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Identification of 129 Micrococcaceae Strains Isolated from Food of Animal Origin C

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Identification of 129 Micrococcaceae Strains Isolated from Food of Animal Origin C Identification of 129 Micrococcaceae strains isolated from food of animal origin C. Delarras, C. Guichaoua and M.-P. Caprais Code words: Staphylococcus- nitrofurantoin- aurease- ID 32 STAPH Tab. 1: List of 26 biochemical tests in the ID 32 STAPH-system - numerical identification Reaction/ Test Reaction/ Test substrate substrate 129 strains of Micrococcaceae novobiocin (5 11g/ml), 13 to 44% Urease URE Cellobiose (F) GEL were isolated from food of animal were identified with negative dis­ Aginin dihydrolase ADH Acetoin (production) VP origin (minced meat, cakes with cordant tests using this mi­ Ornithin decarboxylase ODC Nitrat (reduction) NIT confectioner's custard) on Baird­ cromethod. 44% produced no ac­ Esculin (hydrolysis) ESC B Galactosidase B GAL Glucose (F) GLU Arginin arylamidase ArgA Parker medium. etoin, 35 % had no urease and Fructose (F) FRU Alkaline phosphatase PAL 15 % no arginine dihydrolase. Maltose (F) MAL Pyrrolidonyl Arylamidase PyrA 120 were Staphylococcus and 9 Mannose (F) MNE Novobiocin (Resistance) NOVO Lactose (F) LAC Sucrose (F) SAC Micrococcus, according to the 10 48 Staphylococcus strains (40 %) Trehalose (F) TRE N-Acetyl Glucosamine (F) NAG 32 STAPH-System (1989). The re­ were identified as human coagula­ Mannitol (F) MAN Turanose (F) TUR sults were analyzed using a com­ se negative Staphylococcus spe­ Raffinose (F) RAF Arabinose (F) ARA Ribose (F) RIB B Glucoronidase B GUR puter program. cies (strains: 39; species: 9) or as animal species (strains: 4; spe­ (F) ~ Fermentation 60% of these 120 Staphylococcus cies: 1). 5 were Staphylococcus strains of animal origin were co­ sp S. epidermidis and S. warneri - similar colonies, but without reading of the biochemical tests agulase positive S. aureus (au­ represented 63 % of these Micro­ halos. Some of them con-espond to after 24 hours incubation at 37 °C. rease+, Rapidec Staph tech­ coccaceae with particular bio­ coagulase positive S. aureus (10) nique*). 46% were resistant to chemical characteristics. but most of them are identified A comparison of the identification with other species of staphylococci of staphylococci of food origin and (14, 12) or other bacteria (micro­ of other origins by 3 micromethods cocci: (19)), as a function of the has shown that the ID 32 STAPH Micrococcaceae, particularly sta­ dium from food of animal origin. origin of their strains. system gives the best results (13). phylococci, with strains of coagu­ They have been studied in relation Micrococcaceae strains from inter­ lase positive S. aureus have- been to their biochemical and ecological The colonies above are cultured on national collections which have al­ detected in food products for 20 to properties and on the basis of pre­ trypticase soya agar before being ready been studied with this system 30 years. Many papers have dealt vious studies (10, 12). studied. were not reintroduced into the with their enumeration and identifi­ present work (13). cation (3, 8, 19, 27. 29). S. aureus Material and methods Differentiation tests for isolated from milk, cheese or other Staphylococcus and Numerical identification foods can produce enterotoxins Preparation of the food Micrococcus The results of the 26 biochemical which cause food poisoning in man Food of animal origin (uncooked, The nitrofurantoin test (18) with tests for each strain are expressed (45, 48). French legislation there­ frozen minced meat, cakes with 300 }lg/ml discs (Diagnostics Pas­ in + and - signs and then in a fore requires their detection and · confectioner's custard) are routine­ teur) is prefeiTed to other tests numerical profile with 9 numbers identification in food products. ly analyzed in hygiene laboratories because it givesc good differentia­ in accordance with the manufactur­ in accordance with the Decree of tion of staphylococci and micro­ er's coding system. Other species of staphylococci of the 21st December 1979 (Joumal cocci of food origin (11). It was human origin which are coagulase Officiel Fran9ais of 19-01-80). carried out in Mueller-Hinton They are interpreted with a compu­ negative (15. 