ロ薗■王

第21巻第3号平成5年9月

内
原

容

著

マレーシアにおけるウエステルマン肺吸虫の第二中間宿主（英文）一波部重久，Karen−Lai，吾妻健

137−142

C．K．Ow−Yang，川島健治郎
インドネシア，スラバヤ地区における下痢症の原因菌（英文）

…岩永
Eddy

正明，仲宗根

Bagus

昇，中村

Wasito，Pitono

哲

14H47

Soeparto，

Subijanto

Marto

Sudarmo，Eddy

1．GustiNyomanGde

Soewandojo，Ranuh

免疫ブロット法による糞線虫症治療前後の抗体反応の比較（英文）…………・…一・一島袋

城間

修一，佐藤

良也，小林

潤，島袋

勲，149−155

祥行

症例報告

大分県で発症を見た人体肺犬糸状虫症の一例，特に虫体の形態学的特徴について
一高岡

北川

宏行，堀

悠子，中山

和生，井上

聡一，堀内

厳，青木

千春，157−1肥

芳夫，後藤

純

会報・記録
1993年度日本熱帯医学会第1回幹事会記録……………一・…………・…・…・……………一…・…

1筋一166

1993年度会員名簿…………一・…・一………………・一・……………・………………一・……………

167−189

投稿規定（和文）…・…………・…・……………・・…・…………一・…・・……………一…・……………

192

1■

137

J pn. J. Trop. Med. Hyg., Vol. 21, No. 3, 1993, pp. 137‑142

CRAB HOSTS FOR PARAGONIMUS WESTERMANI(KERBERT, 1878) IN MALAYSIASHIGEHISA HABE1, KAREN P.F. LA12, TAKESHI AGATSUMA3,

C.K. OW‑YANG2 AND KENJIRO KAWASHIMA4
Received May 12 1993/Accepted July 7 1993

Abstract: During July and August 1985, a field survey on the lung fluke was carried out at six localities in
the peninsular Malaysia. A total of 1,009 fresh‑water crabs belonging to eight species were examined andParagonimus westernrani metacercariae were found in four species of the crabs, Parathelphusa maculata from
Kuala Pilah and Ulu Langat, Parathelphusa malaysiana. Johora tahanensis and lrmengardia pilosimana from
Sungai Wa. The three species of crabs except Parathelphusa maculata are recorded as new crustacean hostsof P. westermani. Parathelphusa maculata harboured more than 60% of the total metacercariae in the muscle,
20‑25% in the gills and about 10% in the liver, but no metacercariae were found in the heart. In contrast,
a substantial number of metacercariae parasitized the heart of the crab, Parathelphusa malaysiana.

INTRODUCTION(Figure I and Table l) . They were brought back to the

In Malaysia, three species of fresh‑water crabs,

Institute for Medical Research in Kuala Lumpur andwere examined for Paragonimus infection after being

Johora johorensis (Roux, 1936) , Parathelphusa maculata

classified by species, sex and carapace width. The gills

de Man, 1879 and Potamiscus cognatus (Roux, 1936)

of each crab were examined under a stereomicroscope.

have been reported as the second intermediate hosts of
Paragonimus westermani (Kim, 1967; Lee and Miyazaki,
1965; Miyazaki et al., 1968; Miyazaki and Kwo, 1969).

distribution of metacercariae in the crab tissues. The
examination procedure used is as follows. Gills, Iiver,During July and August 1985, the authors carriedout a field survey on the Malaysian lung fluke. In theinvestigation, three species of the fresh‑water crabs

intestine, heart and gonad were separately pressed
between two glass plates and examined under a ste‑reomicroscope. Muscles of the cephalothorax and legs

were found as new second intermediate hosts of P.

were chopped up into small pieces by scissors and thenwestermani.
We report here the new crab hosts of P. westerneani
in Malaysia, the prevalence of Paragonimus infection in
crabs, and the distribution of its metacercariae in thetransferred into a metal sieve of about 5‑mesh. The
chopped muscle was further ground with a wooden rod
in the sieve immersed in a beaker of tap‑water. The

crab tissues.

then the supernatant was discarded. The sediment waswashed several times by repeating this procedure andthen strained through , Sieve of 30‑mesh. The super‑

MATERIALS AND METHODS

Some of crabs were examined in detail for thefiltrate was allowed to stand still for a few minutes andnatant of the final filtrate 'was poured off and the sedi‑

A total of 1,009 fresh‑water crabs belonging to eight

ment was examined for metacercariae undet a stereomi‑

species were collected from the following six different

localities: Kampong Langkap near Kuala Pilah (Negeri

croscope. The metacercariae recovered from crabs
were measured and used for experimental infection.

Sembilan) , sev. eral streams in Baling District (Kedah) ,ldentification of the crabs was performed by Dr. M.

Sungai Wa and Sungai Kuching at Taman Negara

Takeda of National Science Museum. Tokyo and the(Pahang) , Sungai Lui at Ulu Langat (Selangor) andSungai Lalang (Selangor) in the peninsular Malaysia

results have been published (Ng and Takeda, 1992) .

l

2
3
4

Department of Parasitology, School of Medicine, Fukuoka University, Fukuoka 814‑01, Japan
Division of Parasitology, Institute for Medical Research, 50588 Kuala Lumpur, Malaysia
Department of Parasitology, Kochi Medical School, Nankoku 781‑51, Japan
School of Health Sciences, Kyushu University, Fukuoka 812, Japan

138

Table 2 Relationship between carapace size of crabs,
Parathelphusa maculata, and number of Para‑

gonimus westermani metacercariae in crab
gills

Carapace
Ave. No. ofinsize metacercariae
No. of crabs..t r¥... ¥ ‑.,
d"I' "(in mm) examinedinfaected
positive crabl'/ /'¥‑ J/
O ¥J'

D BalingeTaman Negara

MALAYSIA

25 . 0‑29 . 9

17

6 (35.3%)

1.7

30 . 0‑34 . 9

49

12 (24.5%)

2.3

29 (36.7%)

1.5

35 . 0‑39 . 97940 . 0‑44 . 957

31 (54.4%)

2.5

45 . 0‑49 . 9

60

2.9

50 . 0‑54 . 9

21

45 (75.0%)
18 (85.7%)

44(100.0%)

5.8

55 . o‑2.6

The crabs were collected from Kuala Pilah

also indicates the prevalence rate and the number ofKuala Lumpur

8 IeUlu Langat

metacercariae in the gills.

O sungai Lalang
Keuala Pilah

S I NGAPO RE¥
Figure 1 A map showing the localities surveyed in

Malaysia e: positive for metacercariae ofP. westermani O: negative

145 (50.0%) of 287 Parathelphusa maculaia (cara‑
pace width 25‑57 mm) from Kuala Pilah, were positive.
Although the infection rate was relatively high, thenumber of metacercariae per positive crab was low.
One or two metacercariae per crab were found in many
the positive crabs, and eleven was the maximum numberin the gills. The prevalence rate and number of metacer‑

cariae in gills increased with carapace size, therefore
they are proportional to the age of the crab (Table 2) .

In Ulu Langat, 6 (24.0%) out of 25 Parathelphusa

RESULTS

maculata (carapace width 20‑36 mm) were positive.

The results of the crab examination for Para‑
gonimus infection are shown in Table 1. Paragonimus
metacercariae were recovered from Parathelphusa
maculata collected from Kuala Pilah and Ulu Langat,
Parathelphusa maiaysiana Ng and Takeda, 1992, Irmen‑
gardia pilosimana (Roux, 1936) and Johora tahanensis(Bott, 1966) from Sungai Wa (Photos. 1‑4). The table

The number of metacercariae per positive crab ranged
from I to 7, and 3.0 on the average.

In Sungai Wa (Taman Negara), I (1.0%) of 98
Johora tahanensis, 4 (6.2%) of 65 Irmengardia
pilosimana and 12 (63.2%) of 19 Parathelphusa
malaysiana were found to be infected with Paragonimus

metacercariae. The average number of metacercariae

Table I Prevalence of Paragonimus westernrani metacercariae in crab gills in Malaysia
Locality

Crab hostNo. of crabs

No. of metacercariae

examined

inf ected/positive crab

Kuala Pilah

Parathelphusa maculata287

145 (50.0%)

2.5

Ulu Langat

Parathelphusa maculata25

6 (24.0%)

3.0

Sungai Lalang

Parathelphusa maculata
Johora intermedia
Geosesarma peraccae

14
3
12

O
O
O

BalingSomanniathelphusa sexpunctata
Stoliczia tweedei

213
52

O
O

Sungai Kuching Johora tahanensis

Taman Negara
Sungad Wa

221

OParathe lphusa ma laysiana

19

lrmengardia pilosimana
Johora tahanensis

65
98

12 (63.2%)
4 ( 6.2%)
1 ( 1.0%)

11 . 2

1.0
6.0

139

Photo. I Parathelphusa maculata de Man, 1879

Photo. 2 Parathelphusa malaysiana Ng and Takeda, 1992
.' I

i

s

i
*';i=:ti*;;i;:; ,j; ji 2 1; 3i4
7 *. q̲. =;+Photo. 3 Johora lahanensis (Bott, 1966)

Photo. 4 Irmengardia pilosimana (Roux, 1936)

in the gills per positive crab was 6.0, 1.0 and 11.2, respec‑

single metacercariae each in genital organs. No

tively (Table 1).

metacercariae were found in the heart region of theTable 3 shows the distribution of metacercariae inthe crabs hosts. In 21 (13 males and 8 females) Parathel‑crabs. There was no significant difference between both
sexes of crabs in the infection rate of metacercariae inphusa maculaia from Kuala Pilah, out of 250 metacercar‑
iae counted, 156 (62.4%) were found in the muscles, 58(23.2%) in the gills, 18 (7.2%) in the liver, 4 (1.6%) in

each organ or tissue.

the genital organs and 14 (5.6%) in other parts of the
body (mostly in the inner membrane of the carapace) .
The infection rate of each organ or tissue was follows:
20 (95.2%) out of 21 crabs harboured 1‑31 metacercar‑

metacercariae in the host. The crab (carapace width 35.8

iae (7.8 on the average) in the muscles; 14 (66.7%) , 1‑10

ed from 4 Parathelphusa malaysiana from Sungai Wa.

in the gills; 7 (33.3%), 1‑5 in the liver and 4 (19.0%), a

More than half, 252 (57.7%) , were obtained from mus‑An infected crab, Parathelphusa maculata from Ulu

Langat, was examined for the distribution of themm) harboured 21 metacercariae in the muscles, 7 in thegills and 2 in the liver.

