Effect of Temafloxacin
VOL.41on Intestinal

Flora273

S-5

INFLUENCE OF A NEW-QUINOLONE ANTIBACTERIAL AGENT,TEMAFLOXACIN, ON NORMAL HUMAN INTESTINAL MICROFLORA
Junko Watabe, Jun Mizutani, Hiromi Suzuki and Koichi Wada
Intestinal Flora Laboratory, Calpis Food Industry Co. Ltd,
11-10, 5-chome, Fuchinobe, Sagamihara, Kanagawa 229, Japan
Kunitomo

Watanabeand Kazue Ueno

Institute of Anaerobic Bacteriology, School of Medicine, University

The effect of temafloxacin(TMFX), a newly synthesized

quinoloneof Gifu,

antibacterialagent,

given orally for 7 days to five healthy male volunteers, on intestinal microflora wasinvestigated. Faecal specimens were cultured quantitatively
for aerobic and anaerobic
bacteria before, during
Marked suppressions

and after administration.
of Enterobacteriaceae
and Streptococcus, including Enterococcus,

were observed during TMFX administration.

The anaerobic bacteria, Bacteroides, Eubac-terium, Bifidobacterium, Veillonella and Gram-positive

cocci were also reduced by TMFX

administration in most subjects. Lecithinase-positive
Clostridium was eliminated, though,lecithinase-negative Clostridium was not affected. Recovery of the levels and compositionof the intestinal

microflora

Key words: Intestinal

was not delayed.

flora; Quinolone; Temafloxacin

Introduction

volunteers

Temafloxacin (TMFX) (Abbott Laboratories,
is a newly developed quinolone antibacterialUSA)
agent,

which has good activity against a broad-spectrum of
microorganisms including
anaerobic
bacteria4,6,8).
Therapy with broad spectrum

antimicrobial

agentsinduces changes in the intestinal
flora2) Thesechanges in the faecal microflora may induce diarrhea,

were admitted

to the clinical study units

during the period of TMFX use.
2. Treatments
A dose of 300 mg of TMFX was given orally to fivesubjects followed by subsequent
12 h intervals for 7 days.
3.

MicrobiologicalFaecal samples

doses of 300 mg atstudies

were collected

before administra-

overgrowth of Clostridium difficile")and fungi, andtion of TMFX, on the fourth day of treatment and thenimpairment of colonization resistancein the digestive

tract9).
The present study was performed

on one day (day 8), one week (day 14), 2 weeks (day21) and 4 weeks (day 35) after cessation of treatment.

to investigate

The faecal samples

the influence of TMFX on intestinal

microflora

healthy male volunteers

state(BBL, Microbiology Systems, Cockeysville, Md. USA)and transported
to the laboratory immediately after

under steadyincondi-tions.
1.

were put into Gaspak

defecation into sterile plastic containers.
Materials
Subjects

and Methods

Five healthy male volunteers , aged 30-43years,were studied. None of the volunteers had taken any
antimicrobial drugs or experienced gastrointestinal
disorders for at least
Written informed

1 month before

consentwasthe study .
obtained and thestool was transferredpouches

One gram of

to 9 ml of prereduced

phos-phate buffered saline with 0.1 per cent agar, serial
10-fold dilutions were made and 0.05 ml volumes
were seeded on various media for detection of intestinal bacteria.

The media for detection of intestinalbacteria

and the incubation

scribedpreviously"

methodNon-selective

have been demedia for total

CHEMOTHERAPY

274

aerobes

andtotalbacteria,

Enterobacteriaceae,
phylococcus,

Pseudomonas

Lactobacillus,

yeast,

Bifidobacterium,
fragilis
wereVeillonella,

for

10-3,

10-5

Nonselectivewith

10-6,latedat

total

aerobes
10-7

media
10-7

h

Different
media

were

Gram

stainandtotal

bacteria

jars
onselectiveandandidentified

tostripNo.

1(EikenNeglerand20A

NeomycinTokyo,forJapan).

Enterobac-

werecountedbiochemically

La(Analytab

byperfringens

Tokyo,Product,

APIlevelmorphology.

