🦠 Anaerobic spore-forming bacteria and anaerobic spore-forming sulphite-reducing bacteria in meat and meat products — detection by the culture method with biochemical (and microscopic) confirmation, according to the PKN card also identification of certain species and detection of Clostridium botulinum toxin; PN-A-82055-12 (withdrawn standard)

Microbiology PN-A-82055-12

Overview

WHAT THE STANDARD COVERS. Scope from the PKN catalogue card, in full: “The following test methods are discussed: detection of anaerobic spore-forming bacteria and anaerobic spore-forming sulphite-reducing bacteria; identification of certain species of anaerobic spore-forming bacteria most frequently occurring in meat and meat products; detection of Clostridium botulinum toxin in cultures of the strains tested and in food samples”. Title from the card: “Meat and meat products -- Microbiological examination -- Detection of anaerobic spore-forming bacteria and anaerobic spore-forming sulphite-reducing bacteria”. We have not read the text of the standard — we do not know the procedure of the methods, the media, temperatures or the quantity of sample from the standard.

STATUS (PKN cards as of 08.11.2026). Card PN-A-82055-12:1997 (Polish version) has the label “Withdrawn” and the line “Withdrawal date: 04-07-2007”; there is no “Replaced by” field — the card does not indicate a successor. Edition: published 28-10-1997, 10 pages, price group E, Food, Agriculture and Forestry Sector, KT 93 Meat, Eggs and their Products, ICS 07.100.30, “Replaces: PN-A-82054:1983 - Polish version”. Card PN-A-82054:1983 “Meat and meat products -- Bacteriological examination” — “Withdrawn and replaced by”, withdrawal date 07-01-1999; it was replaced by twelve standards: sheets 2, 3, 4, 6, 7, 8, 9, 10, 11, 12 and 14 of the PN-A-82055 series and PN-ISO 13720:1999 — one standard covering sampling and sample preparation, inoculation and interpretation of many determinations was replaced by sheets devoted to separate topics. Searching “PN-A-82055” in the PKN catalogue (08.11.2026) returns 17 sheets of the series and amendment Az1:2002 to sheet 4; only PN-A-82055-4:1997 (testing the tightness of hermetically sealed canned products) and its amendment are without the “Withdrawn” label — all the others, including sheets 2 (general rules of testing) and 3 (preparation of samples and dilutions), are withdrawn. When withdrawn: 04-07-2007. Replaced by: according to the card — nothing. What changed for the result: with no successor there is nothing to compare; the closest current standard on the topic, PN-EN ISO 15213-1:2023-08 (PKN card without a withdrawal note, read 08.11.2026: enumeration of sulphite-reducing anaerobes and sulphite-reducing Clostridium spp. by the colony-count technique; it replaced PN-ISO 15213:2005), gives a colony count, not a presence result, and the card does not present it as the successor of PN-A-82055-12.

HOW MANY LABORATORIES AND IN WHAT FORM (copy of the accreditation scope database, read 08.11.2026; “Laboratories” tab on labcoda.pl, 08.11.2026; current scope documents on the PCA website, read 08.11.2026). The number 82055-12 appears in the copy in 8 records at 6 Polish laboratories: AB 400 (2), AB 1415 (2), AB 437, AB 462, AB 483, AB 510 (1 each); it does not appear at laboratories of other countries. Counted across all fields of the records and all forms of notation of the number (notations with a colon, slash, full stop and space after the series number were also checked); the other sheets of the PN-A-82055 series are not included in the set. Notations: “PN-A-82055-12” without a year (AB 400 — 2, AB 462), “PN-A-82055-12:1997” (AB 437), “PN-A-82055-12:1997 pkt.2” (AB 483), “PN-A-82055-12:1997 p. 2” (AB 510), “PN-A-82055-12:1997 z wył. pkt 4” (excluding point 4; AB 1415 — 2). Current PCA scope documents: AB 400 — issue 37 of 07.08.2026 (2 records), AB 462 — 31 of 27.04.2026, AB 483 — 26 of 04.05.2026, AB 510 — 25 of 07.08.2026, AB 1415 — 17 of 08.04.2026 (2 records) — the same notations as in the copy; AB 437 no longer has this standard in issue 31 of 10.09.2026. The current PCA documents therefore contain 7 records at 5 laboratories. The “Laboratories” tab (“PN-A-82055-12”) shows 6 laboratories and 8 records — including AB 437, because it reads the same data as the copy (tab query on labcoda.pl, 08.11.2026).