A total number of 437 metacercariae were recover‑140

Table 3 Distribution of Paragonimus westermani metacercariae in the crab hosts
Crab hostNo, ofcrabs

No. of metacercariae recovered fromNo. of Meta‑

examined

cercariae
obtained

Parathelphusa
maculata

21 *

250

Parathelphusa
malaysiana

4t

lrmengard ia
pil osimana

4t

Johora

It

437
18

18

liver

heart

58

18

o

(23 . 2%)

(7.2%)

gills

muscleorgansothertissues

4

156

14

(1.6%)(62 . 3%)

(5.6%)

52

38

68

8

252

19

(11.9%)(15.6%)(8 . 7%)(1 . 8%)(57.7%)(4 . 4%)

o

O

14

O

4
(22 . 2%)

6

(33.3%)

tahanensis

genital(77.8%)
2(11 . 1%)o

9o

(50 . O%)

(5.6%)* Collected from Kuala Pilah. t Collected from Sungai wa, Taman Negara
All crabs harboured metacercariae somewhere in their bodies.

Table 4 Measurements of the inner cyst of metacercariae of Malaysian Paragonimus
westermani (in pm)
Crab hostLocality

Cyst size

No. cysts

measuredmaximamminimamParathelphusa maculata

Kuala Pilah

56

334 . 917 . O

312 . 2 + 17 . O

Parathelphusa maculata

Ulu Langat

14352 . 420 . 8320 . 6

20 . 8Parathelphusa malaysiana

Sungai Wa
Sungai Wa
Sungai Wa

52

342 . 9 i 15 . 1

314 . 715 . 113345 . 5 + 23 . 6

327 . O

16 . 118341 . 0321 . 8 + 13 . 2

Johora tahanensis

lrmengardia pilosimana

15 . 1cles and 68 (15.6%) from heart or pericardium. Only 52

found in the muscles, 6 in the gills, 2 in the liver and 1(11.9% ) of them were distributed in the gills and the rest

elsewhere.

in the liver, genital organs, and other part of the body.

In the examination of 4 infected individuals of
lrmengardia pilosimana from Sungai Wa, only one crab

Another infected crab, Johora tahaleensis, from
Sungai Wa, harboured 18 metacarcariae, 9 of them were

Photo. 5

Metacercariae in the gill of a host crab,
Parathelphusa malaysiana. (Scale=100 pm)

harboured a single metacercaria in the gills but all four

Photo. 6 Metacercariae removed from a host crab,
Parathelphusa malaysiana. (Scale= 100 pm)

141

had 3‑4 metacercariae in the muscles.

P. westermani metacercariae obtained fromdifferent crab hosts in Malaysia were oval in shape andhad two cyst walls, i.e., outer and inner layers. Thick‑ness of outer layer was 2‑3 pm and that of inner layer

varied from 5 to 25 pm. The measurements of theencysted metacercariae from different localities areshown in Table 4. Pinkish granules in the metacercariae
were rarely seen (Photos. 5 and 6).
On the basis of the morphological features of themetacercariae together with those of the adults obtained

from experimentally infected animals and thecytogenetical studies of the flukes, these flukes werecomfirmed as P. westermani (diploid type).

DISCUSSION
Up to the present, the following 3 species of the

fresh‑water crabs have been reported as the second

and the remaining from liver or gills (Habe, 1979., Habe

and Terasaki, 1982). As for Philippine P. westermani.
namely P. w. filipinus Miyazaki 1978, in the crab, Sun‑dathelphusa philippina Martens, 70.0% were found in themuscle, 22.3% in the heart and 4.9% in the gills (Miyaza‑

ki and Habe, 1979). The above mentioned results wereobtained by the same methods as used in the present
study.
In the present investigation on the distribution of P.

westermani metacercariae in the crab, Parathelphusa
maculata harboured more than 60% of the total metacer‑
cariae in the muscles, 20‑25% in the gills and about lO%
in the liver. No metacercariae were found in the heartregion. The distribution of metacercariae showed nofound to be significant difference between the Japanese
P. w stermani in Eriocheir japonicus and the Malaysian
P. westermani in Parathelphusa maculata. The distribu‑
tion of metacercariae in Johora tahanensis and lrmen ‑gardia pilosimana was similar to that in Parathelphusa

intermediate host of P. westermani in Malaysia:

maculata. Lee and Miyazaki (1965) also obtained 23

potamon johorense from Ulu Langat (Lee and Miyazaki,1965), Parathelphusa maculata from Ulu Langat (Miya‑zaki et al., 1968) and Potamiscus cognatus from Baling(Miyazaki and Kwo, 1969) . Kin (1967) also re orted
two species of crabs, Potamon johorense and Parathel‑phusa maculata as second intermediate host. The genus

Paragonimus metacercariae from 4 infected crabs, Jo ‑hora johorensis. 14 were in the muscle, 6 in the gills and

3 in the liver. On the other hand, Parathelphusa
malaysiana showed a high percentage of the metacercar‑
iae in the heart region. This is similar to the Philippine

P. westermani in Sundathelphusa philippina.

Johora (type species: Potamon (Potamon) johorense

In the present study, it became clear that theRoux, 1936) was first established by Bott (1966) as asubgenus of Stoliczia Bott, 1966 and Ng (1987) elevatedit to a full genus. Nowadays, Potamon johorense comesunder the genus Johora. In the present investigation, 4

Malaysian P. westermani shows different metacercarial
distribution in accordance with the host species. The

species of the fresh water crabs, Parathelphusa maculata

distribution pattern of metacercariae in the crab hosts isimportant to know accurate prevalence of Paragonimus.
For example, Table I shows the prevalence rate and thefrom Kuala Pilah and Ulu Langat, Parathelphusa
malaysiana. Johora tahanensis and lrmengardia

number of metacercariae per crab host by examining

pilosimana from Sungai Wa, were found to be infectedwith P. westermani. These crab hosts except Parathel‑phusa maculata are recorded as new cru tacean hosts ofas compared with the whole body examination.

P. westermani.
P. westermani is widely distributed in Asia, and ateach locality the fluke may have many crustaceans as
the second intermediate host and some differences have

only the gill. This value is quite a low rate of infection

ACKNOWLEDGMENTS
We would like to express our gratitude to Dr. M.
Takeda of the National Science Museum, Tokyo, for his
identification of the crab hosts and to Prof. T. Kifune of

the crab hosts. Observations on the crab, Geothelphusa

the Fukuoka University for reviewing the mauscript.
This study was supported by the research grant Nos.

dehaani White, in Japan showed that 86.5% of P.

60041052 and 61043047 under the International Scientificwestermani (diploid type) metacercariae were found in
the muscles, 7.8% in the liver, and 4.0% in the gills(Habe and Miyazaki, 1982) . In the case of the JapaneseReserch Program from the Ministry of Education Sci‑
ence and Culture Japan.

been observed in the distribution of the metacercariae in

P. westermani (triploid type) in Eriocheir japonicus (deHaan) , 76% of metacercariae were found in the muscles,
16% in the gills, and 5% in the liver, while in Geothel‑phusa dehaani. 82.9% were obtained from the muscles

REFERENCES
1 ) Bott, R. (1966) : Potamiden aus Asien (Potamon Saviguy
und Potamiscus Alcock) (Crustacea, Decapoda) , Senck‑

142
enb. Biol., 47; 469‑509, Pls. 16‑21.

2 ) Habe, S. (1979): Distribution of the metacercariae oflung fiukes in their second intermediate hosts, Med. Bull.

Fukuoka Univ., 6, 135‑138.
3 ) Habe, S. and Miyazaki, I. (1982): On the occurrence ofthe bisexual type of Paragonimus westermani (Kerbert,
1878) found in Innai‑machi, Oita Prefecture, Japan, Jpn.
J. Parasitol., 31, 275‑280 (in Japanese with English
abstract) .

4 ) Habe, S. and Terasaki, K. (1982): On the type ofParagonimus westermani from crab hosts in Yaku Island
Kagoshima Prefecture, Jpn. J. Parasitol., 31, 27‑32 (inJapanese with English abstract) .
5 ) Kim, J.S. (1967): Paragonimiasis in Malaysia, Univer‑sity of California: International Center for Medical
Research and Training Annual Report, 45‑51.
6 ) Lee, H.F. and Miyazaki, I. (1965) : Paragonimus wester‑mani infection in wild mammals and crustacean hosts in
Malaysia, Am. J. Trop. Med. Hyg., 14, 581‑585.
7 ) Miyazaki, I. and Habe, S. (1979): Paragonimus wester‑mani filipinus Miyazaki, 1978, stat. n. occurring at Jaro,
Leyte, the Philippines, Med. Bull. Fukuoka Univ., 6, 447‑462 (in Japanese with English abstract) .