Bacteroides

Kagaku,(Analytab

France)cell

onlevelfor

non-selective

media

speciesorgenus

Seiyaku,

andtoinocu-cultures.

andselective

Enterococcus

wereClostridium(Nissuionidentified

STREPTanaerobic

withinoculated

culturesforstrains10-1,

samples.

Platesaerobiccolonies

colonies

Biotestfaecalweredilution.

identified

selectivewith

thepredominant anaerobic bacteria, Eubacterium,Grampositive anaerobic cocci and Bifidobacteriumwere
eliminated during antibacterial agent intakes in all
subjects. In subject A, Eubacterium and Bifidobacterium decreased markedly, and in subject B, Eubacterium and Gram-positive anaerobic cocci decreased,and in subject C, Gram-positive anaerobic coccidecreased. Veillonella also decreased on the fourth dayof treatment, but increased from 7 days to 28 days
after treatment in all subjects. Lecithinase positive
Clostridium was detected in faeces of 3 subjects before treatment and the levels of these organismswere
decreased below the lowest detectable level during
and after treatment. However, lecithinase negative
Clostridium was not affected by TMFX administration. The major anaerobic bacteria had returnedto

F'usobacter-noninoculatedcharacteristics

teriaceae,andandcounteddifferentiation

anddifficileofanaerobic

wereDifferent

Bacteroides

h for

Lecithinase-positive
agar101

Bacteroides,were10_8

albicans,
Clostridium,

Balme

pretreatment levels by one day after treatment.
TMFX eliminated Escherichia coli that was detectedin faeces of most subjects before treatment andenumerated Enterobacteriaceae other than E. coliby
7 days after the last dose of TMFX. E. coli had
returned to the pretreatment levels by 14 days aftertreatment (Table 2). During the TMFX administration

withJapan),

APILesproduct),

Grottes,respective-ly.
Results
Microbiologicalteer

duringmentarechanges

listedaerobesinwereTable

afterweresuppressedonvolunteers

and5levelsday

bacteriaceae
duringdays

2.3

after

creasedontheInsubjectA.
especiallyfourthday

Intreatment.
dayofreturnedtotheofbyofThe

andandafterdays

one

anaerobes

treatmentin3

pretreatment

levelsTMFXofEntero-markedly

hadreturned

totreatment.

The

numbersofBacteroides

oftreatment

in

all

of

Bacter-dayA,theof treatment.

ThewasInaeruginosa

B. fragilisEnterococcus,

total

Total

7

thecustreat-

general,decreasedsubject

involun-

afterP.

fourthdecreasesof eachtreatment

Enterococcus

logs

inducedand1-2.

treatment.

treatment.

oides,

1-1,

fourthlevelsC,

andofadministration

pretreatment
subjectusetheafterandfaeces

duringcessation

one

theagent

eliminateddayofinantibacterial

detectableviablecountgroup,increased

administration,

also

causedbut

theof TMFXEnterobacteriaceae,
and7de-onStreptococa

decrease

1993

anaerobes including the B. fragilis group, Eubacterium, Bifidobacteriumand Gram-positive cocci.However, the levels of total bacteria and total aerobesreturned to the same level as before administrationone
day and 7 days after treatment, respectively. Otherfor
Sta-Candida

dilution

24-48

media

positive

media

forforinC.

Selectiveand37°C

48•`72

aeruginosa,lecithinaseused.

mediumselective
Streptococcus,

Eubacterium,group,iumandEnterococcus,DEC.

inperiod, the level of Enterococcusdecreased markedly
and had returned to normal by one week after cessation of treatment. There was no significant increasein
Enterococcus(Table 3). The composition of Bacteroides
species in the faeces of each subject varied duringthe
period of this study.
B. ruminicola was detected dominantly in faecesof
subjects B, D and E before treatment and decreased
markedly during TMFX use (data not shown).
C. difficile was not recovered from the faecesof anyof the volunteers.
Discussion
Intestinal flora can be influenced by antimicrobial
agents, especially by drugs which are excreted extensively in the bile. The altered gastrointestinal microflora may permit potentialiy invasive pathogensto
overgrow5) or lead to a diminution in colonization
resistance to exogenous microorganisms91.TMFXhas

Effect of Temafloxacin
VOL.41on Intestinal

Flora

S-5275been reported to be excreted mainly in the urine3,8).
However, the concentrations of TMFX in faeces might

