c- Bmed Approach for Food Safely Mdnageme~it Proceeding qf internut tonal Sem I nor Current Issues and Challenges in Food Sqfeiy

. P., Woutersen, R. A., Feron, V. J., Hollanders, V. H., Falke, H. E., & OF SP IN CIary, J. 1. 1989. TWO-Year ~rinking-water study of THE OCCURRENCE Pseudomonas Formaldehyde in Rats. Food Chem. Toxicol., 27(2):77-87. FILLETS (Plectroporna , M., Naito, K., & Kurokawa, Y. 1989. Chronic Toxicity Study on leopordus) AT BONE PANTE Formaldehyde Administered Orally to Rats. Toxicology, 56(1]:79-86. Rieny Sulistijowati* and Royhan Alhasen** * Universitas Muhamrnadiyah Gorontalo **Universitas Negeri Gorontalo [email protected]

ABSTRACT Study on the occurrence of Psetrdomonas sp., associated with grouper fish (PIectroporna leopardus) was done. The research aimed to know the occurrence of Pseudomor~assp it1 fresh grouper fish (P. leopardus) fillets chilled in ice. The fish samples were taken from grouper fish farm located at Bone Pante village (separated into two sampling area 1 and 11). The result showed that both of total plate count [TPC) and total Pseurlomonas (TP) of grouper fish were lower in samples taken from sampling area I than those taken from sampling area I I. The TPC of sample from sampl~ngarea 1 ranged between 2,1 x 1 O4 CFU/g to 2,8 x 1 04,while from sarl~plingarea II it ranged between 1,9 x 10TCC;Ug to 5,Z x 104 CFll/g. Total P~~rrdornonossp. of sample from sampling area I ranged between 1.8 x103 CFIJ/g to 4,4 x103 CFU/g, while from sampling area II, it ranged between 8,O xlO2 CFU/g to 3,7 x lo3 CFU/g. The number of 'JPC and TP varied among samples and the tlutnber was lower than the required number. as suggested by Standirr Nus~onaiIndonesia (SNI) for the quality of rn~crobiologicalof frozen grouper fish. IJs~ngthe Bergey's Manual of Microbiological Ident~hcation,the isolated strai 11 were identified as Pseudumunas caryophylli {6,9%] , P. fluorescens (1 1,l %), P. deiafiedii (23,6%), and P. aeruginosrr (12,s %). They have physiological characteristic as follows : grow well at temperature between O0 - 370 1: with optimum temperature for growth at 37°C: pH range between 4 - 9, with optimum pH of 7, tolerate salinity with NaCl concentration between 3-8 YO. P. caryophylli isolated from grouper fish fitlet demonstrated 13- haemolysis and a- haernolysis for P. aeruginosa. Wh~leP. caryophylli and P. flourecens were agglutir~asepositive. ln general, samples taken From farm at sampling area 1 were better as compared to that of sampling area 11. Sensory values from sampling area I were 24,9 with averages value of more than 8,f while from sampling area 11, the value were between 21- 23,06 with averages sensory of Iess than 7,6.

loster presentar ion poster presentation (345 Science- Based Approach ,for Food Safep Marrugement Procerding ~flnrernu~iun~ri,Seminar C'urrcnl irrues and Chr3iiengt.u rn Food