8 ) Miyazaki, I., Kawashima, K. and Tan. M.H. (1968) :Parathelphusa maculata de Man, 1879, a new crustacean
host record for Paragonimus westermani (Kerbert, 1878)
in Malaysia, J. Parasitol., 54, 178‑179.

9 ) Miyazaki, I. and Kwo, E.H. (1969) : Potamiscus cognatus(Roux, 1936) , a new crab host for Paragonimus wester ‑mani in Malaysia, J. Parasitol., 55, 459.
lO) Ng, P.K.L. (1987) : A revison of the Malayah freshwater
crabs of the genus Johora Bott, 1966 sta. nov. (Decapoda:
Brachyura: Potamidae) , Malayan Nat. J., 41, 13‑44.

ll) Ng, P.K.L. and Takeda, M. (1992): On some freshwater
crabs (Crustacea : Brachyura : Potamidae : Parathel
pusidae and Grapsidae) from Penisular Malaysia, Bull.
Natn. Sci. Mus.. Tokyo, Ser. A, 18 (3), 103‑116.

143

Jpn. J. Trop. Med. Hyg., Vol. 21, No. 3, 1993, pp. 143 147

ETIOLOGIC AGENTS OF DIARRHEAL DISEASESIN SURABAYA, INDONESIAMASAAKI IWANAGA1,2, NoBORU NAKASONEl, SATOSHI NAKAMURA1,
EDDY BAGUS WASIT03, PITONO SOEPART04, SUBIJANTO MARTO SUDARM04,
EDDY SOEWANDOJ05 AND I. GUSTI NYOMAN GDE RANUH6
Received May 28 1993/Accepted July 7 1993

Abstract: Isolation frequency of enteropathogens from diarrheal stools was examined in Surabaya,
Indonesia. A total of 385 patients with diarrhea including 264 children under 2 years of age and 121 adults
were enrolled. Bacterial enteropathogens other than Campylobacter jejuni were positive in 164 cases out of
385 (44%), while C. jejuni was isolated from 13 cases out of 173 examined (7.5%) . Rotavirus was detected
in 91 cases of 184 examined (49%), and 34 cases of these 91 were co‑infected with bacterial pathogens.
Diarrheagenic E cherichia coli, which were isolated from about 30% of all 385 patients, were highly resistantto ampicillin and tetracycline. Vibrio cholerae O1 was isolated in 17 of 121 adult diarrhea cases.

INTRODUCTION

PATIENTS, MATERIALS AND METHODS

Diarrheal disease was the first leading cause ofdeath among young children in developing countries

Patients: A total of 385 patients with diarrhea at
Dr. Soetomo Hospital (Surabaya, Indonesia) during theperiod from August to December in 1992 were examined.
Of 385 patients, 264 were children under 2 years of age

until around 1980. It was estimated in 1976 that 5 to 18

milion people die each year as a direct result of thisdisease (7) . Although the mortality of diarrheal diseasein the past decade appears to be decreasing because of
intensive world wide promotion of Primary Health Care

(PHO activities including the enhancement of oral
rehydration therapy (ORT) , it still remains as a major

cause of death among young children in these areas.
Moreover, about 50% of tourists to the Third Worlddevelop traveler's diarrhea (3) . From these viewpoints,
diarrheal disease should be regarded as a global healthproblem.
Although there have been many reports on etiologic
agents of diarrheal diseases in various countries anddistricts, repeated studies are needed since the isolation

frequency of the enteropathogens differs from place toplace, and the drug sensitivity pattern can change. In
this communication, the isolation frequency of enter‑

opathogens from diarrheal patients in Surabaya, In‑
donesia, and the drug sensitivity of E. coli isolated fromthem are described.

and 121 were adult cases. Of 264 pediatric patients, 137
were admitted to the hospital and 127 went back to theirhome after initial diagnosis and treatment were conduct‑

ed at the outpatient department.
Specimens: Stool samples were taken before giving
antibiotics, Self medication prior to visiting the hospital

could not be completely excluded. An apropriate
amount of stool was collected in a plastic container, but

rectal swab was used for 70 children out of 127 who
returned home directly from the outpatient department.
Microbiological examinations: The target bacteria
for isolation from all patients were Escherichia. Sal‑
monella. Shigella. Vibrio, Aeromonas, and Plesiomonas.
Serial lO‑fold dilutions of stool samples were made innormal saline solution, and 50 pl of the dilution at 10*
was inoculated on to agar plate of modified Drigalsky
medium (Eiken) . The inoculum was spread over half of
the agar plate by a glass bar spreader and then streaked
them to the other side with a wire inoculation loop. In
the same manner, the dilution at 10' was inoculated on
to an agar plate of SS medium (Eiken) , and the dilution

1 Department of Bacteriology and 2 Research Institute of Comprehensive Medicine, University of the Ryukyus School of Medicine.
207 Uehara, Nishihara, Okinawa 903‑01, Japan. 3 Department of Microbiology, 4 Department of Pediatrics, and 5 Department ofInternal Medicine, Faculty of Medicine, 6 Tropical Disease Research Center, Airlangga University. Dharumahusada 47, Surabaya,
Indonesia.

144

at 103 on to TCBS agar (Eiken) . The colonies grown on
agar plates were identified by routine laboratory tests.

(CFDN) , and ofloxacine (OFLX) ; preparations for oral
administration are available for all 5 drugs. The E. coli

Campylobacter jejuni was isolated on "Campylobacterblood free selective agar medium (Oxoid) , and was
identified by confirming the morphology with Gramexamined contained 113 diarrheagenic strains and 160
non‑diarrheagenic strains. Heart infusion agar plates
containing serial 2‑fold increasing concentrations ofeach drug ranging from 0.0125 to 100 pglml were pre‑pared. The organisms were cultured in heart infusion

staining and degradation activities for hippuric acid.

For the detection of rotavirus, the stools were
centrifuged and a drop of the supernatant was mixed

broth at 37'C overnight. The culture was diluted I to 10
with saline solution and inoculated on the drug contain‑

with a drop of anti‑rotavirus antibody sensitized latex(Rota virus detection kit; Denka Seiken Co., Tokyo,

ing plates using a microplanter (Sakuma‑MITP,

Japan) on a slide glass to detect rotavirus. Enterotoxins

Tokyo, Japan) . The growth of organisms was observed
after 24‑hour incubation at 37'C, and minimum inhibi‑
tory concentrations of each drug against each organism
were recorded.

were detected using Biken method (5) for LT, and using

ST detection kit for ST (COLIST EIA; Denka Seiken
Co. Tokyo, Japan) . Verotoxin was examined by thecytopathic effect of E. coli culture supernatant on ver‑RESULTSocyte monolayer.
Drug sensitivity test: Two hundreds arid seventy‑three strains of E. coli isolated from 273 cases were
examined for susceptibility against ampicillin (ABPO ,385 examined (44%). More than half of the isolates

tetracycline (TO , minocycline (MINO) , cefdinir

were diarrheagenic E. coli (DEO . The isolation rate ofTable 1Isolation frequency: Facultative anaerobic enter‑
opathogenic bacteria were isolated from 164 cases out of

Isolation Frequency of Enteropathogens

Total

In (Ped)Out (Ped)

Adult (In/Out)

Number of patients

385

137

127

121

DEC

113

44

39

29

Shigel la

8

Sal monel la

8

V. cholerae O1

non‑Ol

3

5

4

O

4

22

5

O

17

3

2

V. parahemolyt.

O

O

Other vibrios

2

2

O

O

Aeromonas

7

3

4

O

Plesiomonas

O
O

O

O

Total

164 (44% )61(47%)O

48 (38%)55 (45% )In (Ped) : pediatric inpatient, Out (Ped) : pediatric outpatient

DEC: Diarrheagenic E. coli

Table 2

Details of the Isolated Diarrheagenic E. coli (DEO

Total In ( Ped)

Out (Ped) Adult (In/Out)

Cases with DEC

ll3

44

39

29

EPEC

70

30

23

16

EIEC

7

2

ETEC toxin ( + )158serovar

21

5

7

9

VTEC

O

O

o

O

(E. coli‑0157

4

1

2

1)

3
7

EPEC: enteropathogenic E, coli, EIEC: enteroinvasive E. coli. ETEC: enterotox‑
igenic E, coli, VTEC: verotoxin‑producing E. coli. ETEC serovar: characteristic
serovar for ETEC such as 06, 015, 078, etc. but the toxm was not detected in a single

examination.

145

Detection Rate of Campylobacter jejuni and Rotavirus(No. of positive cases/No. of Patients examined, %)Table 3Total

In (Ped)Out (Ped)C, jejuni

13/173, 7.5%

ll/127, 8.7%

Rotavirus
Rotavirus

91/184, 49%

2146, 4.3%
71/137, 52%

26/184, 14%

21/137, 15%5147, 11%

8/184, 4.4%

6/137, 4.4%

2147, 4.3%+
DEC
Rotavirus+

20/47, 43%

otherDEC: Diarrheagenic E. coli

DEC among pathogens isolated was 72% in children and
52% in adults (Table l). Most of the DEC belonged toso called enteropathogenic E. coli (EPEO . Enterotox‑
ins were detected from 15 strains by singe examination(14 strains with ST, one strain with ST/LT, and no
strain with LT only) . There were 4 strains with serovar

0157, but their H antigens were not type 7 (Table 2).
These 4 strains did not produce verotoxin, and verotox‑
in‑coded gene was not detected from them as examinedby polymerase chain reaction. Campylobacter jejuni was
isolated from 13 cases out of 173 children (7.5%) , but
was not isolated from 58 adult cases examined. Rotavir‑
us was detected in 91 pediatric diarrhea cases out of 184

examined (49%). However, bacterial pathogens werealso isolated from 34 cases of the 91 with rotavirus‑

positive diarrhea (Table 3). The serovar of EPEC
showed 15 types, of which serovar 0127a was dominant(Table 4) .

