1411
TABLE II-HCV SEROPOSITIVE RATES BY POSITIVITY OF SPOUSE AMONG 306 COUPLES IN TOWN X, JAPAN (1990)
Differences
in
husbands and
the
positive rates between positive and negative significant (p < 01).).
groups in both
wives
This large study on HCV infection in married couples suggests that HCV is naturally transmissible through sexual contact. Furthermore, the risk for men and women is roughly the same or even a little higher in men. For HTLV-1 the reverse is true. HCV is transmissible by viral particle alone in body fluids whereas HTLV-1 is transmissible through lymphocytes integrated by provirus (eg, lymphocytes in semen) so that the woman is more at risk than the man during sexual intercourse.
Supported by grants from Ministry of Health and Welfare (for a comprehensive 10-year Strategy for Cancer Control) and from Ministry of Education, Science, and Culture. Division of Epidemiology, Aichi Cancer Centre Research Institute,
Nagoya 464, Japan Division of Virology, National Cancer Centre Research Institute, Tokyo
Seirei Health Screening Centre for Disease Prevention, Shizuoka
Virology Laboratory, Laval University, Quebec, Quebec G1 K 7P4, Canada
MICHEL COUILLARD
Regional Virology Laboratory,
KAZUO TAJIMA
St Joseph’s Hospital, Hamilton, Ontario
LOUISE DESCHÊNES
Neurophysiology Laboratory, Hôpital de l’Enfant-Jésus, Quebec
JACQUES P. TREMBLAY
KUNITADA SHIMOTOHNO
SHIMASUKE OKI
1. Watanabe J, Minegishi K, Mitsumori T, et al. Prevalence of anti-HCV antibody in blood donors in the Tokyo area. Vox Sang 1990; 598: 1-3. 2. Choo GL, Kuo G, Weiner AJ, Overby LP, Bradley DW, Houghton M. Isolation of a cDNA clone derived from a blood borne non-A, non-B viral hepatitis genome. Science 1989; 244: 359-62. 3. Muraiso K, Hijikata M, Ohkoshi S, et al. A structural protein of hepatitis C virus expressed in E coli facilitates acccurate detection of hepatitis C virus. Biochem 4. Ito
Therefore, it became important to verify whether myoblasts obtained from seropositive (histocompatible for classes I and IIDR) donors and selected for transplantation in seronegative recipients were indeed infected. The shell vial assay indicated that these myoblasts were not infected. The possibility that these clones and donor muscle might carry latent CMV was explored by the polymerase chain reaction (PCR).8 A pair of primers was used to amplify a 123 base-pairs sequence on the CMV major immediate early gene.9 This experiment showed no evidence of the CMV genome in the material tested. These clones were therefore considered to be safe for transplantation in seronegative recipients. It seems unlikely that an infected myoblast clone could be developed because CMV would probably hinder cell proliferation. However, since CMV infects a large variety of apparently normal cells" the possibility of developing myoblast clones containing a latent CMV infection remains. We know of no evidence for myoblast clones containing latent CMV infection, nevertheless, we recommend PCR on myoblast clones from seropositive donors before transplantation.
Biophys Commun 1990; 172: 511-16. S, Ito M, Shimotohno K, Tajima K. Massive sero-epidemiological study of hepatitis C virus: clustering of carriers on the southwest coast of Tsushima, Japan. Jpn J Cancer Res 1991; 82: 1-3.
Cytomegalovirus and myoblast transplantation SiR,-Identification of the gene implicated in Duchenne dystrophy (DMD) offers new possibilities for finding a treatment. One possibility is to inject cloned myoblasts from histocompatible donors into the limbs of DMD patients, and human myoblast transplantations have been initiated by research groups in Montreal,l Memphis,2 San Francisco,3 and Quebec.’ Cytomegalovirus (CMV) is an important cause of morbidity and mortality in transplant recipients.5 Ethical problems may arise when myoblasts from seropositive donors are to be transplanted in seronegative recipients, especially if recipients are given immunosuppressive medication. We have evaluated the capacity of cultivated myoblasts to be productively or latently infected with
1. Karpati G. The principle and practice of myoblast transfer. J Neurol Sci 1990; S98: 33. 2. Law PK, Bertorini TE, Goodwin TG, et al. Dystrophin production induced by myoblast transfer therapy in Duchenne muscular dystrophy. Lancet 1990; 336: 114-15. 3. Miller RG, Blau H, Almada A, Steinman L. Myoblast implantation in Duchenne muscular dystrophy. J Neurol Sci 1990; S98: 32. 4. Huard J, Bouchard JP, Roy R, et al. Myoblast transplantation produces dystrophin positive muscle fibers in a 16 year old Duchenne patient. Clin Sci (in press). 5. Rubin RH. Impact of cytomegalovirus infection on organ transplant recipients. Rev Infect Dis 1990; 12: S754-66. 6. Roy R, Dansereau G, Tremblay JP, et al. Expression of major histocompatibility complex antigens on human myoblasts. Transpl Proc 1991; 23: 799-801. 7. Gleaves CA, Smith TF, Shuster EA, Pearson GR. Rapid detection of cytomegalovirus in MRC-5 cells inoculated with urine specimens by using low-speed centrifugation and monoclonal antibody to an early antigen. Clin Microbiol 1984; 19: 917-19. J 8. Saiki RK, Gelfand DH, Stoffel S, et al. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 1988; 239: 487-91. 9. Olive DM, Simsek M, Al-Mufti S. Polymerase chain reaction assay for detection of human cytomegalovirus. J Clin Microbiol 1989; 27: 1238-42. 10. Myerson D, Hackman RC, Nelson JA, Ward DC, McDougall JK. Widespread presence of histologically occult cytomegalovirus. Hum Pathol 1984; 15: 430-39.
