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Shwartzman reaction in the brain induced by Haemophilus somnus and Escherichia coli lipopolysaccharide in rabbits
J. Camp.
Path.
1989 Vol.
100
Shwartzman Reaction in the Brain Induced by Htiemophilus somnus and Escherichia coli Lipopolysaccharide in Rabbits Y. Nakajima Tohoku Branch, National
and H. Ueda
Institute of Animal
Health, Shichinohe, Aomori, 039-25, Japan
Summary Intracerebral inoculation of viable Haemophilus somnus resulted in suppurative or fibrino-suppurative meningitis of the brain and spinal cord in rabbits. Multiple fibrin thrombosis complicated with meningitis in the central nervous system was produced by intracerebral inoculation of H. somnus followed by intravenous inoculation with Escherichia coli lipopolysaccharide. The latter reaction may be attributable to a form of Shwartzman reaction. Introduction
Thrombo-embolic meningo-encephalitis (TEME) in cattle is caused by Huemophilus somnus. Characteristics of the nervous lesions include fibrino-suppurative meningo-encephalitis, vasculitis, thrombosis, haemorrhages and metastatic multiple microabscesses. These manifest a septicaemic-like condition complicated with disseminated intravascular coagulation (DIC) (Humphrey and Stephens, 1983; Ishikawa, Tsukuda, Nakajima and Oshima, 1984; Momotani, Yabuki, Miho, Ishikawa and Yoshino, 1985; Stephens, Little, Wilkie and Barnum, 1981). Suppurative meningitis has been produced in mice by infection with H. somnus, but in other studies on mice, rabbits and other rodents, attempts tb produce the disease have failed, suggesting a complex interrelationship between H. somnus and host (Humphrey and Stephens, 1983). DIC has not been produced with H. somnus in an experimental animal model. We report here the formation of multiple fibrin thrombosis and suppurative meningitis in the central nervous system of rabbits, induced by viable H. somnus in combination with Escherichia c.oIi Iipopoiysaccharide. Materials
aqd
Methods
lnocula H. somnus KS-~ was cultured on brain heart inlksion agar supplemented with 0.5 per cent yeast extract and 5 per cent sheep serum at 37°C for 24 h in 5 per cent CO, atmosphere. This strain had been subcultured five times after isolation from a cow with spontaneous TEME. Colonies of KS-~ were 0.1 mm to 0.5 mm in diameter, glistening and pale yellow in colour. For intracerebral inoculation, growth was suspended in 0.2 ml of saline to contain doses ranging from lO* colony-forming units (CFU) to lo5 002 l-9975/89/03023
I + 06 $03.00/O
0
1989 Academic
Press Limited
232
Y. Nakajima
and
H. Ueda
CFU. Eschscherichiacoli lipopolysaccharide (ELPS) (0111: B4, Difco, U.S.A.) was dissolved in 5 ml of 0.85 per cent saline for intravenous inoculation.
(0.5 mg)
Rabbits Thirty-one Japanese albino rabbits were divided into six groups. They weighed 2.5 to 3.5 kg and were neither pregnant nor puerperal. They were intracerebrally inoculated with various doses of H. somnus suspensions (Table 1). Twenty-four hours later, four groups of these rabbits were given ELPS into the ear vein, and the others received 5 ml of 0.85 per cent saline (Table 1).
Thrombosis
in the nervous
Table 1 of groups of rabbits E. coli lipopolysaccharide
system
rnocuta
Animals
died/in
&Yw H. .somnus ix. (0 h) IO’ 10” 10’ 10’ 10b 10’
CFU CFU CFU CFU CFU CFU
inoculated
Multiple* thrombosir/positiue
i.u. (24 h)
Number
H. SOWIPWS followed
ofjibrin
BTlZi?l
l/W
Saline ELPS Salk ELPS ELPS ELPS
O/4 516 o/o 415 213 O/l
316 O/5
O/6 O/3 O/3
CFU: colony forming unit; ELPS: intravenous inoculation. Fibrin thrombi *Number of rabbits with thrombosis tH. somnu was isolated in onr animal.
with
0.29 22.61 0 22.01 1.91 0.13
thrombi
by
per I cm2
Spinal
cord
(1 + ) (4+)
1.20 52.31
(I+) (4+)
t-1 (4+) (I + ) (I + I
0 27.20 9.52 0
C-J (4+ ) (3+) t-1
8. coli lipopolysaccharide; i.c.: intracerebral inoculation; i.v.: is given as mean of group and Kim’s score is given in parentheses. graded by 3 + or 4 +/rabbits with thrombosis.
