Research ArticleMicrobial Profile of FreshBeef Sold in theMarkets ofNgaoundereCameroon and Antiadhesive Activity of a Biosurfactant againstSelected Bacterial Pathogens

Hippolyte T Mouafo 12 Annick M B Baomog1 Jorelle J B Adjele1

Alphonse T Sokamte2 Augustin Mbawala2 and Robert Ndjouenkeu 2

1Centre for Food and Nutrition Research Institute of Medical Research and Medicinal Plants Studies PO Box 6163Yaounde Cameroon2Department of Food Sciences and Nutrition National School of Agroindustrial Sciences University of Ngaoundere PO Box 454Ngaoundere Cameroon

Correspondence should be addressed to Hippolyte T Mouafo hippolytetenegmailcom

Received 26 November 2019 Revised 10 January 2020 Accepted 27 January 2020 Published 19 February 2020

Academic Editor Efstathios Giaouris

Copyright copy 2020 Hippolyte T Mouafo et al is is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work isproperly cited

Owing to its composition meat is recognized as one of the best media for microbial growth leading to meat spoilage and food-borne illnesse ability of microorganisms to adhere to surfaces where meat is deposited during selling is a nonnegligible cause ofmeat contaminationis work was performed to assess themicrobial profile of fresh beef sold in themarkets of Ngaoundere townand evaluate the antiadhesive activity of a biosurfactant derived from Lactobacillus paracasei subsp tolerans N2 against selectedpathogenic strains isolated in fresh beef All fresh beef samples analysed were contaminated with both pathogenic and spoilagemicroorganisms at levels higher than the microbiological criteria set by the European Commission A total of 151 strainsbelonging to 12 species (Pseudomonas putida Pseudomonas aeruginosa Pseudomonas sp Escherichia coli 1 Escherichia coliSalmonella enteritidis Salmonella sp Staphylococcus epidermidis Staphylococcus xylosus Staphylococcus aureus Candidaalbicans andCandida sp) were isolated and identified A specific relationship between themicrobial diversity of fresh beef and thesampling sites was observed Biosurfactant displayed antiadhesive activity against all the tested strains and the complete inhibition(100) of Bacillus sp BC1 S aureus STP1 and S xylosus STP2 was noticed at biosurfactant concentration of 10mgmL isstudy indicates the microbial diversity of fresh beef sold in Ngaoundere markets and suggests the potential use of biosurfactant asan antiadhesive agent in the meat industry

1 Introduction

Meat is a regular part of the human diet worldwide Itcontains proteins (with all essential amino acids) vitamins(A B12 B6 D and E) and minerals (iron and zinc) whichare very important for human growth and well-being [1]Due to its nutrient content meat is a highly perishablefood as it represents good media for the proliferation ofmicroorganisms [1 2] ese microorganisms are essen-tially spoilage organisms (Brochothrix thermosphactaLactobacillus spp Clostridium spp Leuconostoc sppCarnobacterium spp Pseudomonas spp Candida spp

Penicillium Mucor Aspergillus Rhyzopus and Enter-obacteriaceae) responsible for off-odors discolorationdevelopment of slime and production of toxic compoundsand pathogens (Salmonella spp E coli Staphylococcusaureus Listeria monocytogenes and Campylobacter spp)responsible for food-borne diseases [3] Generally meat isconsumed cooked but according to some food habits andtraditional recipes meat may be consumed raw or partiallycooked [4] Under these conditions the consumption ofmeat was many times associated with outbreaks of food-borne diseases [5] According to the World Health Or-ganization 30 of inhabitants in industrialized countries

HindawiJournal of Food QualityVolume 2020 Article ID 5989428 10 pageshttpsdoiorg10115520205989428

suffer every year of food-borne diseases [6] with most ofthe cases attributable to the consumption of meat [7] Insub-Saharan Africa food-borne diseases due to the con-sumption of meat are not uncommon Because of theweakness of bacteriological diagnose resources associatedwith the unawareness and nonenforcement of laws [8] theestimation of food-borne diseases as well as resultinglosses is underevaluated [9]

Nowadays as the demand and consumption of rawmeat have increased in many countries [10] regulationswhich specified the microbiological criteria of raw meathave been established in order to protect consumersrsquohealth and to facilitate its commercialization To this endthe microbial contamination of meat is becoming a publichealth concern worldwide and particularly in Cameroon[11] It has been reported in the literature that microor-ganisms contaminate meat during the production pro-cessing transportation and distribution processes [12]e first step in the control of the microbiological qualityof meat consists of evaluating their initial contaminationlevels and identifying the different microorganisms in-volved e studies assessing the microbiological quality offresh beef have been carried out in several countries suchas Turkey [13] Ethiopia [1] Nigeria [14] Sudan [15]Ghana [16] Rwanda [17] and India [18] It thereforeappears interesting to perform such kind of study in someCameroonian metropolises where fresh beef is highlyconsumed by the whole class of the population

Another important step in the control of the micro-biological quality of meat is the cleaning process of sur-faces where meat is processed [19] A study conducted byZerabruk et al [1] showed the presence at high levels ofdifferent groups of microorganisms on cutting tables inbutcher shops of Addis Ababa In fact microorganismsdisplay the ability of adhering to the surface and formingstructured communities encapsulated within a self-de-veloped polymeric matrix known as biofilms which areresistant to the conventional cleaning processesereforeinhibiting the initial adhesion of microorganisms to sur-faces is a challenging task in the meat industry Utilizationof biosurfactants which deserve antiadhesive properties[20ndash22] appears as an alternative Biosurfactants areamphiphilic compounds produced by a wide variety ofmicroorganisms such as bacteria yeasts and moulds thatcan have some influence on interfaces Opposite to theirsynthetic homologues these compounds have uniquecharacteristics such as being nontoxic biodegradableenvironmentally friendly and selective having structuraldiversity being effective under extreme conditions of pHtemperature and salinity and having unique surface-ac-tive properties [23] To the limit of our knowledge theantiadhesive properties of biosurfactants were not yettested against strains isolated from fresh beef despite theability of these strains of adhering to cutting meat surfacesHence the objective of the present work is to establish themicrobial profile of fresh beef sold in NgaoundereCameroon and evaluate the antiadhesive properties ofbiosurfactants against selected pathogenic strains isolatedin fresh beef

2 Materials and Methods

21 Study Area e study was conducted in the city ofNgaoundere Adamawa region of Cameroon from February2015 to May 2016 e six main markets of the town wherefresh beef is daily sold were chosen ese markets wereBantail market (MBT) Bamnyanga market (MBG) Dangmarket (MD) ldquoGrand Marcherdquo (GM) ldquoPetit Marcherdquo (PM)and ldquoCentre Commercialrdquo (CC)

22 Fresh Beef Samples e normalized method ISO 176042015 [24] which specifies the sampling methods for thedetection and enumeration of microorganisms on the sur-face of carcasses or part of carcasses of slaughtered meat wasused for the sampling procedure 10 samples of 500 g eachwere collected in the morning (between 930 and 1030 AM)from butcher shops in each market using a systematicrandom sampling technique Briefly in eachmarket butchershops were numbered and the corresponding number wasnoted on a paper e papers bearing each number werefolded introduced into a closed container and thoroughlymixed One paper was selected from the container and itsnumber was recorded e paper was reintroduced into thecontainer and the container was thoroughly mixed again Inthe case that the same number was drawn twice the seconddrawing was not considered and the number was returned tothe containere operation was performed until 10 butchershops were selected e collected samples were transferredinto a sterile bag After being labelled the samples werestored in an icebox and transported to the lab for analyses

23 Biosurfactant e biosurfactant used in this study waspreviously produced by Lactobacillus paracasei subsp tol-erans N2 with sugar cane molasses as substrate and char-acterized [25] Biosurfactant was dissolved in methanol andcentrifuged at 15000g 4degC for 10min e supernatant wasfiltered (022 μm) and evaporated to dryness under nitrogen

24 Chemicals Glacial acetic acid ethanol and methanol ofHPLC-grade were purchased from Sigma-Aldrich CoGermany Plate Count Agar (PCA) Eosin Methylene Bleu(EMB) agar Bile Azide Esculin (BEA) agar Mannitol SaltAgar (MSA) Mossel agar Cetrimide agar and Sabouraudagar supplemented with chloramphenicol Tryptone SulfiteNeomycin (TSN) agar Salmonella and Shigella (SS) agarSelenite cystine broth Xylose Lysine Decarboxylate (XLD)agar Triple Iron Sugar (TSI) agar Brain Heart Infusion(BHI) broth and Nutrient Agar (NA) were purchased fromScharlau (Germany) All other reagents used in this studywere of analytical grade

25 Cultures e following reference cultures E coliMTCC 118 Staphylococcus aureusMTCC 1430Micrococcusluteus MTCC 106 and Listeria monocytogenes MTCC 839were used for the antiadhesive test ese Microbial TypeCulture Collection (MTCC) strains were kindly provided bythe Department of Microbiology and Fermentation

2 Journal of Food Quality

Technology CSIR-Central Food Technological ResearchInstitute Mysore India

26 Microbiological Analysis of Fresh Beef Samples

261 Sample Preparation Samples were processedaccording to the method ISO 6887-22017 [26] In theprocedure 500 g of the different samples was weighed andground and 25 g was introduced into an Erlenmeyer con-taining 225mL of sterile peptone water e mixture washomogenized for 2min with a vortex (IKA Vortex Genius 3UK) and allowed to stand for 30min at room temperatureSerial decimal dilutions (10minus 1 to 10minus 6) were prepared

262 Inoculation and Culture Conditions e total mes-ophilic aerobic count was determined using the pour platemethod [27] Briefly 1mL of each dilution was introducedinto a sterile Petri dish followed with the addition of 20mLof sterile PCA e plates were incubated at 37degC for 48 hSpread plate method was used for the enumeration of totaland faecal coliforms [28] faecal Streptococci [29] Staphy-lococcus spp [30] Bacillus spp [31] Pseudomonas spp [32]yeasts and moulds [33] In the procedure 01mL of thedifferent dilutions was surface-inoculated on Petri platescontaining EMB agar BEA agar MSA agar Mossel agarCetrimide agar and Sabouraud agar supplemented withchloramphenicol respectively for total and faecal coliformsfaecal Streptococci Staphylococcus spp Bacillus spp Pseu-domonas spp and yeasts and moulds e plates wereincubated at 37degC for 48 h for total coliforms faecal Strep-tococci Staphylococcus spp Bacillus spp and Pseudomonasspp at 44degC for 48 h for faecal coliforms and at 25degC for 3ndash5days for yeasts and moulds

e presence of Salmonella spp in samples was assessedaccording to the method ISO 6579-12017 [34] After ho-mogenization of 25 g of sample in 225mL of sterile peptonewater the suspension was incubated for 16 h at 37degC forpreenrichment en 1mL of the suspension was transferredinto a tube containing 10mL of sterile selenite cystine brothand incubated for 24 h at 37degC for enrichmentereafter oneloopful of each enrichment broth was streaked onto SS andXLD agar and incubated at 37degC for 24 h Uncolored colonieswith black centers on SS agar and red colonies with blackcenters on XLD agar were considered as Salmonella

e enumeration of sulphite-reducing Clostridium wasdone according to the method ISO 79372004 [35] 2mL ofdilution (10minus 2) was introduced into a tube containing 15mLof sterile TSN agar homogenized and heated at 80degC for10min en the tubes were rapidly cooled and 1mL ofsterile paraffin oil was introduced into each tube followedwith incubation at 37degC for 48 h After incubation blackcolonies were considered as sulphite-reducing Clostridium

263 Plates Reading e colony-forming units (CFU)appearing on the Petri dishes after the incubation periodwere counted Only plates with colony-forming units be-tween 30 and 300 were considered All experiments were

performed with three replications and the results wereexpressed as colony-forming units per gram of fresh beef(CFUg)

27 Identification of Microorganisms Isolated from Fresh BeefSamples From each Petri plate individual colonies werestreaked twice on nutrient agar and characterized throughmacroscopic (colony shape and color) microscopic (Gramstaining) cultural (growth at different temperatures andNaCl concentrations) and standard biochemical tests(catalase motility coagulase oxidase urease citrate utili-zation indole gelatin hydrolysis and TSI test) according toBergeyrsquosManual of Determinative Bacteriology [36] Furthercharacterization of the isolates was performed through theirAnalytical Profile Index (API) e following galleries wereused according to the manufacturerrsquos instructions API 20 EAPI 20 non-E API 20 Strep API 20 Staph and API 20 AUX(BioMerieux SA France) e specificity of these gallerieswas at least 92 e results were recorded and the iden-tification process was performed with Apident 20 (Bio-Merieux France) Identity of isolates was confirmed usingonline API web services (httpsapiwebbiomerieuxcom)and Bergeyrsquos Manual of Determinative Bacteriology forcomparison of fermentation profiles

28 Antiadhesive Activity of Biosurfactants e antiadhesiveactivity of biosurfactant was assessed against reference culturesand selected cultures isolated from fresh beef following themethod of Gudintildea et al [37] with slight modifications In theprocedure the different strains were cultured in BHI broth for16 h at 37degC e culture was centrifuged (10000g 10min4degC) and cells were washed three times with phosphate-buffered saline (PBS pH 72) Cells were suspended in PBS andtheir concentrations were adjusted to 108CFUmL Solutionsof biosurfactant at concentrations ranging from 001 to 10mgmL were prepared in millipore water For the test 200μL ofeach solution of biosurfactant was introduced into the wells ofa sterile 96-well flat-bottomed plastic tissue culture plate(Tarsons India) and the plates were incubated for 1 h at 4degCAfterwards the plates were washed twice with PBS and 200μLof the bacterial suspension was added to each well followedwith incubation at 4degC for 24 h en the plates were washedtwice with PBS and adherent cells were fixed with 200μL ofmethanol 99 After incubation at room temperature for15min methanol was removed and plates were air-driedAdherent cells were stained for 5min with 200μL of 33crystal violete plates were washed three times with PBS andair-dried and 200μL of 33 (vv) glacial acetic acid was in-troduced in each well to resolubilize adherent cellse opticaldensity of the plates was read at 595nm using a Sparkreg 10MMultimode Microplate Reader (Tecan Switzerland) Wellsfilled with PBS without biosurfactants were used as controle inhibition of microbial adhesion was calculated using

Microbial inhibition() A0 minus Ai

A01113888 1113889 times 100 (1)

Journal of Food Quality 3

Technology CSIR-Central Food Technological ResearchInstitute Mysore India

26 Microbiological Analysis of Fresh Beef Samples

261 Sample Preparation Samples were processedaccording to the method ISO 6887-22017 [26] In theprocedure 500 g of the different samples was weighed andground and 25 g was introduced into an Erlenmeyer con-taining 225mL of sterile peptone water e mixture washomogenized for 2min with a vortex (IKA Vortex Genius 3UK) and allowed to stand for 30min at room temperatureSerial decimal dilutions (10minus 1 to 10minus 6) were prepared

262 Inoculation and Culture Conditions e total mes-ophilic aerobic count was determined using the pour platemethod [27] Briefly 1mL of each dilution was introducedinto a sterile Petri dish followed with the addition of 20mLof sterile PCA e plates were incubated at 37degC for 48 hSpread plate method was used for the enumeration of totaland faecal coliforms [28] faecal Streptococci [29] Staphy-lococcus spp [30] Bacillus spp [31] Pseudomonas spp [32]yeasts and moulds [33] In the procedure 01mL of thedifferent dilutions was surface-inoculated on Petri platescontaining EMB agar BEA agar MSA agar Mossel agarCetrimide agar and Sabouraud agar supplemented withchloramphenicol respectively for total and faecal coliformsfaecal Streptococci Staphylococcus spp Bacillus spp Pseu-domonas spp and yeasts and moulds e plates wereincubated at 37degC for 48 h for total coliforms faecal Strep-tococci Staphylococcus spp Bacillus spp and Pseudomonasspp at 44degC for 48 h for faecal coliforms and at 25degC for 3ndash5days for yeasts and moulds

e presence of Salmonella spp in samples was assessedaccording to the method ISO 6579-12017 [34] After ho-mogenization of 25 g of sample in 225mL of sterile peptonewater the suspension was incubated for 16 h at 37degC forpreenrichment en 1mL of the suspension was transferredinto a tube containing 10mL of sterile selenite cystine brothand incubated for 24 h at 37degC for enrichmentereafter oneloopful of each enrichment broth was streaked onto SS andXLD agar and incubated at 37degC for 24 h Uncolored colonieswith black centers on SS agar and red colonies with blackcenters on XLD agar were considered as Salmonella

e enumeration of sulphite-reducing Clostridium wasdone according to the method ISO 79372004 [35] 2mL ofdilution (10minus 2) was introduced into a tube containing 15mLof sterile TSN agar homogenized and heated at 80degC for10min en the tubes were rapidly cooled and 1mL ofsterile paraffin oil was introduced into each tube followedwith incubation at 37degC for 48 h After incubation blackcolonies were considered as sulphite-reducing Clostridium