Drug sensitivity of E. coli revealed that more than
50% of the isolates were highly resistant to ampicillin
and tetracycline (MIC = 100 pg/ml or higher) . Minocy‑cline was better than tetracycline, but the MIC was 6.25
pg/ml or higher against 50% of the isolates. Cefdinirinhibited 95% of the isolates at the concentration of 6.25
pg/ml. Ofloxacin revealed excellent activity against theisolates. It inhibited the growth of all isolates at theconcentration of 0.78 pg/ml except one which was inhib‑

ited at 3.13 pg/ml (Table 5). Drug sensitivity patternsof diarrheagenic E. coli and non‑diarrheagenic E. coli

were essentially the same, as shown in Table 6.
Table 4 Serovar Distributionof EPEC, 70 strains

Serovar No. of strain
018
020
026
044
055
086a
O111
O114
Oll9
0125
0126
0127a
0128ab
0142
0146
0151
0158
0166
Total

DISCUSSION

8

The isolation rate of enteropathogens from diarr‑
heal stools is largely dependent on the effort of the2

person conducting the test, and this should be considered7Oin evaluating reports of isolation frequency. Althoughmore than 10 genuses and many species of enteropath‑ogens are known, it is usually difficult to focus attention

3

on all these pathogens. In well conducted studies, theisolation rates of bacterial enteropathogens from diarr‑

3

heal stools in developing countries have been reported as1

about 40 to 60% (1, 6, 9).
The present study also revealed on isolation rate of

2

8

23

about 50% including Campylobacter. However, it may

4

have been possible to increase the isolation rate by usingO

enrichment media such as Selenite‑F broth for Sal‑
monella, alkaline peptone water for Vibrios, andampicillin‑blood agar for Aeromonas. Adkins et al

O
l

5

70

reported that approximately 60% of Salmonella isolates
were obtained only after enrichment (1), and we also

have the same experience. Four colonies of E. coli

146

Table 5

Antimicrobial Susceptibility of E. coli

Drug con‑centration

Antimicrobials

ABPC

TC

MINO

CFDN

O . 0125

O

O

O

O

O

O . 025

O

O

o

O

4

( p g/mD

OFLX

O . 05

O

O

o

2

72

0.1

O

O

O

12

125

O

0.2

O

O . 39O

O . 78Ol . 56

98

38

7

57

17

4

49

16

16

4

56

39

15

O

3 . 13

42

11

43

16

6 . 25

37

2

58

44

O

12 . 5

19

3

43

12

O

25

3

2

24

50

O

30

7

100

5

62

O

O

200

163

102

O

O

O
O

O

E. coli: 273 strains isolated from diarrheal patients, included ll3 diarrheagenic E. coli.

Numerals indicate number of strains inhibited by the drug concentration.

Table 6 Drug Susceptibility of Diarrheagenic E. coli =DEC (ll3)and Non‑diarrheagenic E. Coli=NDEC (160)

Drug
conc.

( pglmDABPCDEC NDEC

TC

MlNO

DEC

N DEC

DEC NDECCFDN
DEC NDECOFLX
DEC NDEC

O . 0125

O

O

O

O

O

O

O

O

O

O . 025

O

O

O

O

O

O

O

O

2

2

O

24

28

O

O . 05

O

O

O

O

O

O

0.1

O

O

O

O

O

O

3

6

49

43

0.2

O

O

O

O

O

39

33

ll

16

O . 39

O

O

4

1

21

20

6

7

O . 78

O

O

17

19

1

lO

7

5

O

O

O

3

1 . 56

2

24

17

16

14

6

5

3 . 13

14

16

3

5

14

17

3

7

6 . 25

16

12

O

16

24

24

11

O

O

12 . 5

6

8

O

2

19

14

7

O

O

25

1

9

8

O

O

O

50

O

O

10

11

3

2

O

O

O

O

28

19

O

O

O

O

O

30

43

O

O

O

O

O

100

3

200

58

61

O

Numerals indicate percent of strains inhibited at the drug concentration.

isolated from I sample were examined for their enter‑
opathogenicities in the present study, however, enter‑epidemiological feature is in contrast to that in Japan

otoxin production is a delicate phenomenon and therepeated examination may result in higher detection

trated in the first 3 months of the year (4). The reason
for this epidemiological contrast is of great interest in
the field of tropical medicine. More than one third of therate.

Rotavirus was detected in almost 50% of infantile

diarrhea with no definite seasonal variation. This

where the epidemic of rotavirus infection is concen‑

cases with rotavirus infection were also infected withanother enteropathogen. The pathogenic role of micro‑

147organisms in the dual infection is obscure.

Muts. uddy, P., Eusof, A., Haider, K., Islam, S. and Sack

Drug sensitivities of Shigella were not examinedsince the number of isolates was very low. Neverthe‑

J. Diarroeal Dis. Res. 10:79‑86.

less, antibiotic therapy is most important for shigellosis.

H owever, the drug sensitivity pattern of Shigella isconsidered to be simillar to that of E. coli because of thepresence of transmisible plasmid. The drug sensitivity
pattern of diarrheagenic E. coli and non‑diarrheagenic
E. coli are quite similar as shown in the pFesent study(Table 6) .

Prevailing etiologic agents of diarrheal diseases andtheir drug sensitivities vary from place to place, andfrom year to year. The recent emergence of a newserovar of non‑OI Vibrio cholerae and the disappear‑
ance of V. cholerae O1 in India and Bangladesh are good

examples (2, 8) . Continuous monitoring of etiologic
agents of diarrheal diseases is required.

REFERENCES
1 ) Adkind, H.J., Escamilla, J., Santiago, L.T., Ranoa, C.,
Echeverria, P. and Cross, J.H. (1987) : Two year surveyof etiologic agents of diarrheal diaease at San Laaro
Hospital, Manila, Republic of the Philippines. J. Clin.
Microbiol. 25:1143‑1147.
2 ) Alberto, M.J., Siddique, A.K., Islam, M.S., Faruque, A.S.

G., Ansaruzaman, M., Faruque, S.M. and Sack, R.B.
(1993) : Large out break of clinical cholera due to Vibrio

cholerae non‑OI in Bangladesh. Lancet 341:704.
3 ) Echeverria, P., Blacklow, N.R., Sanford, L.B. and Cukor,

G. (1981): Traveller's diarrhea among American peace
corps volunteers in rural Thailand. J. Infect. Dis. 143:767‑771.

4 ) Fujita, Y. (1990): Rotavirus infection ‑ Clinicalsymtoms and influence of climate. J. Jpn. Assoc. Infect.
Dis. 64:1255‑1263.

5 ) Honda, T., Taga, S., Takeda, Y. and Miwatani, T.
(198D : Modified Eleck test for detection of heat‑1abile
enterotoxin of Enterotoxigenic Escherichia coli. J. Clin.
Microbiol. 13:1‑5.
6 ) Kain, K.C., Barteluk, R.L., Kelly, M.T., Xin. H., Hua, G.
D., Yu. an, G., Proctor, E.M., Byrne, S. and Stiver. H. G.
(19,91) : Etiology of childhood diarrhea in Beijing, China.
J. Clin. Microbiol. 29:90‑95.

7 ) Nichols, B.L. and Soriano, H.A. (1976) : A critique oforal therapy of dehydration due to diarrheal syndrome.
Am. J. Clin. Nutr. 30:1457‑1472.
8 ) Ramamurthy, T., Garg, S., Sharma, R., Bhattacharya, S.
K., Nair, G.B., Shimada. T., Takeda, T., Karasawa, T.,

Kurazono, H., Pal, A. and Takeda, Y. (1993): Emer‑
gence of novel strain of Vibrio chelerae with epidemic
potential in southern and eastern India. Lancet 341:703‑704.

9 ) Siddique, A.K. Zaman. K., Baqui, A.H., Akram, K.,

R.B. (1992) : Cholera epidemic in Bangladesh: 1985‑1991.

149

Jpn. J. Trop. Med. Hyg., Vol. 21, No. 3, 1993, pp. 149 155

IMMUNOBLOT ANALYSIS OF ANTIBODYRESPONSE BEFORE AND AFTER TREATMENTOF HUMAN STRONGYLOIDIASISSHUICHI SHIMABUKURO1, YOSHlYA SATO1,2, JUN KOBAYASHI*,ISAO SHIMABUKUROI AND YOSHIYUKI SHIROMA3
Received July 1 1993/Accepted July 31 1993

Abstract: Qualitative analysis of antibody response before and 12 months after treatment of humanstrongyloidiasis was performed to identify the antigenic component which may be useful for serologicalevaluation of the effectiveness of postchemotherapy. The immunoblotting patterns changed significantly
after successful treatment in almost all patients; some responses decreased in their intensity and some
disappeared. Whereas, the pattems of reactivity after treatment could not be distinguished from those
before treatment in patients who were interpreted to be equivocal for complete cure because their antibody
levels did not show a significant decrease after treatment. The immunoblotting patterns, however, varied
considerably for each patient and it was difficult to identify the antigenic component which may be effective

to detect reduced antibody responses early after treatment.

I N TRODUCTION
Strongyloidiasis, which is relatively common inafter treatment by the immunoblotting method to iden‑
tify the antigenic component which may be useful for
serological evaluation of successful treatment.