WHAT THE LABORATORIES TEST (database copy, complete items — 8). Presence of anaerobic spore-forming bacteria and anaerobic spore-forming sulphite-reducing bacteria in one record: AB 483 — meat and meat products, culture method; AB 510 — meat and meat products, poultry, fish and fish products, food concentrates, fruit and vegetable products and vegetable-meat products, delicatessen products, food additives, culture method with biochemical confirmation; AB 437 — meat and meat products, poultry and poultry products, fish and fish products, frozen products, canned products, delicatessen products and meat-vegetable products, culture method with biochemical and microscopic confirmation; AB 400 (two records) — food in a flexible scope, culture method with biochemical confirmation; AB 462 — food and feed in a flexible scope, worded “anaerobic spore-forming bacilli”, method with biochemical confirmation. AB 1415 splits the test into two records — presence of anaerobic spore-forming bacteria and separately of sulphite-reducing ones — in meat and meat products, by the culture method with microscopic and biochemical confirmation, excluding point 4 of the standard. No record has a numerical range or gives the quantity of sample in which the bacteria are detected. The list of laboratories approved by the Chief Veterinary Officer of 2018 (GIWlab decisions of 2018) lists this standard at two laboratories: for meat and meat products with point 2 and for food together with the laboratory’s own instruction, worded “detection of … in a specified mass/volume of sample”.

WHAT SUCH DETECTION LOOKS LIKE (instructions for food microbiology classes, University of Warmia and Mazury, 2012 — this is not the text of the standard and we do not give figures from it). Spores are determined in the sample after heating, which destroys vegetative cells. For anaerobic spore-forming sulphite-reducing bacilli, the inoculum in tubes is poured over with liquefied and cooled differential medium with sodium sulphite and ferric ammonium citrate, after solidification overlaid with a layer of water agar and incubated, checking growth daily; black colonies (iron sulphide from the reduction of sulphite) in the replicates indicate the presence of spores of sulphite-reducing anaerobes in the tested quantity of material. Whether PN-A-82055-12 provides for the same media, conditions and quantities we do not know.

WHERE A SEEMINGLY CORRECT RESULT IS EASY TO GET. Standard: a result according to a standard withdrawn in 2007 without a successor has no current equivalent in the PKN catalogue — the 1997 edition must be stated in the report; five laboratories keep this standard in their current scopes, one (AB 437) removed it in the issue of 10.09.2026, and the “Laboratories” tab still shows it. Sampling and preparation: a qualitative result refers to a specified mass or volume of sample — “absent in 0.1 g” and “absent in 1 g” are different results, and no record in the scopes gives this quantity; sheets 2 and 3 of the series (general rules of testing, preparation of samples and dilutions) are also withdrawn. Measurement: “anaerobic spore-forming bacteria” and “anaerobic spore-forming sulphite-reducing bacteria” are two different results — the result for the first group says nothing about the second and vice versa; AB 1415 records them as two records, the others as one. The card lists three methods (detection, identification of species, Clostridium botulinum toxin); AB 1415 excludes point 4, AB 483 and AB 510 cite point 2 — which method is contained in which point we have not read; a result “according to PN-A-82055-12” without a point does not say whether it includes identification of the species or of the toxin. Scope of the test object: the title of the standard concerns meat and meat products; AB 400 and AB 462 apply it to food in a flexible scope (AB 462 also to feed), AB 510 and AB 437 — also to fish, concentrates, fruit and vegetable products and food additives — whether the standard allows products other than meat does not follow from the card. Calculation: a qualitative result is not converted into a number — it is not comparable with the colony count of sulphite-reducing Clostridium spp. according to PN-EN ISO 15213-1 (separate entry in the Knowledge Base), even when both results appear in one report (AB 400 has both standards for food, PN-A-82055-12 and PN-EN ISO 15213-1).