263 Plates Reading e colony-forming units (CFU)appearing on the Petri dishes after the incubation periodwere counted Only plates with colony-forming units be-tween 30 and 300 were considered All experiments were

performed with three replications and the results wereexpressed as colony-forming units per gram of fresh beef(CFUg)

27 Identification of Microorganisms Isolated from Fresh BeefSamples From each Petri plate individual colonies werestreaked twice on nutrient agar and characterized throughmacroscopic (colony shape and color) microscopic (Gramstaining) cultural (growth at different temperatures andNaCl concentrations) and standard biochemical tests(catalase motility coagulase oxidase urease citrate utili-zation indole gelatin hydrolysis and TSI test) according toBergeyrsquosManual of Determinative Bacteriology [36] Furthercharacterization of the isolates was performed through theirAnalytical Profile Index (API) e following galleries wereused according to the manufacturerrsquos instructions API 20 EAPI 20 non-E API 20 Strep API 20 Staph and API 20 AUX(BioMerieux SA France) e specificity of these gallerieswas at least 92 e results were recorded and the iden-tification process was performed with Apident 20 (Bio-Merieux France) Identity of isolates was confirmed usingonline API web services (httpsapiwebbiomerieuxcom)and Bergeyrsquos Manual of Determinative Bacteriology forcomparison of fermentation profiles

28 Antiadhesive Activity of Biosurfactants e antiadhesiveactivity of biosurfactant was assessed against reference culturesand selected cultures isolated from fresh beef following themethod of Gudintildea et al [37] with slight modifications In theprocedure the different strains were cultured in BHI broth for16 h at 37degC e culture was centrifuged (10000g 10min4degC) and cells were washed three times with phosphate-buffered saline (PBS pH 72) Cells were suspended in PBS andtheir concentrations were adjusted to 108CFUmL Solutionsof biosurfactant at concentrations ranging from 001 to 10mgmL were prepared in millipore water For the test 200μL ofeach solution of biosurfactant was introduced into the wells ofa sterile 96-well flat-bottomed plastic tissue culture plate(Tarsons India) and the plates were incubated for 1 h at 4degCAfterwards the plates were washed twice with PBS and 200μLof the bacterial suspension was added to each well followedwith incubation at 4degC for 24 h en the plates were washedtwice with PBS and adherent cells were fixed with 200μL ofmethanol 99 After incubation at room temperature for15min methanol was removed and plates were air-driedAdherent cells were stained for 5min with 200μL of 33crystal violete plates were washed three times with PBS andair-dried and 200μL of 33 (vv) glacial acetic acid was in-troduced in each well to resolubilize adherent cellse opticaldensity of the plates was read at 595nm using a Sparkreg 10MMultimode Microplate Reader (Tecan Switzerland) Wellsfilled with PBS without biosurfactants were used as controle inhibition of microbial adhesion was calculated using

Microbial inhibition() A0 minus Ai

A01113888 1113889 times 100 (1)

Journal of Food Quality 3

where A0 is the absorbance of the control and Ai is theabsorbance of the sample at biosurfactant concentration iFor each tested microorganism and biosurfactant concen-tration essays were performed in triplicate

29 Statistical Analysis All experiments were carried out intriplicate Microbial loads were transformed into logarithmsand expressed as meansplusmn standard deviation DuncanrsquosMultiple Range test was performed to compare microbialloads of samples with the microbiological criteria of theEuropean Commission [38] and to compare the anti-adhesive activity of biosurfactant against the tested micro-organisms e statistical significance was set at plt 005 andStatgraphics centurion XVI version 16118 (StatPointTechnologies Inc USA) was used to perform these analysesPrincipal component analysis was done with XLSTAT 2018(Addinsoft Inc New York USA) in order to visualize theassociation between the microbial strains identified and thesampling sites

3 Results and Discussion

31Microbiological Contamination of Samples Total aerobiccount shows the wide variability of microorganisms whichcan be found in a product As observed in Table 1 the totalaerobic count of fresh beef samples ranges significantly(plt 005) from 538plusmn 001 (GM) to 593plusmn 0022 LogCFUg(PM) e high level of contamination of fresh beef sampleswas also reported by Obeng et al [39] and Goja et al [15] intheir studies ey respectively found a total aerobic countranging from 482 to 692 LogCFUg of fresh beef sold inGhanaian markets and from 467 to 553 LogCFUg of freshbeef sold in Sudanese markets Zerabruk et al [1] notified intheir study a total aerobic count of fresh beef sold in butchershops of Addis Ababa Ethiopia which ranged from 5 to712 LogCFUg Compared to the microbiological criteria[38] all samples analysed in this study presented a totalaerobic count lower than the recommended values(7 LogCFUg) However it does not mean that there is nopathogen because it is not necessary to have a high con-centration of the total aerobic flora to have safety issuesAccording to Emele et al [40] the total aerobic count of afood product does not necessarily determine its sanitaryquality rather it is the range of its different group of mi-croorganisms which determined if the product is safe or note safety criteria of fresh beef placed on the market duringtheir shelf life deal with the presence of Salmonella in thesamples analysed [38] Another important criterion for freshbeef placed on themarket during their shelf life is the processhygiene criteria It concerns the presence of microorganismslike Enterobacteria and E coli in the samples analysed [38]In this light pathogenic microorganisms were sought

Faecal Streptococci which are indicative of faecal con-tamination were found in all samples at loads ranging from433plusmn 002 (PM) to 511plusmn 014 LogCFUg (MBT) e co-liforms another pathogenic group of microorganisms whosepresence in raw meat is indicative of faecal contaminationwere assessed e results (as shown in Table 1) show that

total and faecal coliforms were present in the fresh beefsamples at loads higher than the values specified by themicrobiological criteria (3 and 27 LogCFUg resp) of theEuropean Commission [38] e highest level of contami-nation was observed at the site CC for both total(593plusmn 004 LogCFUg) and faecal (538plusmn 001 LogCFUg)coliforms e presence of coliforms in the fresh beefsamples could result from the nonrespect of good slaugh-tering and evisceration practicese unwholesome cleaningprocess of equipment and cutting meat surfaces could alsoexplain the high contamination Moreover the presence ofthese Enterobacteria in fresh beef samples could originatefrom the deplorable levels of hygiene and sanitary practicesof butchers as observed during this study e nonrespect ofhygiene during the production distribution and selling ofmeat was notified in the literature as the source of coliformsrsquocontamination [39] Afnabi et al [19] have justified thepresence of coliforms in beef sold in Northern Cameroon bythe poor application of the principles of cleaning and dis-infection and the lack of supervision of staff on hygiene

According to safety criteria of the European Commis-sion Regulation [38] Salmonella must be absent in 25 g offresh beef intended to human consumption HoweverSalmonella spp were present in all samples analysedprobably as the result of improper sanitary practices ofbutchers Hence the meat was consequently unsafe forconsumption Staphylococci the germs which indicate thelevel of hygiene of fresh beef handlers were found in allsamples at loads which significantly (plt 005) vary from onesite to another e high level of Staphylococcus spp wasobserved in the site MD (497plusmn 003 LogCFUg) while theleast level was noticed in the site MBT (430plusmn 013 LogCFUg) Poor hygienic practices of meat handlers duringslaughtering evisceration cutting and selling (such ascoughing and sneezing) could explain the presence ofStaphylococcus spp in the analysed samples at level higherthan the 3 LogCFUg recommended by the EuropeanCommission Regulation [38] A high contamination level(457plusmn 087 LogCFUg) of fresh beef sold in butcher shopsof Addis Ababa Ethiopia was also noticed by Zerabruk et al[1]

However despite the poor sanitary quality of fresh beefobserved in this study sulphite-reducing Clostridium wasnot found in any sample is result was very surprisingconsidering the fact that slaughtering evisceration dressingand cleaning of the carcasses are carried out on the groundRegarding Pseudomonas spp they were present in allsamples at loads which significantly (plt 005) vary from475plusmn 006 (CC) to 505plusmn 003 LogCFUg (MD) eirpresence in the fresh beef samples is indicative of a high riskof spoilage In fact bacteria belonging to Pseudomonas genusare generally recognized to actively contribute to fresh meatspoilage owing to their ability to degrade sugars lipids andproteins even at refrigeration temperature [3 41] As ob-served in Table 1 Bacillus spp were found in all samples witha contamination level which significantly varies from onesite to another e high contamination level was observedin the site MBT (543plusmn 007 LogCFUg) Yeasts and mouldswere enumerated in the different samples and results

4 Journal of Food Quality

obtained show that yeasts were present in these samples withloads which range from 405plusmn 002 (GM) to467plusmn 011 LogCFUg (CC) Mouldsrsquo count varies from146plusmn 005 (GM) to 229plusmn 008 LogCFUg (MD) epresence of these microorganisms in fresh beef samplescould result from the contamination during slaughteringdressing and evisceration processes Moreover air exposi-tion of meat during selling could lead to meat contaminationby mould spores

32 Identification of Microorganisms Isolated from Fresh BeefSamples e strains isolated from the different fresh beefsamples were purified and identified by their macroscopicmicroscopic cultural and biochemical properties e re-sults obtained show that all the 151 strains isolated belongedto five genera Pseudomonas Staphylococcus SalmonellaEscherichia and Candida ese isolates were identified atspecies level using API galleries Table 2 presents theidentification percentages gathered from the fermentativeprofile of the different isolates on API 20 E API 20 NE API20 Staph and API 20 AUX respectively Twelve species (12)were identified with at least 97 of identity as followsPseudomonas putida (12 strains) Pseudomonas aeruginosa(13 strains) Pseudomonas sp (26 strains) Escherichia coli 1(5 strains) Escherichia coli (14 strains) Salmonella enteritidis(9 strains) Salmonella sp (15 strains) Staphylococcus epi-dermidis (8 strains) Staphylococcus xylosus (12 strains)Staphylococcus aureus (18 strains) Candida albicans (5strains) and Candida sp (14 strains)

e different isolates identified in this study were groupedby genus species and sampling sites (Table 2) e highestmicrobial diversity (10 species) was observed in the samplingsites PM and MBG while the lowest diversity was noticed inthe site GM (6 species) Accordingly the high proportion ofstrains (2052 and 2582) was isolated from fresh beef soldin the sites MBG and PM respectively 1523 1258 1788 and794 of strains were respectively obtained from fresh beefsold in the sites CC MD MBT and GM

For the genus Pseudomonas 51 strains belonging to threespecies P putida (12) P aeruginosa (13) and Pseudomonas

sp (26) were isolated (Table 2) e proportion of strainsisolated varies with the species identified and the samplingsites e high proportions of P putida (3333) andPseudomonas sp (2307) were noticed in the site CC andthat of P aeruginosa (3846) was observed in the site MBGe high proportions of strains belonging to the genusPseudomonas observed in this study (3377) could bejustified by the fact that they are versatile bacterium ubiq-uitous in nature and are mostly found in water and soilsDuring slaughtering evisceration and dressing processingwhich are generally carried out on the ground they couldeasily contaminate the meat Strains of Pseudomonas sp andP aeruginosa were also identified in fresh beef sold inCalabar metropolis Nigeria [14] and in Kenyan markets[42]

e second most important proportion of strains iso-lated in this study was those belonging to the genusStaphylococcus (2516) A total of 38 strains regrouped in 3species S xylosus (12) S epidermidis (8) and S aureus (18)were isolated (Table 2) e high proportion of S aureus(2777) and S epidermidis (50) was isolated from the siteMBT while S xylosus was isolated only in the sites MBG(6666) and PM (3333) It is important to highlight thatthe strain S xylosus is useful in meat processing products assausage due to its ability to degrade biogenic amines Saureus considered to be a well-known pathogen to humanswas the predominant strain of the genus Staphylococcuswith4736 Its predominance could result from contaminationthrough skin mouth hand and noses of butchers as thisstrain colonized at 40 to 60 the nasal mucosa and skin ofhealthy human being S xylosus S epidermidis and Saureus were isolated by Goja et al [15] in fresh beef sold inmarkets of Khartoum Omdurman and Bahri cities Sudan

In this study 19 strains of E coli belonging to the speciesE coli (14) and E coli 1 (5) were isolated (Table 2) ehighest proportion of E coli strains (2857) was observedin the sites CC MBG and PM Strains of E coli 1 werenoticed only in the sites MD (40) and MBT (60) eresults obtained in the present study corroborate with thosementioned by Balcha et al [43] who found on fresh beef soldin Mekelle Quiha and Wukro markets of Ethiopia strains

Table 1 Mean microbial loads in LogCFUg of fresh beef samples collected from different markets of the city of Ngaoundere

Microbial types PM (n 10) GM (n 10) MBT (n 10) MBG (n 10) MD (n 10) CC (n 10) NormsTotal aerobic count 593plusmn 002d 538plusmn 001a 582plusmn 006c 584plusmn 001c 551plusmn 012b 580plusmn 013cd 700Total coliforms 581plusmn 001d 526plusmn 008b 502plusmn 002a 563plusmn 005c 529plusmn 001b 593plusmn 004e 300Faecal coliforms 520plusmn 001d 508plusmn 002b 495plusmn 003a 508plusmn 011abc 519plusmn 001c 538plusmn 001e 270Faecal Streptococci 433plusmn 002a 465plusmn 007b 511plusmn 014c 470plusmn 012b 439plusmn 008a 476plusmn 014b mdashStaphylococcus spp 465plusmn 002b 435plusmn 006a 430plusmn 013a 482plusmn 011cd 497plusmn 003d 487plusmn 003c 300Salmonella spp + + + + + + 0SR-Clostridium minus minus minus minus minus minus 0Bacillus spp 536plusmn 009d 526plusmn 005c 543plusmn 007e 509plusmn 001b 529plusmn 002c 448plusmn 002a mdashPseudomonas spp 481plusmn 002a 493plusmn 006c 482plusmn 010abc 487plusmn 003b 505plusmn 003d 475plusmn 006a 200Yeasts 431plusmn 001c 405plusmn 002a 420plusmn 010bc 426plusmn 004b 444plusmn 002d 467plusmn 011e mdashMoulds 176plusmn 001d 146plusmn 005a 157plusmn 003b 163plusmn 001c 229plusmn 008f 183plusmn 002e mdashnnumber of samples per site minus absence + presence PM ldquoPetit Marcherdquo GM ldquoGrand Marcherdquo MD ldquoDang marketrdquo CC ldquoCentre CommercialrdquoMBG ldquoBamnyanga marketrdquo MBT ldquoBantail marketrdquo SR-Clostridium sulphite-reducing Clostridium values with different letters within a line aresignificantly different (plt 005) according to Duncanrsquo multiple range test normsmicrobiological criteria of fresh beef established by the EuropeanCommission Regulation (EC 2005)

Journal of Food Quality 5

of E coli at proportions ranging from a market to anotherey also highlighted that some strains of E coli isolatedfrom fresh beef were identified as Enterohemorrhagic E coli(E coliO157 H7) Giving that some strains of E coli such asEnterohemorrhagic E coli (EHEC) are recognized as food-borne pathogens associated with several outbreaks of dis-eases [44] its presence in fresh beef as observed in this studyis worrisome Molecular identification of the strains isolatedis required in order to bring more information about theirpathogenicity

e main reservoir of zoonotic Salmonella is food fromanimal origin and the main sources of infections incrimi-nated Salmonella in industrialized and developing countriesare animal-derived products notably fresh meat products[45] In this study 24 strains of Salmonella were isolatedamong which there were 3750 of S enterica serovarEnteritidis and 6250 of Salmonella sp (Table 2) RegardingS enterica serovar Enteritidis it was found only in sites MDMBG and PM at the proportion of 3333 Salmonella spwas absent in the site MD and present in the other sites SitesMBG and PM rank first with 2666 each Ukut et al [14]also identified Salmonella spp in fresh beef sold in Calabarmarket Nigeria In another study conducted by Obeng et al[39] Salmonella spp were isolated and identified in freshmeat sold in Tolon and Kumbungu districts of the NorthernRegion of Ghana