Okinawa, Japan, is a parasitic disease resulting from aninfection with a nematode Strongyloides stercoralis. Oneof the unique properties of the parasite is its ability to

propagate in a host by internal autoinfection. The
parasite is usually nonpathogenic in immunocompetent

MATERIALS AND METHODSPatients

hosts, but due to the autoinfection, the asymptomatic

Thirty‑three individuals were found to be harboring
the parasite at a mass screem ng by stool examination in

infection often progresses to a severe or fatal infection

Sashiki Town, Okinawa Prefecture, Japan. They were

under immunosuppressed conditions.
The chemotherapy of such severe cases is known to

12 males and 21 females ages 49 to 76 years (mean = 68.
3 years) . Two months after the initial diagnosis, they

be difficult and it is essential to treat the patients during

were treated with pyrvinium pamoate at a dosage of 5

their chronic infection to prevent a severe infection. On

mg/kg daily for 3 days and subsequently received follow‑up faecal examination 12 months after the treatment todetermine the effectiveness of the therapy.

the other hand, assessment of therapeutic efficacy bycoprological examination is difficult because stron‑
gyloidiasis patients frequently fail to respond to anthel‑minthic treatment and also because the stool examina‑

Experimental grouption is not sensitive enough to detect continuous chronic

Out of the 33 individuals treated, stool examination

infection after unsuccessful treatment. In the previous

revealed that 10 had not been cured. The remaining 23
were negative in the follow‑up examination, but 6 ofthem were considered to be equivocal for complete curebecause their antibody titers did not significantly
decrease after treatment. The immunoblotting patterns

study, the authors have demonstrated a significant
decrease in ELISA antibody levels a year after treat‑ment, indicating that the serologic testing is useful for
postchemotherapy evaluation (Kobayashi et al., 1993) .
In the present study, the antibody responses specificto Strongyloides were qualitatively compared before andbefore and after treatment were analyzed on the 3groups of therapeutic efficacy.

1. Department of Parasitology, School of Medicine, University of the Ryukyus, 207 Nishihara, Okinawa 903‑01, Japan

Research Center of Comprehensive Medicine, School of Medicine, University of the Ryukyus, Okinawa 903‑01, Japan
3. lzumizaki Hospital, Naha, Okinawa 901, Japan

150

Stool examination
Stool examination after treatment was performed
daily for 3 consecutive days by three different methods;

absorbancy (OD) at 500 nm. The change in antibody
titer after treatment was expressed as an antibody ratio

direct smear, formalin‑ether concentration method andagainst antibody value before treatment, calculating by
division of the antibody values at the follow‑up exami‑

faecal culture with an agarpiate (Arakaki et al., 1988) .

nation by those at the diagnosis. On the basis of a

The details of the follow‑up faecal examination were
the same as in the previous report (Kobayashi et al.,previous study (Kobayashi et al., 1993) , the individuals

1993) .

with an antibody ratio of over O.6 were interpreted to beequivocal for effective treatment, regardless of negative

results in the follow=up faecal examination.

Anti‑Strongyloides antibody
Serum antibodies to S, stercoralis were compared
before and after treatment to assess serologically theefficacy of treatment. Three serum samples were col‑lected from each patient on the same times of theirdiagnosis, treatment and follow‑up examination. The
antigen used was prepared from S. stercoralis filariform
larvae collected from faeces of strongyloidiasis patientsand the antibodies were measured by an enzyme‑linked
immunosorbent assay (ELISA) (Sato et al., 1985) . The
sera were tested at a single dilution of I : 50 and theintensity of antibody response was measured as the

Immunoblot analysis
The polyacrylamide gel electrophoresis in sodium

dodecyl sulphate (SDS‑PAGE) was carried out with
lO% polyacrylamide slab gels (Laemmli, 1970) . Sam‑
ples for electrophoresis were applied at a protein con‑centration of 1.0 mg/ml in the presence of 5% 2‑mer‑captoethanol (2=ME) and electrophoresis was perfor‑
med at a constant voltage of 120 V for 4 hr.
After electrophoresis, the proteins separated weretransferred electrophoretically to a nitrocellulose mem‑brane (Towbin et al., 1979) . The blotted membrane was
ii

i

li6‑
92.5 ‑

66 ‑

45

3i ‑
2i .5 ‑

i 4.4>'' ""*
̲ .=*,. :**..=*,,*=..*.

Ab

0.41
ratiaO.9ei.08
O.18 O.OO 0.42

Cured ･ i Cured ‑ 2 Cured ‑ 3

Figure 1

Immunoblotting patterns in three patients who were coprologically andserologically considered to be successfully treated. Three serum samples
were collected before and after treatment and the immunoblotting pattems
of the sera were compared. For each patient, the left‑hand lane shows thepattern of serum samples collected when first diagn08ed, the middle laneshows the pattern during treatment and the right‑hand lane shows thepattern at the follow‑up examination. The numbers at the bottom of eachlane show antibody ratio (Ab ratio) against antibody value at the initial
diaguosis. The position and molecular weight (KDa) are represented at the
far left.

151

then treated with a l/lO dilution of patients' sera for 2

hr at 37'C and subsequently overnight at 4'C. The lgG

1 t6 ‑

antibodies which reacted with antigens on the mem‑brane were detected by autoradiography using 1251

92.5 ‑‑labelled protein A. The above method was described indetail in the previous report (Sato et al., 1990). Autor‑

adiograms were scanned with a Dual‑Wavelength Fly‑
ing‑Spot Scanner (Shimazu CS‑9000; Shimazu Co. Ltd.,Tokyo, Japan) . The area values for each peak were
calculated by a personal computer (IBM PC‑AT) Iin‑

66 ‑

45 ‑

ked to the densitometer.

RESULTS31 ‑Immunoblotting patterns before and after success‑ful treatment in three patients are illustrated in Fig. 1.

21 .5 ‑The patterns of antibody response in serurn samples
collected at their initial diaguosis could not be distin‑

guished from those at the time of treatment. The

1 4.4 ‑

reactivities, however, decreased significantly after treat‑" *"(' ";' =** +'ment in all of the patients. Especially, the positive bands

before treatment disappeared almost completely after
treatment in a case (Cured‑2) in which antibody ratio
was only 0.18 after treatment. In the remaining twocases, positive reactivities remained to the bands in theregion of relatively low molecular masses from 20 to 30
KDa where strong reactions were detected before treat‑
ment. On the other hand, no significant difference ofpattern was observed in 6 patients who were interpreted

to be equivocal for complete cure. The difference inreactivity could also not be recoguized in 10 patients
who were unsuccessfully treated. The representative

Ab ratio

O .90 0.91

Equivocal
Figure 2* " 'O.98 0.96
Not cured

Immunoblotting patterns of patterns of twopatients. Three patterns with serum samples
collected at the initial diagnosis, treatment andfollowup examination are compared for apatient who was negative in the follow‑up
faecal･ examination but serologically equivocal
for complete cure (Equivocal) , and for a patient

who was unsuccessfully treated (Not cured)The other remarks are. the same as in Figure I .

patterns of reactivity before and after treatment in an
equivocal and an unsuccessful cases are shown in Fig. 2.

Densitometric trace of autoradiograms in Figs. lequivocal case and contrarily 8% increase in the case ofand 2 are shown in Fig. 3 and 4. As seen in Fig. 3, someunsuccessful treatment.

peaks in the relatively high molecular weight region
were found to decrease significantly or to disappear
after treatment. Total area values of peaks detectedcessful treatment in the remaining 14 cases are re‑decreased 66% after treatment in the Cured･l patient,as much as 85% decr,ease in the Cured‑2 patient and
48% decrease in the Cured‑3 patient, as compared to thearea values before treatment. These results were wellconsistent with those of the ELISA assay, in which the

ELISA values are 59% decrease in the Cured‑1, 82%

The immunoblotting pa･tterns before and after suc‑presented in Fig. 5. The patterns of reactivity varied
considerably for each patient; some bands differed intheir intensity and some were absent. The reactivities

decreased noticeably after treatment in all but twopatients (Cases 5 and 12) whose antibody ratios were0.58 and 0.56, respectively. The patterns shown after

decrease in the Cured‑2 and 58% decrease in the Cured

treatment also displayed significant diversity among thepatients and it was difficult to determine which bands‑ 3. On the other hand, the densitometric patterns before

generally became negative in response with antibodies

and after treatment did not change so much in anafter successful treatment.

equivocally cured patient and in a patient of unsuccess‑ful treatment (Fig. 4). The total area values of thesecases were only 25% decrease after treatment in the

152
E

0.80

A

A

A

c

,U

::

g
0.00

(30106)B{33 77 ‑(444 1 3̲).̲

B

B

o.so

0.00

(30585)c(305 1 3)(.354. 57)

c

c

0.80

0.00

̲

,̲Q 64) , (5 1 14) ̲= ̲̲̲̲̲(..2̲ 2̲1̲o),.=.̲.

so,o80.0tum')( sd.oo,o 50:oi 80.0 tun;)
Cured ‑ I Cured ‑ 2 Mobiuty
Figure 3 Densitometric patterns of autoradiograms in Figure 1. For each patient, the
d.o

jti.o

o,o

Cured ‑ 3patterns at their diagnosis(A), treatment (B) and follow‑up examination (O are shown. The
number in parentheses are a total area value of peaks detected.
E
c

o
o

A

A

(]̲80

JO

oOO

(395̲q7)

B(3009 l)B

0.80 *

0,00(43833)

c

̲=J 2

J +

c

o, so

0.00 " ‑{22?‑P? 1 = ' (32556‑)
o:o 5cto 80,0 (') ti.o 50:o so.o (iEquivqcal Mobuity
Not curedFigure 4 Densitometric patterns of autoradiograms in Figure 2.
The remarks are the same as in Figure 3.

