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IMMUNOPEROXIDASE STAINING OF TROPHOZOITES IN PRIMARY AMŒBIC MENINGOENCEPHALITIS
479
repaired. The concentration of airborne bacteria ranged from 104 to 107 colonies/m3. All samples contained a predominance of gram-negative bacteria. 30 plant workers and 16 controls were given a complete nlz0ical examination. Immunoglobulins were measured by sinzle radial immunodiffusion. C-reactive protein (C.R.P.) was measured by rocket electrophoresis and fibrinogen degradation products in urine were determined by the latex agglutination test. Antibodies against Escherichia coli endotoxins were analvsed by the enzyme-linked immunosorbent assay. About halt the workers in the sewage plant occasionally had attacks of high fever which appeared a few hours after work. 50% occasionally had a pustulant discharge from the eyes consistiiig exclusively of polymorphonuclear leucocytes. About 20% reported efflorescences of the skin. In the controls, no cases of fever and only one case of eye trouble or skin symptoms were found. No chest symptoms were reported in the exposed or control groups. The
of
amounts
exposed population IMMUNOGBULINS
IgG, IgA, and IgM were than in the control group
higher in the (table). If the
(g/1) AMONG BLOOD-DONORS AND PERSONS WORKING
IN THE SEWAGE TREATMENT PLANT
hgures in parentheses represent standard deviation.
IMMUNOPEROXIDASE STAINING OF TROPHOZOITES IN PRIMARY AMŒBIC MENINGOENCEPHALITIS
SIR,-In 1968 four people developed fatal primary amoebic
meningoencephalitis (P.A.M.), soon after swimming in thermal pools in the North Island of New Zealand. Mandal et al.’ attributed all these cases to invasion by a slime mould, whose identification was subsequently questioned.23 In an attempt to clarify the situation Cursons and Brown,4using Mandal’s original cultures, set out to clone and re-identify the organisms taken from the cerebrospinal fluid of the victims. They were unable to isolate any myxomycetes or pathogenic Ncegleria fowleri and concluded that, since the original cultures were not clones’ and probably contained both N. fowleri and N. gruberi, the former, having a longer generation time, was diluted out by serial subculture, leaving only the faster-growing non-pathogenic N. gruberi. Post-mortem brain tissue of two of the victims, acquired from Hamilton Hospital, New Zealand, was stained by the immunoperoxidase method.5 Using anti-N. fowleri HB-1 serum and anti- Acanthamœba culbertsoni A-1 serum, it was demonstrated unequivocally that the anti-N. fowleri reacted positively with the amoehae in the brain tissue, whereas the anti-A. culbertsoni did not. With the reservation that,some day some immunological cross-reaction might be shown between Na-gleria and slime-mould amoeba (though this seems unlikely because their GC buoyant densities are distinctly different6), we conclude that the 1968 New Zealand cases of P.A.M. were the result of invasion by Ncsgleria fowleri, as were all other reported cases in New Zealand. Department of Microbiology and Genetics, Massey University, Palmerston North,
02
New Zealand
subdivided into those who were never, to the sewage dust, a trend to a continuous increase was found for IgG. The mean number of thrombocytes and leucocytes in peri-
exposed
group
was
sometimes, or often exposed
pheral blood was increased in the exposed population. The thrombocyte count was 219 vs 187 cells x 103/µ1 blood and thatofleucocytes 10.2 vs 7.5 cells x 103/µl blood (P≼0.05 and 0 01, respectively). The titres of IgM anti-endotoxin were significantly increased in the exposed group (10 log 2.79 vs 3 10, P≼0.01). C.R.P. showed values exceeding 1 mg/l in 13 cases out of 30 in the exposed group as compared with 2 out of 16 in the control group. Determinations of F.D.P. in the urine demonstrated that 14 out of 30 in the exposed group had a value exceeding 10 mg/1 compared with 2 out of 16 in the control group. The specific agent in the work environment studied was an aerosol of sewage sludge or a dust of dried sludge. Sewage sludge contains a variety of antigenic materials of microbiological origin. It is therefore difficult to connect with certainty the clinical findings in the exposed population group to any specific agent in the sludge. The symptoms and biochemical findings are, however, expected effects from an exposure to endotoxins. It is difficult to judge the risk for development of some type of chronic disease after an exposure to endotoxin lasting several years. However, any exposure at work which causes both acute clinical symptoms and systemic effects must be carefully investigated before a risk of chronic effects can be ruled out. Such studies are under way.