INTRODUCTION used were chilled grouper fish fillets, aquadest, Natrium medium, Cetrimide Agar and NaCl, APW (enrichment), alcahol. Background Grouper fish [PIectropoma leopardus) is a potential export Sampling commodity and can contribute to national foreign exchange. The fish Grouper fish fillets were sampled from processing unit at is also a source of protein, lipid and minerals essential for human. Pante I and Bone Pante 11. The fish fillets samples were 250 - 5 However, grouper fish fish fillet is a perishable product. Currerltly weight x 2 pieces, Sampling was taken triplicate in each locatior some studies focused on the occurrence of bacteria in grouper fish frequency of sampling was every one month after previous sam are conducted. From the studies, it was known that bacteria Live grouper fish was used as test controls. The controls were commonly found in the tjsh are Pseudomonas sp.. Salmonellu, V~brio, from water each time the fish fillet and sea water was sampled. Sthaphylococcus, and some more. Pseudomonas sp is one of fish contaminating bacterium that may cause diseases. Base on these Microbiology analysis findings, the writer want to research more abou "the occurrence of Pseudomonas sp, in the grouper fish fillet (Plectropoma leopardus). It Fish fillet was analyzed for its Total Plnte Courrt (?'PC) and is expected that the dominant species of Pseudomonas sp. associated Pseudomonas [TP). with the fish can be identifled. Isolation and identification of Pseudomonas The objectives of research Selection and isolation steps of Pseudomoncrs consist of sf The research aims are as follows. physiology and biochemical tests such as: Gram coloration motility test, oxidase enzyme test, catalase enzyme test, ferment 1. To undel-stand the occurrence of Pseudomonas sp. in freshly test, indol test, methyl red test, voges-proskouer test, and citric chilled fillet of grouper fish taken from Bone Pante area I and 11. test. 2. To identify the species of Pseudomonas sp. dominantly associated To examine factors influences the growth of Pseudomont: with grouper fish fish fillet. several growth conditions were tested, i.e. growth at six difl temperatures; 00 C, 20 C, 40 C, 60 C, 25O C, 37" for 24 hours; at 5,6, 7,8,9, and 10; in NaCI 0, 3,5, 7,9% THE METHODOLOGY OF RESEARCH Assay of pathogenic characteristics of Pse{~domonnssp. in Place and Time of Research agglutination and haemolysis tests.

This research was conducted in Microbiology laboratory at Sensory test Faculty of Agriculture NationaI University of Gorontalo. The research was completed in three months from October to December 2008. Sensory test was performed according to SN I

Material and Methods The equipment utilized were outaclave, incubators, petridisc, ovens, blenders, pipette, analytical balances, volumetric glasses, beaker glass, reaction tubes, magnetic stirrers, microscopes and object glasses, ose needles, pH meters, and durharn tubes. The material

346lposter presentation poster prescntatio nce- Based Approach for Food Safe@ Mmagemenr Proceeding of lnrernaliclnul .Tcminar Current issue.^ and Challeng~sin 1;ood SuIep I i 1

'RODUCTION used were chilled grouper fish fillets, aquadest, Natrium Agar medium, Cetrimide Agar and NaC1, APW (enrichment), alcohol. :kground

Grouper fish [Plectropornn leopardus) is a potential export Sampling lmodity and can contribute to national foreign exchange. The fish Grouper fish fillets were sampled from processing unit at Bone Iso a source of protein, lipid and minerals essential for human. ?ante I and Bone Pante 11. The fish fillets samples werc 250 - 500 gr- vcver, grouper fish fish fillet is a perishable product. Currently weigh1 x 2 pieces. San~plingwas taken triplicate in each location. The le studies focused on the occurrence of bacteria in grouper fish frequency of sampling was every one month after previous sampling. conducted. From the studies, it was known that bacteria I.ive grouper fish was used as test controls. The controls were taken 1111onlyfound in the fish are Pseudomonas sp., Salmonella, Vibrio, from water each time the fish fillet and sea water was sampled. rphylococcus, and some more. Pseudornonas sp is one of fish !amhating bacterium that may cause diseases. Base on these Microbiology analysis ings, the writer want to research more abou "the occurrence of rdomonos sp. in the grouper fish fillet (Plectropoma leopardus]. It Fish fillet was analyzed for its Total Place Count (I'I'C) arid Total rpected that the dominant species of Pseudomonas sp. associated Pseudomonas(TP). I the fish can be identified. Isolation and Identification of Pseudomonas objectives of research Selection and isolation steps of Pseudomonas consist of several research aims are as follows. physiology and biochemical tests such as: Cram colol.ation test, motiIity test, oxidase enzyme test, catalase enzyme tcst, fermentation To understand the occurrence of Pseudomonas sp. in freshly test, indol test, methyl red test, voges-proskauer test, and citric acid chilled fillet of grouper fish taken from Bone Pante area I and 11. test. To identify the species of Pseudomonas sp. dominantly associated To examine factors influences the growth of Pseudomonas sp., with grouper fish Fish fillet. several growth conditions were tested, i.e, growth at six different temperatures; 00 C, 20 C, 40 C, GW, 25O C, 37O C for 24 hours; at pH 4, 5,6,7, 8,9,and 10; in NaC1 0,3, 5, 7,9% METHODOLOGY OF RESEARCH Assay of pathogenic characteristics of Pseudomonas sp. include e and Time of Research agglutination and haemolysis tests.

