Laporan Praktikum
Hari/Tgl : Senin, 12 Oktober 2015
Analisis Mutu Mikrobiologi Pangan
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: Ai !mas " Stp#Ms$
Asis Asiste ten n
: in inaa %ro %ro&n &nia ia,, Am' (e)ita Permata Hati, Am'
UJI MIKROBIOLOGI SUSU isusun ole*: +elompok / -P2
%e$ep Sumantri
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Arista Hanu'iana
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Tina ameria S
.1103
Al4ina Saikani
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PROGRAM KEAHLIAN SUPERVISOR JAMINAN MUTU PANGAN PROGRAM DIPLOMA INSTITUT PERTANIAN BOGOR 2015
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BAB I PENDAHULUAN 1.1 Latar Latar Belaka! Belaka!
-a*an pangan maupun pro'uk ola*anna umumna 'apat 'itumbu*i ole* satu atau lebi* 6enis mikroba# A'ana mikroba pa'a pangan kemungkinan 'apat menebab menebabkan kan kerusa kerusakan kan pangan pangan atau atau ba*kan ba*kan 'apat 'apat memba*a memba*aak akan an kese*at kese*atan an konsumen# -a*an ang 'igunakan pa'a praktikum kali ini a'ala* susu# Susu a'ala* suat suatu u sekr sekres esii ang ang komp kompos osis isin ina a sanga sangatt berb berbe' e'aa 'ari 'ari kompos komposis isii 'ara* 'ara* ang ang merupak merupakan an asal asal susu# susu# Misaln Misalnaa lemak lemak susu, susu, casein, casein, laktos laktosaa ang ang 'isint 'isintesa esa ole* ole* alveoli 'alam alveoli 'alam kambing, 'an ti'ak ter'apat 'i tempat lain manapun 'alam 'alam tubu* sapi 7Mu$*ta'i 'kk 20108# Menuru Menurutt Mu$*ta' Mu$*ta'ii 'kk 720108 720108 susu a'ala* a'ala* $airan $airan ang ang 'i*asi 'i*asilka lkan n ole* ole* kelen6ar susu 7mammae 7mammae88 baik 'ari binatang maupun 'ari seorang ibu# Air susu ibu 'ikenal 'engan AS!, AS!, se'angkan susu *e&an atau susu tiruan sebagai pengganti pengganti susu ibu 'isebut PAS! 7Pengganti Air Susu !bu8# Susu memiliki aroma ang ringan 'an umumna 'apat 'iterima orang9orang 'i &ilaa* perkotaan# i pe'esaan, sebagian pen'u'uk memang menolak untuk minum susu 'engan alasan berbau amis 'an ti'ak biasa# amun susu 'engan nilai gi;ina tinggi merupakan makanan penting bagi mereka ang 'alam kea'aan sakit 7-e$k 1<<8# =6i mikrobiologi ter*a'ap susu perlu 'ilakukan untuk mengeta*ui mutu susu sebelum 'iola*# Meto'e ang 'ilakukan untuk mutu mikrobiologi susu, aitu meto'e Mikr Mikros osko kopi pik k
Lang Langsu sung ng
7M% 7M%88
'igu 'iguna naka kan n
untu untuk k
meng mengan anal alis isis is
susu susu
ang ang
mengan'ung bakteri 'alam 6umla* tinggi, meto'e M-(T 7 Methylene Blue Reduction Redu ction Test 8 untuk memberikan gambaran perkiraan 6umla* bakteri ang ter'apat 'alam susu, susu, meto'e meto'e (esa;u (esa;urin rin,, meto'e meto'e pengu6i pengu6ian an menggun menggunakan akan alko*ol alko*ol,, 'an pengu6i pengu6in n se$ara )isual#
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1.2 T"#"a