For yeasts belonging to the genus Candida Table 2 showsthat 19 strains were isolated and identified as the species Calbicans (5 strains) and Candida sp (14 strains) Strains of Calbicans were found only in sites MBG MBT and PM atproportions of 40 20 and 40 respectively Strains ofCandida sp were identified in fresh beef samples collectedfrom the different sites and the site GM ranks first with 2857of strainse results of this study are in accordance with thosereported byComi andCantoni [46] who showed that strains ofCandida spp are dominant yeasts of fresh beef

In order to visualize the association between the mi-crobial species identified in this study and the samplingsites a principal component analysis was carried outFigure 1 presents the distribution of microbial speciesisolated from fresh beef and sampling sites on the axissystem F1 times F2 As seen in Figure 1 three main groupswere formed e first group shows that the strains be-longing to the species C albicans Salmonella sp Paeruginosa P putida S xylosus and S enteritidis areassociated with the sites PM and MBG e second groupshows that the strains belonging to the species Pseudo-monas sp E coli 1 S epidermis and S aureus are as-sociated with site MBT Finally the third group shows thatthe strains belonging to the species Candida sp are mainlyassociated with the sites MD CC and GM is principalcomponent analysis pointed out the specific relationship

Table 2 Genus species identification percentages and proportions of strains isolated from fresh beef samples collected from differentmarkets of the city of Ngaoundere

Genus Species ID percentages()

Number ofstrains

Numbers and proportions of strains per sampling sitesCC MD MBG MBT GM PM

Pseudomonas

Pseudomonasputida 9850 12 4

(3333)2

(3333)4

(1666)0

(0)0

(0)2

(1666)Pseudomonasaeruginosa 9980 13 2

(1538)3

(2307)5

(3846)0

(0) 1 (769) 2(1538)

Pseudomonas sp 9740 26 6(2307)

3(1153)

3(1153)

8(3076) 2 (769) 4

(1538)

EscherichiaEscherichia coli 9960 14 4

(2857)0

(0)4

(2857)0

(0)2

(1428)4

(2857)

Escherichia coli 1 9940 5 0(0)

2(4000)

0(0)

3(6000)

0(0)

0(0)

Salmonella

S enterica serEnteritidis 9860 9 0

(0)3

(3333)3

(3333)0

(0)0

(0)3

(3333)

Salmonella sp 9910 15 2(1333)

0(0)

4(2666)

3(2000)

2(1333)

4(2666)

Staphylococcus

Staphylococcusaureus 9860 18 2

(1111)4

(2222)3

(1666)5

(2777) 1 (555) 3(1666)

Staphylococcusxylosus 10000 12 0

(0)0

(0)8

(6666)0

(0)0

(0)4

(3333)Staphylococcus

epidermis 9830 8 0(0)

0(0)

2(2500)

4(5000)

0(0)

2(2500)

CandidaCandida albicans 9860 5 0

(0)0

(0)2

(4000)1

(2000)0

(0)2

(4000)

Candida sp 9830 14 3(2142)

2(1428)

1(714)

3(2142)

4(2857)

1(714)

Total 151 23(1523)

19(1258)

39(2582)

27(1788)

12(794)

31(2052)

ID identification percentages of the strains from API data ser serovar PM ldquoPetit Marcherdquo GM ldquoGrandMarcherdquo MD ldquoDang marketrdquo CC ldquoCentreCommercialrdquo MBG ldquoBamnyanga marketrdquo MBT ldquoBantail marketrdquo

6 Journal of Food Quality

between the microbial diversity of fresh beef and thesampling sites It enables to observe associations betweenthe sampling sites and the microbial profile of fresh beefsold in Ngaoundere e use of 16S rRNA gene sequencingand a high number of meat samples will improve this PCArepresentation through an increase of the microbialdiversity

33 Antiadhesive Activity of Biosurfactant Food spoilageand disease transmission due to microorganisms attachedinto surfaces are one of the main problems faced by foodindustries particularly in the meat industry In this studythe antiadhesive property of biosurfactant was tested againstsome selected bacterial and fungal strains isolated from freshbeef and against some reference cultures e biosurfactantsderived from L paracasei subsp tolerans N2 inhibited theadhesion of all the tested strains in a dose-dependentmanner (Table 3) Biosurfactant was active even at 001against all the tested microorganisms At that concentration3396plusmn 236 and 2770plusmn 075 of inhibition were recordedagainst S aureusMTCC 1430 and S aureus STP1 Completeinhibition (100) of microbial adhesion was observedagainst Bacillus sp BC1 S aureus STP1 and S xylosus STP2at biosurfactant concentration of 10mgmL e leastantiadhesive activity (5612plusmn 016) of biosurfactant wasnoticed against the yeast strain C albicans LV1 is anti-adhesive activity of biosurfactants observed in this studycould be due to their ability to change the hydrophobicity ofthe interface between the strain and the surface of thepolystyrene plate Hence microbial adhesion to the surfacewill decrease as biosurfactant concentration increasesSharma and Saharan [21] have explained the antiadhesiveactivity of biosurfactants of L helveticus by their ability toform a thin film which modifies the wettability of the surfaceand thus decreases the adhesive properties of microorgan-isms e inhibition percentages observed in Table 3 are notdifferent from the values reported byMerghni et al [22] with

biosurfactants from L casei ATCC 393 LZ9 and L caseiATCC 393 LBl ey noticed inhibition percentages rangingfrom 6884 to 8486 against S aureus 6538 S aureus 9Pand S aureus 29P at biosurfactant concentration of 125mgmL As shown in Table 3 Gram-negative bacteria were moreresistant to the antiadhesive activity of biosurfactant Asimilar observation was noticed by Sambanthamoorthy et al[47] with biosurfactants from L jensenii and L rhamnosusis resistance of Gram-negative bacteria could result intheir ability to produce once adhered to surfaces homo-serine lactone acyl (HLA) molecules which increase theirresistance to antiadhesive compounds [48]

Besides the adhesive properties of microorganismsisolated from fresh beef samples as shown in this studysuggest that cutting meat surfaces represent an ecologicalkennel of continuous contamination of meat Hence theantiadhesive activity of biosurfactant from L paracaseisubsp tolerans N2 against these strains shows that thesemolecules could be useful in the reduction of meat con-tamination through cutting surfaces

4 Conclusion

is study showed that fresh beef sold in the markets ofNgaoundere contained both pathogenic and spoilage mi-croorganisms at levels higher than the threshold valuesrecommended by the European Commission Twelve mi-crobial species colonized these fresh beef P putida Paeruginosa Pseudomonas sp E coli 1 E coli S enteritidisSalmonella sp S epidermidis S xylosus S aureus Calbicans and Candida sp with Pseudomonas sp as the mostabundant strains e presence of Salmonella in all samplesmakes no doubt on the safety status of the meat In the sameway the presence of microorganisms like P putida and Paeruginosa which can lead to a quick spoilage of fresh beefsuggests a reduced shelf life of the product Howeveridentification of these strains through 16S rRNA gene an-alyses needs to be done to confirm their identity e results

CC

MD

MBG

MBT

GM

PM

P putidaP aeruginosa

Pseudomonas sp

E coli

E coli 1

S enteritidis

Salmonella sp

S aureus

S xylosus

S epidermis

C albicans

Candida sp

ndash6

ndash4

ndash2

0

2

4

6

8

ndash10 ndash8 ndash6 ndash4 ndash2 0 2 4 6 8 10

F2 (2

876

)

F1 (5455)

Biplot (axis F1 amp F2 8331)

Figure 1 Distribution of microbial species isolated from fresh beef and sampling sites on F1times F2 axis PM ldquoPetit Marcherdquo GM ldquoGrandMarcherdquo MD ldquoDang marketrdquo CC ldquoCentre Commercialrdquo MBG ldquoBamnyanga marketrdquo MBT ldquoBantail marketrdquo

Journal of Food Quality 7

Tabl

e3

Microbial

inhibitio

npercentages(

)ob

tained

from

theantia

dhesiveassays

with

thebiosurfactantfrom

Lparacaseisub

sptoleransN2

Test

organism

sBiosurfactantc

oncentratio

n(m

gmL)

001

01

05

125

575

10S

aureus

MTC

C1430

3396plusmn236

aG4915plusmn248

bI6903plusmn006

cH7626plusmn038

dI8251plusmn143

eG9209plusmn023

fG9734plusmn109

gH9918plusmn010

hI

MluteusMTC

C106

2259plusmn259

aD4238plusmn218

bH6573plusmn147

cG7890plusmn333

dI8717plusmn308

eH9263plusmn041

fG9411plusmn051

gF9559plusmn071

hG

Lmon

ocytogenes

MTC

C839

2051plusmn119

aD3241plusmn009

bE4150plusmn043

cE5094plusmn019

dF5829plusmn148

eF6222plusmn024

fE6840plusmn155

gE7515plusmn053

hF

EcoliMTC

C118

1995plusmn032

aD3024plusmn069

bD3877plusmn240

cD4862plusmn194

dE5465plusmn146

eD6008plusmn016

fD6578plusmn048

gD7214plusmn033

hE

Paerugino

saPS

B21546plusmn075

aC2164plusmn225

bB3130plusmn100

cB4072plusmn010

dB4790plusmn095

eB5170plusmn055

fB5680plusmn015

gB6170plusmn055

hB

Ppu

tidaPS

J11310plusmn010

aB2315plusmn181

bB3071plusmn152

cB4030plusmn040

dB4695plusmn150

eB5271plusmn052

fB5722plusmn082

gB6071plusmn050

hB

SenteritidisSL

21445plusmn065

aC2680plusmn110

bC3473plusmn014

cC4210plusmn020

dC5040plusmn090

eC5821plusmn005

fC6120plusmn010

gC6610plusmn050

hC

EcoliE2

B1932plusmn090

aD2718plusmn045

bC3610plusmn040

cD4590plusmn015

dD5350plusmn035

eE5910plusmn075

fD6415plusmn150

gD7010plusmn045

hD

Calbicans

LV1

578plusmn035

aA1219plusmn230

bA2480plusmn211

cA3678plusmn191

dA4310plusmn066

eA4790plusmn025

fA5362plusmn031

gA5612plusmn016

hA

Bacillu

sspB

C1

2425plusmn090

aE4182plusmn114

bH6770plusmn067

cG7030plusmn050

dG8190plusmn022

eG9062plusmn020

fH9878plusmn050

gH10000plusmn000

hJ

Sau

reus

STP1

2770plusmn075

aF4001plusmn023

bH6617plusmn180

cG7338plusmn070

dH8221plusmn082

eG9330plusmn075

fI9925plusmn082

gH10000plusmn000

hJ

SxylosusST

P22132plusmn210

aD3841plusmn025

bG6570plusmn150

cG7313plusmn070

dH8192plusmn031

eG9240plusmn021

fI9816plusmn110

gH10000plusmn000

hJ

SepidermisST

P32180plusmn229

aD3711plusmn031

bF6020plusmn166

cF7176plusmn050

dG8068plusmn179

eG8818plusmn013

fF9594plusmn005

gG9870plusmn0050h

H

Negativec

ontrolsw

eresetat0

toindicatethea

bsence

ofbiosurfactantPo

sitivep

ercentagesindicatethereductio

nsinmicrobialadhesio

nwhencomparedto

thec

ontrolR

esultsaree

xpressed

asmeansplusmnstandard

deviationof

results

from

triplicateexperiments

8 Journal of Food Quality

of this study demonstrated the antiadhesive activity ofbiosurfactant against microbial strains isolated from freshbeef Efforts made by the Government to improve the mi-crobiological quality of fresh beef should deal not only withthe respect of good slaughtering evisceration and hygienicpractices but also with the cleaning process of surfaces wheremeat is cut and sold

Data Availability

e data used in this study are available from the corre-sponding author upon request

Conflicts of Interest

e authors declare no conflicts of interest

Acknowledgments

e authors thank the Director of the National School ofAgroindustrial Sciences of the University of NgaoundereCameroon and the Department of Microbiology and Fer-mentation Technology CSIR-Central Food TechnologicalResearch Institute of Mysore India for providing necessaryfacilities for the successful completion of this research worke authors gratefully acknowledge butchers of the differentmarkets of Ngaoundere for their kind cooperation duringthe sampling process

References

[1] K Zerabruk N Retta D Muleta and A T Tefera ldquoAs-sessment of microbiological safety and quality of minced meatand meat contact surfaces in selected butcher shops of AddisAbaba Ethiopiardquo Journal of Food Quality vol 2019 pp 1ndash92019

[2] A Holck L Axelsson A McLeod T M Rode and E HeirldquoHealth and safety considerations of fermented sausagesrdquoJournal of Food Quality vol 2017 Article ID 975389425 pages 2017

[3] G-J E Nychas P N Skandamis C C Tassou andK P Koutsoumanis ldquoMeat spoilage during distributionrdquoMeat Science vol 78 no 1-2 pp 77ndash89 2008

[4] G anigaivel and A S Anandhan ldquoIsolation and charac-terization of microorganisms from raw meat obtained fromdifferent market places in and around Chennairdquo Journal ofPharmaceutical Chemical and Biological Sciences vol 3 no 2pp 295ndash301 2015

[5] Centre for Disease Control (CDC) Multistate Outbreak ofSalmonella Enteritidis Infections Linked to Ground BeefCentre for Disease Control (CDC) Atlanta GA USA 2012

[6] J D Bailly H Brugere and H Chadron ldquoMicroorganismes etparasites des viandes les connaıtre pour les maıtriser delrsquoeleveur au consommateur CIVrdquo 2012 httpwwwciv-Viandeorg

[7] C Salifou K Boko G Ahounou et al ldquoDiversite de lamicroflore initiale de la viande et securite sanitaire desconsommateursrdquo International Journal of Biological andChemical Sciences vol 7 no 3 pp 1351ndash1369 2013

[8] S Datta A Akter I G Shah et al ldquoMicrobiological qualityassessment of raw meat and meat products and antibioticsusceptibility of isolated Staphylococcus aureusrdquo Agriculture

Food and Analytical Bacteriology vol 2 no 3 pp 187ndash1942012

[9] E Abayneh D Nolkes and B Asrade ldquoReview on commonfoodborne pathogens in Ethiopiardquo African Journal of Mi-crobiology Research vol 8 no 53 pp 4027ndash4040 2014

[10] T Kebede B Afera H Taddele and A Bsrat ldquoAssessment ofbacteriological quality of sold meat in the butcher shops ofAdigrat Tigray Ethiopiardquo Applied Journal of Hygiene vol 3no 3 pp 38ndash44 2014

[11] A J Ilboudo A Savadogo S Samandoulougou M AbreM Seydi and A Traore ldquoQualite bacteriologique des car-casses de viandes porcines et bovines produites a lrsquoabattoir deOuagadougou Burkina Fasordquo Revue Microbiologie Industri-elle Sanitaire et Environnementale vol 10 no 1 pp 33ndash352016

[12] N M Sachindra P Z Sakhare K P Yashoda andD Narasimha Rao ldquoMicrobial profile of buffalo sausageduring processing and storagerdquo Food Control vol 16 no 1pp 31ndash35 2005

[13] B Siriken ldquoe microbiological quality of ground beef inAydin and Afyon Provinces Turkeyrdquo Revue de MedecineVeterinaire vol 155 no 12 pp 632ndash636 2004

[14] I O E Ukut I O Okonko I S Ikpoh et al ldquoAssessment ofbacteriological quality of fresh meats sold in Calabar me-tropolis Nigeriardquo Electronic Journal of Environmental Ag-ricultural and Food Chemistry vol 9 no 1 pp 89ndash100 2010

[15] A M Goja T A A Ahmed S A M Saeed and H A DirarldquoIsolation and identification of Staphylococcus spp in freshbeefrdquo Pakistan Journal of Nutrition vol 12 no 2 pp 114ndash1202013

[16] P Antwi-Agyei and B K Maalekuu ldquoDetermination ofmicrobial contamination in meat and fish products sold in theKumasi metropolis (a case study of Kumasi central marketand the Bantama market)rdquo Merit Research Journal of Agri-cultural Science and Soil vol 2 pp 038ndash046 2014

[17] E Niyonzima D Bora and M P Ongol ldquoAssessment of beefmeat microbial contamination during skinning dressingtransportation and marketing at a commercial abattoir inKigali city Rwandardquo Pakistan Journal of Food Sciencesvol 23 pp 133ndash138 2013

[18] V K Singh J Udit J K Yadav and B Basanti ldquoAssessmentof bacterial quality of raw meat samples (carabeef chevonpork and poultry) from retail meat outlets and local slaughterhouses of Agra Region Indiardquo Journal of Foodborne andZoonotic Diseases vol 2 no 1 pp 15ndash18 2014