WHAT WE DO NOT GIVE. We have not read the text of PN-A-82055-12:1997, and in open access we found neither the text nor a publication describing its procedure. We do not give: the quantity of sample, the way of heating, the media, incubation temperatures and times, the way of confirmation (biochemical, microscopic), the species covered by identification, the procedure for detecting Clostridium botulinum toxin, the numbering of the points of the standard or the way of expressing the result. The procedure described above comes from the university instructions and is marked as such. We have not established why PKN withdrew the standard without a successor. We have not read separately the status of PN-ISO 13720:1999 and the cards of the other sheets of the series (status of the sheets — from the PKN search list, 08.11.2026). We have not checked citations of the standard in legislation in force. We have not reviewed the current PCA scope documents of laboratories outside the copy.

Method principle

Qualitative test by the culture method: in a specified quantity of a sample of meat or a meat product the presence of anaerobic spore-forming bacteria is detected and — as a separate result — of those among them that reduce sulphite, with biochemical confirmation (in some scopes also microscopic); according to the PKN card the standard also describes the identification of certain species and the detection of Clostridium botulinum toxin (PKN card; we have not read the text of the standard — we do not give the procedure, media or conditions).

Applications

• Meat and meat products — presence of anaerobic spore-forming bacteria and anaerobic spore-forming sulphite-reducing bacteria (PKN card)
• According to the PKN card also: identification of certain species of anaerobic spore-forming bacteria and detection of Clostridium botulinum toxin in cultures and food samples
• In accreditation scopes: 6 Polish laboratories in the copy (5 in current PCA documents) — meat and meat products, poultry, fish, canned products, delicatessen products, and in flexible scopes food and feed
• Standard withdrawn 04-07-2007 without a successor — the result has to be described with the 1997 edition

Key parameters

Standard

Standard number

PN-A-82055-12:1997 (Polish version; withdrawn 04-07-2007 with no successor indicated)

Title (PL)

Mięso i przetwory mięsne — Badania mikrobiologiczne — Wykrywanie obecności beztlenowych bakterii przetrwalnikujących i beztlenowych bakterii przetrwalnikujących redukujących siarczany(IV)

Title (EN)

Meat and meat products — Microbiological examination — Detection of anaerobic spore-forming bacteria and anaerobic spore-forming sulphite-reducing bacteria

Step-by-step procedure

1. Sample and tested quantity

   Sample of meat or a meat product; the quantity in which the bacteria are detected determines the result — according to the standard (not given) and according to the order; the sheets of the series on general rules of testing and sample preparation (2 and 3) are withdrawn (PKN cards, 08.11.2026).

2. Heating and inoculation

   Heating that destroys vegetative cells, deep inoculation in the medium in tubes, agar overlay (general procedure according to the university instructions; we have not read the conditions of the standard).

3. Incubation under anaerobic conditions

   Checking of growth; black colonies indicate sulphite-reducing bacteria (university instructions; we do not give the time and temperature according to the standard).

4. Confirmation

   Biochemical confirmation, in some scopes also microscopic (accreditation scopes); according to the PKN card the standard also describes the identification of certain species and the detection of Clostridium botulinum toxin.

5. Result

   Presence or absence of each of the two groups of bacteria in the tested quantity of sample, stating the edition of the standard and the points applied. PN-A-82055-12:1997 — WITHDRAWN 04-07-2007, with no successor indicated (PKN card as of 08.11.2026).

Required equipment and apparatus

Reagents, media and consumables

Health and safety (OHS)

• Cultures of anaerobic spore-forming bacteria from food may contain pathogenic strains, and according to the PKN card one of the methods of the standard concerns Clostridium botulinum toxin — work under biosafety conditions appropriate for such microorganisms and their toxins (general rule; we have not read the text of the standard)
• TECHNICAL NOTE: a qualitative result is not interchangeable with the colony count of sulphite-reducing Clostridium spp. according to PN-EN ISO 15213-1; the standard was withdrawn without a successor — the 1997 edition in the report

Frequently asked questions