Humanised monoclonal
antibody to respiratory syncytial virus
muscular
CMV.
Myoblast clones were obtained from the donor’s muscle biopsy samples by the method of Roy et al.6 Permissivity of three cloned myoblast monolayers on coverslips was tested with an inoculum of the AD-169 strain of human CMV and with a clinical specimen known to be positive for CMV (throat specimen from a 44-year-old man). Infected cells were sought by the shell vial assay with a monoclonal antibody against the 72 kD CMV early antigen .7 Coverslips were stained with a fluorescein-conjugated immunoglobulin. Fluorescent foci were seen after 2-3 days of incubation, whereas uninoculated myoblasts showed no sign of infection. Evidence of secondary infection foci after 7 days of incubation was obtained for one infected clone tested. These results suggest that myoblasts are permissive cells for CMV.
SIR,-Respiratory syncytial virus (RSV) is a major cause of bronchiolitis and pneumonia in children less than 2 years of age.1 Epidemics of RSV infection result in substantial morbidity and some mortality, especially in those who are immunocompromised or have congenital heart disease. There is no vaccine and treatment for severe RSV infections is mainly supportive care. Ribavirin has been approved for the treatment of infants in hospital but prolonged aerosol administration is required to produce any significant clinical improvement and reduction in virus shedding.2 Animal studies indicate that passively transferred antibody may be useful in the control of RSV infection,3,4 and clinical trials show that human gammaglobulin containing high levels of neutralising antibody to RSV produce significant reductions in virus shedding and clinical improvement.s However, large amounts (2 g/kg body weight of ’Sandglobulin’) were required to reduce virus shedding. Certain murine monoclonal antibodies (mAbs) to the fusion glycoprotein of RSV inhibits virus-induced cell fusion and are highly effective against RSV infections in mice.6 However, their clinical use is limited by the immune response to mouse protein, and the production of human mAbs from human antibody-producing cells has proved difficult. One solution is to transfer the antigenbinding regions (complementarity-determining regions from
a mouse
mAb to
a
or
CDR)
human mAb.7 We have used this CDR
1412
grafting technique to reshape a murine mAb, specific for the fusion protein of RSV, as a human IgG mAb.8 This "humanised" mAb recognised all of 24 clinical isolates of RSV tested (representing subgroup A and B strains), neutralised RSV in vitro, inhibited virus-induced cell fusion, and prevented and cleared RSV infections in mice as effectively as did the original mouse
mAb:
MAb19 Property Concentration giving 50% reduction in 1-7 Ilgjml virus plaques in vitro Concentration giving 50% reduction in 6-3 Ilgjml giant cells in vitro Dose giving > 105 reduction m virus titre in lung 5 mg/kg
Hu 19 0-4
(ig/ml
4-0
Ilg/ml
5
mg/kg
A
single dose of 1-25 mg/kg body weight of humanised mAb prevented infection when given intraperitoneally 24 h before intranasal inoculation of BALB/c mice with 104 plaque-forming units of RSV and eliminated virus from the lungs when given on day 4 of infection. A dose of 5 mg/kg body weight was required to eliminate RSV when the virus challenge was increased 10-fold. These studies raise hopes that this humanised antibody will provide an effective treatment of this serious childhood disease. AFRC Institute for Animal Health, Compton, Newbury RG160NN, UK
G. TAYLOR J. FURZE P. R. TEMPEST P. BREMNER
Scotgen Ltd, Aberdeen
F.J. CARR W. J. HARRIS
1. Stott EJ, Taylor G. In: Dimmock NJ, Minor PD, eds. Immune responses, virus infections and disease. London. IRL Press, 1989: 85-104. 2. Conrad DA, Christenson JC, Waner JL, Marks MI Aerosolised ribavirin treatment of respiratory syncytial virus infection in infants hospitalised during an epidemic. Pediatr Infect Dis 1987; 6: 152-58. 3. Prince GA, Horswood RL, Camargo E, Koenig D, Chanock RM. Immunoprophylaxis and immunotherapy of respiratory syncytial virus infection of cotton rats. Infect Immun 1985; 42: 81-87. 4. Hemming VG, Prince GA, Horswood RL, et al. Studies of passive immunotherapy for infections of respiratory syncytial virus in the respiratory tract of a primate
model. J Infect Dis 1985; 152: 1083-87. 5. Hemming VG, Rodriguez W, Kim HW, et al. Intravenous immunoglobulin treatment of respiratory syncynal virus infections in infants and young children. Antimicrobial Ag Chemother 1987; 31: 1882-86. 6. Taylor G, Stott EJ, Bew M, et al. Monoclonal antibodies protect against respiratory syncytial virus infection in mice. Immunology 1984; 52: 137-42. 7. Jones PT, Dear PH, Foote J, Neuberger M, Winter G. Replacing the complementarity determining regions in a human antibody with those of a mouse. Nature 1986; 321: 522-25. 8. Tempest PR, Bremner P, Lambert M, et al. Reshaping a human monoclonal antibody to inhibit human respiratory syncytial virus infection in vivo. Bio Technol 1991; 9: 266-71.