[19] R B Afnabi R P Nameni S S Kamdem J J Essia Ngangand R B Alambedji ldquoMicrobial load of beef sold in thetraditional slaughterhouse and butcher shops in northernCameroonrdquo International Journal of Veterinary Science vol 4no 4 pp 183ndash189 2015

[20] E J Gudintildea E C Fernandes J A Teixeira andL R Rodrigues ldquoAntimicrobial and anti-adhesive activities ofcell-bound biosurfactant from Lactobacillus agilisCCUG31450rdquo RSC Advances vol 5 no 110 p 90960 2015

[21] D Sharma and B S Saharan ldquoFunctional characterization ofbiomedical potential of biosurfactant produced by Lactoba-cillus helveticusrdquo Biotechnology Reports vol 11 pp 27ndash352016

[22] A Merghni I Dallel E Noumi et al ldquoAntioxidant andantiproliferative potential of biosurfactants isolated fromLactobacillus casei and their anti-biofilm effect in oralStaphylococcus aureus strainsrdquo Microbial Pathogenesisvol 104 pp 84ndash89 2017

Journal of Food Quality 9

obtained show that yeasts were present in these samples withloads which range from 405plusmn 002 (GM) to467plusmn 011 LogCFUg (CC) Mouldsrsquo count varies from146plusmn 005 (GM) to 229plusmn 008 LogCFUg (MD) epresence of these microorganisms in fresh beef samplescould result from the contamination during slaughteringdressing and evisceration processes Moreover air exposi-tion of meat during selling could lead to meat contaminationby mould spores

32 Identification of Microorganisms Isolated from Fresh BeefSamples e strains isolated from the different fresh beefsamples were purified and identified by their macroscopicmicroscopic cultural and biochemical properties e re-sults obtained show that all the 151 strains isolated belongedto five genera Pseudomonas Staphylococcus SalmonellaEscherichia and Candida ese isolates were identified atspecies level using API galleries Table 2 presents theidentification percentages gathered from the fermentativeprofile of the different isolates on API 20 E API 20 NE API20 Staph and API 20 AUX respectively Twelve species (12)were identified with at least 97 of identity as followsPseudomonas putida (12 strains) Pseudomonas aeruginosa(13 strains) Pseudomonas sp (26 strains) Escherichia coli 1(5 strains) Escherichia coli (14 strains) Salmonella enteritidis(9 strains) Salmonella sp (15 strains) Staphylococcus epi-dermidis (8 strains) Staphylococcus xylosus (12 strains)Staphylococcus aureus (18 strains) Candida albicans (5strains) and Candida sp (14 strains)

e different isolates identified in this study were groupedby genus species and sampling sites (Table 2) e highestmicrobial diversity (10 species) was observed in the samplingsites PM and MBG while the lowest diversity was noticed inthe site GM (6 species) Accordingly the high proportion ofstrains (2052 and 2582) was isolated from fresh beef soldin the sites MBG and PM respectively 1523 1258 1788 and794 of strains were respectively obtained from fresh beefsold in the sites CC MD MBT and GM

For the genus Pseudomonas 51 strains belonging to threespecies P putida (12) P aeruginosa (13) and Pseudomonas

sp (26) were isolated (Table 2) e proportion of strainsisolated varies with the species identified and the samplingsites e high proportions of P putida (3333) andPseudomonas sp (2307) were noticed in the site CC andthat of P aeruginosa (3846) was observed in the site MBGe high proportions of strains belonging to the genusPseudomonas observed in this study (3377) could bejustified by the fact that they are versatile bacterium ubiq-uitous in nature and are mostly found in water and soilsDuring slaughtering evisceration and dressing processingwhich are generally carried out on the ground they couldeasily contaminate the meat Strains of Pseudomonas sp andP aeruginosa were also identified in fresh beef sold inCalabar metropolis Nigeria [14] and in Kenyan markets[42]

e second most important proportion of strains iso-lated in this study was those belonging to the genusStaphylococcus (2516) A total of 38 strains regrouped in 3species S xylosus (12) S epidermidis (8) and S aureus (18)were isolated (Table 2) e high proportion of S aureus(2777) and S epidermidis (50) was isolated from the siteMBT while S xylosus was isolated only in the sites MBG(6666) and PM (3333) It is important to highlight thatthe strain S xylosus is useful in meat processing products assausage due to its ability to degrade biogenic amines Saureus considered to be a well-known pathogen to humanswas the predominant strain of the genus Staphylococcuswith4736 Its predominance could result from contaminationthrough skin mouth hand and noses of butchers as thisstrain colonized at 40 to 60 the nasal mucosa and skin ofhealthy human being S xylosus S epidermidis and Saureus were isolated by Goja et al [15] in fresh beef sold inmarkets of Khartoum Omdurman and Bahri cities Sudan

In this study 19 strains of E coli belonging to the speciesE coli (14) and E coli 1 (5) were isolated (Table 2) ehighest proportion of E coli strains (2857) was observedin the sites CC MBG and PM Strains of E coli 1 werenoticed only in the sites MD (40) and MBT (60) eresults obtained in the present study corroborate with thosementioned by Balcha et al [43] who found on fresh beef soldin Mekelle Quiha and Wukro markets of Ethiopia strains

Table 1 Mean microbial loads in LogCFUg of fresh beef samples collected from different markets of the city of Ngaoundere

Microbial types PM (n 10) GM (n 10) MBT (n 10) MBG (n 10) MD (n 10) CC (n 10) NormsTotal aerobic count 593plusmn 002d 538plusmn 001a 582plusmn 006c 584plusmn 001c 551plusmn 012b 580plusmn 013cd 700Total coliforms 581plusmn 001d 526plusmn 008b 502plusmn 002a 563plusmn 005c 529plusmn 001b 593plusmn 004e 300Faecal coliforms 520plusmn 001d 508plusmn 002b 495plusmn 003a 508plusmn 011abc 519plusmn 001c 538plusmn 001e 270Faecal Streptococci 433plusmn 002a 465plusmn 007b 511plusmn 014c 470plusmn 012b 439plusmn 008a 476plusmn 014b mdashStaphylococcus spp 465plusmn 002b 435plusmn 006a 430plusmn 013a 482plusmn 011cd 497plusmn 003d 487plusmn 003c 300Salmonella spp + + + + + + 0SR-Clostridium minus minus minus minus minus minus 0Bacillus spp 536plusmn 009d 526plusmn 005c 543plusmn 007e 509plusmn 001b 529plusmn 002c 448plusmn 002a mdashPseudomonas spp 481plusmn 002a 493plusmn 006c 482plusmn 010abc 487plusmn 003b 505plusmn 003d 475plusmn 006a 200Yeasts 431plusmn 001c 405plusmn 002a 420plusmn 010bc 426plusmn 004b 444plusmn 002d 467plusmn 011e mdashMoulds 176plusmn 001d 146plusmn 005a 157plusmn 003b 163plusmn 001c 229plusmn 008f 183plusmn 002e mdashnnumber of samples per site minus absence + presence PM ldquoPetit Marcherdquo GM ldquoGrand Marcherdquo MD ldquoDang marketrdquo CC ldquoCentre CommercialrdquoMBG ldquoBamnyanga marketrdquo MBT ldquoBantail marketrdquo SR-Clostridium sulphite-reducing Clostridium values with different letters within a line aresignificantly different (plt 005) according to Duncanrsquo multiple range test normsmicrobiological criteria of fresh beef established by the EuropeanCommission Regulation (EC 2005)

Journal of Food Quality 5

of E coli at proportions ranging from a market to anotherey also highlighted that some strains of E coli isolatedfrom fresh beef were identified as Enterohemorrhagic E coli(E coliO157 H7) Giving that some strains of E coli such asEnterohemorrhagic E coli (EHEC) are recognized as food-borne pathogens associated with several outbreaks of dis-eases [44] its presence in fresh beef as observed in this studyis worrisome Molecular identification of the strains isolatedis required in order to bring more information about theirpathogenicity

e main reservoir of zoonotic Salmonella is food fromanimal origin and the main sources of infections incrimi-nated Salmonella in industrialized and developing countriesare animal-derived products notably fresh meat products[45] In this study 24 strains of Salmonella were isolatedamong which there were 3750 of S enterica serovarEnteritidis and 6250 of Salmonella sp (Table 2) RegardingS enterica serovar Enteritidis it was found only in sites MDMBG and PM at the proportion of 3333 Salmonella spwas absent in the site MD and present in the other sites SitesMBG and PM rank first with 2666 each Ukut et al [14]also identified Salmonella spp in fresh beef sold in Calabarmarket Nigeria In another study conducted by Obeng et al[39] Salmonella spp were isolated and identified in freshmeat sold in Tolon and Kumbungu districts of the NorthernRegion of Ghana

For yeasts belonging to the genus Candida Table 2 showsthat 19 strains were isolated and identified as the species Calbicans (5 strains) and Candida sp (14 strains) Strains of Calbicans were found only in sites MBG MBT and PM atproportions of 40 20 and 40 respectively Strains ofCandida sp were identified in fresh beef samples collectedfrom the different sites and the site GM ranks first with 2857of strainse results of this study are in accordance with thosereported byComi andCantoni [46] who showed that strains ofCandida spp are dominant yeasts of fresh beef

In order to visualize the association between the mi-crobial species identified in this study and the samplingsites a principal component analysis was carried outFigure 1 presents the distribution of microbial speciesisolated from fresh beef and sampling sites on the axissystem F1 times F2 As seen in Figure 1 three main groupswere formed e first group shows that the strains be-longing to the species C albicans Salmonella sp Paeruginosa P putida S xylosus and S enteritidis areassociated with the sites PM and MBG e second groupshows that the strains belonging to the species Pseudo-monas sp E coli 1 S epidermis and S aureus are as-sociated with site MBT Finally the third group shows thatthe strains belonging to the species Candida sp are mainlyassociated with the sites MD CC and GM is principalcomponent analysis pointed out the specific relationship

Table 2 Genus species identification percentages and proportions of strains isolated from fresh beef samples collected from differentmarkets of the city of Ngaoundere

Genus Species ID percentages()

Number ofstrains

Numbers and proportions of strains per sampling sitesCC MD MBG MBT GM PM

Pseudomonas

Pseudomonasputida 9850 12 4

(3333)2

(3333)4

(1666)0

(0)0

(0)2

(1666)Pseudomonasaeruginosa 9980 13 2

(1538)3

(2307)5

(3846)0

(0) 1 (769) 2(1538)

Pseudomonas sp 9740 26 6(2307)

3(1153)

3(1153)

8(3076) 2 (769) 4

(1538)

EscherichiaEscherichia coli 9960 14 4

(2857)0

(0)4

(2857)0

(0)2

(1428)4

(2857)

Escherichia coli 1 9940 5 0(0)

2(4000)

0(0)

3(6000)

0(0)

0(0)

Salmonella

S enterica serEnteritidis 9860 9 0

(0)3

(3333)3

(3333)0

(0)0

(0)3

(3333)

Salmonella sp 9910 15 2(1333)

0(0)

4(2666)

3(2000)

2(1333)

4(2666)

Staphylococcus

Staphylococcusaureus 9860 18 2

(1111)4

(2222)3

(1666)5

(2777) 1 (555) 3(1666)

Staphylococcusxylosus 10000 12 0

(0)0

(0)8

(6666)0

(0)0

(0)4

(3333)Staphylococcus

epidermis 9830 8 0(0)

0(0)

2(2500)

4(5000)

0(0)

2(2500)

CandidaCandida albicans 9860 5 0

(0)0

(0)2

(4000)1

(2000)0

(0)2

(4000)

Candida sp 9830 14 3(2142)

2(1428)

1(714)

3(2142)

4(2857)

1(714)

Total 151 23(1523)

19(1258)

39(2582)

27(1788)

12(794)

31(2052)

ID identification percentages of the strains from API data ser serovar PM ldquoPetit Marcherdquo GM ldquoGrandMarcherdquo MD ldquoDang marketrdquo CC ldquoCentreCommercialrdquo MBG ldquoBamnyanga marketrdquo MBT ldquoBantail marketrdquo

6 Journal of Food Quality

between the microbial diversity of fresh beef and thesampling sites It enables to observe associations betweenthe sampling sites and the microbial profile of fresh beefsold in Ngaoundere e use of 16S rRNA gene sequencingand a high number of meat samples will improve this PCArepresentation through an increase of the microbialdiversity

33 Antiadhesive Activity of Biosurfactant Food spoilageand disease transmission due to microorganisms attachedinto surfaces are one of the main problems faced by foodindustries particularly in the meat industry In this studythe antiadhesive property of biosurfactant was tested againstsome selected bacterial and fungal strains isolated from freshbeef and against some reference cultures e biosurfactantsderived from L paracasei subsp tolerans N2 inhibited theadhesion of all the tested strains in a dose-dependentmanner (Table 3) Biosurfactant was active even at 001against all the tested microorganisms At that concentration3396plusmn 236 and 2770plusmn 075 of inhibition were recordedagainst S aureusMTCC 1430 and S aureus STP1 Completeinhibition (100) of microbial adhesion was observedagainst Bacillus sp BC1 S aureus STP1 and S xylosus STP2at biosurfactant concentration of 10mgmL e leastantiadhesive activity (5612plusmn 016) of biosurfactant wasnoticed against the yeast strain C albicans LV1 is anti-adhesive activity of biosurfactants observed in this studycould be due to their ability to change the hydrophobicity ofthe interface between the strain and the surface of thepolystyrene plate Hence microbial adhesion to the surfacewill decrease as biosurfactant concentration increasesSharma and Saharan [21] have explained the antiadhesiveactivity of biosurfactants of L helveticus by their ability toform a thin film which modifies the wettability of the surfaceand thus decreases the adhesive properties of microorgan-isms e inhibition percentages observed in Table 3 are notdifferent from the values reported byMerghni et al [22] with

biosurfactants from L casei ATCC 393 LZ9 and L caseiATCC 393 LBl ey noticed inhibition percentages rangingfrom 6884 to 8486 against S aureus 6538 S aureus 9Pand S aureus 29P at biosurfactant concentration of 125mgmL As shown in Table 3 Gram-negative bacteria were moreresistant to the antiadhesive activity of biosurfactant Asimilar observation was noticed by Sambanthamoorthy et al[47] with biosurfactants from L jensenii and L rhamnosusis resistance of Gram-negative bacteria could result intheir ability to produce once adhered to surfaces homo-serine lactone acyl (HLA) molecules which increase theirresistance to antiadhesive compounds [48]

Besides the adhesive properties of microorganismsisolated from fresh beef samples as shown in this studysuggest that cutting meat surfaces represent an ecologicalkennel of continuous contamination of meat Hence theantiadhesive activity of biosurfactant from L paracaseisubsp tolerans N2 against these strains shows that thesemolecules could be useful in the reduction of meat con-tamination through cutting surfaces

4 Conclusion

is study showed that fresh beef sold in the markets ofNgaoundere contained both pathogenic and spoilage mi-croorganisms at levels higher than the threshold valuesrecommended by the European Commission Twelve mi-crobial species colonized these fresh beef P putida Paeruginosa Pseudomonas sp E coli 1 E coli S enteritidisSalmonella sp S epidermidis S xylosus S aureus Calbicans and Candida sp with Pseudomonas sp as the mostabundant strains e presence of Salmonella in all samplesmakes no doubt on the safety status of the meat In the sameway the presence of microorganisms like P putida and Paeruginosa which can lead to a quick spoilage of fresh beefsuggests a reduced shelf life of the product Howeveridentification of these strains through 16S rRNA gene an-alyses needs to be done to confirm their identity e results

CC

MD

MBG

MBT

GM

PM

P putidaP aeruginosa

Pseudomonas sp

E coli

E coli 1

S enteritidis

Salmonella sp

S aureus

S xylosus

S epidermis

C albicans

Candida sp

ndash6

ndash4

ndash2

0

2

4

6

8

ndash10 ndash8 ndash6 ndash4 ndash2 0 2 4 6 8 10

F2 (2

876

)

F1 (5455)

Biplot (axis F1 amp F2 8331)

Figure 1 Distribution of microbial species isolated from fresh beef and sampling sites on F1times F2 axis PM ldquoPetit Marcherdquo GM ldquoGrandMarcherdquo MD ldquoDang marketrdquo CC ldquoCentre Commercialrdquo MBG ldquoBamnyanga marketrdquo MBT ldquoBantail marketrdquo