Bacteroides more markedly than did sparfloxacin 10),
Although the antibacterial activity of TMFX against

be sufficient to affect both aerobes and anaerobes

faecal aerobes

inwas almost same as those of otherthe intestine. None of volunteers complained of abdominal pain, nausea or diarrhoea.

quinolone drugs, the levels of faecal anaerobes, especially the B. fragilis group, decreased during TMFX

In vitro, TMFX was more active against, the B.

use and recovered promptly after administration.
Thus, TMFX administration
might not evoke sidefragilis group than other new quinolone agents suchas ciprofloxacin, ofloxacin and enoxacin 6) With invivo human testing, TMFX decreased the level ofreactions

such as diarrhoea.

Taken together, these results show that TMFX isTable 1-1. Counts* of microorganisms in faecal flora of subjects A, B and C receiving 300 mg of temafloxacin
orally at 12 h intervals for 7 days*Countsare expressed―: Do organisms

as log
10 per gramof<2detected(counts

wet faeces.
10910 per gram).

Table 1-2. Counts* of microorganisms in faecal flora of subjects D and E receiving 300 mg of temafloxacin
orally at 12 h intervals for 7 days*Countsare expressed as―: no orgahisms

detectedlog
lo per gram(countsof<2wet faeces,log 10 per gram).

CHEMOTHERAPY

276

DEC.

Table 2. Levels* of Enterobacteriaceae
in faeces of volunteers
orally at 12 h intervals for 7 days*Countsof

d血ferentcoloniesonEnterobacteriaceae

selectivereceiving 300 mg of temafloxacin

medium (DHL)expressedaslog
lo per―:noorganisms

detected(countsof<2log 10 pergram)

gram

wetfaeces.

.

Table 3. Levels* of Enterococcus spp. in faeces of volunteers receiving 300 mg of temafloxacin
orally at 12 h intervals for 7 days*Countsofdifferentcoloniesonenterococci

selective

mediumexpressed

as log
10 per―:noorganisms

detected(countsof<2log 10 pergram).

gram

wet

faeces.

1993

Effect of Temafloxacin
VOL.41

activerods,

againstafter

aerobic

Enterococcus

Microbial

on Intestinalandandlevels

anaerobic

werecessation

anaerobic

Nutrition

Gram-negative

Gram-positive

almostnormal6)

cocci.

withinone

weekof administration.
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inCHEMOTHERAPY

278

新 しい ニ ュ ー キ ノ ロ ン系 抗 菌 剤,
temafloxaCinの ヒ ト腸 内 細 菌 叢 に 及 ぼ す 影 響
渡 部 怐 子.水谷

潤 ・鈴 木 宏 美 ・和 田 光 一

カ ル ピス 食 品工 梁 株 式 会社 腸 内 フ ロー ラ ラボ ラ トリー*渡辺 邦 友 ・上 野一 恵
岐阜 大学医学部附属嫌気性菌実験施般

新 し く合 成 さ れ た キ ノロ ン系 抗 菌 剤 で あ るtemafloxacinを5人 の健 常 成 人男 子 に経 口 で7日 間
投 与 し,腸 内 細 菌叢 へ の影 騨 を綱 べ た 。 投 与 前,投 与 中お よ び投 与 終 了 後 の 糞便 サ ンプ ル を培
養 し,糞 便 中 の 好気 性 菌 及 び嫌 気 性 菌 の 菌 数 を測定 した と こ ろ,投 与 期 間 中,大 腸 菌 群 お よび
腸 球 菌 の 著 しい 減少 が観 察 され た 。 バ クテ ロ イ デ ス,ビ フ ィ ドバ クテ リウ ム お よ びユ ウバ クテ
リウム 等 の腸 内優 勢 嫌 気 性 菌 も,投 与 期 間 中 多 くの 被 験 者 で 減 少 した 。 レ シチ ナ ーゼ 陽 性 クロス トリジ ウム も減 少 したが,レ シ チ ナ ー ゼ陰 性 ク ロス トリ ジ ウム の 菌 数 に変 化 はな か った。 菌数 お よ び菌 叢 の構 成 は投 与 終 了後 す み や か に 回復 した。

*〒229相模 原 市 淵 野 辺5‑11‑10DEC.

1993