Observations on the growth requirements of non-pathogenic Neisseria in supplemented casein hydrolysate and chemically defined media

216 76 6MB

English Pages 70

Report DMCA / Copyright

DOWNLOAD FILE

Polecaj historie

Observations on the growth requirements of non-pathogenic  Neisseria in supplemented casein hydrolysate and chemically defined media

Citation preview

m u n itto m

o i m tactfnt l i i m s i w of mmsbsu x i m r as© m m a m s . •

cash*

anrXMBD » $ I A

%

rV"'

J o N l £»* ^00811

’ShhiIo oubwjtttod to th* foooltsr of Mrttteata School of itwi %S*«rBltgr of taaryCUral Is partial. fuXillliwmfc of tlio rm ^drm m ^B for the 4*gr«» of Sfoetor of IM&wophy X®50 i> s

UMI Number: DP70506

All rights reserved INFO RM ATION TO ALL USERS The quality o f this reproduction is dependent upon the quality of the copy subm itted. In the unlikely event that the author did not send a com plete m anuscript and there are m issing pages, these will be noted. Also, if m aterial had to be removed, a note will indicate the deletion.

Dissertation Publishing

UMI DP70506 Published by ProQ uest LLC (2015). C opyright in the Dissertation held by the Author. M icroform Edition © ProQ uest LLC. All rights reserved. This w ork is protected against unauthorized copying under Title 17, United States Code

P r o Q u e st ProQ uest LLC. 789 East Eisenhower Parkway P.O. Box 1346 Ann Arbor, Ml 48106 1346

Umi jKHttar. Motes to express ht» otoeere agproeioftUii to Br# Miobael «l* fm km m § J r m$ f o r #«§;f**«t$ti$ ^

M b m m tm m *

t i r e eritic iasa# tiurlng th e course o f th i* •mmtk a«! M e to d ies* p e tie e e e l a eoriclisg wttfc th e etsbjeet aad th e stu d e n t toward* th e o o epletion o f t h i s th eo to f g ra titu d e to Dr, 1* % l a t e s t* f o r h i# under Btarsding end counsel during th e program mi th e work end th e s is ppepersttoBii end

to %* d , B* ?«her;f dr** fo r

texig th e f a o i l t t t e # ' o f th e d e p ert^ e o t atsS Mo ptti«ft«% l i M t l e s s a t tin#** during th e ten u re o f t h i s p ro jec t*

t m m o f c a im a n s P & f#

chaxts&

t* m sm iB A i, i m f t o r a r a o i i ,,,,.

i

.

GBAJPTRK IX. CMLTORlt AMD BtOCBB&CAL CHARACTERISTICS OF TOE OKWS a jOSSKBXA. ........................ ............. A, iforphologiesi Ch a r a c t e r i s t i c s , , , , . . . . . . . . . 8 Methods o f i s o l a t i o n , 10 Stock c u l t u r e s * , . . 10 P reparation o f ln o e u lt,* * ..................... 11 Colony Morphology and sta in in g r e a c t i o n s . * 11 Pigm entation. ................................................ 11 14 Figment i s o l a t i o n * . * . * «•••*«• *• 14 B. BioohoMic&l C h a ra c te ris tic s Carbohydrate fe rm e n ta tio n s. C&talase t e s t , * Oxidase t e s t R e s u l t s . . . . . ............. ............

....................................... 16 • • « . . . » . , • 1? ............. . . . 17 . * . . 1? 17

€-. D irect Olaaceharid# FSnseiiteticm .

c m ? m i n , n m m wAmmmms Prop&ration P reparation P reparation Composition R e s

of of of of u

of

..........

h i mmm m is s m u

22

* .* ..* ,* * a?

i m e c m l n s a * * , 27 t e s t m e d i u m . . . . , , . , . . . . . . .......... 27 irltaadn s o l u t i o n s , . , , . . . . . . 2 6 b asal medium**,*... . . . . . . . . . • • * « • • « • * * # 29 l t s . . . . • . . . *• *. *** 30

CHAPm i t . m m acib m gm nm M n or mu n u ssm u ........ mAFfm ? , mrmnoma, sb b s tiw tfs urn m o m iw m u m of A. B. C. B, B*