Journal of Food Quality 7

Tabl

e3

Microbial

inhibitio

npercentages(

)ob

tained

from

theantia

dhesiveassays

with

thebiosurfactantfrom

Lparacaseisub

sptoleransN2

Test

organism

sBiosurfactantc

oncentratio

n(m

gmL)

001

01

05

125

575

10S

aureus

MTC

C1430

3396plusmn236

aG4915plusmn248

bI6903plusmn006

cH7626plusmn038

dI8251plusmn143

eG9209plusmn023

fG9734plusmn109

gH9918plusmn010

hI

MluteusMTC

C106

2259plusmn259

aD4238plusmn218

bH6573plusmn147

cG7890plusmn333

dI8717plusmn308

eH9263plusmn041

fG9411plusmn051

gF9559plusmn071

hG

Lmon

ocytogenes

MTC

C839

2051plusmn119

aD3241plusmn009

bE4150plusmn043

cE5094plusmn019

dF5829plusmn148

eF6222plusmn024

fE6840plusmn155

gE7515plusmn053

hF

EcoliMTC

C118

1995plusmn032

aD3024plusmn069

bD3877plusmn240

cD4862plusmn194

dE5465plusmn146

eD6008plusmn016

fD6578plusmn048

gD7214plusmn033

hE

Paerugino

saPS

B21546plusmn075

aC2164plusmn225

bB3130plusmn100

cB4072plusmn010

dB4790plusmn095

eB5170plusmn055

fB5680plusmn015

gB6170plusmn055

hB

Ppu

tidaPS

J11310plusmn010

aB2315plusmn181

bB3071plusmn152

cB4030plusmn040

dB4695plusmn150

eB5271plusmn052

fB5722plusmn082

gB6071plusmn050

hB

SenteritidisSL

21445plusmn065

aC2680plusmn110

bC3473plusmn014

cC4210plusmn020

dC5040plusmn090

eC5821plusmn005

fC6120plusmn010

gC6610plusmn050

hC

EcoliE2

B1932plusmn090

aD2718plusmn045

bC3610plusmn040

cD4590plusmn015

dD5350plusmn035

eE5910plusmn075

fD6415plusmn150

gD7010plusmn045

hD

Calbicans

LV1

578plusmn035

aA1219plusmn230

bA2480plusmn211

cA3678plusmn191

dA4310plusmn066

eA4790plusmn025

fA5362plusmn031

gA5612plusmn016

hA

Bacillu

sspB

C1

2425plusmn090

aE4182plusmn114

bH6770plusmn067

cG7030plusmn050

dG8190plusmn022

eG9062plusmn020

fH9878plusmn050

gH10000plusmn000

hJ

Sau

reus

STP1

2770plusmn075

aF4001plusmn023

bH6617plusmn180

cG7338plusmn070

dH8221plusmn082

eG9330plusmn075

fI9925plusmn082

gH10000plusmn000

hJ

SxylosusST

P22132plusmn210

aD3841plusmn025

bG6570plusmn150

cG7313plusmn070

dH8192plusmn031

eG9240plusmn021

fI9816plusmn110

gH10000plusmn000

hJ

SepidermisST

P32180plusmn229

aD3711plusmn031

bF6020plusmn166

cF7176plusmn050

dG8068plusmn179

eG8818plusmn013

fF9594plusmn005

gG9870plusmn0050h

H

Negativec

ontrolsw

eresetat0

toindicatethea

bsence

ofbiosurfactantPo

sitivep

ercentagesindicatethereductio

nsinmicrobialadhesio

nwhencomparedto

thec

ontrolR

esultsaree

xpressed

asmeansplusmnstandard

deviationof

results

from

triplicateexperiments

8 Journal of Food Quality

of this study demonstrated the antiadhesive activity ofbiosurfactant against microbial strains isolated from freshbeef Efforts made by the Government to improve the mi-crobiological quality of fresh beef should deal not only withthe respect of good slaughtering evisceration and hygienicpractices but also with the cleaning process of surfaces wheremeat is cut and sold

Data Availability

e data used in this study are available from the corre-sponding author upon request

Conflicts of Interest

e authors declare no conflicts of interest

Acknowledgments

e authors thank the Director of the National School ofAgroindustrial Sciences of the University of NgaoundereCameroon and the Department of Microbiology and Fer-mentation Technology CSIR-Central Food TechnologicalResearch Institute of Mysore India for providing necessaryfacilities for the successful completion of this research worke authors gratefully acknowledge butchers of the differentmarkets of Ngaoundere for their kind cooperation duringthe sampling process

References

[1] K Zerabruk N Retta D Muleta and A T Tefera ldquoAs-sessment of microbiological safety and quality of minced meatand meat contact surfaces in selected butcher shops of AddisAbaba Ethiopiardquo Journal of Food Quality vol 2019 pp 1ndash92019

[2] A Holck L Axelsson A McLeod T M Rode and E HeirldquoHealth and safety considerations of fermented sausagesrdquoJournal of Food Quality vol 2017 Article ID 975389425 pages 2017

[3] G-J E Nychas P N Skandamis C C Tassou andK P Koutsoumanis ldquoMeat spoilage during distributionrdquoMeat Science vol 78 no 1-2 pp 77ndash89 2008

[4] G anigaivel and A S Anandhan ldquoIsolation and charac-terization of microorganisms from raw meat obtained fromdifferent market places in and around Chennairdquo Journal ofPharmaceutical Chemical and Biological Sciences vol 3 no 2pp 295ndash301 2015

[5] Centre for Disease Control (CDC) Multistate Outbreak ofSalmonella Enteritidis Infections Linked to Ground BeefCentre for Disease Control (CDC) Atlanta GA USA 2012

[6] J D Bailly H Brugere and H Chadron ldquoMicroorganismes etparasites des viandes les connaıtre pour les maıtriser delrsquoeleveur au consommateur CIVrdquo 2012 httpwwwciv-Viandeorg

[7] C Salifou K Boko G Ahounou et al ldquoDiversite de lamicroflore initiale de la viande et securite sanitaire desconsommateursrdquo International Journal of Biological andChemical Sciences vol 7 no 3 pp 1351ndash1369 2013

[8] S Datta A Akter I G Shah et al ldquoMicrobiological qualityassessment of raw meat and meat products and antibioticsusceptibility of isolated Staphylococcus aureusrdquo Agriculture

Food and Analytical Bacteriology vol 2 no 3 pp 187ndash1942012

[9] E Abayneh D Nolkes and B Asrade ldquoReview on commonfoodborne pathogens in Ethiopiardquo African Journal of Mi-crobiology Research vol 8 no 53 pp 4027ndash4040 2014

[10] T Kebede B Afera H Taddele and A Bsrat ldquoAssessment ofbacteriological quality of sold meat in the butcher shops ofAdigrat Tigray Ethiopiardquo Applied Journal of Hygiene vol 3no 3 pp 38ndash44 2014

[11] A J Ilboudo A Savadogo S Samandoulougou M AbreM Seydi and A Traore ldquoQualite bacteriologique des car-casses de viandes porcines et bovines produites a lrsquoabattoir deOuagadougou Burkina Fasordquo Revue Microbiologie Industri-elle Sanitaire et Environnementale vol 10 no 1 pp 33ndash352016

[12] N M Sachindra P Z Sakhare K P Yashoda andD Narasimha Rao ldquoMicrobial profile of buffalo sausageduring processing and storagerdquo Food Control vol 16 no 1pp 31ndash35 2005

[13] B Siriken ldquoe microbiological quality of ground beef inAydin and Afyon Provinces Turkeyrdquo Revue de MedecineVeterinaire vol 155 no 12 pp 632ndash636 2004

[14] I O E Ukut I O Okonko I S Ikpoh et al ldquoAssessment ofbacteriological quality of fresh meats sold in Calabar me-tropolis Nigeriardquo Electronic Journal of Environmental Ag-ricultural and Food Chemistry vol 9 no 1 pp 89ndash100 2010

[15] A M Goja T A A Ahmed S A M Saeed and H A DirarldquoIsolation and identification of Staphylococcus spp in freshbeefrdquo Pakistan Journal of Nutrition vol 12 no 2 pp 114ndash1202013

[16] P Antwi-Agyei and B K Maalekuu ldquoDetermination ofmicrobial contamination in meat and fish products sold in theKumasi metropolis (a case study of Kumasi central marketand the Bantama market)rdquo Merit Research Journal of Agri-cultural Science and Soil vol 2 pp 038ndash046 2014

[17] E Niyonzima D Bora and M P Ongol ldquoAssessment of beefmeat microbial contamination during skinning dressingtransportation and marketing at a commercial abattoir inKigali city Rwandardquo Pakistan Journal of Food Sciencesvol 23 pp 133ndash138 2013

[18] V K Singh J Udit J K Yadav and B Basanti ldquoAssessmentof bacterial quality of raw meat samples (carabeef chevonpork and poultry) from retail meat outlets and local slaughterhouses of Agra Region Indiardquo Journal of Foodborne andZoonotic Diseases vol 2 no 1 pp 15ndash18 2014

[19] R B Afnabi R P Nameni S S Kamdem J J Essia Ngangand R B Alambedji ldquoMicrobial load of beef sold in thetraditional slaughterhouse and butcher shops in northernCameroonrdquo International Journal of Veterinary Science vol 4no 4 pp 183ndash189 2015

[20] E J Gudintildea E C Fernandes J A Teixeira andL R Rodrigues ldquoAntimicrobial and anti-adhesive activities ofcell-bound biosurfactant from Lactobacillus agilisCCUG31450rdquo RSC Advances vol 5 no 110 p 90960 2015

[21] D Sharma and B S Saharan ldquoFunctional characterization ofbiomedical potential of biosurfactant produced by Lactoba-cillus helveticusrdquo Biotechnology Reports vol 11 pp 27ndash352016

[22] A Merghni I Dallel E Noumi et al ldquoAntioxidant andantiproliferative potential of biosurfactants isolated fromLactobacillus casei and their anti-biofilm effect in oralStaphylococcus aureus strainsrdquo Microbial Pathogenesisvol 104 pp 84ndash89 2017

Journal of Food Quality 9

of E coli at proportions ranging from a market to anotherey also highlighted that some strains of E coli isolatedfrom fresh beef were identified as Enterohemorrhagic E coli(E coliO157 H7) Giving that some strains of E coli such asEnterohemorrhagic E coli (EHEC) are recognized as food-borne pathogens associated with several outbreaks of dis-eases [44] its presence in fresh beef as observed in this studyis worrisome Molecular identification of the strains isolatedis required in order to bring more information about theirpathogenicity

e main reservoir of zoonotic Salmonella is food fromanimal origin and the main sources of infections incrimi-nated Salmonella in industrialized and developing countriesare animal-derived products notably fresh meat products[45] In this study 24 strains of Salmonella were isolatedamong which there were 3750 of S enterica serovarEnteritidis and 6250 of Salmonella sp (Table 2) RegardingS enterica serovar Enteritidis it was found only in sites MDMBG and PM at the proportion of 3333 Salmonella spwas absent in the site MD and present in the other sites SitesMBG and PM rank first with 2666 each Ukut et al [14]also identified Salmonella spp in fresh beef sold in Calabarmarket Nigeria In another study conducted by Obeng et al[39] Salmonella spp were isolated and identified in freshmeat sold in Tolon and Kumbungu districts of the NorthernRegion of Ghana

For yeasts belonging to the genus Candida Table 2 showsthat 19 strains were isolated and identified as the species Calbicans (5 strains) and Candida sp (14 strains) Strains of Calbicans were found only in sites MBG MBT and PM atproportions of 40 20 and 40 respectively Strains ofCandida sp were identified in fresh beef samples collectedfrom the different sites and the site GM ranks first with 2857of strainse results of this study are in accordance with thosereported byComi andCantoni [46] who showed that strains ofCandida spp are dominant yeasts of fresh beef

In order to visualize the association between the mi-crobial species identified in this study and the samplingsites a principal component analysis was carried outFigure 1 presents the distribution of microbial speciesisolated from fresh beef and sampling sites on the axissystem F1 times F2 As seen in Figure 1 three main groupswere formed e first group shows that the strains be-longing to the species C albicans Salmonella sp Paeruginosa P putida S xylosus and S enteritidis areassociated with the sites PM and MBG e second groupshows that the strains belonging to the species Pseudo-monas sp E coli 1 S epidermis and S aureus are as-sociated with site MBT Finally the third group shows thatthe strains belonging to the species Candida sp are mainlyassociated with the sites MD CC and GM is principalcomponent analysis pointed out the specific relationship

Table 2 Genus species identification percentages and proportions of strains isolated from fresh beef samples collected from differentmarkets of the city of Ngaoundere

Genus Species ID percentages()

Number ofstrains

Numbers and proportions of strains per sampling sitesCC MD MBG MBT GM PM

Pseudomonas

Pseudomonasputida 9850 12 4

(3333)2

(3333)4

(1666)0

(0)0

(0)2

(1666)Pseudomonasaeruginosa 9980 13 2

(1538)3

(2307)5

(3846)0

(0) 1 (769) 2(1538)

Pseudomonas sp 9740 26 6(2307)

3(1153)

3(1153)

8(3076) 2 (769) 4

(1538)

EscherichiaEscherichia coli 9960 14 4

(2857)0

(0)4

(2857)0

(0)2

(1428)4

(2857)

Escherichia coli 1 9940 5 0(0)

2(4000)

0(0)

3(6000)

0(0)

0(0)

Salmonella

S enterica serEnteritidis 9860 9 0

(0)3

(3333)3

(3333)0

(0)0

(0)3

(3333)

Salmonella sp 9910 15 2(1333)

0(0)

4(2666)

3(2000)

2(1333)

4(2666)

Staphylococcus

Staphylococcusaureus 9860 18 2

(1111)4

(2222)3

(1666)5

(2777) 1 (555) 3(1666)

Staphylococcusxylosus 10000 12 0

(0)0

(0)8

(6666)0

(0)0

(0)4

(3333)Staphylococcus

epidermis 9830 8 0(0)

0(0)

2(2500)

4(5000)

0(0)

2(2500)

CandidaCandida albicans 9860 5 0

(0)0

(0)2

(4000)1

(2000)0

(0)2

(4000)

Candida sp 9830 14 3(2142)

2(1428)

1(714)

3(2142)

4(2857)

1(714)

Total 151 23(1523)

19(1258)

39(2582)

27(1788)

12(794)

31(2052)

ID identification percentages of the strains from API data ser serovar PM ldquoPetit Marcherdquo GM ldquoGrandMarcherdquo MD ldquoDang marketrdquo CC ldquoCentreCommercialrdquo MBG ldquoBamnyanga marketrdquo MBT ldquoBantail marketrdquo

6 Journal of Food Quality

between the microbial diversity of fresh beef and thesampling sites It enables to observe associations betweenthe sampling sites and the microbial profile of fresh beefsold in Ngaoundere e use of 16S rRNA gene sequencingand a high number of meat samples will improve this PCArepresentation through an increase of the microbialdiversity

33 Antiadhesive Activity of Biosurfactant Food spoilageand disease transmission due to microorganisms attachedinto surfaces are one of the main problems faced by foodindustries particularly in the meat industry In this studythe antiadhesive property of biosurfactant was tested againstsome selected bacterial and fungal strains isolated from freshbeef and against some reference cultures e biosurfactantsderived from L paracasei subsp tolerans N2 inhibited theadhesion of all the tested strains in a dose-dependentmanner (Table 3) Biosurfactant was active even at 001against all the tested microorganisms At that concentration3396plusmn 236 and 2770plusmn 075 of inhibition were recordedagainst S aureusMTCC 1430 and S aureus STP1 Completeinhibition (100) of microbial adhesion was observedagainst Bacillus sp BC1 S aureus STP1 and S xylosus STP2at biosurfactant concentration of 10mgmL e leastantiadhesive activity (5612plusmn 016) of biosurfactant wasnoticed against the yeast strain C albicans LV1 is anti-adhesive activity of biosurfactants observed in this studycould be due to their ability to change the hydrophobicity ofthe interface between the strain and the surface of thepolystyrene plate Hence microbial adhesion to the surfacewill decrease as biosurfactant concentration increasesSharma and Saharan [21] have explained the antiadhesiveactivity of biosurfactants of L helveticus by their ability toform a thin film which modifies the wettability of the surfaceand thus decreases the adhesive properties of microorgan-isms e inhibition percentages observed in Table 3 are notdifferent from the values reported byMerghni et al [22] with

biosurfactants from L casei ATCC 393 LZ9 and L caseiATCC 393 LBl ey noticed inhibition percentages rangingfrom 6884 to 8486 against S aureus 6538 S aureus 9Pand S aureus 29P at biosurfactant concentration of 125mgmL As shown in Table 3 Gram-negative bacteria were moreresistant to the antiadhesive activity of biosurfactant Asimilar observation was noticed by Sambanthamoorthy et al[47] with biosurfactants from L jensenii and L rhamnosusis resistance of Gram-negative bacteria could result intheir ability to produce once adhered to surfaces homo-serine lactone acyl (HLA) molecules which increase theirresistance to antiadhesive compounds [48]