Waterborne outbreak of Escherichia coli 0157 SIR,-4 patients with Escherichia coli 0157 infection from the village of Tarves (population 1700) in Grampian presented during the very hot summer of 1990. All 4 cases (3 boys aged 4, 8, and 9 years; 1 woman aged 20 years) had faecal isolates of verotoxinproducing E coli 0157 H7 phage-type 2. In addition, 2 cases (both boys aged 8 and 9 years) also had Campylobacterjejuni 5/23 isolated from their faeces. All cases gave a history of haemorrhagic colitis with cramping abdominal pain followed by bloody diarrhoea; nausea and vomiting were reported by 3 patients, in 2 of whom a low-grade pyrexia developed. A food history pointed to meat from two barbecue parties as possible causes in 2 cases. However, no pathogens were grown from either left-over food or food obtained from retail outlets that sold the same meat. Some children had been "off sick" from school with diarrhoea for a day or two during the same period as 2 cases, but they had not sought medical advice and no faecal specimen had been submitted for culture from these other cases. Analysis of the drinking water supply to patient’s homes revealed heavy contamination with faecal E coli, but E coli 0 157 was not isolated. Tarves obtains its water supply from a reservoir that is located on a hillock in the village. This supply was beginning to become inadequate for local needs because of heavy demand during the hot
dry weather. A subsidiary water supply was opened up from two nearby reservoirs that had not been used for some time. The supply to one of the subsidiary reservoirs that augmented the main reservoir came from a source that both resembled a field-drain system and may have been contaminated by cattle slurry. The subsidiary reservoir has now been cut off permanently. No further cases of haemorrhagic colitis have been reported. The identification of 4 cases of E coli 0157 H7 as phage type 2, and Cjejuni as both Lior-type 5 and Penner-type 23 confirmed the belief that this outbreak came from a single source. Type 2 is one of eight E coli 0157 H7 phage types commonly seen in Grampian. The serotype of Cjejuni is rarely isolated in Grampian (about one isolate per month). If this outbreak was waterbome, it is disappointing, although not surprising, that we failed to isolate either pathogen from the water supply. Such isolation has been achieved previously for Cjejunil but only once for E coli 0157 H7.12 Recovery of these organisms from environmental samples is often technically difficult because of the altered physiological state that bacteria must exist in to survive a hostile environmentMoreover, any contamination of the water supply is likely to have been intermittent. We thank Dr B. Rowe, Division of Enteric Pathogens, Colindale, for further identification of E coli 0157 and Dr D. Jones, Manchester, for
campylobacter typing. Department of Public Health Medicine, Grampian Health Board, Aberdeen AB9 8QP, UK
V.
Department of Environmental Health, Gordon District Council, Ellon, Aberdeenshire
M. MAIN
Department of Microbiology, City Hospital, Aberdeen
I. GOULD
J. DEV
Melby K, Dahl OP, Crisp L, Penner JL. Epidemiology, clinical, and serological manifestations in patients during a waterborne epidemic due to Campylobacter jejum. J Infect 1990; 21: 309-16. 2. McGowan K, Wickersham E, Strockbine N. Escherichia coli O157:H7 from water Lancet 1989; i. 967-68. 3. Roszak DB, Colwell RR. Survival strategies of bacteria in the natural environment. Micobiol Rev 1987; September: 365-79. 1.