Patholqpy Rabbits were necropsied two days after the intracerebral inoculation. Organs were fixed in buffered 10 per cent formalin or were cultured for bacterial isolation. Specimens, including five portions from the brain and three from the spinal cord, were embedded in paraffin wax and stajned with haematoxylin and eosin (HE). Selected sections from the central nervous system were stained with phosphotungstic-acid haematoxylin (PTAH I. Calculation
of:jibrin
thrombi
P’l’AH stained sections were pro.jetted on a digitizer (WT-4000, Wacom, Japan), and areas of the sections were measured with the aid of a personal computer (PC9801, KEC, *Japan). These areas included transverse sections of two spinal cords and one involving the midbrain and occipital lobes. All fibrin thrombi in these sections werr countrd and their mean number in 1 cm’ of section was calculated. ‘I’hey were scored according to Kim’s standard as fbllows: ,l or 2 as 1-t ; 3 or 4 as 2 + : 5 to 10 as 3 + : more than 10 as 4+ (Kim, Suzuki, l.ie and Titus. 1976: Momotani PI ul., 19851. Results Lesions caused h_y Gable organisms Focal brain
petechiae or spinal
were cord
occasionally of rabbits
and ELPS observed inoculated
on the surface or cut surface with more than 10” CFU
of the of the
Shwartzman
Reaction
in Brain
233
organisms followed by ELPS. Rabbits given 10” CFU of the organisms followed by ELPS had suppurative or fibrino-suppurative meningitis, multiple fibrin thrombosis and haemorrhages in the leptomeninges (Figs 1 and 2). Mural fibrinoid degeneration of small blood vessels was occasionally seen, their endothelial cells being pyknotic or desquamated. The effect of PTAH was to stain the fibrin thrombi, the walls of the degenerate blood vessels, fibrin exudate and the cytoplasm of infiltrated granular leucocytes (Fig. 3). In the parenchyma, annular and focal haemorrhages were often seen, but fibrin thrombi were less frequent. Nervous tissue occasionally showed ischaemic changes or spongy degeneration. The ventricles were often involved in suppurative meningitis. Rabbits given IO7 CFU of the organism followed by ELPS also manifested multiple thrombosis in the leptomeninges; however, the meningitis was less severe than that in the above rabbits. Meningitis and multiple fibrin thrombosis frequently involved the occipital lobes, midbrain, cerebellum, medulla oblongata and spinal cord. In rabbits that were given lo6 or lo5 CFU of the organisms followed by ELPS, macrophages and granular leucocytes were scattered in the oedematous leptomeninges. Formation of fibrin thrombosis was less frequent (Table 1). Besides lesions in the central nervous system, .a generalized Shwartzman reaction was induced in seven of 18 rabbits which were given ELPS. The lesions included multiple haemorrhages in the thymus, lungs or heart, multiple microthrombosis in the alveolar capillaries, glomeruli or splenic red pulp and focal necrosis of the hepatic tissue or cardiac muscles. H. somnus was isolated from the brain of only one rabbit given lo8 CFU of the organism followed by ELPS. Lesions caused by viable organisms without ELPS The brains of rabbits revealed oedematous and cloudy meninges. Microscopic lesions were similar to those caused by H. somnus and ELPS, except that haemorrhages were not observed and fibrin thrombosis was rare (Fig. 4). In the rabbits that were given lo8 CFU of the organisms, mural fibrinoid degeneration was occasionally seen in blood vessels. The cerebral cortex was occasionally infiltrated with granular leucocytes. Macrophages and granular leucocytes were present in the oedematous leptomeninges in rabbits given lo7 CFU of the organisms. Involvement of the cerebral cortex was less frequent. H. somnus was not isolated from any of these rabbits.