Tu6uan 'ari praktikum ini a'ala* untuk mengeta*ui kualitas susu 'engan $ara melakukan pengu6ian se$ara mikrobiologi aitu u6i meto'e breed , pengu6ian alko*ol, u6i M-(T (Methylene Blue Reduction Test), u6i (esa;urin pa'a susu segar 'an susu ang su'a* 'itamba*kan mikroba 'an u6i )isual#
BAB II METODOLOGI 2.1 Ba$a %a Alat 2.1.1 Ba$a
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Alko*ol Minak imersi Pe&arna biru metilen (esa;urin Pe&arna biru metilen tiosianat
• • • • •
2.1.2 Alat
Alat9alat ang 'igunakan: -unsen .arum ose Mikroskop Mikro pipet ukuran 0#01 ml +artu penolong berukuran 1 $m2 >elas Ob6ek Tabung Tabung reaksi Penangas air bersu*u 3 ? %
• • • • • • • •
2.2
Pr&'e%"r Ker#a
Meto'e Breed Meto'e Breed 0,01 ml
0,01 ml
susu
susu
1 $m
1 $m 1
1
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ibuang kelebi*an ;at &arna 'engan kertas serap
ibiarkan kering
ibilas 'engan air
ikeringkan 'iu'ara
iamati 'i ba&a* mikroskop
itamba*kan minak imersi
i*itung 6umla* bakteri per areal pan'ang
Pengu6ian Alko*ol 5 ml susu
@ alko*ol 0
iamati penggumpalan
=6i M-(T (Methylene Blue Reduction Redu ction Test) Test) 10 ml susu
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ibalikan sebanak kali
itempatkan Water bath 3 o%, 'i$atat &aktu mulaina
Setela* 5 menit 'ibalikkan untuk men$ampur ;at &arna
iamati peruba*an &arna setiap 0 menit
i$atat M-(T 7s/' kali pengamatan8
=6i (esa;urin 10 ml susu
@ 1 ml (esa;urin
iko$ok
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BAB III HASIL DAN PEMBAHASAN (.1 Ha')l U#) V)'"al
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U#) Re*a*"r) Kel&+,&k Ga#)l -Sa+,el '"'" UHT E.coli /
+elompok
Hasil Pengamatan Sebelum 'itamba* Sesu'a* 'itamba* (e;a;urin
1 5
Puti* Puti* Puti* Puti*
Setela* 'i Water
(e;a;urin Ciolet Ciolet Ciolet Ciolet
Pink Pink Pink Pink
batch
Kel&+,&k Ga#)l -Sa+,el '"'" UHT + E.coli /
+elompok
Hasil Pengamatan Sebelum 'itamba* Sesu'a* 'itamba* (e;a;urin
1 5
Puti* Puti* Puti* Puti*
Setela* 'i Water
(e;a;urin Ciolet Ciolet Ciolet Ciolet
batch -iru -iru -iru -iru
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+elompok
Su*u susu 7kontrol8
Su*u D%
Hasil Pengamatan
D% 2
D 0D D D D D D D 0D D D D D D D 0D D D D D D D
0D ke9 0D ke91 0D ke92 0D ke9 0D ke9 0D ke95 0D ke93 0D ke9 0D ke91 0D ke92 0D ke9 0D ke9 0D ke95 0D ke93 0D ke9 0D ke91 0D ke92 0D ke9 0D ke9 0D ke95 0D ke93 0D ke9
-iru memu'ar -iru lebi* pekat -iru se'ikit memisa* -iru se'ikit memisa* -iru memu'ar -iru memu'ar -iru memu'ar -iru memu'ar -iru lebi* pekat -iru lebi* pekat -iru memu'ar -iru memu'ar -iru memu'ar -iru memu'ar -iru memu'ar -iru lebi* pekat -iru -iru -iru memu'ar -iru memu'ar -iru memu'ar -iru memu'ar
Kel&+,&k Gea,
+elompok 5
Su*u susu 7control 8 0D
Su*u 0D ke 1
Hasil Pengamatan -iru se'ikit memisa*
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+elompok
(
Su*u susu 7control 8 D 0D D D D D D D 0D D D D D D D
Su*u 0D ke9 0D ke91 0D ke92 0D ke9 0D ke9 0D ke95 0D ke93 0D ke9 0D ke91 0D ke92 0D ke9 0D ke9 0D ke95 0D ke93 0D ke9