This research was conducted in Microbiology laboratory at Sensory test Ity of Agriculture National University of Gorontalo. The research zompleted in three months from October to December 2008. Sensory test was performed according to SNI

!rial and Methods The equipment utilized were outoclave, incubators, petridisc, s, blenders, pipette, analytical balances, volumetric glasses, ~rglass,reaction tubes, magnetic stirrers, microscopes and object es, ose needles, pH meters, and durharn tubes. The material

-- ~osterpresentation postpr prl:sl:n~ation1347 Science- Baed Approach for Food Safiety Manugemenf Proceeding qf In~ernationalSeminar Currenf 1.r.rtre.rand Chullen,qc.~in Food 5

RESULT AND DISCUSSION According to Huss (1995), the number of totai bacteria in f~ products was 103 - lo7. The Occurrence of Bacteria in Grouper fish fillets The average of total Pseudomonas value in samples is show The average of TPC value are shown in Table 1 and 2. Table 3 and in control can be seen at Table 4.

Table 1. TPC of samples Table 3. Total Pseudomonas in samples

Replication Average of Tot Location ". ..* Pseudomona~ I 11 111 value (TVC/gr ce-Based Approach fm Food Safev Muvugement Proceding of International Brrn~nrtrCurrent Issues und Challenges in Food Sujer).

ULT AND DlSCUSSION According to Huss [1995), the number of total bacteria in fresh seafood products was lo3- lo7. Occurrence of Bacteria in Grouper fish fillets The average of total Pseudomonas value in samples is shown in average of TPC value are shown in Table I and 2. Table 3 and in control can be seen at Table 4. e 1. TPC of samples Table 3. Total Pseudomonos in samples

Replicates Replication Average of Total 'ation The average of TPC values I I I Ill CCFU/grl Location Pseudomonas I 11 III value (TVC jgr] 1 1.8 x 103 4.4 x 103 3.6 x 103 3.3 x 10" lav c 7 .. I I 8.0 x 102 2.4 x 103 3,7 x 103 z,3 x 103

.- TVC: Total Viable Count Colony Forming Unit

Tabel 4. Total Pseudomonas in control fish and seawater 81 2. TPC of control fish and sea water Ulangan Average of Total

Replication Average of control l IT Pseudomonas value Control I 11 11 TPC values (CFU/p,r] Live Grouper e Grouper fish 8,2 x 103 7,9 xlo3 8,l x 103 fish Sea water 1,6 x 104 1,8 X lo4 1,7 x 103 Sea water - 1,2 x lo4 1,2 x lo4 Colony Forming Unit TVC: TotaJ Viable Count

TPC values varied between 1,8 x lo4 CFU/gr and 2,5 x lo4 From table above it can be seen that the total number of :r with average were 2,2 x 104CFU/gr. The average of TPC value nple from Iocation 1 were 2,5 x 104 CFU/gr. This value was pseudonlonas in samples from location I was 3.3 x 10VVC/gr. This was than found in II r as compared to the sample from location II with average was number higher those samples from location which was 2,3 x 103 TVC/gr. The results also showed that t the LO4 CFU/gr. average of total pseudomonas in samples from location I and II were rPC values of sample from both location were at the range of higher as compared to controls of live-grouper fish. However the 104 CFU/gr to 2,5 x 104 CFU/gr. Whereas TPC values of control average of total Pseudornonas in sea water was higher. There was grouper fish and sea water ranged from 1,7 X 103 to 8,l X 103 possibility that the bacteria were washed away during the ;r, The TPC values of control were higher than TPC values of processing of grouper fish fillets. 11. Based on quality requirements from Standar Nasional ?sia [SNl 01-2696-1992) the maximum number for bacteria in er fish fillets were 5x10s colonies / gr (Anonymous, 19921.

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The result of methyl red-tests was varied. There were 1 1 strains The growth temperature of the 4 isolates at 0°, 20, 40,6T, 2! showed positive result and another 11 strains showed negative and 37% is shown in Table 6. result. Voges Proskauer tests revealed that 10 strains had positive result. This test was conduct to show the ability of bacteria to ferment carbohydrate and produce acetylmetyl carbinol. The citric acid tests showed variation in results. There were positive and negative results. The result of the test can be seen in