Departments of Hygiene, Immunology, Institute of Medical Micro biology, and Section of Allergol ogy and Department of Medicine I University of Gothenburg,
RAGNAR RYLANDER KJELL ANDERSSON LARS BELIN GUNILLA BERGLUND RIKARD BERGSTRÖM LARS-ÅKE HANSON MONICA LUNDHOLM INGER MATTSBY
RAY T. M. CURSONS TIM J. BROWN
Lilly Laboratory for Clinical Research, Wishard Memorial Hospital, 1001 West 10th Street, Indianapolis, Indiana 46202, U.S.A.
CLYDE G. CULBERTSON
1. Mandal, B. N. et al. N.Z. med. J. 1970, 71, 16. 2. Culbertson, C. G. A. Rev. Microbiol. 1971, 25, 231. 3. Chang, S. L. C.R.C. crit. Rev. Microbiol. 1974, 3, (2), 135. 4. Cursons, R. T. M., Brown, T. J. N.Z. med J. 1975, 82, 123. 5. Culbertson, C. G. Am. J. clin. Path. 1975, 63, 475. 6. Laskin, A. I., Lechevalier, H. A. (editors) CRC Handbook of Microbiology ii, Microbial Decomposition, p. 703. Ohio, 1973.
Notes and News CHILEAN DOCTORS The Lancet is anxious to get in touch with two doctors in Chile with whom contact has apparently been lost. They are Dr Ivan Inzunza and Dr Carlos Godoy. DrInzunza,who is 42 and married with one child, is a gastroenterologist. He qualified in 1960, and was director of the National Health Service for Employees during the Allende Administration. He was working as a doctor until Aug. 4 this year, when he disappeared. Dr Godoy, who is 39, and is married with three children, is an obstetrician and gynaecologist. He too qualified in 1960 and worked in the National Health Service during the Allende Administration. He was working in the Hospital de San Bernado until Aug. 4; he left the hospital on that day to visit a health centre, but never arrived there. Inquiries have so far produced no information about their whereabouts.
repaired. The concentration of airborne bacteria ranged from 104 to 107 colonies/m3. All samples contained a predominance of gram-negative bacteria. 30 plant workers and 16 controls were given a complete nlz0ical examination. Immunoglobulins were measured by sinzle radial immunodiffusion. C-reactive protein (C.R.P.) was measured by rocket electrophoresis and fibrinogen degradation products in urine were determined by the latex agglutination test. Antibodies against Escherichia coli endotoxins were analvsed by the enzyme-linked immunosorbent assay. About halt the workers in the sewage plant occasionally had attacks of high fever which appeared a few hours after work. 50% occasionally had a pustulant discharge from the eyes consistiiig exclusively of polymorphonuclear leucocytes. About 20% reported efflorescences of the skin. In the controls, no cases of fever and only one case of eye trouble or skin symptoms were found. No chest symptoms were reported in the exposed or control groups. The
of
amounts
exposed population IMMUNOGBULINS
IgG, IgA, and IgM were than in the control group
higher in the (table). If the
(g/1) AMONG BLOOD-DONORS AND PERSONS WORKING
IN THE SEWAGE TREATMENT PLANT
hgures in parentheses represent standard deviation.