Hasil Pengamatan -iru memu'ar -iru se'ikit memisa* -iru lebi* pekat -iru memu'ar -iru memu'ar -iru memu'ar -iru memu'ar -iru memu'ar -iru pekat se'ikit memisa* -iru se'ikit memisa* -iru se'ikit memisa* -iru memu'ar -iru memu'ar -iru memu'ar -iru memu'ar
U#) Bree%
+elompok 1 5
Sampel ang 'igunakan Susu =HT @ #$oli
.umla* bakteri per ml 1#0 E 10 # E 1093 1# E 10 3
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terli*at ber&ana kekuningan# Susu memiliki &arna puti* kekuningan atau bisa 6uga 'isebut 'isebut puti* tulang pekat ang 'isebabkan 'isebabkan ole* a'ana kan'ungan lemak 'an beta karoten karoten ang menebabkan menebabkan susu itu ber&arna kuning, kuning, se'angkan &arna puti*na puti*na merupak merupakan an *asil *asil 'isper 'ispersi si $a*aa $a*aa 'ari 'ari butira butiran9bu n9butir tiran an lemak, lemak, protein, protein, 'an mineral mineral ang a'a 'i 'alam susu 7+*osman 2008# Susu se'ikit manis 'an asin 7guri*8 ang 'isebabkan a'ana kan'ungan gula laktosa gula laktosa 'an garam 'an garam mineral 'i 'alam susu tersebut# Arom Aromaa amis amis ini ini 'ise 'iseba babka bkan n karen karenaa pa'a pa'a susu susu bubuk bubuk 4ull 4ull $ream $ream lebi lebi* * ban banak ak mengan'ung lemak# 5.2. 5.2.2 2
Pe! Pe!"# "#)a )a Alk& Alk&$& $&ll Prinsip 'asar pa'a u6i alko*ol merupakan kestabilan si4at koloi'al protein
susu tergantung pa'a selubung atau mantel air ang menelimuti butir9butir protein terutama kasein# =6i alko*ol bertu6uan memeriksa 'engan $epat 'era6at keasaman susu# -ila alko*ol 7bersi4at 'e*i'rasi8 'i$ampurkan 'engan susu, maka akan ter6a'i koagula koagulasi si protei protein n se*ing se*ingga ga pe$a*an pe$a*an9pe$ 9pe$a*a a*an n susu susu akan akan tampak tampak## Semaki Semakin n tinggi tinggi 'era 'era6a 6att asam asam susu susu,, sema semaki kin n bana banak k 6uml 6umla* a* alko alko*ol *ol ang ang berk berkur urang ang 'eng 'engan an konsent konsentars arsii alko*ol alko*ol ang ang sama sama 'alam 'alam meme$a* meme$a*kan kan susu susu ang ang sama sama banakn banakna# a# Menuru Menurutt Ma*es& Ma*es&ari ari 72008 72008 susu susu ang ang mengan' mengan'ung ung lebi* lebi* 'ari 'ari 0,21 0,21 asam asam atau atau kalsium 'an magnesium 'alam 6umla* tinggi, akan terkoagulasi 'engan penamba*an alko*ol# =6i alko*ol alko*ol sampel sampel susu susu A ang 'itamb 'itamba*ka a*kan n alko*o alko*oll 0 ti'ak ti'ak ter6a' ter6a'ii penggumpalan# Menurut Su'ono 'kk 720058 susu ang positi4 pa'a u6i alko*ol ini menan'a menan'akan kan susu susu mulai mulai masam, masam, ter'apa ter'apatt kolost kolostrum rum 'an kemungk kemungkina inan n a'ana a'ana