3501poster presentation poster presentation ( -Based Approachfor Food Safety Management PruL>cedinguf Inrmuiionr11 Seminar Current Issues and Chalkengus in Food Sqf;?r),

logical and biochemical characteristic of the isolated Table 5. The positive result showed that the bacteria could use citric omonas sp. acid as carbon source for its cell to produce energy. logical and biochemical characteristics Tabel 5. Composition of Pseudolnonas sp. In Grouper fish Fillet 1e result of gram coloration into 72 bacteria strains isolated Tetrimide Agar, showed gram negative characteristic with red Genus/species Strains Amount ,od. ("/o) be result of motility test showed that there were 48 bacteria P. caryophylli FK.la-1, FK.la-2, FK.la-3, FK.ld-5, 6.9 ; gave positive results and the other 24 bacteria gave negative E'K-la-6 ;. Positive result means that tested bacteria had ability to make P. fluorescens FK.la-8, FK.13-9, FK.la-10, FK.la- 11.1 nent or had flagella to move. 12, FK.Ia-13, FK.lb-3, FK.lc-9, le result of oxidase test showed that thet-e are 61 bacteria FK.lc-10 ; had positive result, while I1 other bacteria strains had P, dela Jieldii FK.lb-1, FK.lb-6, FK.lb-7, FK.lb-9, 23.6 ve result. Positive reaction means that thcse bacteria had FK.lb-12, FK.Ic-1, FK.lc-2, FK.lc-3, .om oxidclse enzyme that take apart in aerobic respiration. FK.lla-1, FK.IIa-2, FK.IIa-3, FK.lla- 8, FK.IIa-9, FK.llb-9, FK.Ilc-4, talase test showed that 57 bacteria strains gave pos~tiveresult, FK.I[Ic-11, FK.IIc-12 15 bacteria strains were negative. Positive t-esult means that P. aeruginosa FK.1c-5, FK.Ic-7, FK.Ic-14, FK.IIa- 12.5 :teria produce catalase enzyme. 6, FK.IIb-1, FK.IIb-2, FK.Ilb-3, rmentation tests were done on several sugar medium such as e, glucose, ma1 tose and mannitol. The result from thc tests was There were 14 bacteria strains fermented glucose and :ed air bubble. Whereas 17 bacteria strains fermented glucose The growth characteristic of Pseudomonas sp. ~t produced air bubble, and another 14 strains gave uncertain Todar, (2004) stated that the optimum growth of Pseudomonas were influenced by temperature, salitlity and pH. Base on biochemical lo1 tests of 19 strains showed positive result. This means the tests. 4 strains that had been identified were selected. They were P. s could produce triptophanase enzyme as catalyst in separating Jrrorescens (FK. [a-91, P. aeruginoso, [FK.Ic-53, P. delafieldii, (FK.1 ia-9) loety From triptophan. and P, caryophylli (FK.Ia-1) to represent 72 strains. e result of methyl red-tests was varied. There were 11 strains The growth temperature of the 4 isolates at 00,20, 40, 6") 25") 1 positive result and another 11 strains showed negative and 37" Cis shown in Table 6.

ges Proskauer tests revealed that 10 strains had positive result. !st was conduct to show the ability of bacteria to ferment ydra te and produce acetylme~icarbinol. e citric acid tests showed variation in results. There were e and negative results. The result of the test can be seen in

~sterpresentation poster presentation (351 Scirncc-Bused Approuch for FoodSafev Management Proceeding r$fnternationaf Semifiar Ctrrrcnr Isvues and C*hullrnge,..~in Fooa

Table 6. Growth Temperature of Pseudomonas sp. The growth of Pseudomonas at pH range 5-9 can be seen in Ta' Table 8 shows that Pseudomonas had optimum growth at pH I Temperahare T~me from 5 to 8. No growth was observed at pH 4 dan 10 lsolatcs 00 2" 4- 6" 2Smm 370 [hour) GI U/o GI Yn GI ? GI %r GI '?i, GI 4'0 P. tluorescens 24 8 80 10 100 10 100 10 100 10 100 LO 1017 FK.I~-8 Table 8. Growth of Pseudomonas sp at different pH values P. aerunlnosa 24 7 70 10 100 10 100 10 100 10 100 10 lull FK.IO-SY I P. delafieldii 24 9 90 10 100 10 100 10 100 10 100 10 100 FK.Iia-9 P. caryophylli 24 8 80 10 100 10 1011 10 100 10 100 10 100 FK.la- L GI = Growth lndex

Table 6 and Figure 1 showed that Pseudornonas could grow well at temperature 37" when incubated for 24 hour. The optimum temperature for the growth of microbe was between 2- 37°C. The effect of various levels NaCI on the growth of Pseudornonas Patogen i ty characteristics of Pseudomonas sp. In grouper is presented in Table 7. fillets Haemolysis Test Tabel 7. Growth of Pseudomot~ossp. at different NaCl concentration The results from the test varied. There were beta haemc type (no blood surrounded colony), alpha hemolysis (several t I IT..-... NaCl A - cells found at haemolysis zone or several greenish char surrounded colony] and gamma haemolysis(No haernolysis).