Besides the adhesive properties of microorganismsisolated from fresh beef samples as shown in this studysuggest that cutting meat surfaces represent an ecologicalkennel of continuous contamination of meat Hence theantiadhesive activity of biosurfactant from L paracaseisubsp tolerans N2 against these strains shows that thesemolecules could be useful in the reduction of meat con-tamination through cutting surfaces

4 Conclusion

is study showed that fresh beef sold in the markets ofNgaoundere contained both pathogenic and spoilage mi-croorganisms at levels higher than the threshold valuesrecommended by the European Commission Twelve mi-crobial species colonized these fresh beef P putida Paeruginosa Pseudomonas sp E coli 1 E coli S enteritidisSalmonella sp S epidermidis S xylosus S aureus Calbicans and Candida sp with Pseudomonas sp as the mostabundant strains e presence of Salmonella in all samplesmakes no doubt on the safety status of the meat In the sameway the presence of microorganisms like P putida and Paeruginosa which can lead to a quick spoilage of fresh beefsuggests a reduced shelf life of the product Howeveridentification of these strains through 16S rRNA gene an-alyses needs to be done to confirm their identity e results

CC

MD

MBG

MBT

GM

PM

P putidaP aeruginosa

Pseudomonas sp

E coli

E coli 1

S enteritidis

Salmonella sp

S aureus

S xylosus

S epidermis

C albicans

Candida sp

ndash6

ndash4

ndash2

0

2

4

6

8

ndash10 ndash8 ndash6 ndash4 ndash2 0 2 4 6 8 10

F2 (2

876

)

F1 (5455)

Biplot (axis F1 amp F2 8331)

Figure 1 Distribution of microbial species isolated from fresh beef and sampling sites on F1times F2 axis PM ldquoPetit Marcherdquo GM ldquoGrandMarcherdquo MD ldquoDang marketrdquo CC ldquoCentre Commercialrdquo MBG ldquoBamnyanga marketrdquo MBT ldquoBantail marketrdquo

Journal of Food Quality 7

Tabl

e3

Microbial

inhibitio

npercentages(

)ob

tained

from

theantia

dhesiveassays

with

thebiosurfactantfrom

Lparacaseisub

sptoleransN2

Test

organism

sBiosurfactantc

oncentratio

n(m

gmL)

001

01

05

125

575

10S

aureus

MTC

C1430

3396plusmn236

aG4915plusmn248

bI6903plusmn006

cH7626plusmn038

dI8251plusmn143

eG9209plusmn023

fG9734plusmn109

gH9918plusmn010

hI

MluteusMTC

C106

2259plusmn259

aD4238plusmn218

bH6573plusmn147

cG7890plusmn333

dI8717plusmn308

eH9263plusmn041

fG9411plusmn051

gF9559plusmn071

hG

Lmon

ocytogenes

MTC

C839

2051plusmn119

aD3241plusmn009

bE4150plusmn043

cE5094plusmn019

dF5829plusmn148

eF6222plusmn024

fE6840plusmn155

gE7515plusmn053

hF

EcoliMTC

C118

1995plusmn032

aD3024plusmn069

bD3877plusmn240

cD4862plusmn194

dE5465plusmn146

eD6008plusmn016

fD6578plusmn048

gD7214plusmn033

hE

Paerugino

saPS

B21546plusmn075

aC2164plusmn225

bB3130plusmn100

cB4072plusmn010

dB4790plusmn095

eB5170plusmn055

fB5680plusmn015

gB6170plusmn055

hB

Ppu

tidaPS

J11310plusmn010

aB2315plusmn181

bB3071plusmn152

cB4030plusmn040

dB4695plusmn150

eB5271plusmn052

fB5722plusmn082

gB6071plusmn050

hB

SenteritidisSL

21445plusmn065

aC2680plusmn110

bC3473plusmn014

cC4210plusmn020

dC5040plusmn090

eC5821plusmn005

fC6120plusmn010

gC6610plusmn050

hC

EcoliE2

B1932plusmn090

aD2718plusmn045

bC3610plusmn040

cD4590plusmn015

dD5350plusmn035

eE5910plusmn075

fD6415plusmn150

gD7010plusmn045

hD

Calbicans

LV1

578plusmn035

aA1219plusmn230

bA2480plusmn211

cA3678plusmn191

dA4310plusmn066

eA4790plusmn025

fA5362plusmn031

gA5612plusmn016

hA

Bacillu

sspB

C1

2425plusmn090

aE4182plusmn114

bH6770plusmn067

cG7030plusmn050

dG8190plusmn022

eG9062plusmn020

fH9878plusmn050

gH10000plusmn000

hJ

Sau

reus

STP1

2770plusmn075

aF4001plusmn023

bH6617plusmn180

cG7338plusmn070

dH8221plusmn082

eG9330plusmn075

fI9925plusmn082

gH10000plusmn000

hJ

SxylosusST

P22132plusmn210

aD3841plusmn025

bG6570plusmn150

cG7313plusmn070

dH8192plusmn031

eG9240plusmn021

fI9816plusmn110

gH10000plusmn000

hJ

SepidermisST

P32180plusmn229

aD3711plusmn031

bF6020plusmn166

cF7176plusmn050

dG8068plusmn179

eG8818plusmn013

fF9594plusmn005

gG9870plusmn0050h

H

Negativec

ontrolsw

eresetat0

toindicatethea

bsence

ofbiosurfactantPo

sitivep

ercentagesindicatethereductio

nsinmicrobialadhesio

nwhencomparedto

thec

ontrolR

esultsaree

xpressed

asmeansplusmnstandard

deviationof

results

from

triplicateexperiments

8 Journal of Food Quality

of this study demonstrated the antiadhesive activity ofbiosurfactant against microbial strains isolated from freshbeef Efforts made by the Government to improve the mi-crobiological quality of fresh beef should deal not only withthe respect of good slaughtering evisceration and hygienicpractices but also with the cleaning process of surfaces wheremeat is cut and sold

Data Availability

e data used in this study are available from the corre-sponding author upon request

Conflicts of Interest

e authors declare no conflicts of interest

Acknowledgments

e authors thank the Director of the National School ofAgroindustrial Sciences of the University of NgaoundereCameroon and the Department of Microbiology and Fer-mentation Technology CSIR-Central Food TechnologicalResearch Institute of Mysore India for providing necessaryfacilities for the successful completion of this research worke authors gratefully acknowledge butchers of the differentmarkets of Ngaoundere for their kind cooperation duringthe sampling process

References

[1] K Zerabruk N Retta D Muleta and A T Tefera ldquoAs-sessment of microbiological safety and quality of minced meatand meat contact surfaces in selected butcher shops of AddisAbaba Ethiopiardquo Journal of Food Quality vol 2019 pp 1ndash92019

[2] A Holck L Axelsson A McLeod T M Rode and E HeirldquoHealth and safety considerations of fermented sausagesrdquoJournal of Food Quality vol 2017 Article ID 975389425 pages 2017

[3] G-J E Nychas P N Skandamis C C Tassou andK P Koutsoumanis ldquoMeat spoilage during distributionrdquoMeat Science vol 78 no 1-2 pp 77ndash89 2008

[4] G anigaivel and A S Anandhan ldquoIsolation and charac-terization of microorganisms from raw meat obtained fromdifferent market places in and around Chennairdquo Journal ofPharmaceutical Chemical and Biological Sciences vol 3 no 2pp 295ndash301 2015

[5] Centre for Disease Control (CDC) Multistate Outbreak ofSalmonella Enteritidis Infections Linked to Ground BeefCentre for Disease Control (CDC) Atlanta GA USA 2012

[6] J D Bailly H Brugere and H Chadron ldquoMicroorganismes etparasites des viandes les connaıtre pour les maıtriser delrsquoeleveur au consommateur CIVrdquo 2012 httpwwwciv-Viandeorg

[7] C Salifou K Boko G Ahounou et al ldquoDiversite de lamicroflore initiale de la viande et securite sanitaire desconsommateursrdquo International Journal of Biological andChemical Sciences vol 7 no 3 pp 1351ndash1369 2013

[8] S Datta A Akter I G Shah et al ldquoMicrobiological qualityassessment of raw meat and meat products and antibioticsusceptibility of isolated Staphylococcus aureusrdquo Agriculture

Food and Analytical Bacteriology vol 2 no 3 pp 187ndash1942012

[9] E Abayneh D Nolkes and B Asrade ldquoReview on commonfoodborne pathogens in Ethiopiardquo African Journal of Mi-crobiology Research vol 8 no 53 pp 4027ndash4040 2014

[10] T Kebede B Afera H Taddele and A Bsrat ldquoAssessment ofbacteriological quality of sold meat in the butcher shops ofAdigrat Tigray Ethiopiardquo Applied Journal of Hygiene vol 3no 3 pp 38ndash44 2014

[11] A J Ilboudo A Savadogo S Samandoulougou M AbreM Seydi and A Traore ldquoQualite bacteriologique des car-casses de viandes porcines et bovines produites a lrsquoabattoir deOuagadougou Burkina Fasordquo Revue Microbiologie Industri-elle Sanitaire et Environnementale vol 10 no 1 pp 33ndash352016

[12] N M Sachindra P Z Sakhare K P Yashoda andD Narasimha Rao ldquoMicrobial profile of buffalo sausageduring processing and storagerdquo Food Control vol 16 no 1pp 31ndash35 2005

[13] B Siriken ldquoe microbiological quality of ground beef inAydin and Afyon Provinces Turkeyrdquo Revue de MedecineVeterinaire vol 155 no 12 pp 632ndash636 2004

[14] I O E Ukut I O Okonko I S Ikpoh et al ldquoAssessment ofbacteriological quality of fresh meats sold in Calabar me-tropolis Nigeriardquo Electronic Journal of Environmental Ag-ricultural and Food Chemistry vol 9 no 1 pp 89ndash100 2010

[15] A M Goja T A A Ahmed S A M Saeed and H A DirarldquoIsolation and identification of Staphylococcus spp in freshbeefrdquo Pakistan Journal of Nutrition vol 12 no 2 pp 114ndash1202013

[16] P Antwi-Agyei and B K Maalekuu ldquoDetermination ofmicrobial contamination in meat and fish products sold in theKumasi metropolis (a case study of Kumasi central marketand the Bantama market)rdquo Merit Research Journal of Agri-cultural Science and Soil vol 2 pp 038ndash046 2014

[17] E Niyonzima D Bora and M P Ongol ldquoAssessment of beefmeat microbial contamination during skinning dressingtransportation and marketing at a commercial abattoir inKigali city Rwandardquo Pakistan Journal of Food Sciencesvol 23 pp 133ndash138 2013

[18] V K Singh J Udit J K Yadav and B Basanti ldquoAssessmentof bacterial quality of raw meat samples (carabeef chevonpork and poultry) from retail meat outlets and local slaughterhouses of Agra Region Indiardquo Journal of Foodborne andZoonotic Diseases vol 2 no 1 pp 15ndash18 2014

[19] R B Afnabi R P Nameni S S Kamdem J J Essia Ngangand R B Alambedji ldquoMicrobial load of beef sold in thetraditional slaughterhouse and butcher shops in northernCameroonrdquo International Journal of Veterinary Science vol 4no 4 pp 183ndash189 2015

[20] E J Gudintildea E C Fernandes J A Teixeira andL R Rodrigues ldquoAntimicrobial and anti-adhesive activities ofcell-bound biosurfactant from Lactobacillus agilisCCUG31450rdquo RSC Advances vol 5 no 110 p 90960 2015

[21] D Sharma and B S Saharan ldquoFunctional characterization ofbiomedical potential of biosurfactant produced by Lactoba-cillus helveticusrdquo Biotechnology Reports vol 11 pp 27ndash352016

[22] A Merghni I Dallel E Noumi et al ldquoAntioxidant andantiproliferative potential of biosurfactants isolated fromLactobacillus casei and their anti-biofilm effect in oralStaphylococcus aureus strainsrdquo Microbial Pathogenesisvol 104 pp 84ndash89 2017

Journal of Food Quality 9

between the microbial diversity of fresh beef and thesampling sites It enables to observe associations betweenthe sampling sites and the microbial profile of fresh beefsold in Ngaoundere e use of 16S rRNA gene sequencingand a high number of meat samples will improve this PCArepresentation through an increase of the microbialdiversity

33 Antiadhesive Activity of Biosurfactant Food spoilageand disease transmission due to microorganisms attachedinto surfaces are one of the main problems faced by foodindustries particularly in the meat industry In this studythe antiadhesive property of biosurfactant was tested againstsome selected bacterial and fungal strains isolated from freshbeef and against some reference cultures e biosurfactantsderived from L paracasei subsp tolerans N2 inhibited theadhesion of all the tested strains in a dose-dependentmanner (Table 3) Biosurfactant was active even at 001against all the tested microorganisms At that concentration3396plusmn 236 and 2770plusmn 075 of inhibition were recordedagainst S aureusMTCC 1430 and S aureus STP1 Completeinhibition (100) of microbial adhesion was observedagainst Bacillus sp BC1 S aureus STP1 and S xylosus STP2at biosurfactant concentration of 10mgmL e leastantiadhesive activity (5612plusmn 016) of biosurfactant wasnoticed against the yeast strain C albicans LV1 is anti-adhesive activity of biosurfactants observed in this studycould be due to their ability to change the hydrophobicity ofthe interface between the strain and the surface of thepolystyrene plate Hence microbial adhesion to the surfacewill decrease as biosurfactant concentration increasesSharma and Saharan [21] have explained the antiadhesiveactivity of biosurfactants of L helveticus by their ability toform a thin film which modifies the wettability of the surfaceand thus decreases the adhesive properties of microorgan-isms e inhibition percentages observed in Table 3 are notdifferent from the values reported byMerghni et al [22] with

biosurfactants from L casei ATCC 393 LZ9 and L caseiATCC 393 LBl ey noticed inhibition percentages rangingfrom 6884 to 8486 against S aureus 6538 S aureus 9Pand S aureus 29P at biosurfactant concentration of 125mgmL As shown in Table 3 Gram-negative bacteria were moreresistant to the antiadhesive activity of biosurfactant Asimilar observation was noticed by Sambanthamoorthy et al[47] with biosurfactants from L jensenii and L rhamnosusis resistance of Gram-negative bacteria could result intheir ability to produce once adhered to surfaces homo-serine lactone acyl (HLA) molecules which increase theirresistance to antiadhesive compounds [48]

Besides the adhesive properties of microorganismsisolated from fresh beef samples as shown in this studysuggest that cutting meat surfaces represent an ecologicalkennel of continuous contamination of meat Hence theantiadhesive activity of biosurfactant from L paracaseisubsp tolerans N2 against these strains shows that thesemolecules could be useful in the reduction of meat con-tamination through cutting surfaces

4 Conclusion

is study showed that fresh beef sold in the markets ofNgaoundere contained both pathogenic and spoilage mi-croorganisms at levels higher than the threshold valuesrecommended by the European Commission Twelve mi-crobial species colonized these fresh beef P putida Paeruginosa Pseudomonas sp E coli 1 E coli S enteritidisSalmonella sp S epidermidis S xylosus S aureus Calbicans and Candida sp with Pseudomonas sp as the mostabundant strains e presence of Salmonella in all samplesmakes no doubt on the safety status of the meat In the sameway the presence of microorganisms like P putida and Paeruginosa which can lead to a quick spoilage of fresh beefsuggests a reduced shelf life of the product Howeveridentification of these strains through 16S rRNA gene an-alyses needs to be done to confirm their identity e results

CC

MD

MBG

MBT

GM

PM

P putidaP aeruginosa

Pseudomonas sp

E coli

E coli 1

S enteritidis

Salmonella sp

S aureus

S xylosus

S epidermis

C albicans

Candida sp

ndash6

ndash4

ndash2

0

2

4

6

8

ndash10 ndash8 ndash6 ndash4 ndash2 0 2 4 6 8 10

F2 (2

876

)

F1 (5455)

Biplot (axis F1 amp F2 8331)

