioned here:

In the case of activities involving biological agents of risk group 1 and non-specificactivities that pose very little or no risk to employees, the occurrence of an infectious disease is unlikely. It is therefore sufficient to ensure that the laboratory is operated as intended:

(1) Protection level 1 laboratories should consist of separate, sufficiently large rooms. Sufficient working space must be provided for each employee in accordance with the activity.

(2) Surfaces (work surfaces, floors) should be easy to clean and must be imperviousand resistant to the substances and cleaning agents used.

(3) A wash basin should be available in the work area.

(4) The basic rules of good microbiological techniquesmust be observed (see Annex: "Basic rules of good microbiological techniques"). Point 9 of the GMT only applies to specificactivities.

(5) Biological agents in risk group 1 can only be disposed of without pre-treatment if the outcomeof the risk assessment or other regulations (e.g. water, waste or genetic engineering legislation) do not prevent this.

(6) When working with biological agents of risk group 1 with a sensitising or toxic effect, measures must be taken to minimise employee exposure. This may involve, for example, the use of a safety cabinet, the use of suitable respiratory protection or the avoidance of spore-forming development phases in fungi or actinomycetes.

The "basic rules of good microbiological techniques" (GMT) are as follows in accordance with TRBA 100:

(1) Windows and doors in the work areas should remainclosed during activities.

(2) It is prohibitedto drink, eat or smoke in the work areas. Food may not be stored in the work area.

(3) Lab coats or other protective clothing must be worn in the work area.

(4) Mouth pipetting is prohibited; pipetting aids must be used.

(5) Syringes and cannulas should only be used if absolutely necessary.

(6) All activities must beperformed as carefully as possible to prevent the formation of aerosols.

(7) After finishing work and before leaving the work area, hands must be carefully washed, disinfected if necessary and re-moisturised(skin protection plan).

(8) Work areas should be kept clean and tidy. Only the equipment and materials actually required should be presenton the work surfaces. Supplies should only be stored in areas and cupboards provided for this purpose.

(9) The identity of the biological agents used must be checked regularly, provided thisis necessary to assess the hazard potential. The intervals depend on the hazard potential.

(10) When working with biological agents, employees must be instructed verbally and at the workplace before starting work and at least once a year thereafter.

(11) Employees who are inexperienced in microbiology, virology, cell biology or parasitology must be given particularly comprehensive instruction, careful guidance and supervision.

(12) If necessary, pests must be controlled regularly and professionally.

In addition, the following general advice applies tothe laboratoryset-up:

Clear separation from other work areas and clear labelling of the area

Self-closing doors with a viewing window

Floors and work surfaces in the sterile area should be seamless and easy to clean

Avoid dust traps (pipework, suspended lamps, cupboards that do not end at ceiling height, open shelves, etc.).

Walls and ceilings must also besmooth and washable

Avoid floor drains, air conditioning and ventilation systems wherever possible

The sterile room should contain the bare minimum of equipment

The number of people in the sterile area must be kept to a minimum

If possible, provide separate rooms or clearly demarcated areas for:

Sample receipt

Sample preparation

Sample analysisincluding incubation

Storage of the reference organisms

Preparationof media and the corresponding equipmentincluding their sterilisation

Sterility check

Decontamination

To prevent accidental cross-contamination, laboratory equipment should not be routinely exchanged between areas.

Further information on laboratory equipment can be found in the literature.

In order to test the influence and effects of filter aids in a routine production situation, it is recommended that a filtration trial be conducted under standard operating conditions.

The odor and flavor of the filtered beer is also tested as part of this trial.

The test should be conducted against a control sample that has not been in contact with the material to be tested.

This test provides information regarding any odors transferred by filter aids to wort and beer. A minimum of three samples should be collected from each lot of the product delivered.

Alkaline cleaning agents and disinfectants containing chlorine can be determined by their alkalinity concentration. The chlorine present destroys most indicators by oxidation. Before adding the indicator, the chlorine must be eliminated by adding sodium thiosulphate to the sample.

Determination of the quicklime (CaO) or sodium hydroxide (NaOH) content of the detergent solution with an acid solution (HCl or H₂SO₄) with corresponding equivalent concentration (normality) up to the color change of phenolphthalein (pH 8.2).

Quicklime

Sodium hydroxide

If the alkaline solution is likely to be strongly influenced by carbonic acid or if the cleaning solutions are used over a longer period of time ('batch cleaning'), the degree of carbonation must be taken into account to determine the exact concentration or pH value, as the resulting soda has a greatly reduced cleaning effect, but simulates an excessively high concentration of the active cleaning agent.

Determine the sodium hydroxide (NaOH) content of the detergent solution with an acid solution (HCl or H₂SO₄) with corresponding normality up to the color change of phenolphthalein (pH 8.2).

   NaOH + HCl           →         NaCl      +   H₂O             colorless against phenolphthalein

2 NaOH + H₂SO₄      →         Na₂SO₄ + 2 H₂O

Determination of the soda ash (Na₂CO₃) content of the detergent solution with an acid solution (HCl or H₂SO₄) with corresponding normality up to the color change from methyl orange (pH 4.3).

   Na₂CO₃  + HCl        →       NaHCO₃  + NaCl

   NaHCO₃  + HCl        →      NaCl      + H₂O + CO₂    yellowish brown against methyl orange

2 Na₂CO₃   + H₂SO₄   →   2 NaHCO₃ + Na₂SO₄

2 NaHCO₃  + H₂SO₄    →  2 Na₂SO₄ + H₂O + 2 CO₂