Agglutination test Agglutination test showed varied results. There were poi resut ts and negative results.

GI :Growth lndex Sensory test of Grouper fish fillets

Table 7 showed the effect of NaCl to the growth of Pseudomonas. Sensory tests on mucus, odors, texture showed that groupe All Pseudvmonos isolates could grow well in the absence of salt. from location I showed that they had value of 24,9 and the avt P.delaflelddi, P-caryophylii grew at NaCl concentration of 0-7%. value was > 8. This meant that grouper fish samples from local P.J7uorecensJ grew at NaCl concentration of 0-3%) and inhibited at were categorized as class I (excellent). Their freshness scores qi concentration 5-9%. At level 9% NaCl all isolated were inhibited. were between 21 and 27 with no average values of less thi Therefore it can be concluded that Pseudomonas need 0-7% salt for Meanwhile, grouper fish samples from location 11 had scores fro their growth. to 23,06 with average value was of > 7.

352Iposter presentation poster presentarion I 'usedApproach fbr Food Safeiy Management Proceeding r~lnternaiionalSeminar Current Issues and Chollengw in Food Safiry

Table 6. Growth Temperature of Pseudomonas SP. The growth of Pseudomonas at pH range 5-9 can be seen in Table 8. Table 8 shows that Pseudomonos had optimum growth at pH range from 5 to 8. No growth was observed at pH 4 dan 10 I Table 8. Growth of Pseudornonas sp at different pH values

: Growth Index rable 6 and Figure 1 showed that Pseudornonas could grow temperature 37% when incubated for 24 hour. The optimum 3 ture for the growth of microbe was between 2- 37". le effect of various levels NaCl on the growth of Pseudomona~ Patogenity characteristics of Pseudomonas sp. In grouper fish nted in Table 7. fillets Hoemolysis Test 7. of concentration Growth Pseudomonas sp, at different NaCl The results from the test varied. There were beta haemolysis type (no blood surrounded colony), alpha hemolysis (several blood cells found at haernolysis zone or several greenish changing surrounded colony) and gamma haemolysis [No haemolysis).

Agglutination test Agglutination test showed varied results. There were positive results and negative results. I I I I I I I I I I I i Index Sensory test of Grouper fish fillets ible 7 showed the effect of NaCl to the growth of Pseudomonas.. Sensory tests on mucus, odors, texture showed that grouper fish ~domonasisolates could grow well in the absence of salt. from location I showed that they had value of 24,9 and the average elddi, P.caryophylii grew at NaCl concentration of 0-7%. value was > 8. This meant that grouper fish samples from location I cens, grew at NaCl concentration of 0-3%, and inhibited at were categorized as class I (excellent). Their freshness scores quality :ration 5-9%. At level 9% NaCl all isolated were inhibited. were between 21 and 27 with no average values of less than 8. )re it can be concluded that Pseudomonas need 0-7% salt for Meanwhile, grouper fish samples from location I1 had scores from 21 owth. to 23,06 with average value was of > 7.

ster presentation poster prese~~tation1353 Science- Bmed Appr ouch for Food Safe[v Managen~ent Pmcc.eding oflniurnu~runuISeminor Current Issues unnd c7hullen,y~..sin Foou

CONCLUSION AND RECOMMENDATlON Safety and Applied Nutrition, Office of Seafood, Wnshir DC. Conclusion ht tp:~~seafood.ucdavis.edu/haccp!com~endi~n~/chapt20. The total Pseudomonas found in Grouper fish fillets from Vibrio%ZOsp.p. location 1 was higher than those from location [I. The Pseudomonas ,2002. Cetrimide Agar. EMD. Merck KgaA. Darm content in Grouper fish fillets from both locations was higher than Germany. those in control-live grouper fish. The high content was caused by: handling techniques in processing fillets that had not applied a htt~://www.emdchernicals.com/anaIytics/cro Manual1 correct cold chain system and low hygiene and sanitation process. Sdata/prods/l 10263 0500.html The dominant species of Pseudomonas associated with Grouper ,2003a. Food Safety And Shelfish Program. fish fillets was P. delafieldii with percentage 23,6%.