Figure 1 Distribution of microbial species isolated from fresh beef and sampling sites on F1times F2 axis PM ldquoPetit Marcherdquo GM ldquoGrandMarcherdquo MD ldquoDang marketrdquo CC ldquoCentre Commercialrdquo MBG ldquoBamnyanga marketrdquo MBT ldquoBantail marketrdquo

Journal of Food Quality 7

Tabl

e3

Microbial

inhibitio

npercentages(

)ob

tained

from

theantia

dhesiveassays

with

thebiosurfactantfrom

Lparacaseisub

sptoleransN2

Test

organism

sBiosurfactantc

oncentratio

n(m

gmL)

001

01

05

125

575

10S

aureus

MTC

C1430

3396plusmn236

aG4915plusmn248

bI6903plusmn006

cH7626plusmn038

dI8251plusmn143

eG9209plusmn023

fG9734plusmn109

gH9918plusmn010

hI

MluteusMTC

C106

2259plusmn259

aD4238plusmn218

bH6573plusmn147

cG7890plusmn333

dI8717plusmn308

eH9263plusmn041

fG9411plusmn051

gF9559plusmn071

hG

Lmon

ocytogenes

MTC

C839

2051plusmn119

aD3241plusmn009

bE4150plusmn043

cE5094plusmn019

dF5829plusmn148

eF6222plusmn024

fE6840plusmn155

gE7515plusmn053

hF

EcoliMTC

C118

1995plusmn032

aD3024plusmn069

bD3877plusmn240

cD4862plusmn194

dE5465plusmn146

eD6008plusmn016

fD6578plusmn048

gD7214plusmn033

hE

Paerugino

saPS

B21546plusmn075

aC2164plusmn225

bB3130plusmn100

cB4072plusmn010

dB4790plusmn095

eB5170plusmn055

fB5680plusmn015

gB6170plusmn055

hB

Ppu

tidaPS

J11310plusmn010

aB2315plusmn181

bB3071plusmn152

cB4030plusmn040

dB4695plusmn150

eB5271plusmn052

fB5722plusmn082

gB6071plusmn050

hB

SenteritidisSL

21445plusmn065

aC2680plusmn110

bC3473plusmn014

cC4210plusmn020

dC5040plusmn090

eC5821plusmn005

fC6120plusmn010

gC6610plusmn050

hC

EcoliE2

B1932plusmn090

aD2718plusmn045

bC3610plusmn040

cD4590plusmn015

dD5350plusmn035

eE5910plusmn075

fD6415plusmn150

gD7010plusmn045

hD

Calbicans

LV1

578plusmn035

aA1219plusmn230

bA2480plusmn211

cA3678plusmn191

dA4310plusmn066

eA4790plusmn025

fA5362plusmn031

gA5612plusmn016

hA

Bacillu

sspB

C1

2425plusmn090

aE4182plusmn114

bH6770plusmn067

cG7030plusmn050

dG8190plusmn022

eG9062plusmn020

fH9878plusmn050

gH10000plusmn000

hJ

Sau

reus

STP1

2770plusmn075

aF4001plusmn023

bH6617plusmn180

cG7338plusmn070

dH8221plusmn082

eG9330plusmn075

fI9925plusmn082

gH10000plusmn000

hJ

SxylosusST

P22132plusmn210

aD3841plusmn025

bG6570plusmn150

cG7313plusmn070

dH8192plusmn031

eG9240plusmn021

fI9816plusmn110

gH10000plusmn000

hJ

SepidermisST

P32180plusmn229

aD3711plusmn031

bF6020plusmn166

cF7176plusmn050

dG8068plusmn179

eG8818plusmn013

fF9594plusmn005

gG9870plusmn0050h

H

Negativec

ontrolsw

eresetat0

toindicatethea

bsence

ofbiosurfactantPo

sitivep

ercentagesindicatethereductio

nsinmicrobialadhesio

nwhencomparedto

thec

ontrolR

esultsaree

xpressed

asmeansplusmnstandard

deviationof

results

from

triplicateexperiments

8 Journal of Food Quality

of this study demonstrated the antiadhesive activity ofbiosurfactant against microbial strains isolated from freshbeef Efforts made by the Government to improve the mi-crobiological quality of fresh beef should deal not only withthe respect of good slaughtering evisceration and hygienicpractices but also with the cleaning process of surfaces wheremeat is cut and sold

Data Availability

e data used in this study are available from the corre-sponding author upon request

Conflicts of Interest

e authors declare no conflicts of interest

Acknowledgments

e authors thank the Director of the National School ofAgroindustrial Sciences of the University of NgaoundereCameroon and the Department of Microbiology and Fer-mentation Technology CSIR-Central Food TechnologicalResearch Institute of Mysore India for providing necessaryfacilities for the successful completion of this research worke authors gratefully acknowledge butchers of the differentmarkets of Ngaoundere for their kind cooperation duringthe sampling process

References

[1] K Zerabruk N Retta D Muleta and A T Tefera ldquoAs-sessment of microbiological safety and quality of minced meatand meat contact surfaces in selected butcher shops of AddisAbaba Ethiopiardquo Journal of Food Quality vol 2019 pp 1ndash92019

[2] A Holck L Axelsson A McLeod T M Rode and E HeirldquoHealth and safety considerations of fermented sausagesrdquoJournal of Food Quality vol 2017 Article ID 975389425 pages 2017

[3] G-J E Nychas P N Skandamis C C Tassou andK P Koutsoumanis ldquoMeat spoilage during distributionrdquoMeat Science vol 78 no 1-2 pp 77ndash89 2008

[4] G anigaivel and A S Anandhan ldquoIsolation and charac-terization of microorganisms from raw meat obtained fromdifferent market places in and around Chennairdquo Journal ofPharmaceutical Chemical and Biological Sciences vol 3 no 2pp 295ndash301 2015

[5] Centre for Disease Control (CDC) Multistate Outbreak ofSalmonella Enteritidis Infections Linked to Ground BeefCentre for Disease Control (CDC) Atlanta GA USA 2012

[6] J D Bailly H Brugere and H Chadron ldquoMicroorganismes etparasites des viandes les connaıtre pour les maıtriser delrsquoeleveur au consommateur CIVrdquo 2012 httpwwwciv-Viandeorg

[7] C Salifou K Boko G Ahounou et al ldquoDiversite de lamicroflore initiale de la viande et securite sanitaire desconsommateursrdquo International Journal of Biological andChemical Sciences vol 7 no 3 pp 1351ndash1369 2013

[8] S Datta A Akter I G Shah et al ldquoMicrobiological qualityassessment of raw meat and meat products and antibioticsusceptibility of isolated Staphylococcus aureusrdquo Agriculture

Food and Analytical Bacteriology vol 2 no 3 pp 187ndash1942012

[9] E Abayneh D Nolkes and B Asrade ldquoReview on commonfoodborne pathogens in Ethiopiardquo African Journal of Mi-crobiology Research vol 8 no 53 pp 4027ndash4040 2014

[10] T Kebede B Afera H Taddele and A Bsrat ldquoAssessment ofbacteriological quality of sold meat in the butcher shops ofAdigrat Tigray Ethiopiardquo Applied Journal of Hygiene vol 3no 3 pp 38ndash44 2014

[11] A J Ilboudo A Savadogo S Samandoulougou M AbreM Seydi and A Traore ldquoQualite bacteriologique des car-casses de viandes porcines et bovines produites a lrsquoabattoir deOuagadougou Burkina Fasordquo Revue Microbiologie Industri-elle Sanitaire et Environnementale vol 10 no 1 pp 33ndash352016

[12] N M Sachindra P Z Sakhare K P Yashoda andD Narasimha Rao ldquoMicrobial profile of buffalo sausageduring processing and storagerdquo Food Control vol 16 no 1pp 31ndash35 2005

[13] B Siriken ldquoe microbiological quality of ground beef inAydin and Afyon Provinces Turkeyrdquo Revue de MedecineVeterinaire vol 155 no 12 pp 632ndash636 2004

[14] I O E Ukut I O Okonko I S Ikpoh et al ldquoAssessment ofbacteriological quality of fresh meats sold in Calabar me-tropolis Nigeriardquo Electronic Journal of Environmental Ag-ricultural and Food Chemistry vol 9 no 1 pp 89ndash100 2010

[15] A M Goja T A A Ahmed S A M Saeed and H A DirarldquoIsolation and identification of Staphylococcus spp in freshbeefrdquo Pakistan Journal of Nutrition vol 12 no 2 pp 114ndash1202013

[16] P Antwi-Agyei and B K Maalekuu ldquoDetermination ofmicrobial contamination in meat and fish products sold in theKumasi metropolis (a case study of Kumasi central marketand the Bantama market)rdquo Merit Research Journal of Agri-cultural Science and Soil vol 2 pp 038ndash046 2014

[17] E Niyonzima D Bora and M P Ongol ldquoAssessment of beefmeat microbial contamination during skinning dressingtransportation and marketing at a commercial abattoir inKigali city Rwandardquo Pakistan Journal of Food Sciencesvol 23 pp 133ndash138 2013

[18] V K Singh J Udit J K Yadav and B Basanti ldquoAssessmentof bacterial quality of raw meat samples (carabeef chevonpork and poultry) from retail meat outlets and local slaughterhouses of Agra Region Indiardquo Journal of Foodborne andZoonotic Diseases vol 2 no 1 pp 15ndash18 2014

[19] R B Afnabi R P Nameni S S Kamdem J J Essia Ngangand R B Alambedji ldquoMicrobial load of beef sold in thetraditional slaughterhouse and butcher shops in northernCameroonrdquo International Journal of Veterinary Science vol 4no 4 pp 183ndash189 2015

[20] E J Gudintildea E C Fernandes J A Teixeira andL R Rodrigues ldquoAntimicrobial and anti-adhesive activities ofcell-bound biosurfactant from Lactobacillus agilisCCUG31450rdquo RSC Advances vol 5 no 110 p 90960 2015

[21] D Sharma and B S Saharan ldquoFunctional characterization ofbiomedical potential of biosurfactant produced by Lactoba-cillus helveticusrdquo Biotechnology Reports vol 11 pp 27ndash352016

[22] A Merghni I Dallel E Noumi et al ldquoAntioxidant andantiproliferative potential of biosurfactants isolated fromLactobacillus casei and their anti-biofilm effect in oralStaphylococcus aureus strainsrdquo Microbial Pathogenesisvol 104 pp 84ndash89 2017

Journal of Food Quality 9

Tabl

e3

Microbial

inhibitio

npercentages(

)ob

tained

from

theantia

dhesiveassays

with

thebiosurfactantfrom

Lparacaseisub

sptoleransN2

Test

organism

sBiosurfactantc

oncentratio

n(m

gmL)

001

01

05

125

575

10S

aureus

MTC

C1430

3396plusmn236

aG4915plusmn248

bI6903plusmn006

cH7626plusmn038

dI8251plusmn143

eG9209plusmn023

fG9734plusmn109

gH9918plusmn010

hI

MluteusMTC

C106

2259plusmn259

aD4238plusmn218

bH6573plusmn147

cG7890plusmn333

dI8717plusmn308

eH9263plusmn041

fG9411plusmn051

gF9559plusmn071

hG

Lmon

ocytogenes

MTC

C839

2051plusmn119

aD3241plusmn009

bE4150plusmn043

cE5094plusmn019

dF5829plusmn148

eF6222plusmn024

fE6840plusmn155

gE7515plusmn053

hF

EcoliMTC

C118

1995plusmn032

aD3024plusmn069

bD3877plusmn240

cD4862plusmn194

dE5465plusmn146

eD6008plusmn016

fD6578plusmn048

gD7214plusmn033

hE

Paerugino

saPS

B21546plusmn075

aC2164plusmn225

bB3130plusmn100

cB4072plusmn010

dB4790plusmn095

eB5170plusmn055

fB5680plusmn015

gB6170plusmn055

hB

Ppu

tidaPS

J11310plusmn010

aB2315plusmn181

bB3071plusmn152

cB4030plusmn040

dB4695plusmn150

eB5271plusmn052

fB5722plusmn082

gB6071plusmn050

hB

SenteritidisSL

21445plusmn065

aC2680plusmn110

bC3473plusmn014

cC4210plusmn020

dC5040plusmn090

eC5821plusmn005

fC6120plusmn010

gC6610plusmn050

hC

EcoliE2

B1932plusmn090

aD2718plusmn045

bC3610plusmn040

cD4590plusmn015

dD5350plusmn035

eE5910plusmn075

fD6415plusmn150

gD7010plusmn045

hD

Calbicans

LV1

578plusmn035

aA1219plusmn230

bA2480plusmn211

cA3678plusmn191

dA4310plusmn066

eA4790plusmn025

fA5362plusmn031

gA5612plusmn016

hA

Bacillu

sspB

C1

2425plusmn090

aE4182plusmn114

bH6770plusmn067

cG7030plusmn050

dG8190plusmn022

eG9062plusmn020

fH9878plusmn050

gH10000plusmn000

hJ

Sau

reus

STP1

2770plusmn075

aF4001plusmn023

bH6617plusmn180

cG7338plusmn070

dH8221plusmn082

eG9330plusmn075

fI9925plusmn082

gH10000plusmn000

hJ

SxylosusST

P22132plusmn210

aD3841plusmn025

bG6570plusmn150

cG7313plusmn070

dH8192plusmn031

eG9240plusmn021

fI9816plusmn110

gH10000plusmn000

hJ

SepidermisST

P32180plusmn229

aD3711plusmn031

bF6020plusmn166

cF7176plusmn050

dG8068plusmn179

eG8818plusmn013

fF9594plusmn005

gG9870plusmn0050h

H

Negativec

ontrolsw

eresetat0

toindicatethea

bsence

ofbiosurfactantPo

sitivep

ercentagesindicatethereductio

nsinmicrobialadhesio

nwhencomparedto

thec

ontrolR

esultsaree

xpressed

asmeansplusmnstandard

deviationof

results

from

triplicateexperiments

8 Journal of Food Quality

of this study demonstrated the antiadhesive activity ofbiosurfactant against microbial strains isolated from freshbeef Efforts made by the Government to improve the mi-crobiological quality of fresh beef should deal not only withthe respect of good slaughtering evisceration and hygienicpractices but also with the cleaning process of surfaces wheremeat is cut and sold

Data Availability

e data used in this study are available from the corre-sponding author upon request

Conflicts of Interest

e authors declare no conflicts of interest

Acknowledgments

e authors thank the Director of the National School ofAgroindustrial Sciences of the University of NgaoundereCameroon and the Department of Microbiology and Fer-mentation Technology CSIR-Central Food TechnologicalResearch Institute of Mysore India for providing necessaryfacilities for the successful completion of this research worke authors gratefully acknowledge butchers of the differentmarkets of Ngaoundere for their kind cooperation duringthe sampling process

References

[1] K Zerabruk N Retta D Muleta and A T Tefera ldquoAs-sessment of microbiological safety and quality of minced meatand meat contact surfaces in selected butcher shops of AddisAbaba Ethiopiardquo Journal of Food Quality vol 2019 pp 1ndash92019

[2] A Holck L Axelsson A McLeod T M Rode and E HeirldquoHealth and safety considerations of fermented sausagesrdquoJournal of Food Quality vol 2017 Article ID 975389425 pages 2017

[3] G-J E Nychas P N Skandamis C C Tassou andK P Koutsoumanis ldquoMeat spoilage during distributionrdquoMeat Science vol 78 no 1-2 pp 77ndash89 2008

[4] G anigaivel and A S Anandhan ldquoIsolation and charac-terization of microorganisms from raw meat obtained fromdifferent market places in and around Chennairdquo Journal ofPharmaceutical Chemical and Biological Sciences vol 3 no 2pp 295ndash301 2015

[5] Centre for Disease Control (CDC) Multistate Outbreak ofSalmonella Enteritidis Infections Linked to Ground BeefCentre for Disease Control (CDC) Atlanta GA USA 2012

[6] J D Bailly H Brugere and H Chadron ldquoMicroorganismes etparasites des viandes les connaıtre pour les maıtriser delrsquoeleveur au consommateur CIVrdquo 2012 httpwwwciv-Viandeorg

[7] C Salifou K Boko G Ahounou et al ldquoDiversite de lamicroflore initiale de la viande et securite sanitaire desconsommateursrdquo International Journal of Biological andChemical Sciences vol 7 no 3 pp 1351ndash1369 2013

[8] S Datta A Akter I G Shah et al ldquoMicrobiological qualityassessment of raw meat and meat products and antibioticsusceptibility of isolated Staphylococcus aureusrdquo Agriculture