153

3

7

4

1

2.

O . 26o.44

0.31 0.57 o.58

0.31

o . 36

9

10

13

14

Ab ratio 0.59 0.58 0.54 0.54 0.56 0.52

o . 28N. P' '

116‑
92.5

66 ‑

45 ‑

31 ‑
21 ,5 ‑
1 4.4 ‑Ab ratio

No. 8

1 16‑

92.5‑

11

12

****';** { ":i:t'{* ;:===;,=*:'( ;='*:;":' i*

66 ‑

45 ‑

31 ‑
21 .5‑
1 4.4 ‑

Figure 5 Immunoblotting patterns of 1 4 patients who were successfully treated. For
each patient, the patterns of response at the initial diagnosis (before treat‑ment) are shown on the left lane and those at the follow‑up examination(after treatment) are on the right lane. The numbers at the bottom of eachlane represents the antibody ratio after treatment. The numbers at the fa,rleft represent the position and moleculam weight (KDa) of marker proteins.

154

DISCUSSION
Postchemotherapy evaluation of strongyloidiasis is
an important clinical problem because strongyloidiasis
patients frequently fail to respond to chemotherapy andespecially because it is difficult to confirm successful
trea,tment by coprological examination. In a previous
study in which 123 persons with chronic Strongyloides
infection were followed for several months without any
treatment, faecal larvae were reconfirmed in only 15%of them by the direct smear, 24.4% by faecal concentra‑tion method and 16.7% by faecal culture (Harada‑Morifaecal culture) , if the faecal examinations were perfor‑

med only once (Sato et al., 1993). At the present time,several samples collected on different days ought to be
examined repeatedly to obtain a correct diagnosis andeven after repeated examinations, failure to demon‑
strate larvae cannot be unequivocally interpreted asabsence of the infection (Jones and Abadie, 1954; Grove,1980). Thus, the postchemotherapy evaluation may be
more difficult, if complete cure is not achieved and theinfection progressed to a latent infection after treat‑patients who were negative in the follow‑up faecal
examination. However, on the basis of clinical andserological observations he considered that perhaps one‑third of the patients were still infected with the para‑site. These results indicate that the serological testing

may effectively complement faecal examination .in the
postchemotherapy evaluation.
In the present study, the authors further compared
qualitatively the antibody responses before and after
treatment by the immunoblotting method to determine

the antigenic component to which positive response

predominantly diminish after treatment.

The

reactivities to the antigenic bands significantly de‑creased after treatment in almost all of the patients
successfully treated. Whereas, the reactivities after
treatment did not differ from those before treatment in
the patients who were equivocal for complete cure, as
well as in patients who were unsuccessfully treated.
The authors have shown in a previous study in whichantibody responses in human strongyloidiasis were anal‑
yzed by immunoblotting method, that the patterns ofment.
In Okinawa, where there are many patients withconcurrent HTLV‑1 infection, an unnegligible number

antigenic recognition by patients' antibodies varied sig‑
nificantly for each patient (Sato et al., 1990). In thepresent study, the immunoblotting patterns also varied
considerably for each patient and due to the diversity inof the patients progress to the severe, often fatal, hyper‑

the reactivity, it was difficult to identify the antigenic

infection state due to the depressed immune competence
condition caused by the viral infection (Nakada et al.,1984, 1987; Takara et al., 1987; Sato & Shiroma, 1989).
Most recently, patients with concurrent viral infections
were found to be intractable to chemotherapy for stron‑
gyloidiasis (Takara et al., 1992; Sato et al., 1992).
Therefore, an effective therapy nd exact evaluation
following the therapy are necessary to prevent such acomponent which will be useful for serological assess‑ment of successful treatment. At the present time, it
should be better to assess the responses to whole extract
before and after treatment for the effective evaluationof chemotherapy.

REFEREN CES

severe infection.

l ) Arakaki, T., Hasegawa, H., Asato, R.. Ikeshiro, T.,

In the previous study, the authors monitored thechanges in the ELISA antibody levels against Stron‑
gyloides before and 1 2 months after pyrvinium pamoate
treatment and demonstrated that the antibody levels

Kinjo, F., Saito, A., Iwanaga, M. (1988) : A new method
ot detect Strongyloides stercoralis from human stool. J.
Trop. Med., 16, 11‑172 ) Fujita, K.. Tajima, K., Tominaga, S., Tsukidate. S.,

significantly decreased after treatment in patients who

become nagative for faecal larvae by the treatment.
Among the patients, however, the antibody response did
not significantly decrease in about 40% of them after
treatment, suggesting that they might be equivocal forcomplete cure. When further faecal examination was
performed on the equivocal cases, about 20% of themwere additionally found to be unsuccessfully treated(Kobayashi et al., 1993). In the previous study, Grove(1982) also followed 43 patients for 6 months after
thiabendazole treatment and detected a significant
decrease in anti‑Strongyloides antibody levels in many

Nakada, K., Imai, J. and Hinuma, Y. ( 1985): Sero‑epidemiological studies of Stron yloides infection in
adult T‑cell leukemia virus carriers in Okinawa Island,
Trop. Med., 27, 203‑209
3 ) Grove. D.1. (1980): Strongyloidiasis in A1lied ex‑pris‑
oners of war in Southeast Asia, Brit. Med. J., 280, 598‑6014 ) Grove. D.1. (1982): Treatment of strongyloidiasis withthiabendazole: an analysis of toxicity and effectiveness,

Trans. Roy. Soc. Trop. Med. Hyg., 76, 114‑118
5 ) Jones, C.A. and Abadie, S.H. (1954): Studies in hurnan
strongyloidiasis. II. A comparison of the efficacy of
diagnosis by examination of feces and duodenal fluid,
Am. J. Clin. Pathol., 24, 1154‑1158

1556 ) Kobayashi, J., Sato. Y., Toma, H., Takara, M. and
Shiroma, Y. (1993): Application of enzyme‑linked im‑
munoassay for postchemotherapy evaluation of humanstrongyloidiasis, Diag. Microbiol. Inf. Dis. (in press)

7 ) Laemmli, U.K. (1970) : Cleavage of structural proteinsduring the assembly of the head of bacteriohpage T4,
Nature, 227, 680‑6858 ) Nakada, K., Kohakura, M., Komoda, H. and Hinuma, Y.
(1984) : High incidence of HTLV antibody in carriers ofStrongyloides stercoralis, Lancet, 1, 633

9 ) Nakada, K.. Yamaguchi, K., Furugen, S., Nakasone, T.,
Nakasone, K., Oshiro, Y., Kohakura, M., Hinuma. Y.,
Seiki, M., Yoshida, M., Matutes. E., Catovsky. D., Ishii,
T. and Takatsuki, K. (1987) : Monoclonal integration of

HTLV‑1 proviral DNA in patients with strongyloidiasis,
Int. J. Cancer, 40, 145‑148

10) Sato, Y., Inoue, F., Matsuyama. R. and Shiroma, Y.
(1990) : Immunoblot analysis of antibodies in humanstrongyloidiasis, Trans. Roy. Soc. Trop. Med. Hyg., 84,
403‑406
1D Sato, Y. and Shiroma, Y. (1989): Concurrent infections
with Strongyloides and T‑cell leukemia virus and their
possible effect on immune responses of host, Clin. Im‑
munol. Immunopathol., 52, 214‑224
12) Sato, Y., Shiroma, Y., Kiyuna. S., Toma, H. and Kobaya‑
shi, J. (1992) : Reduced effect of chemotherapy of stron‑
gyloidiasis in patients with concurrent HTLV‑1 infec‑
tion in Okinawa, Japan, Jpn. J. Trop. Med. Hyg., 20, 183‑19213) Sato, Y.. Shiroma, Y. and Otsuru, M. (1993): Stron‑

gyloidiasis and ATL ‑High concurrency of stron‑
gyloidiasis and HTLV‑1 infection and its background‑,Ryukyu Med. J., 13, 3‑12 (in Japanese)
14) Sato, Y.. Takara, M. and Otsuru. M. (1985) : Detectionof antibodies in strongyloidiasis by enzyme‑linked im‑

munosorbent assay (ELISA) , Trans. Roy. Soc. Trop.
Med. Hyg., 79, 51‑55

15) Takara, M., Hirata, R., Maeshiro, H., Nakamura, M.,
Shiroma, Y., Akaboshi, N., Nakaji, S. and Murata, S.