Discussion Among the characteristic nervous lesions of TEME, meningitis, multiple fibrin thrombosis and haemorrhages were induced by our combined inoculation of H. somnus followed by ELPS in rabbits. Though endotoxin may be detected in the peripheral blood of cattle with TEME by the Limulus test (Hakogi, Shimada, Kume and Tabuchi, 1984), the role of endotoxin in TEME is still unclear. Our study suggests that the effects of endotoxin are involved, at least partly, in the formation of DIC during infection with H. somnus. The
234
Y. Nakajima
and H. Ueda
suppurative meningitis caused by H. somnus was similar to that reported in mice by Humphrey and Stephens (1983). H owever, it is suggested that a single inoculation of H. somnus into the brain of rabbits does not provide an adequate experimental model of TEME, since bacterial isolations were negative. Administration of H. somnus, followed after a 24 h interval by ELPS, resulted in lesions (fibrin thrombosis and haemorrhage) which were comparable with those of a localized Shwartzman reaction in the skin. A similar reaction, termed “univisceral Shwartzman reaction” , has been induced in certain organs (Mori, 1981), and it has been suggested that such a reaction possibly triggers Gram-negative infections (Nakajima, Ueda, Takeuchi and Fujimoto, 19871. Characteristic of the univisceral Shwartzman reaction are lesions restricted to a single organ with minimal involvement of other organs. Our results may be attributable to the univisceral Shwartzman reaction; however, involvement of fibrin thrombosis in organs other than the central nervous system was not minimal. This may have been due to the procedure of intracerebral inoculation, in which some of the inoculum may flow into blood vessels of the brain due to sudden high pressure and rupture of capillaries CMims, 1960). Concerning the factors that induced the present fibrin thrombosis, suppurative meningitis caused by H. somnus might be involved. In cattle, the primary pathological process of TEME may be vasculitis (Humphrey and Stephens, 1983). When ELPS is intravenously inoculated, endothelial injury during vascular degeneration might trigger blood coagulation (Culbertson and Osburn, 1980; Elin and Wolff, 1976). Granular leucocytes may have participated in induction of the Shwartzman reaction; acid mucosubstances and lysosomal cationic proteins of granular leucocytes may interact with soluble fibrin-monomer complexes or related derivatives (Horn, 1973). PTAH, a special stain for fibrin, also stained degenerate blood vesselsand cytoplasm of the infiltrated granular leucocytes. Another factor involved in the thrombosis may have been tissue thromboplastin, an important factor in blood coagulation. It is abundant in the brain and might be released during suppurative inflammation. Single and double intravenous inoculation of endotoxin into rabbit brain does not result in the formation of fibrin thrombi, except in the choroid plexus (Kojima, Shimamura and Kanari, 1980). Intracerebral inoculation of ELPS followed by intra\‘enous inoculation of- ELPS did not consistently result in the development of‘ multiple fibrin thrombosis or suppurati\.e meningitis in rabbit brain (Naka,jima. 1988). The exact mechanism causing fibrin thrombosis remains to 1~ discovered.
Fig.
I.
hlultiplr ml/ LPS.
fibrin PI‘AH
Fig.
2.
Suppurative 10” CFU
meningitis of H. somnu
Fig.
3.
Positivrly stained fibrin thrombus. arterial rabbit given IO” CFC of N. ~‘ornnu~ followed
Fig. 4.
Artwial in thr
thrombosis x 18.
irt the ccrcbrllunr
of ;i rahbit
given
associated with tibrin thrombosis and followed by E. co/i LPS. HE x 90.
IO” (XT
of’ff.
rontnu.~ follrwcd
fo( al harmorrhages
wall and exudates in the cerrbral by E. co/r LPS. PTAH x 360.
in a rabbit
h\ E.
give
lrptomeninges
degeneration accompanied by subrndothehal nuclear debris and perivascular cerrbral Icptomcningrs of a rabbit given IO’ UT of H. .wnnus. HE x 360.