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ren'a* 'an sebalikna 6ika &arna susu tetap ber&arna biru berarti susu tersebut memiliki mutu mutu 'an kualitas ang bagus 'an memiliki se'ikit 6umla* bakteri ba*kan ti'ak a'a bakteri ang mere'uksi resa;urin tersebut# Pengu6ian 'ilakukan 'engan memasukkan 10 ml sampel susu ke 'alam tabung reaksi kemu'ian 'itamba*kan 1 ml larutan (esa;urin 'an 'ipanaskan 'i 'alam water bath 'engan bath 'engan su*u 3o% selama 15 menit kemu'ian 'i ko$ok# Setela* itu 'ipanaskan kembal kembalii 'i 'alam water bath selama bath selama 1 6am kemu'ian amati peruba*an &arna ang ter6a'i pa'a masing9masing sampel per 15 menit selama 1 6am# -er'asarkan *asil ang a'a, 'ari ke'ua sampel 'iketa*ui ba*&a mutu sampel A 7susu segar8 lebi* bagus 'iban'ingkan sampel - 7@ E.coli). Hal E.coli). Hal ini 'apat 'ili*at 'ari peruba*an &arna ang an g ter6a'i, Sampel A &arnana tetap atau ti'ak beruba* 'ari a&al per$obaan aitu &arna biru# Se'angkan S e'angkan untuk sampel - mengalami peruba*an &arna 'ari biru pink men6a'i men6a'i &arna mera* mu'a 'i permukaan permukaan atau ba*kan men6a'i &arna puti*# Menurut &i'6oseputro 72008 sampel ang ti'ak mengalami peruba*an &arna apabila 'itamba*kan pereaksi (esa;urin berarti sampel tersebut memiliki kualitas
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methylene methylene blue akan blue akan beruba* men6a'i methylene white white 'imana menan'akan 6umla* akti4itas mikroba ang meningkat 'engan $epat# Ter'apat beberapa 4aktor ang 'apat mempeng mempengaru aru*i *i *asil *asil ak*ir ak*ir 'ari 'ari analis analisis is resa;u resa;urin rin ini, ini, antara antara lain lain kesala kesala*an *an pa'a pereaksi, 'an pengaturan su*u pa'a water bath# bath# Test -MBRT/ 5.2. 5.2. Met& Met&%e %e Methyl Blue Reduction Test Sala* satu meto'e pengu6ian mikrobiologi susu selan6utma a'ala* u6i biru metilen atau M-(T# Meto'e ini 'ilakukan untuk memberikan gambaran perkiraan 6umla* bakteri ang ter'apat 'alam susu# alam u6i ini 'itamba*kan se6umla* ;at &arna ke 'alam susu, susu, kemu'ian kemu'ian 'iamati &aktu &aktu ang 'ibutu*kan ole* ole* bakteri untuk melakukan akti)itas ang 'apat menebabkan peruba*an &arna tersebut# Semakin tinggi 6umla* bakteri ang ter'apat 'alam susu, semakin $epat ter6a'i peruba*an &arn &arna# a# +ele +elema ma*a *an n meto' eto'ee ini ini ti'a ti'ak k prak prakti tiss 'ila 'ilaku kuka kan n ter* ter*a' a'ap ap susu susu ang ang mengan'ung bakteri 'alam 6umla* se'ikit, meto'e ini ti'ak 'apat membe'akan 6enis bakteri ang ter'apat 'alam susu 'an analisis membutu*kan &aktu ang relati4 lama# -er'asarkan -er'asarkan *asil pengamatan, pengamatan, ba*&a sampel susu setela* setela* 'iamati 'iamati ter6a'i ter6a'i peruba*an &arna 'ari &arna biru pekat men6a'i &arna &a rna biru memu'ar# Prinsip 'ari u6i
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alat9alat pemera*an ang kurang bersi* 'an tempat9tempat penimpanan ang kurang bersi*, 'ebu, u'ara, lalat 'an penanganan ole* manusia 7-u$kle et# al 2008# Se'angkan pa'a sapi ang ter6angkit ra'ang 7mastitis8 'apat 'i6umpai bakteri 'ari 6enis
Cory Coryne neba bact cter eriu ium, m,
Sta Stahy hylo loco cocc ccus us
s. s.,,
Str Stretoc tococ occu cuss
s. s.,,
'an