Pseudornonas was aIso found in control-live Grouper fish and in ,2003b. Bacteriological Analytical Manual , Pseudor sea water samples taken from both locations. The existence of auruginosahttp://www.cfsan.fda.~iov~bam-9.htd Pseudomonas in control and sea water can be used as early warning on the quality of the water in two locations because Pseudomonas had ,2004. Bacteriological A~lalytical Manual On been proven as indicator of polluted water. Pseudomonas. Chap 9. CFSAN-Food And Drugs Administr Washington DC, USA.

References ,2005a. Pseudomonas Infection Afrianto, E. dan E. Liviawaty., 1989. Pengawetan dan Pengolahan Ikan. Kanisius. logyakarta. - ,2005b. Bio 203, General Microbiology & Infection COJ Anonimous, 1992. Standar Nasional Indonesia. 01-0729- 1992: Ikan http://www. rrjcrobio@iubi~.bio.indiana.edu cmicrobi Segar. Pusat Standarisasi Industri. Departemen Perindustrian Jakarta. I ,2005c, Pseudomonas. .1998. Fact Sheeton Pseudomonas sp.. National Food -http:/!www . Universtv of Texas! Houston Medical S Processors Associotion.http://www.nfa- DPALM MEDIC, food.0-c- food.0-c- Austin, B., 1993. Marine Microbiology. Cambridge U~~iversityPrc -200la. Bacteriological Analytical Manual On i.ine, Baumann, P. dan L.R.H.W . Schubert, 1984. Bergey's man^ Pseudornonas Cholcrae, V. parahoemolyticus, K Vuyuniccrs, and Syaternatic Bacteriology. Vol. 1. The Williams and Wilkir Other Pseudomonas Sp.p. Chapter 9. Center For food Safety and Baltimore. Applied Nutrition. Washington, D.C, USA. Berhimpon, S., 1993. Mikrobiologi Pangan Ikani. Laborat Pengolahan Dan Pembinaan Mutu Hasil Perikanan. Fa .2001b. Fish and Products Hazards and Controls Perikanan Dan Ilmu Kelautan. Unsrat. Manado Guide, 3ded. Department of Health and Human Services, Public Boel, T. 2004, Infeksi Saluran Kemih Dan Kelamin, Fa Health Service, Food and Drug Administration, Center for Food Kedokteran Sumatera Utara. Medan.

354Iposter presentation poster presentatlo -Based Approachfor Food Safe@ Management Pro~.eedingof lnrernurional Seminar Current Issues und Chl11Ienge.sin Food Safety

LUSION AND RECOMMENDATION Safety and Applied Nutrition, Office of Seafood, Washington, DC. usion http:f /seafood,ucdavis.edu/hacc_p/compendium/chapt20.htm# The total Pseudomonas found in Grouper fish fillets from VibrioO/o20sp.p. In I was higher than those from location 11. The Pseudomonas ,2002. Cetrimide Agar. EMD. Merck KgaA. Darmstadt, ~t in Grouper fish fillets from both locations was higher than Germany. in cotltrol-live grouper fish. The high content was caused by: ,ng techniques in processing fillets that had not applied a :t cold chain system and low hygiene and sanitation process. The dominant species of Pseudomonas associated with Grouper ,2003a. Food Safety And Shelfish Program. .lets was P. delafieldii with percentage 23,6%. http:~/www.doh.wa.~ov/chp!sf/biotoxinprogrc?m.html Dseudomonas was also found in control-live Grouper fish and in ,2UU3b. Bacteriological Analytical Manual , Pseudomonas rater samples taken from both locations. The existence of auruginosahttp://www.cfsan.fda.~ov/-ebam-9.html pmonas in control and sea water can be used as early warning ,2004. Bacteriological Analytical Manual On Line, r quality of the water in two locations because Pseudomonas had lroven as indicator of polluted water. Pseudomonas. Chap 9. CFSAN-Food And Drugs Administration. Washington DC, USA. ~:!/Jwww.cfsan.fda.gov/-ebam/bam-9.html ences ,2005a. Pseudomonas Infection to, E. dan E. Liviawaty., 1989. Pengawetan dan Pengolahan -.co.id/aurelia at.adu.wash in~ton Ikan. Kanisius. logyakarta. ,2005b. Bio 203, General Microbiology & Infection Control mous, 1992. Standar Nasional Indonesia. 01-0729-1992: Ikan httg://www, [email protected] cmicrobi Segar. Pusat Standarisasi Industri. Departemen Perindustrian Jakarta. ,2005~.Pseudomonas. -.1998. Fact Sheeton Pseudornonas sp.. National Food http://www. Universtv af Texas! Houston Medical School. Processors Associotion.http://www.nfa- DPALM MEDIC. food.Om/mem bers,/science/fact-vibrio.htm1 Austin, B., 1993. Marine Microbiology. Cambridge University Press. .ZOO la. Bacteriological Analytical Manual On Line, Baumann, P. dan L.R.H.W . Schubert, 1984. Bergey's Manual Of Pseudomonas Cholerael V. parahaemolyticus, V. Vu!funicus, and Syaternatic Bacteriology. Vol. 1. The Williams and Wilkins Co., Other Pseudomonas Sp.p. Chapter 9. Center For food Safety and Baltimore. Applied Nutrition. Washington, D.C. USA. Berhirnpon, S., 1993. Mikrobiologi Pangan Ikani. 1,aboratorium http://vm.cfsan.fda.mv!-e barn/m-9.html Pengolahan Dan Pembinaan Mutu Hasil Perikanan. Fakultas .2001b.Fish and Fishery Products Hazards and Controls Perikanan Dan Ilrnu Kelautan. Unsrat. Manado Guide, 3d ed. Department of Health and Human Services, Public Boel, T. 2004, Infeksi Saluran Kemih Dan Kelamin, Fakultas Health Service, Food and Drug Administration, Center for Food Kedokteran Sumatera Utara. Medan.