infiltration
of J
Shwartzman
Reaction
in Brain
235
236
Y. Nakajima
and
H. Ueda
Acknowledgments We thank
Mr M. Wada
for technical
assistance. References
Culbertson, Jr, R. and Osburn, B. I. (1980). The biologic effects of bacterial endotoxin: a short review. Veterinary Science Communication, 4, 3-14. Elin, R. J. and Wolff, S. M. (1976). Biology ofendotoxin. Annual Review of Medicine, 27, 127-141. Hakogi, E., Shimada, Y., K ume, T. and Tabuchi, K. ( 1984). Perchloric acid treatment and use of chromogenic substrate in the Limulus test: application to veterinary diagnosis. Veterinary Microbiology, 10, 33-42. Horn, R. G. (1973). Evidence for participation of granulocytes in the pathogenesis of the generalized Shwartzman reaction: a review. Journal of Infectious Disease, 128 (suppl), s134-s143. Humphrey, J. D. and Stephens, L. R. (1983). “Haemophilus somnus”: a review. Velerinary Bulletin, 53, 987-1004. Ishikawa, Y., Tsukuda, S., Nakajima, Y. and Oshima, K. (1984). Atypical nervous lesion in Haemophilus somnus infection of cattle. Cornell Veterinarian, 74, 349-353. Kim, H. S., Suzuki, M., Lie, J. T. and Titus, J. L. (1976). Clinically unsuspected disseminated intravascular coagulation (DIC) . An autopsy survey. American Journal of Clinical Pathology, 66, 31-39. Kojima, M., Shimamura, K. and Kanari, T. (1980). Endotoxin and disseminated intravascular coagulation (DIC). In Readings on DIC and Allied Condition, N. Aoki, T. Matsuda, W. Mori and M. Tamaguma, Eds, Ishiyaku-shuppan, Tokyo, pp. 66-72 (in Japanese). Mims, C. A. (1960). Intracerebral injections and the growth of viruses in the mouse brain. British Journal of Experimental Pathology, 41, 52-59. Momotani, E., Yabuki, H., Miho, H., Ishikawa, Y. and Yoshino, T. ( 19851. Histopathological evaluation of disseminated intravascular coagulation in Haemophilus somnus infection in cattle. Journal of Comparative Pathology, 95, 15-23. Mori, W. (1981). The Shwartzman reaction: a review including clinical manifestations and proposal for a univisceral or single organ third type. Histopathology, 5, 113126. Nakajima, Y. (1988). Shwartzman reaction in the brain induced by fractions of Fusobacterium nectrophorum and Escherichia coli lipopolysaccharide in rabbits. L4cta Pathologica Japonica, 38, 541-547. Nakajima, Y., Ueda, H., Takeuchi, S. and Fujimoto, Y. (1987). The effects of Escherichiu coli endotoxin as a trigger for hepatic infection of rabbits with Fusobacterium necrophorum. Journal of Comparative Pathology, 97, 207-2 15. Stephens, L. R., Little, P. B., Wilkie, B. N. and Barnum, D. A. ( 1981). Infectious thromboembolic meningoencephalitis in cattle: a review. Journal of the American Veterinary Association, 178, 378-384. [Received Jbr publication.
September 15th. 19871
Path.
1989 Vol.
100
Shwartzman Reaction in the Brain Induced by Htiemophilus somnus and Escherichia coli Lipopolysaccharide in Rabbits Y. Nakajima Tohoku Branch, National
and H. Ueda
Institute of Animal
Health, Shichinohe, Aomori, 039-25, Japan
Summary Intracerebral inoculation of viable Haemophilus somnus resulted in suppurative or fibrino-suppurative meningitis of the brain and spinal cord in rabbits. Multiple fibrin thrombosis complicated with meningitis in the central nervous system was produced by intracerebral inoculation of H. somnus followed by intravenous inoculation with Escherichia coli lipopolysaccharide. The latter reaction may be attributable to a form of Shwartzman reaction. Introduction
Thrombo-embolic meningo-encephalitis (TEME) in cattle is caused by Huemophilus somnus. Characteristics of the nervous lesions include fibrino-suppurative meningo-encephalitis, vasculitis, thrombosis, haemorrhages and metastatic multiple microabscesses. These manifest a septicaemic-like condition complicated with disseminated intravascular coagulation (DIC) (Humphrey and Stephens, 1983; Ishikawa, Tsukuda, Nakajima and Oshima, 1984; Momotani, Yabuki, Miho, Ishikawa and Yoshino, 1985; Stephens, Little, Wilkie and Barnum, 1981). Suppurative meningitis has been produced in mice by infection with H. somnus, but in other studies on mice, rabbits and other rodents, attempts tb produce the disease have failed, suggesting a complex interrelationship between H. somnus and host (Humphrey and Stephens, 1983). DIC has not been produced with H. somnus in an experimental animal model. We report here the formation of multiple fibrin thrombosis and suppurative meningitis in the central nervous system of rabbits, induced by viable H. somnus in combination with Escherichia c.oIi Iipopoiysaccharide. Materials
aqd
Methods
lnocula H. somnus KS-~ was cultured on brain heart inlksion agar supplemented with 0.5 per cent yeast extract and 5 per cent sheep serum at 37°C for 24 h in 5 per cent CO, atmosphere. This strain had been subcultured five times after isolation from a cow with spontaneous TEME. Colonies of KS-~ were 0.1 mm to 0.5 mm in diameter, glistening and pale yellow in colour. For intracerebral inoculation, growth was suspended in 0.2 ml of saline to contain doses ranging from lO* colony-forming units (CFU) to lo5 002 l-9975/89/03023
I + 06 $03.00/O
0
1989 Academic
Press Limited
232
Y. Nakajima
and
H. Ueda
CFU. Eschscherichiacoli lipopolysaccharide (ELPS) (0111: B4, Difco, U.S.A.) was dissolved in 5 ml of 0.85 per cent saline for intravenous inoculation.