Enterobacteriaceae ang 6umla*n 6umla*naa 'apat 'apat men$apa men$apaii 105 %"=/ %"=/ ml susu# susu# Apabila Apabila kontaminasi berasal 'ari lingkunganna 7tana*, rumput, 'an pakan8, 6umla* mikroba 'alam susu 'apat men$apai lebi* 'ari 103 %"=/ ml#. ml#. 7Biniati P# (a*au 'an %#%# ur&itri, 20128 Susu 6uga memiliki ka'ar air 'an protein ang tinggi se*ingga memungkinkan mikroba tumbu*# Susu ang 'isimpan 'ingin pa'a su*u kurang 'ari 50 ˚% selama 192 *ari, kontaminasi mikroba 'isominasi ole* bakteri ta*an 'ingin atau psikrotropik# -akteri psikotropik ini ter'iri 'ari bakteri >ram negati4 'an >ram positi4# %onto* bakteri psikrotropik >ram negati4 ang mengontaminasi susu, antara lain !seudomanas, "lavobacterium, #lcali$enes, #lcali$enes, 'a ' an #cinetobacter. Sement Sementara ara itu, itu, bakteri psikrotropik >ram Positi4
mengontaminasi
pa'a
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bakteri patogen# Sapi ang terserang mastitis, susuna susun a biasana mengan'ung sel9sel 'ara 'ara* * puti puti* * 'alam 'alam 6uml 6umla* a* ting tinggi# gi# +eunt +euntun ungan gan meng menggu gunak nakan an meto meto'e 'e ini ini ait aitu u per*itungan $epat 'an bakteri langsung 'i*itung menggunakan
mikroskop#
Se'angkan Se'angkan kelema*an meto'e ini aitu ti'ak 'apat 'ilakukan 'ilakukan ter*a'ap susu ang 'ipasteurisasi karena se$ara mikroskopik ti'ak 'apat 'ibe'akan antara sel9sel bakteri ang masi* *i'up atau ang tela* mati karena perlakuan pasteurisasi# alam praktikum u6i analisis mikrobiologi susu 'engan meto'e M% atau meto'e breed ini ini 'igunakan 2 sampel, aitu sampel susu segar 7sampel A8 'an susu ang 'itamba* bakteri 7sampel 7sampel -8# -er'asarkan -er'asarkan *asil pengamatan, pengamatan, 'i'apat 6umla* bakteri pa'a pa' a sampel A a'ala* 0 sel/ml, se'angkan untuk pengamatan sampel - ang 'ilakukan ole* kelompok gan6il aitu kelompok 1,,5, 'an 6umla* bakteri ang tumbu* berturut9turut 1#0 E 10, , E 103, 1# E 10 'an 1# E 103# -er'asarkan *asil per*itungan 'apat 'ili*at ba*&a sampel A ti'ak mengan'ung bakteri karena sampel A merupakan susu *asil proses =HT# Susu =HT 7ultra 7 ultra hi$h temerature8 temerature8 merupakan susu ang 'iola* menggunakan pemanasan 'engan su*u tinggi 'an 'alam &aktu ang
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BAB IV KESIMPULAN DAN SARAN .1 Ke')+,"la
=6i mikrobiologi mikrobiologi susu menggunakan menggunakan 5 meto'e meto'e aitu u6i )isual, u6i alko*ol,
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breed atau atau meto'e *itungan langsung 7M%8 aitu 'engan $ara meng*itung 6umla* mikroba ang bera'a pa'a suatu ruang ang 'apat 'ili*at 'engan mikroskop# Hasil 'ari meto'e *itungan langsung pa'a sampel A 7susu segar8 ti'ak ter'apat 6umla* mikroba ang teramati se'angkan pa'a sampel - 7susu @ E.coli @ E.coli88 6umla* bakteri ang tumbu* berturut9turut 1#0 E 10, , E 103, 1# E 10 'an 1# E 103
4.2 Saran
alam alam pengu6i pengu6ian an mikrob mikrobiol iologi ogi susu, susu, analisi analisiss 'ilakuk 'ilakukan an sebaik sebaikna na 'alam 'alam kea'aan *igienis agar 'i'apatkan *asil ang baik 'an ter*in'ar 'ari kemungkinan
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