(1980) : Five cases of malignant lymphoma became
overt during the course of strongyloidiasis. Okinawa
Med. J., 18, 129‑131 (in Japanese)16) Takara, M., Toma. H., Kobayashi, J. and Sato, Y.
(1992) : Effect of concurrent HTLV‑1 infection on theefficacy of pyrvinium pamoate treatment of stron‑
gyloidiasis, Jpn. J. Parasitol., 41, 202‑212

17) Towbin, H., Staehlin, T. and Gordon, J. (1979) : Electro‑

phoretic transfer of proteins from polyacrylamide gelsto nitrocellulose sheets: procedures and some applica‑
tions, Proc. Natl. Acad. Sci. USA, 76, 4350‑4354

157

Jpn．J．Trop．Med．Hyg．，VoL21，No．3，1993，pp。157−163

症例報告

大分県で発症を見た人体肺犬糸状虫症の一例，
特に虫体の形態学的特徴について（例えば，吉田ら，1984）。

本症例は，これまでの多くの症例と同様に，胸部X線写
真のcoinlesionに端を発し，術後肺組織中に見いだされた虫体によって診断された典型的な肺犬糸状虫症である。本例では，病変部の肺組織から約25mmの長さに切断された虫体自体が得られた他，肺病変の連続切片標本に虫体の食道部位を含む前方部分と腸部位を含む中間の部分の横断像が得られたので参考に供したい。

2︐3︐
巌
一

（1990）の集計によると1990年5月の段階で85例に達する。
これらの多くは近年になって報告された症例である。犬糸状虫の人体症例では無症状の場合が多く，大半は肺寄生であるが，肺以外に寄生する例（主に皮下に寄生し腫瘤を形成する）も知られている（吉村，1988）。肺寄生の場合は，
胸部X線撮影によりいわゆる銭型陰影coin lesionを呈す
ることが多く，開胸手術によって摘出された病理組織から寄生虫の横断像が認められることから，本症と診断されることが少なくない。肺組織中に見いだされる犬糸状虫は，成虫またはそれに近い段階まで発育している場合もあるが，多くは未成熟虫である。また，しばしば変性過程にあって正常の虫体と形態が異なることがある。虫体の組織や形態の保存性が良ければ，角皮の形状などの特徴によって同定が可能であるが，虫体の切断面や変性の度合などによっては形態学的に同定することが困難な場合もみられている

15

により本寄生虫の第一例が発見されているが，真喜屋

聡 理
山上縛暖

による人体寄生例はこれまで米国南部をはじめ世界各地から報告されている。我国からは，Nishimura8彪1．（1964）

中井 月
ク
2︐
子
生3︐藤
5年

日

3︐付
夫受

本来犬の寄生線虫である犬糸状虫Z）伽φ伽吻加吻挑

悠和轟

〜

堀北芳肋
徹剤内鰯
堀廊
宏千 平

岡木

高青

はじめに

し。犬猫などのペットは飼っていない。

現病歴
1992年8月頃，右前胸下部に軽い鈍痛を覚えたが，2〜3日で軽快したため放置。同年9月下旬に行われた住民検診において胸部X線撮影に異常陰影を指摘される。検査のため，同年11月18日国立大分病院外来を受診。初診時，自覚症状なし。胸部理学所見も異常なし。
胸部X線撮影で，右肺中葉に比較的輪郭の明瞭な浸潤陰影（Photos．1＆2）が存在した。経気管支肺生検では，慢性気管支炎の所見はあるが，悪性腫瘍の所見は認められなかった。

精密検査の目的で同年12月17日同病院に入院，入院時，身体所見異常なし。血液，尿の生化学検査異常なし。WBC

7180。EOS4．1％。IgG1480。IgM231。IgE検査せず。
ツ反17×16mm。CRP O．36mg／dl。治療のため抗生剤｛セフォペラゾン・スルバクタム（1：1）2g＋クリンダマイ
シン1200mg／日，7日間1およびステロイド剤（コハク酸ヒドロコルチゾンナトリウム400mg／日，4日間）投与す
るも変化なし。

1993年1月18日，同病院にて右肺中・下葉部分切除手術。

手術所見：右肺中葉末梢側に約2cm大の肉芽腫様病変を認めた。病変の内部は乾酪壊死様に変性。肺胸膜は一部壊れ，中・下葉間に限局した腔洞が形成され，隣接する下葉の一部にも肉芽腫様病変が認められた。中・下葉の病変部を一塊として切除。

同年1月31日退院。

同年1月29日および3月5日に採血した血清は免疫電気泳動法により犬糸状虫抗原に陽性を示した。

病理学的所見病変部より得られた切片をH・E染色およびEVG染色症

例

患者：39歳。日本人女性。大分県臼杵市在住。主婦。家
族歴および既往歴には特記すべきことなし。海外渡航歴なにより観察した。主病変は壊死組織と虫体の混在であり，その周囲には，線維組織の増生，炎症細胞の浸潤を認める。
EVG染色では，この病変の周囲には弾性線維が輪状に配列する（Photo．3）。すなわち，上記病変は，虫体による動脈

大分医科大学医動物学教室（〒879−55大分県大分郡挾間町医大ケ丘1−1）
大分医科大学病理学講座（同上）
国立大分病院呼吸器科（〒870−02大分県大分市大字横田1000番地1）

158

Photosほa葺d2．Preoperativecoin1ε＄ionまnpostero鋤teでior

middle

robe

of

the

rentgenogram（1）and
right

塞栓である。一つの動脈あたり1〜3個の虫体断面が認められた。

寄生虫学的検索病変部から取り出された虫体は切断された4つの部分か

com卿tedtomogram（2）showin暮alung，

らなり，総計約25mmの長さで，直径約0，670mmの細い紐状を呈した（Photo．4）。実体顕微鏡下では虫体は乳白色で，ほぼ全長を通じて内部に黄褐色の縦縞が透けて見えた。
ラクトフェノール液で透徹し普通顕微鏡で観察した結果，体壁は厚い角皮（27〜47μm）からなり，外側の表面は平滑で，5〜12μmの間隔で平行する横紋線tr鋤sverse stria−
tionが認められた（Photo．5）。また，角皮には互いに直角に交わる2方向に斜走する線維状構造が見られた（Photo，5）。切口の横断面の観察では，角皮の内側に部分的に剥離した筋肉層と偽体腔中に腸管（直径約90μm）と2個の生殖管（直径約0、320mmおよび0．250mm）が存在した。側索は変性し，輪郭が不明であったが，側索部の角皮の内層に，縦走する隆起intemaUongitudinalridge（以下，i．1，r．と鱗

Photo．3，

Obstructedpulmonaryartery曲ow玉ngthree
sectionsofworm，sarrou磁ed

fibroustissue，Arrowthepulmonary

thebyproll琵r＆t磁indicat銘elastic

arteryfibersof（EVG一＄taining）．

Scaleヲ500μm，Photo．5，

En1獄rge揃ent

femalePhoto．4．

1）ズ名瞬惚短αぎ吻魏ゴ彦s，P｛）rtionsofmidbody

of

the

body

w翫11

worm（Photo、4）showingofanstri＆tioねso臓t簸esmoot｝嵐outer＆dult female wo蹴taken from the vessels
the騰sected pulmonary tissue，Sc敬1e，1m瓢

of

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at

159

6

7

8

g

10

11

Photos. 6‑11 .

Dirofilaria immitis, transverse sections of adult male in the pulmonary artery. 6, At
anterior tip, showing transverse striations of the smooth cuticle. 7, Near anterior end. 8,
Between anterior end and nerve ring. 9, At level of nerve ring, showing small dark spotsat the internal longitudinal ridges of the cuticle, and distinct dorsal and ventral muscularlayers, 10 and ll, Between the nerve ring and esophagus‑intestine junction, showing distinct
dark spots at the internal longitudinal ridges, and one ot two reproductory tubes, as weli
as layered cuticular body wall, and muscular layers. Scale, 100pm.

160

41枚目に1本の大きな管（直径0．180mm）として出現し略す）が認められた。

病理観察のための切片標本とは別に，肺病変部の一部を・（Photo．10），47枚目で2本となり（PhotO。11），49枚目

5μmの厚さで連続切片を作製し，2つの血管に存在した虫体の断面像をH・E染色およびPAS染色で観察した。第から6！枚目では5〜8本となった。また，それ以降は3本の断面として認められた。生殖管はPAS染色でよく染色される2重の管壁からなり，内部は生殖細胞で充満してい1の血管内の虫体像は，虫の前端部から食道の部分に相当
し，第2のものは，食道・腸分岐部より後方の一部分と思われる。第1の食道部を含む虫体部分は切片標本で71枚，また第2の中腸部を含む部分は83枚について観察できた。

た（P薮otos．10＆11）。第2の虫体部分はどの標本でも，厚い角皮，発達したi．1．r．，多数の筋細胞，1本の腸管お第1の虫体は前端から約0．355mmに相当し，横断像は部位により違いが見られた。最初の11枚の標本では，横紋をもった角皮のみが見られた（Photo、6）。虫体の横断像は12枚目から出現したが，17枚目までは断面の直径も小さくよび2本の生殖管が認められた（Photo．12）。側索や背腹の中央線は一部を除いて不明瞭であった。虫体断面の直径は大部分が0．670〜O．680m燃であったが，最後の10枚の標本では0，720〜0．891mmと幾分大きくなっていた。角皮は20〜55μmの厚さで4層からなり，中2層は特に厚く斜走

く，三放射対称をなすが，すぐに背腹に偏平で不整形になった。食道断面の直径は64〜76μmであった。29枚目前後に食道を囲んだ神経輪が観察された（Photo．9）。また，側索は全体にわたって不明瞭であったが，神経輪およびその前後の部位では中心部に向かって突出していた（Photo．9）。角
皮は3〜4層から成り，10〜30μmの厚さであった。i．1．r，は20枚目から微かに認められるようになるが，71枚目でも構造が認められた。両側索に対応する部位の角皮が最も厚く，その内層から概ね二等辺三角形をなしてi．1．r．が15〜27μmの高さで内側に突出していた（Photo，13）。〜部の標本ではi．1．r．が角皮内層から遊離したり，全くなくなっていた。第1の虫体部分の標本で認められたi，1．r．の濃染スポットは見られなかった。筋細胞数は4分の1区画
で52〜75であった。腸管は断面の直径が70〜85μmと小さく，内部は非細胞物で詰まっていた。2本の生殖管は大きさが異なり，断面の直径は，大きい管で0，350〜0．450mm，小さい管で0．245〜0．320mmであった。2つの生殖管の大きさの比はL2〜L7と虫体の部位によって違いがあった。
生殖管は管壁が厚さの異なる2重の層からなり，内腔は大部分が中空で，8〜10μmの大きさの楕円形をした生殖細それほど顕著ではなかった。ただ，神経輪より後方ではi．1、胞が散在していた。

r．に濃く染まるスポットが現れるので，位置の確認は容易であった（Photo鼠10＆11）。体壁筋層は側索部を境に背腹に二分され，ほぼ等しい高さの多数の細い細胞からなる，
いわゆる多筋細胞型polymy3rian−coelomyarian mus・
culaturetypeを示した。しかし，神経輪より前方では個々の筋細胞の区別は出来なかった。後方の虫体断面では4分の1区画当たり35から44の筋細胞が認められた。生殖管は病理観察に用いた標本に見られた虫体の断面像（Photo．
3）も，大きさ（直径約0．700mm）および内腔に2つの生殖管と1つの腸管を持つ点，基本的には上記の第2の連続切片に見られた虫体部分と同じ形態であった。
第2の虫体部分の横断面に見られる層状の厚い角皮，角（約0、130搬m），角皮も約套μmと薄かった（Photo，7）。