42

......................................................................... 49 B to tin Replacement by P ssth ld b to ttiw • • • • • • 5 1 Hon-replacement of B io tin by O leic A c i d , . , , . . . * , . . . , . . 52 Boiv-replaeensnt of Bio t i n by Plm ellc A dd and B rea*.,* 53 9Lotln S u b s titu tio n by O leic Add plus B r e a . 55 l i o t i n S u b s titu tio n by O leic .Acid and O leic A dd plus C reati n o . , . . . , ......... $4

m m .m 0:1 c o n clu sio n s,

,* .* .* ..........

* 58

BIBUOlus BioUnt 1 Coaplate Medina rise IboflaTln and Folic Acid, and in a natural M m . . . . * . . . . * . . . . . . . . . * . . * 33

Table onorrhoea# osat&iniaf glutathione, gltiiaalc a c id and Htatid&n# a# t& tregenoas sonro## bat indicated 'ft# media* was f a r froa optical f o r the growth o f ft® gonococcae* W«ltoaf Stoidnger and Carpenter (1944) jsrepared a secHim fo r ft# c u l t l v a tic * o f J|» m m m rfam m , u sin g , a# n itro g en eetareee, g ly cin e, c y s tin e , indol#»3«aoctlc a d d * glu tertic a c id , le w d n e , a rg in in e , h i s t id i n e , c o ftlo o ln # and pralin e*

vjpen occasion.

«TP**» ©?*«*&«*•

as

and eholin® and all strains

mm

jc sapw^s w»tiw

? I 11

thin a*citat was supplanted vlth gl^mdm

J» s&m#*'* s

i

% bmm noted tbovt* H alted .growth

hm m w r, is finrailsfeXe an the

o f the »eaH3oHi#g€nlt le ls s e ria epeetes •* species

other the® M# m m s lm m « * I*

*M m ** the rep o rt! in

lite ra tu re iq p lied a wore eoopilsx growth r«*plres*#ni of these epeeies to o M ltleo to e look of i n t e r e s t In thw o nm*»3*thages&e Qeieaerla* la view iif1 the H alted la fm a itio a im tllabl* as the $renfth lofniifeaeot* o f the gmmm ialsso g ta- ether than M. ^oaarifcosoo uni I* aealiiflfttldisf the preeent etadljr taw

with these ehjeotiwso*

{1} to fe fia e tho

mit&ml eltasd a rm^idLrmsm^m of the m>e~p&thogeftie lslaa#rl& anO (2) to ftttospt to eeltlwmto these iljB&soagha species la a e se p le te l/ sjjm thetle a M Io r*

m A P tm

m vnm m

aw

12

m txm m C A t C M M e m is fic s m

we

m m s mmmm&

A# sferpbeto^eal Characrterlott## tli# o lo s s lf la a tlo a o f toe p*«a negative ##®**A o f th e normal iwe®~ h m received cmmidMmbl® a tte n tio n in the p in t knd to e divergent miew# have a tta in e d promimmm*

%« f i r s t view 1# m btidled l a the

mtrrewt AarpMf.!# iiamml g f I&toradj^&tl##. Bafttorlailafpr and represent# m

a t t e s t to olaeeifjr its# p s negatlv# cooei, th e H#i###riar a# se p ara te e n titie s w ith spaed## d if f e r e n tia tto n m m g th e e x is tin g and rep o rted ettlttsr##*

% e m w m d r im mm suggested tgr Wilson (1928) a# .follow#!