22, 25, 32) are not (D.P.) medium, the antibiogramme ter program which gives infonna­ researched in food. They are poten­ 30 g of food (after thawing out) are technique being used for these 129 tion such as: the number and nature tially pathogenic and this finds ground in a grinder-homogenizer in strains. of the discordant tests with the expression in various infections in buffed peptone water (Bacto-pep­ percentages of positive results for man (6, 7, 30). Some of them may tone 20 g, NaCl 5 g, monopotassi­ After 24 hours incubation at 37 °C the relevant species. also produce enterotoxins (31 ). um phosphate 1,5 g, dis odium the diameter of the zone of inhibi­ phosphate 9 g, distilled water tion is measured. If it is more than Aurease test for S. aureus Staphylococci species of animal 1000 ml, pH 7,2). It is obligatory to 15 mm the strain is sensitive (Sta­ strains origin have been described during lase positive S. treat frozen food for 30 to 45 min phylococcus) but if it is less than the past ten years, i.e. S. sciuri The Micrococcaceae identified as the growth of at 20 ac. 15 mm it is resistant (Micrococ­ (23), S. intermedius (16), S. arlet­ S. aureus by the ID 32 STAPH were unknown cus). derived from tae, S. klosii and S. equorum (33). Staphylococci isolation system were examined for free co­ Some may also be pathogenic to Biochemical study agulase by a standardized mi­ man. This was done according to the cromethod: Rapidec Staph (bi­ AFNOR NF-V-08-014 standard for A micromethod was used here. The oMerieux s.a.). Aurease or free In food control laboratories, coagu­ the enumeration of S. aureus or­ new system ID 32 STAPH (5) with coagulase staphylococci were de­ lase positive S. aureus organisms ganisms in food. 0.1 ml of food its 26 biochemical tests (tab. 1) tected by a U.V. light fluorescent are usually isolated on Baird-Park­ suspension was spread on Baird­ intended mainly for the identifica­ reaction after 2 hours incubation at er medium (2) but many other Parker medium in a Petri dish. tion of the genera Staphylococcus 37 oc. media have been suggested from After 24 and 48 hours incubation 2 and Micrococcus is the one in Zebovitz medium (35) to modified types of colonies were harvested: question. Results Baird-Parker media (26) and com­ parative studies have been made of - circular, convex, black and Each strip was inoculated with a The 129 Micrococcaceae strains in the media (3). shiny colonies with a clear halo of suspension into 2 ml of sterile dis­ the present study were found, when eggyolk colour. These are consid­ tilled water, the opaqueness of submitted to a 300 )..lg/ml nitro­ During obligatory bacteriological ered as characteristic of coagulase which is equal to 0.5 MacFarland furantoin test, to be divided as controls Micrococcaceae have positive S. aureus and are routinely units. Th~ procedures recommend­ follows: been isolated on Baird-Parker me- enumerated after confirmation by a ed by the manufacturer are fol­ *) bioMerieux s.a., Marcy !'Etoile, F- free coagulase test. lowed for inoculation and for the - 123 sensitive strains are staphy· 69752 Charbonnieres-les-Bains Ce­ locci, but 3 of them are confinned dex, Frankreich Fleischwirtsch. 74 (1 0), 1084-1086 (1994) as micrococci (tab. 4) 1084 Fleischwirtsch. 7 4 (1 0), i994 - 6 resistant strains are micrococ­ Tab. 2: Identification of 120 Staphylococcus strains isolated from food of animal origin - 8 species are rarely found in these foods: S. capitis, S. cohnii, S. ci Number of strains Staphylococcus Nb. T.P. I.P./ I.P./ biochemical haemolyticus, S. hominis 2, S. Biochemical identification of the species T.A. T.A. particu- simulans, S. ~cylosus and S. schleif­ larities 129 Micrococcaceae strains using 1 2a4 eri. They were originally isolated the ID 32 STAPH system S. aureus 72 4 22 46 ct. Table 3 from the human skin (15, 22, 32) S. capitis 1 1 but can also be found in milk S. chromogenes 4 4 120 strains isolated from these S. cohnii 1 1 1 products and meat products (8, 9, foods and classified as Staphyloco­ S. epidermidis 1 8 3 5 ADH-/5 27) and in cheese (29). ccus strains are identified with var­ S. epidermidis 2 1 1 S. haemolyticus 1 1 ious species of the data base (tab. 2 S. hominis 2 2 2 They are identified in the ID 32 and 3) and only 9 strains with S. schleiferi 2 1 1 STAPH system by 1 to 4 atypical micrococci (tab. 4). S. simulans 1 1 tests. The 8th species S. chromoge­ S. warneri 21 2 9 10 PAL+/4 VP-/7 nes ( 17) of animal origin (pigs, The results show the number of S. xylosus 1 1 cows with bovine mastitis, poultry) strains with a typical biochemical S. species 5 5 T.A. > 4 is represented by 4 strains which profile (discordant tests: 0), the Total strains 120 7 36 77 can be identified by atypical posi­ strains with profile identified by 1 tive tests. discordant tests (1, 2 to 4) and their Nb. =Number T.P. =typical biochemical profile biochemical details. They are ana­ I.P. = identified profile - 2 other species: S. epidermidis 1 lyzed and discussed. T.A. 1, T.A. 2 to 4 = tests against 1, 2 to 4 and > 4 and 2 (9 strains) and particularly S. Tab. 3: Identification of 72 S. aureus strains isolated from food of animal origin warneri (21 strains) make up 63 % Discussion of these staphylococci of food ori­ Number of strains gin other than S. aureus. S. aureus Nb. T.P. I.P./ I.P./ biochemical 129 i'vlicrococcaceae strains iso­ T.A.
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