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Pritchard AE, Vasal ML. 1990. Possible insertion sequences Ruckle,K. A., R. A. Edward$. H.Fltet,M.Wooton.l987.11muPangan. mosaic genomeorganization upstream of the exotoxin A ge. Penerajemah Hari Purnomo clan Adiono.UI Press. Jakarta. Pseudornonas aeruginosa. J Bacteriol 172:2020. Cappucino and Sherman., 1992. Microbiology: A Laboratory Manual. Singleton, P., dan D. Sainsbury, 1987. Dictionary of Microbioiog 3'6 ed. The Benjamin/Cummings Publishing, Inc. California. Molecular Biology. Znd ed. John Wiley and Song. Ncw York, I Droop, MR dan H.W. Jannasch 1977, Advance In Aquatik Todar K, 2004, Text Book of Mikrobiology, Pseudomonos aerugi Microbiologi,Vol I. Academic Press,Inc New York Department of Bacteriology. University of Wisconsin-Mad Feliatra, Irwan Efendi, Edwar Suryadi, 2004. lsolasi dan Identifikasi http://www.textbookofmicrobiolo~v Bakteri Probioti k dari Ikan Grouper fish Macan (Ephinephelus fuscogatus) dalam Upaya Efisiensi Pakan Ikan. Jurnal Natur Varnam, A.H. dan Evans, M.C., 1991. Food Born Patogen Indonesia 6[2): 75-80 (2004) ISSN 1410-9379.Jakarta. Illustarted Text. Moosby Year Book, Inc. Toron to, Canada. Zdzislaw, E.S., 1989. Seafood: Resources, N utt-itional Compo: Frazier , W.C. dan Westhof, D.C., 1998. Food Microbiology. 4th Edition. Mc Gram Hill Book co. New York. And Preservation. Departement of Food Preservation Technical M icrobiologi. echnical University Plythcni ka Gda Fix, D. F. 2004. Pseudomonas, h ttp://www, ches,siu.edu/fix/medmicro Poland. yseud.htm. 22 Juli 2004 Prrkui 13.00 wita. Hadiwiyoto, S., 1993. Teknologi Refrigrasi Hasil Peri kanan. Penerbit Liberti. Yogyakarta. Huss, H., 1995. Quality and Change in Fresh Fish. FAO. Technical Paper. Fisheries Library. Denmark. Ijong, F.G,, 2002. Mikrobiologi Dasal-. La boratorium Mikrohiologi Hasil Perikat~an.Jurusan Pengolahan Hasil Perikanan. Fakultas Perikanan dan IImu Kelautan. UNSRAT. Manado. I Ilyas, S., 1983. Teknologi Pendinginan Ikan. jilid I. Penerbi t Yayasan Wijaya Kusuma. Jakarta. lawetz, E., Melnick, J.L., Adelberg, E.A. 1986. Mikrobiologi. Edisi 16. Peuerbit Buku Kedokteran. Jakarta. Lay, B., 1994. Analisis Mikroba di Laboratoriurn. PT. Raja Grafinclu Persada. Jakarta. Matthysee,A.G, 1992. Ad. (Translated from japan) A.A. Ralkerna. Roterdam.Adhesion Bacterial. Encyclopedia of Microbiology Volume l.A-C .Academic Press,lnc Harcourt Brace lovanovich, Publishers, San Diego,California. Moningka, M.R., 2005. Efek Pencucian Terhadap Daya Lekat Escherichia coii Pada Beberapa Produk Perikanan. Skripsi. FPIK IJNSRAT. Manado.