***** tfeiero l# f a t p re s e n t, l i t t l e ju s tlf ie a iie i* fo r the r#co£?» s&ilon o f sep arata opeeie# th# gram n egative eo#et*«*wlth the p o ssib le exeaptU * o f & emM r r h a lis . * • shomld, th # r# fsr# 9 oog&lii# tli# ra a a ia la g type® in to a sin g le species, with some apgaropriai# xssm auch a# £• I b is v ia # -ban considered tb s w t a t l a n # in ooleiiiiil ftsrpfeelogy and pt#» enwifcctie# rep o rted % f l e e r and Mwiteoii (1909)f Gordo# (1925) m d i ll# # # C1920-) and had adopted th e een##rvati** s t i lt e d # In taxaoMQr* % # organ!*## stu d ied i n te l# i m a t l g o t l a B wore o f Vm gtm m

flei#mrpftf shrew#ganto and

i®# oorbble* no&'pathegenie a id

characterised a# ^growing wall on ordinary mil tors media*?f Altooogh thro# of to# strain# (JJ* flavgseens 974fe, 15.5# 157) w w iootetod ftw d»flt&to oases of meningitis# th# entire group of organism® fttedtod war# considered I*

mud m stra in s o f e ith e r £* mMSEhiMM. ®r war# iaolndod*

th # '.« altn re 4m®tgn&temm

of the oulter# # a re l i s t e d i n W l s 'l #

toe aatrooo

tm e 1

of Qv&tonm

Scwroa rad txVtti£4 r ito liie h g fd r^ lo rid e td a e in M a tllle d w ater to

. 4 .0 10O0.0 1005*0 1000*0 1000*0 100*0

Merograis© aierograine n tereg ran * M eregraa* ntereg ran* ml

pH 7 .1

^HSSeir* hpdwii^^ laltljm re,

^fairy Keeearoh Mfeoraiortm f Ttomtgjemtmd m le t #6*

* Halt eelntion contained dtaodlwa phosphate* 5 g$ petaaeimi ©felorldef 0*09 g | aodttm dtdbleride* 0 .0 gf mgsseelmB stslfete, 0 .0 6 g$ w ater to ISO wSU

Hm m & lw mm

m follesef

Hie easetn

fin e st* * opeteiiie end tryptophan* w e dinsolved in lose than Urn t o ta l w®tmw o f v o ter*

th e s a l t

t© the ossein hydralysate Mixture*

« n prepared separately m i aided

The aedtae urns nad# op to m \v m

w ith m tlm m nm imp the mXwmm of sltaMin* to b® added la te r* ad ju ste d to the denlsed degr©# o f a e i ii i y tar esene o f a M

m

p i ootor and

s te r ilis e d % f tllr a tts t* thooogt fyrex sin te re d g la ss f 11 tors*

nsdlan a t ib is p e ta l was c a lle d H is toot m dtwn*

the

s te r ile so lu tio n # of

tho wltae&isa s ir s added o o optloall/ to His toot astftoi sad the esaplete aedlen

6m afttr»tod mmimm

with a d u s ts cKMtntratlofi of I t 10® storopoao of nUmLtu 'Cmtmm 4 ro o «itl* tlm o dot* aro utifwm to tigw m 3*

a*

* > '" ru-fr \ ■? A. ■

K.i: ..

50-

.r!t-- :r

40-

30-

-4

MICROGRAMS OF NIACIN

fig u re 3 .

Response o f fiv e s tr a in s o f N eisseria to graded amounts o f n ia c in in th e presenee o f b io tin in a c ase in h y dro ly sate Kseditw,

m kF fm i f

*mmmm

or m wragau

th e aaino ao id req u trw M fts o f th e g q liM fts o f th e *»ww*I naso­ pharynx I i n been o n rsp srted exoept f o r th e r « t o w i o f O rdal h Bunch (2946)*

K epertlag on jj|* sic g a r th ese au th o rs noted*

***** t l i t bjfdrolyaed o ssein wee s e s e s a s fe lly re p la se d with a e ia te r e o f I f endue a c id s (th e asnlxis m lM m i su sslm hydrfsl^s a t* w ith th e ex eo p iio a o f hyriEms^ppollm) • A vvsrsl o f th e s tr a in s v e rs th an c u ltu re d m m s e r ie s mi nedia* each o f th e a e d ia la sh in g mm o f th e 19 m & m a sid e * Urn o ile e io n o f m& on* a n la e s e td d id no4:s a a te rla ll^ affoot- th e ijrowth o f th ese srsflsd ess# * '1

Ijm silgtiilG n on th e c lo se ly te ls te d evgsnlsn* J|» M S83& W *. and f|_ ^«**»