(0.5 mg)
Rabbits Thirty-one Japanese albino rabbits were divided into six groups. They weighed 2.5 to 3.5 kg and were neither pregnant nor puerperal. They were intracerebrally inoculated with various doses of H. somnus suspensions (Table 1). Twenty-four hours later, four groups of these rabbits were given ELPS into the ear vein, and the others received 5 ml of 0.85 per cent saline (Table 1).
Thrombosis
in the nervous
Table 1 of groups of rabbits E. coli lipopolysaccharide
system
rnocuta
Animals
died/in
&Yw H. .somnus ix. (0 h) IO’ 10” 10’ 10’ 10b 10’
CFU CFU CFU CFU CFU CFU
inoculated
Multiple* thrombosir/positiue
i.u. (24 h)
Number
H. SOWIPWS followed
ofjibrin
BTlZi?l
l/W
Saline ELPS Salk ELPS ELPS ELPS
O/4 516 o/o 415 213 O/l
316 O/5
O/6 O/3 O/3
CFU: colony forming unit; ELPS: intravenous inoculation. Fibrin thrombi *Number of rabbits with thrombosis tH. somnu was isolated in onr animal.
with
0.29 22.61 0 22.01 1.91 0.13
thrombi
by
per I cm2
Spinal
cord
(1 + ) (4+)
1.20 52.31
(I+) (4+)
t-1 (4+) (I + ) (I + I
0 27.20 9.52 0
C-J (4+ ) (3+) t-1
8. coli lipopolysaccharide; i.c.: intracerebral inoculation; i.v.: is given as mean of group and Kim’s score is given in parentheses. graded by 3 + or 4 +/rabbits with thrombosis.
Patholqpy Rabbits were necropsied two days after the intracerebral inoculation. Organs were fixed in buffered 10 per cent formalin or were cultured for bacterial isolation. Specimens, including five portions from the brain and three from the spinal cord, were embedded in paraffin wax and stajned with haematoxylin and eosin (HE). Selected sections from the central nervous system were stained with phosphotungstic-acid haematoxylin (PTAH I. Calculation
of:jibrin
thrombi
P’l’AH stained sections were pro.jetted on a digitizer (WT-4000, Wacom, Japan), and areas of the sections were measured with the aid of a personal computer (PC9801, KEC, *Japan). These areas included transverse sections of two spinal cords and one involving the midbrain and occipital lobes. All fibrin thrombi in these sections werr countrd and their mean number in 1 cm’ of section was calculated. ‘I’hey were scored according to Kim’s standard as fbllows: ,l or 2 as 1-t ; 3 or 4 as 2 + : 5 to 10 as 3 + : more than 10 as 4+ (Kim, Suzuki, l.ie and Titus. 1976: Momotani PI ul., 19851. Results Lesions caused h_y Gable organisms Focal brain
petechiae or spinal
were cord
occasionally of rabbits
and ELPS observed inoculated
on the surface or cut surface with more than 10” CFU
of the of the
Shwartzman
Reaction
in Brain
233
organisms followed by ELPS. Rabbits given 10” CFU of the organisms followed by ELPS had suppurative or fibrino-suppurative meningitis, multiple fibrin thrombosis and haemorrhages in the leptomeninges (Figs 1 and 2). Mural fibrinoid degeneration of small blood vessels was occasionally seen, their endothelial cells being pyknotic or desquamated. The effect of PTAH was to stain the fibrin thrombi, the walls of the degenerate blood vessels, fibrin exudate and the cytoplasm of infiltrated granular leucocytes (Fig. 3). In the parenchyma, annular and focal haemorrhages were often seen, but fibrin thrombi were less frequent. Nervous tissue occasionally showed ischaemic changes or spongy degeneration. The ventricles were often involved in suppurative meningitis. Rabbits given IO7 CFU of the organism followed by ELPS also manifested multiple thrombosis in the leptomeninges; however, the meningitis was less severe than that in the above rabbits. Meningitis and multiple fibrin thrombosis frequently involved the occipital lobes, midbrain, cerebellum, medulla oblongata and spinal cord. In rabbits that were given lo6 or lo5 CFU of the organisms followed by ELPS, macrophages and granular leucocytes were scattered in the oedematous leptomeninges. Formation of fibrin thrombosis was less frequent (Table 1). Besides lesions in the central nervous system, .a generalized Shwartzman reaction was induced in seven of 18 rabbits which were given ELPS. The lesions included multiple haemorrhages in the thymus, lungs or heart, multiple microthrombosis in the alveolar capillaries, glomeruli or splenic red pulp and focal