IMMUNOPEROXIDASE STAINING OF TROPHOZOITES IN PRIMARY AMŒBIC MENINGOENCEPHALITIS
SIR,-In 1968 four people developed fatal primary amoebic
meningoencephalitis (P.A.M.), soon after swimming in thermal pools in the North Island of New Zealand. Mandal et al.’ attributed all these cases to invasion by a slime mould, whose identification was subsequently questioned.23 In an attempt to clarify the situation Cursons and Brown,4using Mandal’s original cultures, set out to clone and re-identify the organisms taken from the cerebrospinal fluid of the victims. They were unable to isolate any myxomycetes or pathogenic Ncegleria fowleri and concluded that, since the original cultures were not clones’ and probably contained both N. fowleri and N. gruberi, the former, having a longer generation time, was diluted out by serial subculture, leaving only the faster-growing non-pathogenic N. gruberi. Post-mortem brain tissue of two of the victims, acquired from Hamilton Hospital, New Zealand, was stained by the immunoperoxidase method.5 Using anti-N. fowleri HB-1 serum and anti- Acanthamœba culbertsoni A-1 serum, it was demonstrated unequivocally that the anti-N. fowleri reacted positively with the amoehae in the brain tissue, whereas the anti-A. culbertsoni did not. With the reservation that,some day some immunological cross-reaction might be shown between Na-gleria and slime-mould amoeba (though this seems unlikely because their GC buoyant densities are distinctly different6), we conclude that the 1968 New Zealand cases of P.A.M. were the result of invasion by Ncsgleria fowleri, as were all other reported cases in New Zealand. Department of Microbiology and Genetics, Massey University, Palmerston North,
02
New Zealand
subdivided into those who were never, to the sewage dust, a trend to a continuous increase was found for IgG. The mean number of thrombocytes and leucocytes in peri-
exposed
group
was
sometimes, or often exposed
pheral blood was increased in the exposed population. The thrombocyte count was 219 vs 187 cells x 103/µ1 blood and thatofleucocytes 10.2 vs 7.5 cells x 103/µl blood (P≼0.05 and 0 01, respectively). The titres of IgM anti-endotoxin were significantly increased in the exposed group (10 log 2.79 vs 3 10, P≼0.01). C.R.P. showed values exceeding 1 mg/l in 13 cases out of 30 in the exposed group as compared with 2 out of 16 in the control group. Determinations of F.D.P. in the urine demonstrated that 14 out of 30 in the exposed group had a value exceeding 10 mg/1 compared with 2 out of 16 in the control group. The specific agent in the work environment studied was an aerosol of sewage sludge or a dust of dried sludge. Sewage sludge contains a variety of antigenic materials of microbiological origin. It is therefore difficult to connect with certainty the clinical findings in the exposed population group to any specific agent in the sludge. The symptoms and biochemical findings are, however, expected effects from an exposure to endotoxins. It is difficult to judge the risk for development of some type of chronic disease after an exposure to endotoxin lasting several years. However, any exposure at work which causes both acute clinical symptoms and systemic effects must be carefully investigated before a risk of chronic effects can be ruled out. Such studies are under way.
Departments of Hygiene, Immunology, Institute of Medical Micro biology, and Section of Allergol ogy and Department of Medicine I University of Gothenburg,
RAGNAR RYLANDER KJELL ANDERSSON LARS BELIN GUNILLA BERGLUND RIKARD BERGSTRÖM LARS-ÅKE HANSON MONICA LUNDHOLM INGER MATTSBY
RAY T. M. CURSONS TIM J. BROWN
Lilly Laboratory for Clinical Research, Wishard Memorial Hospital, 1001 West 10th Street, Indianapolis, Indiana 46202, U.S.A.
CLYDE G. CULBERTSON
1. Mandal, B. N. et al. N.Z. med. J. 1970, 71, 16. 2. Culbertson, C. G. A. Rev. Microbiol. 1971, 25, 231. 3. Chang, S. L. C.R.C. crit. Rev. Microbiol. 1974, 3, (2), 135. 4. Cursons, R. T. M., Brown, T. J. N.Z. med J. 1975, 82, 123. 5. Culbertson, C. G. Am. J. clin. Path. 1975, 63, 475. 6. Laskin, A. I., Lechevalier, H. A. (editors) CRC Handbook of Microbiology ii, Microbial Decomposition, p. 703. Ohio, 1973.
Notes and News CHILEAN DOCTORS The Lancet is anxious to get in touch with two doctors in Chile with whom contact has apparently been lost. They are Dr Ivan Inzunza and Dr Carlos Godoy. DrInzunza,who is 42 and married with one child, is a gastroenterologist. He qualified in 1960, and was director of the National Health Service for Employees during the Allende Administration. He was working as a doctor until Aug. 4 this year, when he disappeared. Dr Godoy, who is 39, and is married with three children, is an obstetrician and gynaecologist. He too qualified in 1960 and worked in the National Health Service during the Allende Administration. He was working in the Hospital de San Bernado until Aug. 4; he left the hospital on that day to visit a health centre, but never arrived there. Inquiries have so far produced no information about their whereabouts.