内部には食道内壁をなすと思われる角皮が濃染して認められた。筋細胞やi、1、r．は見られなかった。18枚目で虫体断面の直径が0．330mmと急に大きくなるが，後は徐々に大きさを増し，最大0、400mmに達した。食道は18枚目から厚い筋質部で囲まれた管状を呈する（Photo．8）。内腔は狭Photos．12and13．

1万猶φ勉枷

」窺￠髭鵜traぬsverse

皮内の斜走繊維構造，角皮内層に発達したi．1．r．，および

多筋細胞型の筋層，さらに摘出された虫体部分に見られた

sections

of

ad質1t

femaleinthepulmonaryartery．12，

Through midbody，showing thick cuticular body wall with marked intemal longitudin鼠1
ridges，two reproductory ducts，intestine，and muscl蕊Sc凱1e，200μm．13，Portion of tr哀蔦s・
verse

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161

平滑で横紋線を有する角皮の表面などの形態的特徴から犬

糸状虫D〃⑳肱吻珈吻漉と同定した。我国からは別の犬糸状虫n z砂6郷の人体例が1例だけ沖縄から報告されているが（MacLeaneta1．，1979），この種は角皮の表面に多数の縦走する隆起列があるので本症例の虫体とは異なる。
またn名の6窩の場合，人の皮下に寄生する例がほとんどであり，我国の犬や猫からはまだ見いだされていない（吉田ら，1984）。我国の熊にはn％獅の寄生が知られているが，n名勿6ns同様，角皮表面の縦走隆起を持つ点より，今回の虫体とは区別される。今回の虫体は断面の大きさと大きな生殖管を備えることから，既に成虫にまで発育していたものと思われる。また，断面に腸管を持っていた摘出虫
体部と連続切片にした第2の虫体部分は大きな生殖管を2本持っていることから雌であると思われる。一方，虫体断面に食道が見られた第1の虫体部分は神経輪のすぐ後方に2本以上（最大8本）の生殖管が認められることなどから，
雄の虫体と思われる。すなわち，神経輪のすぐ後ろで，後方から伸びてきた1本の輸精管がUターンし，その先端部分の精巣が何度か折り重なり合っている状態が推測される。

考うかは今後検討する必要がある。

犬糸状虫の宿主である犬における感染は北海道から沖縄まで全国的に見られ，九州では平均25．3％の感染率が示されている（真喜屋，1990）。犬から犬，および犬から人への媒介は蚊によって行われ，アカイエカC鋸忽ρ珈召ns勿1−」召ns，ヒトスジシマカノ16碗sα160が諺欝，コガタアカイエカ

C％忽加惚痂oη解h郷など数種が重要な媒介種である（末永，伊藤，1973）。本症例の発生した臼杵市を含め，大分県
における犬の感染状況や媒介者についてはまとまった資料がなく，今後の調査を待たなければならない。ただ，大分市で1985年に我々が行った予備的調査では，検査した8頭のうち4頭の犬がn伽吻痂sの仔虫を血中に保有しており，また1988年9月に人囮法で採集したヒトスジシマカからも低率（279雌個体中1個体陽性または0．4％）ではあるがZ）．魏郷漉sの幼虫が検出されているので（未発表），人への感染が起きる状況は十分整っていると思われ，予防の面で注意を喚起する必要がある。

うことは困難である（吉村，1988）。

手術によって摘出された肺組織中の寄生虫の種の鑑別に関しては，BeaverandOrihel（1965），OrihelandBeaver（1965）およびNeafieandPiggott（1971）などに詳しく検討され，横断面で他の糸状虫からDlzφ肱7宛属を同定す
る基準も指摘された。これらの形態的特徴の一つとして側

語

結

察

本症例は，胸部X線検査により陰影が指摘され，術後摘出された肺組織内の虫体から診断された人体肺犬糸状虫症である。病理学的には，病変は虫体による動脈塞栓であった。大分からは，血清診断による人体肺犬糸状虫症が1例報告されているので（明石ら，1983），今回の症例が2例目であるが，前述したように，我国における犬糸状虫による人体寄生例，特に今回の症例に似た肺への寄生例の報告が近年増加しているので（真喜屋，1990），人獣共通寄生虫症の一つとして注目する必要がある。犬糸状虫が肺に寄生した場合，肺癌との鑑別が重要であるが，胸部X線で陰影が見つからない腫瘤摘出例も報告されており（真喜屋ら，
1988），開胸術前に免疫学的診断法などだけで確定診断を行

索基部に突出したi．L

置にH・EおよびPAS染色で濃く染まる部位が認められた。このような性質はn加卿痂sだけに特異的なものかど

大分県臼杵市において発症をみた人体肺犬糸状虫症の一例を報告した。本症例は，胸部X線撮影で右肺中葉下部に銭型陰影を認め，術後，肺動脈より摘出された虫体によって診断されたものである。本報告では，摘出虫体の他に，組織切片標本中に種々の虫体断面像が比較的多数得られたので，犬糸状虫の形態学的特徴についての検討もおこなっ
た。

謝

辞

本校を終えるにあたり，患者血清の免疫診断において御世話になった杏林大学医学部寄生虫学講座辻守康教授および貴重な文献を提供いただいた産業医科大学医動物学講座
真喜屋清助教授に感謝します。最後に，原稿の校閲を賜った九州大学医学部寄生虫学講座多田功教授に御礼申し上げ
ます。

献

文

r．の存在が重要視されている。今回の虫体の横断面では，ほとんどの標本で顕著なi．1．r．が認

1）明石光伸，田代隆良，後藤純，那須勝，糸賀敬，坪井峯男，められたが，虫体の前端部近くでは全くないか，あってもそれほど発達していなかった。また，後方の断面像で腸管虫症の1例，日本胸部疾患学会雑誌，21（12）：1228−1232が見られる部位の一部の標本では，発達したi．1．r．が離れかかったり，完全に離れてなくなっていた。これは，成長した虫体でも部位によってi．1．r．の発達の度合に差があること，また虫体の変性過程が進めばi．1．r．がなくなる場合もあることを示唆しており，少ない断面像から酬zφ廊吻属を同定するような場合には留意すべきであろう。また，
食道を含む虫体の幾つかの切片標本において，i．1．r．の位

荒木国興，1983。興味ある胸部X線経過を示した肺犬糸状2）Beaver，P．C．andwith

filariae

ofOrihe1，T．C．，1965．Human
animalsintheUnitedinfection

States，Am．J．

Trop．Med．Hyg．，14（6）：1010−1029
3）MacLean，J．D．，Beaver，P．C．and Michalek，H．，1979．
Subcutaneous dirofilariasis in Okinawa，Japan，Am．J．
Trop．Med．Hyg．，28（1）：45−48

4）真喜屋清，1990。身近な生活環境の中で感染する寄生虫病

一犬糸状虫症，Medico，21（10）：14−221625）真喜屋清，塚本増久，真鍋英夫，岩田康，1988。胸腺摘出時に偶然発見された肺犬糸状虫症の1例，産業医科大学雑誌10（3）：325−330
6）Neafie，R C．and Piggott，J．，197L Human pulmonary
dirofilariasis，Arch．Path．，92：342−349

7）Nishimura，T．，Kondo，K．and
infection

withasubcutaneous

Shoho，C．，1964．Human
Z万名瞬吻万o伽吻痂s，Biken

J．，7：1−88）Orihel

T．C．andrelationshipofBeaver，P．C．，1965．Morphology

Z万名瞬旋z吻彪犯z孤商andand

Z万勉φ血吻oo勿％つzo・

蜘鷹，Am．J．Trop．Med．Hyg．，14（6）：1030−1043

9）末永敷，伊藤達也，1973。長崎市内における犬糸状虫の浸淫状況と伝播蚊に関する研究5。長崎市内における犬糸状
虫の伝搬蚊，熱帯医学，15（3）：131−140
10）吉村裕之，1988。人イヌ糸状虫症とその免疫診断，臨外
43

（11）

：1675−167911）吉田幸雄，山田稔，塩田恒三，作本台五郎，堤啓，中西洋
二，1984。人体肺寄生犬糸状虫症の1例とくに組織切片に現れた形態と虫種について，日本寄生虫学雑誌，33（5）：435−442

163

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CONTENTS
Originalarticle
Habe，S．，Karen

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Iwanaga，M．，Nakasone，N．，Nakamura，S．，E．B．Wasito，P．Soeparto，S．M．Sudarmo，E．Soewandojo

and

I．G．N．G．Ramh
Etiologic

AgentsofDiarrheal

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AnalysisofAntibody

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Beforeand

AfterTreatmentofStrongyloidiasis一・一・一……………・……・………………・……………・………

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Human

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PulmonaryMorphological

Goto，J．

Dirofilariasis

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・・157−163