necrosis of the hepatic tissue or cardiac muscles. H. somnus was isolated from the brain of only one rabbit given lo8 CFU of the organism followed by ELPS. Lesions caused by viable organisms without ELPS The brains of rabbits revealed oedematous and cloudy meninges. Microscopic lesions were similar to those caused by H. somnus and ELPS, except that haemorrhages were not observed and fibrin thrombosis was rare (Fig. 4). In the rabbits that were given lo8 CFU of the organisms, mural fibrinoid degeneration was occasionally seen in blood vessels. The cerebral cortex was occasionally infiltrated with granular leucocytes. Macrophages and granular leucocytes were present in the oedematous leptomeninges in rabbits given lo7 CFU of the organisms. Involvement of the cerebral cortex was less frequent. H. somnus was not isolated from any of these rabbits.
Discussion Among the characteristic nervous lesions of TEME, meningitis, multiple fibrin thrombosis and haemorrhages were induced by our combined inoculation of H. somnus followed by ELPS in rabbits. Though endotoxin may be detected in the peripheral blood of cattle with TEME by the Limulus test (Hakogi, Shimada, Kume and Tabuchi, 1984), the role of endotoxin in TEME is still unclear. Our study suggests that the effects of endotoxin are involved, at least partly, in the formation of DIC during infection with H. somnus. The
234
Y. Nakajima
and H. Ueda
suppurative meningitis caused by H. somnus was similar to that reported in mice by Humphrey and Stephens (1983). H owever, it is suggested that a single inoculation of H. somnus into the brain of rabbits does not provide an adequate experimental model of TEME, since bacterial isolations were negative. Administration of H. somnus, followed after a 24 h interval by ELPS, resulted in lesions (fibrin thrombosis and haemorrhage) which were comparable with those of a localized Shwartzman reaction in the skin. A similar reaction, termed “univisceral Shwartzman reaction” , has been induced in certain organs (Mori, 1981), and it has been suggested that such a reaction possibly triggers Gram-negative infections (Nakajima, Ueda, Takeuchi and Fujimoto, 19871. Characteristic of the univisceral Shwartzman reaction are lesions restricted to a single organ with minimal involvement of other organs. Our results may be attributable to the univisceral Shwartzman reaction; however, involvement of fibrin thrombosis in organs other than the central nervous system was not minimal. This may have been due to the procedure of intracerebral inoculation, in which some of the inoculum may flow into blood vessels of the brain due to sudden high pressure and rupture of capillaries CMims, 1960). Concerning the factors that induced the present fibrin thrombosis, suppurative meningitis caused by H. somnus might be involved. In cattle, the primary pathological process of TEME may be vasculitis (Humphrey and Stephens, 1983). When ELPS is intravenously inoculated, endothelial injury during vascular degeneration might trigger blood coagulation (Culbertson and Osburn, 1980; Elin and Wolff, 1976). Granular leucocytes may have participated in induction of the Shwartzman reaction; acid mucosubstances and lysosomal cationic proteins of granular leucocytes may interact with soluble fibrin-monomer complexes or related derivatives (Horn, 1973). PTAH, a special stain for fibrin, also stained degenerate blood vesselsand cytoplasm of the infiltrated granular leucocytes. Another factor involved in the thrombosis may have been tissue thromboplastin, an important factor in blood coagulation. It is abundant in the brain and might be released during suppurative inflammation. Single and double intravenous inoculation of endotoxin into rabbit brain does not result in the formation of fibrin thrombi, except in the choroid plexus (Kojima, Shimamura and Kanari, 1980). Intracerebral inoculation of ELPS followed by intra\‘enous inoculation of- ELPS did not consistently result in the development of‘ multiple fibrin thrombosis or suppurati\.e meningitis in rabbit brain (Naka,jima. 1988). The exact mechanism causing fibrin thrombosis remains to 1~ discovered.