Food and Analytical Bacteriology vol 2 no 3 pp 187ndash1942012

[9] E Abayneh D Nolkes and B Asrade ldquoReview on commonfoodborne pathogens in Ethiopiardquo African Journal of Mi-crobiology Research vol 8 no 53 pp 4027ndash4040 2014

[10] T Kebede B Afera H Taddele and A Bsrat ldquoAssessment ofbacteriological quality of sold meat in the butcher shops ofAdigrat Tigray Ethiopiardquo Applied Journal of Hygiene vol 3no 3 pp 38ndash44 2014

[11] A J Ilboudo A Savadogo S Samandoulougou M AbreM Seydi and A Traore ldquoQualite bacteriologique des car-casses de viandes porcines et bovines produites a lrsquoabattoir deOuagadougou Burkina Fasordquo Revue Microbiologie Industri-elle Sanitaire et Environnementale vol 10 no 1 pp 33ndash352016

[12] N M Sachindra P Z Sakhare K P Yashoda andD Narasimha Rao ldquoMicrobial profile of buffalo sausageduring processing and storagerdquo Food Control vol 16 no 1pp 31ndash35 2005

[13] B Siriken ldquoe microbiological quality of ground beef inAydin and Afyon Provinces Turkeyrdquo Revue de MedecineVeterinaire vol 155 no 12 pp 632ndash636 2004

[14] I O E Ukut I O Okonko I S Ikpoh et al ldquoAssessment ofbacteriological quality of fresh meats sold in Calabar me-tropolis Nigeriardquo Electronic Journal of Environmental Ag-ricultural and Food Chemistry vol 9 no 1 pp 89ndash100 2010

[15] A M Goja T A A Ahmed S A M Saeed and H A DirarldquoIsolation and identification of Staphylococcus spp in freshbeefrdquo Pakistan Journal of Nutrition vol 12 no 2 pp 114ndash1202013

[16] P Antwi-Agyei and B K Maalekuu ldquoDetermination ofmicrobial contamination in meat and fish products sold in theKumasi metropolis (a case study of Kumasi central marketand the Bantama market)rdquo Merit Research Journal of Agri-cultural Science and Soil vol 2 pp 038ndash046 2014

[17] E Niyonzima D Bora and M P Ongol ldquoAssessment of beefmeat microbial contamination during skinning dressingtransportation and marketing at a commercial abattoir inKigali city Rwandardquo Pakistan Journal of Food Sciencesvol 23 pp 133ndash138 2013

[18] V K Singh J Udit J K Yadav and B Basanti ldquoAssessmentof bacterial quality of raw meat samples (carabeef chevonpork and poultry) from retail meat outlets and local slaughterhouses of Agra Region Indiardquo Journal of Foodborne andZoonotic Diseases vol 2 no 1 pp 15ndash18 2014

[19] R B Afnabi R P Nameni S S Kamdem J J Essia Ngangand R B Alambedji ldquoMicrobial load of beef sold in thetraditional slaughterhouse and butcher shops in northernCameroonrdquo International Journal of Veterinary Science vol 4no 4 pp 183ndash189 2015

[20] E J Gudintildea E C Fernandes J A Teixeira andL R Rodrigues ldquoAntimicrobial and anti-adhesive activities ofcell-bound biosurfactant from Lactobacillus agilisCCUG31450rdquo RSC Advances vol 5 no 110 p 90960 2015

[21] D Sharma and B S Saharan ldquoFunctional characterization ofbiomedical potential of biosurfactant produced by Lactoba-cillus helveticusrdquo Biotechnology Reports vol 11 pp 27ndash352016

[22] A Merghni I Dallel E Noumi et al ldquoAntioxidant andantiproliferative potential of biosurfactants isolated fromLactobacillus casei and their anti-biofilm effect in oralStaphylococcus aureus strainsrdquo Microbial Pathogenesisvol 104 pp 84ndash89 2017

Journal of Food Quality 9

of this study demonstrated the antiadhesive activity ofbiosurfactant against microbial strains isolated from freshbeef Efforts made by the Government to improve the mi-crobiological quality of fresh beef should deal not only withthe respect of good slaughtering evisceration and hygienicpractices but also with the cleaning process of surfaces wheremeat is cut and sold

Data Availability

e data used in this study are available from the corre-sponding author upon request

Conflicts of Interest

e authors declare no conflicts of interest

Acknowledgments

e authors thank the Director of the National School ofAgroindustrial Sciences of the University of NgaoundereCameroon and the Department of Microbiology and Fer-mentation Technology CSIR-Central Food TechnologicalResearch Institute of Mysore India for providing necessaryfacilities for the successful completion of this research worke authors gratefully acknowledge butchers of the differentmarkets of Ngaoundere for their kind cooperation duringthe sampling process

References

[1] K Zerabruk N Retta D Muleta and A T Tefera ldquoAs-sessment of microbiological safety and quality of minced meatand meat contact surfaces in selected butcher shops of AddisAbaba Ethiopiardquo Journal of Food Quality vol 2019 pp 1ndash92019

[2] A Holck L Axelsson A McLeod T M Rode and E HeirldquoHealth and safety considerations of fermented sausagesrdquoJournal of Food Quality vol 2017 Article ID 975389425 pages 2017

[3] G-J E Nychas P N Skandamis C C Tassou andK P Koutsoumanis ldquoMeat spoilage during distributionrdquoMeat Science vol 78 no 1-2 pp 77ndash89 2008

[4] G anigaivel and A S Anandhan ldquoIsolation and charac-terization of microorganisms from raw meat obtained fromdifferent market places in and around Chennairdquo Journal ofPharmaceutical Chemical and Biological Sciences vol 3 no 2pp 295ndash301 2015

[5] Centre for Disease Control (CDC) Multistate Outbreak ofSalmonella Enteritidis Infections Linked to Ground BeefCentre for Disease Control (CDC) Atlanta GA USA 2012

[6] J D Bailly H Brugere and H Chadron ldquoMicroorganismes etparasites des viandes les connaıtre pour les maıtriser delrsquoeleveur au consommateur CIVrdquo 2012 httpwwwciv-Viandeorg

[7] C Salifou K Boko G Ahounou et al ldquoDiversite de lamicroflore initiale de la viande et securite sanitaire desconsommateursrdquo International Journal of Biological andChemical Sciences vol 7 no 3 pp 1351ndash1369 2013

[8] S Datta A Akter I G Shah et al ldquoMicrobiological qualityassessment of raw meat and meat products and antibioticsusceptibility of isolated Staphylococcus aureusrdquo Agriculture

Food and Analytical Bacteriology vol 2 no 3 pp 187ndash1942012

[9] E Abayneh D Nolkes and B Asrade ldquoReview on commonfoodborne pathogens in Ethiopiardquo African Journal of Mi-crobiology Research vol 8 no 53 pp 4027ndash4040 2014

[10] T Kebede B Afera H Taddele and A Bsrat ldquoAssessment ofbacteriological quality of sold meat in the butcher shops ofAdigrat Tigray Ethiopiardquo Applied Journal of Hygiene vol 3no 3 pp 38ndash44 2014

[11] A J Ilboudo A Savadogo S Samandoulougou M AbreM Seydi and A Traore ldquoQualite bacteriologique des car-casses de viandes porcines et bovines produites a lrsquoabattoir deOuagadougou Burkina Fasordquo Revue Microbiologie Industri-elle Sanitaire et Environnementale vol 10 no 1 pp 33ndash352016

[12] N M Sachindra P Z Sakhare K P Yashoda andD Narasimha Rao ldquoMicrobial profile of buffalo sausageduring processing and storagerdquo Food Control vol 16 no 1pp 31ndash35 2005

[13] B Siriken ldquoe microbiological quality of ground beef inAydin and Afyon Provinces Turkeyrdquo Revue de MedecineVeterinaire vol 155 no 12 pp 632ndash636 2004

[14] I O E Ukut I O Okonko I S Ikpoh et al ldquoAssessment ofbacteriological quality of fresh meats sold in Calabar me-tropolis Nigeriardquo Electronic Journal of Environmental Ag-ricultural and Food Chemistry vol 9 no 1 pp 89ndash100 2010

[15] A M Goja T A A Ahmed S A M Saeed and H A DirarldquoIsolation and identification of Staphylococcus spp in freshbeefrdquo Pakistan Journal of Nutrition vol 12 no 2 pp 114ndash1202013

[16] P Antwi-Agyei and B K Maalekuu ldquoDetermination ofmicrobial contamination in meat and fish products sold in theKumasi metropolis (a case study of Kumasi central marketand the Bantama market)rdquo Merit Research Journal of Agri-cultural Science and Soil vol 2 pp 038ndash046 2014

[17] E Niyonzima D Bora and M P Ongol ldquoAssessment of beefmeat microbial contamination during skinning dressingtransportation and marketing at a commercial abattoir inKigali city Rwandardquo Pakistan Journal of Food Sciencesvol 23 pp 133ndash138 2013

[18] V K Singh J Udit J K Yadav and B Basanti ldquoAssessmentof bacterial quality of raw meat samples (carabeef chevonpork and poultry) from retail meat outlets and local slaughterhouses of Agra Region Indiardquo Journal of Foodborne andZoonotic Diseases vol 2 no 1 pp 15ndash18 2014

[19] R B Afnabi R P Nameni S S Kamdem J J Essia Ngangand R B Alambedji ldquoMicrobial load of beef sold in thetraditional slaughterhouse and butcher shops in northernCameroonrdquo International Journal of Veterinary Science vol 4no 4 pp 183ndash189 2015

[20] E J Gudintildea E C Fernandes J A Teixeira andL R Rodrigues ldquoAntimicrobial and anti-adhesive activities ofcell-bound biosurfactant from Lactobacillus agilisCCUG31450rdquo RSC Advances vol 5 no 110 p 90960 2015

[21] D Sharma and B S Saharan ldquoFunctional characterization ofbiomedical potential of biosurfactant produced by Lactoba-cillus helveticusrdquo Biotechnology Reports vol 11 pp 27ndash352016

[22] A Merghni I Dallel E Noumi et al ldquoAntioxidant andantiproliferative potential of biosurfactants isolated fromLactobacillus casei and their anti-biofilm effect in oralStaphylococcus aureus strainsrdquo Microbial Pathogenesisvol 104 pp 84ndash89 2017

Journal of Food Quality 9

[23] R S Makkar S S Cameotra and I M Banat ldquoAdvances inutilization of renewable substrates for biosurfactant pro-ductionrdquo AMB Express vol 1 no 5 pp 1ndash19 2011

[24] International Organization for Standardization (ISO) 17604Microbiology of the Food ChainmdashCarcass Sampling for Mi-crobiological Analysis ISO Geneva Switzerland 2015

[25] T H Mouafo A Mbawala R Ndjouenkeu R Ndjouenkeuand D Somashekar ldquoApplication of response surfacemethodology to improve the production of antimicrobialbiosurfactants by Lactobacillus paracasei Subsprdquo BioMedResearch International vol 2018 no 48 pp 1ndash15 2018

[26] International Organization for Standardization 6887-2 Mi-crobiology of the Food ChainmdashPreparation of Test SamplesInitial Suspension and Decimal Dilutions for MicrobiologicalExaminationmdashPart 2 Specific Rules for the Preparation ofMeat and Meat Products ISO Geneva Switzerland 2017

[27] International Organization for Standardization (ISO) 4833-1Microbiology of the Food ChainmdashHorizontal Method for theEnumeration of MicroorganismsmdashPart 1 Colony Count at30degC by the Pour Plate Technique 2013

[28] International Organization for Standardization (ISO) 4832Microbiology of Food and Animal Feeding StuffsmdashHorizontalMethod for the Enumeration of ColiformsmdashColony-countTechnique ISO Geneva Switzerland 2006

[29] International Organization for Standardization (ISO) 21528-2Microbiology of the Food ChainmdashHorizontal Method for theDetection and Enumeration of EnterobacteriaceaemdashPart 2Colony-count Technique ISO Geneva Switzerland 2017

[30] International Organization for Standardization (ISO) 6888-2Microbiology of Food and Animal Feeding StuffsmdashHorizontalMethod for the Enumeration of Coagulase-Positive Staphylo-cocci (Staphylococcus aureus and Other Species)mdashPart 2Technique Using Rabbit Plasma Fibrinogen Agar MediumISO Geneva Switzerland 1999

[31] International Organization for Standardization (ISO) 7932Microbiology of Food and Animal Feeding StuffsmdashHorizontalMethod for the Enumeration of Presumptive Bacillus Cer-eusmdashColony-count Technique at 30degC ISO Geneva Swit-zerland 2004

[32] International Organization for Standardization (ISO) 13720Meat and Meat ProductsmdashEnumeration of PresumptivePseudomonas Spp ISO Geneva Switzerland 2010

[33] International Organization for Standardization (ISO) 21527-1 Microbiology of Food and Animal FeedingStuffsmdashHorizontal Method for the Enumeration of Yeasts andMouldsmdashPart 1 Colony Count Technique in Products withWater Activity Greater than 095 ISO Geneva Switzerland2008

[34] International Organization for Standardization (ISO) 6579-1Microbiology of the Food ChainmdashHorizontal Method for theDetection Enumeration and Serotyping of SalmonellamdashPart 1Detection of Salmonella Spp ISO Geneva Switzerland 2017

[35] International Organization for Standardization (ISO) 7937Microbiology of Food and Animal Feeding StuffsmdashHorizontalMethod for the Enumeration of Clostridium Per-fringensmdashColony-count Technique ISO Geneva Switzerland2004

[36] D H Bergey J G Holt and N R Krieg Bergeyrsquos Manual ofDeterminative Bacteriology Lippincott Williams amp WilkinsEast Lansing MI USA 9th edition 1994

[37] E J Gudintildea V Rocha J A Teixeira and L R RodriguesldquoAntimicrobial and antiadhesive properties of a biosurfactantisolated from Lactobacillus paracasei ssp paracasei A20rdquo

Letters in Applied Microbiology vol 50 no 4 pp 419ndash4242010

[38] European Commission Regulation (EC) 20732005 ldquoMicro-biological criteria for foodstuffsrdquo Official Journal of the Eu-ropean Union vol 338 pp 1ndash26 2005

[39] A K Obeng F S Johnson and S O Appenteng ldquoMicrobialquality of fresh meat from retail outlets in Tolon andKumbungu districts of the northern region of Ghanardquo In-ternational Journal of Science and Technology vol 2 no 6pp 423ndash428 2013

[40] F E Emele W Pasculle R H Glew and N Mbrey ldquoInci-dence and prevalence of Aeromonas hydrophila in meat soldin Enugu main marketrdquo Nigerian Journal Microbiologyvol 10 pp 77ndash80 1995

[41] K Koutsoumanis A Stamatiou P Skandamis andG-J E Nychas ldquoDevelopment of a microbial model for thecombined effect of temperature and pH on spoilage of groundmeat and validation of the model under dynamic temperatureconditionsrdquo Applied and Environmental Microbiology vol 72no 1 pp 124ndash134 2006

[42] K B Ronoh ldquoDetermination of bacteriological quality offresh beef post-harvesting in Nyagacho Slumrdquo Master thesisSchool of Pure and Applied Sciences of Kenyatta UniversityKericho Kenya 2013

[43] E Balcha K Ashwani and T Habtamu ldquoEvaluation of safetyof beef sold in and around mekelle with special reference toenterohaemorrhagic Escherichia coli O157 H7rdquo GlobalVeterinaria vol 12 no 4 pp 569ndash572 2014

[44] H Karch P I Tarr and M Bielaszewska ldquoEnter-ohaemorrhagic Escherichia coli in human medicinerdquo Inter-national Journal of Medical Microbiology vol 295 no 6-7pp 405ndash418 2005

[45] M Helms S Ethelberg and K Moslashlbak ldquoInternational Sal-monella typhimurium DT104 infections 1992ndash2001rdquoEmerging Infectious Diseases vol 11 no 6 pp 859ndash867 2005

[46] G Comi and C Cantoni ldquoLieviti e carnirdquo Industrie Ali-mentari vol 24 pp 683ndash687 1985

[47] K Sambanthamoorthy X Feng R Patel S Patel andC Paranavitana ldquoAntimicrobial and antibiofilm potential ofbiosurfactants isolated from Lactobacilli against multi-drug-resistant pathogensrdquo BMCMicrobiology vol 14 no 1 pp 1ndash92014

[48] P Biswa and M Doble ldquoProduction of acylated homoserinelactone by gram-positive bacteria isolated from marine wa-terrdquo FEMS Microbiology Letters vol 343 no 1 pp 34ndash412013

10 Journal of Food Quality