1 3561poster presentation Proceeding uflnrernuriutrill Seminar Ctrrre~~tIssues and Chall~ngcsin Food Suj.er)r - Bust-d Approuch for Food Salery Management

Pritchard AE, Vasal ML. 1990. Possible insertion sequences in a :,K. A., R. A. Edward&. H.Fleet,M.Wooton.l987.11mu Pangan. mosaic genorneorganization upstream of the exotoxin A gene in Penerjemah Hari Pu rnomo dan Adiono.UI Press. Jakarta. Pseudomonas aeruginosa. j Bacterial 172:2020. cino and Sherman., 1992. Microbiology: A Laboratory Manual. Singleton, P., dan D. Sainsbury, 1987. Dictionary of Microbiology and 3rd ed. The Benjamin/Cummings Publishing, Inc. California. Molecular Biology. 2nd ed. John Wiley and Song. New York, USA. MR , dan H.W. Jannasch 1977, Advance In Aquatik M Microbiologi,Vol 1. Academic Press,lnc New York Todar K, 2004, Text Book of ikrobiology, Pseudomonas ueruginosa, Department of Bacteriology. University of Wisconsin-Madison. ,a, lrwan Efendi, Edwar Suryadi, 2004. Isolasi dan ldentifikasi httn: textbookofmicr of microbiology 3akteri Probiotik dari Ikan Grouper fish Macan (Ephinephelus Varnam, A.H. dan Evans, M.G., 1991. Food Born Patogens an uscogatus) dalam Upaya Efisiensi Pakan Ikan. jurnal Natur Illustarted Text. Moosby Book, Inc. Toron to, Canada. ndonesia 612): 75-80 (20041 ISSN 1410-9379. Jakarta. Year Zdzislaw, E.S., 1989. Seafood: Resources, Nutritional Composition r , W.C. dan Westhof, D.C., 1998. Food Microbiology. 4th Edition. And Preservation. Departement of Food Preservation and ulc Gram Hill Book co. New York. Technical Microbiologi. echnical University Plythenika Gdanska. F. 2 004. Pseudomonas. A ttp://www,ches.siu.edu~fix/medmicro Poland. 1seud.htm. 22Juli2004 Pukul13.00 wita. iyoto, S., 1993. Teknologi Refrigrasi Hasil Perikanan. Penerbit Aerti. Yogyakarta. H., 1995. Quality and Change in Fresh Fish. FAO. Fisheries 'echnical Paper. Fisheries Library. Denmark. F.G., 2002. Mi krobiologi Dasar. Laboratorium Mi krubin!ogi iasil Perikanan. Jurusan Pengolahan Hasil Perikanan. Fakultas 'erikanan dan Ilmu Kelautan. UNSRAT. Manado. ;., 1983. Teknologi Pendinginan Ikan. Jilid 1. Penerbit Yayasan Vijaya Kusuma. Jakarta.

, E., Melnick, J.L., Adelberg, E.A. 1986. Mikrobiologi. Edisi 16. 'enerbit Buku Kedokteran. Jakarta. , 1994. Analisis Mikroha di Laboratorium. PT. Raja Grafindo 'ersada. Jakarta. see,A.G, 1992. Ad. (Translated from Japan) A.A. Balkema. .oterdam.Adhesion Bacterial. Encyclopedia of Microbiology 'olurne l.A-C .Academic Press,lnc Harcourt Brace jovanovich, ublishers, San Diego,California. ;ka, M.R., 2005. Efek Pencucian Terhadap Daya Lekat scherichia coli Pada Beberapa Produk Perikanan. Skripsi. FPIK -NSRAT. Manado.

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