Fig.
I.
hlultiplr ml/ LPS.
fibrin PI‘AH
Fig.
2.
Suppurative 10” CFU
meningitis of H. somnu
Fig.
3.
Positivrly stained fibrin thrombus. arterial rabbit given IO” CFC of N. ~‘ornnu~ followed
Fig. 4.
Artwial in thr
thrombosis x 18.
irt the ccrcbrllunr
of ;i rahbit
given
associated with tibrin thrombosis and followed by E. co/i LPS. HE x 90.
IO” (XT
of’ff.
rontnu.~ follrwcd
fo( al harmorrhages
wall and exudates in the cerrbral by E. co/r LPS. PTAH x 360.
in a rabbit
h\ E.
give
lrptomeninges
degeneration accompanied by subrndothehal nuclear debris and perivascular cerrbral Icptomcningrs of a rabbit given IO’ UT of H. .wnnus. HE x 360.
infiltration
of J
Shwartzman
Reaction
in Brain
235
236
Y. Nakajima
and
H. Ueda
Acknowledgments We thank
Mr M. Wada
for technical
assistance. References
Culbertson, Jr, R. and Osburn, B. I. (1980). The biologic effects of bacterial endotoxin: a short review. Veterinary Science Communication, 4, 3-14. Elin, R. J. and Wolff, S. M. (1976). Biology ofendotoxin. Annual Review of Medicine, 27, 127-141. Hakogi, E., Shimada, Y., K ume, T. and Tabuchi, K. ( 1984). Perchloric acid treatment and use of chromogenic substrate in the Limulus test: application to veterinary diagnosis. Veterinary Microbiology, 10, 33-42. Horn, R. G. (1973). Evidence for participation of granulocytes in the pathogenesis of the generalized Shwartzman reaction: a review. Journal of Infectious Disease, 128 (suppl), s134-s143. Humphrey, J. D. and Stephens, L. R. (1983). “Haemophilus somnus”: a review. Velerinary Bulletin, 53, 987-1004. Ishikawa, Y., Tsukuda, S., Nakajima, Y. and Oshima, K. (1984). Atypical nervous lesion in Haemophilus somnus infection of cattle. Cornell Veterinarian, 74, 349-353. Kim, H. S., Suzuki, M., Lie, J. T. and Titus, J. L. (1976). Clinically unsuspected disseminated intravascular coagulation (DIC) . An autopsy survey. American Journal of Clinical Pathology, 66, 31-39. Kojima, M., Shimamura, K. and Kanari, T. (1980). Endotoxin and disseminated intravascular coagulation (DIC). In Readings on DIC and Allied Condition, N. Aoki, T. Matsuda, W. Mori and M. Tamaguma, Eds, Ishiyaku-shuppan, Tokyo, pp. 66-72 (in Japanese). Mims, C. A. (1960). Intracerebral injections and the growth of viruses in the mouse brain. British Journal of Experimental Pathology, 41, 52-59. Momotani, E., Yabuki, H., Miho, H., Ishikawa, Y. and Yoshino, T. ( 19851. Histopathological evaluation of disseminated intravascular coagulation in Haemophilus somnus infection in cattle. Journal of Comparative Pathology, 95, 15-23. Mori, W. (1981). The Shwartzman reaction: a review including clinical manifestations and proposal for a univisceral or single organ third type. Histopathology, 5, 113126. Nakajima, Y. (1988). Shwartzman reaction in the brain induced by fractions of Fusobacterium nectrophorum and Escherichia coli lipopolysaccharide in rabbits. L4cta Pathologica Japonica, 38, 541-547. Nakajima, Y., Ueda, H., Takeuchi, S. and Fujimoto, Y. (1987). The effects of Escherichiu coli endotoxin as a trigger for hepatic infection of rabbits with Fusobacterium necrophorum. Journal of Comparative Pathology, 97, 207-2 15. Stephens, L. R., Little, P. B., Wilkie, B. N. and Barnum, D. A. ( 1981). Infectious thromboembolic meningoencephalitis in cattle: a review. Journal of the American Veterinary Association, 178, 378-384. [Received Jbr publication.
September 15th. 19871