Frozen
biopsy specimens and culture rates of Helicobacter pylori
SllR,-Culture of Helicobacter pylori from gastric biopsies is the reference method to show its colonisation of the gastroduodenal tract; this bacterium is thought to be a major cause of gastritis in man.1 Biopsy specimens are routinely frozen at -80°C when immediate innoculation is not possible, but the influence of freezing on subsequent isolation rates has not been evaluated. Over 15 months, we examined prospectively 177 consecutive patients to investigate the influence of freezing. Most patients had either gastroduodenal ulcers or non-ulcer dyspepsia. Three antralmucosal biopsy specimens were taken from each patient during endoscopy; one was fixed for histopathogical examination, the second was placed in a polystyrene tube ("dry" biopsy), and the third was placed in 0-5 ml of BSK-II medium (commonly used for culture of Borrelia burgdorferP’), which we have previously found to be satisfactory as a transport medium ("wet" biopsy). The wet and dry biopsy specimens were sent to the laboratory for bacteriological examination and were randomly either inoculated within four hours after sampling or frozen at - 80°C. 91 (51 %) of 177 paired biopsy samples were frozen at - 80°C. Our results showed that the isolation rates decreased after freezing, irrespective of whether the biopsy was placed (p < 0-001) or not (p < 0-01) in transport medium: Number positive/number tested Frozen Without transport medium With transport medium p
Non-frozen
19/91(2.?%) 36/86 (42%) 34/91 (37%) 54/86 (63%)
TABLE II-HCV SEROPOSITIVE RATES BY POSITIVITY OF SPOUSE AMONG 306 COUPLES IN TOWN X, JAPAN (1990)
Differences
in
husbands and
the
positive rates between positive and negative significant (p < 01).).
groups in both
wives
This large study on HCV infection in married couples suggests that HCV is naturally transmissible through sexual contact. Furthermore, the risk for men and women is roughly the same or even a little higher in men. For HTLV-1 the reverse is true. HCV is transmissible by viral particle alone in body fluids whereas HTLV-1 is transmissible through lymphocytes integrated by provirus (eg, lymphocytes in semen) so that the woman is more at risk than the man during sexual intercourse.
Supported by grants from Ministry of Health and Welfare (for a comprehensive 10-year Strategy for Cancer Control) and from Ministry of Education, Science, and Culture. Division of Epidemiology, Aichi Cancer Centre Research Institute,
Nagoya 464, Japan Division of Virology, National Cancer Centre Research Institute, Tokyo
Seirei Health Screening Centre for Disease Prevention, Shizuoka
Virology Laboratory, Laval University, Quebec, Quebec G1 K 7P4, Canada
MICHEL COUILLARD
Regional Virology Laboratory,
KAZUO TAJIMA
St Joseph’s Hospital, Hamilton, Ontario
LOUISE DESCHÊNES
Neurophysiology Laboratory, Hôpital de l’Enfant-Jésus, Quebec
JACQUES P. TREMBLAY
KUNITADA SHIMOTOHNO
SHIMASUKE OKI
1. Watanabe J, Minegishi K, Mitsumori T, et al. Prevalence of anti-HCV antibody in blood donors in the Tokyo area. Vox Sang 1990; 598: 1-3. 2. Choo GL, Kuo G, Weiner AJ, Overby LP, Bradley DW, Houghton M. Isolation of a cDNA clone derived from a blood borne non-A, non-B viral hepatitis genome. Science 1989; 244: 359-62. 3. Muraiso K, Hijikata M, Ohkoshi S, et al. A structural protein of hepatitis C virus expressed in E coli facilitates acccurate detection of hepatitis C virus. Biochem 4. Ito
Therefore, it became important to verify whether myoblasts obtained from seropositive (histocompatible for classes I and IIDR) donors and selected for transplantation in seronegative recipients were indeed infected. The shell vial assay indicated that these myoblasts were not infected. The possibility that these clones and donor muscle might carry latent CMV was explored by the polymerase chain reaction (PCR).8 A pair of primers was used to amplify a 123 base-pairs sequence on the CMV major immediate early gene.9 This experiment showed no evidence of the CMV genome in the material tested. These clones were therefore considered to be safe for transplantation in seronegative recipients. It seems unlikely that an infected myoblast clone could be developed because CMV would probably hinder cell proliferation. However, since CMV infects a large variety of apparently normal cells" the possibility of developing myoblast clones containing a latent CMV infection remains. We know of no evidence for myoblast clones containing latent CMV infection, nevertheless, we recommend PCR on myoblast clones from seropositive donors before transplantation.
Biophys Commun 1990; 172: 511-16. S, Ito M, Shimotohno K, Tajima K. Massive sero-epidemiological study of hepatitis C virus: clustering of carriers on the southwest coast of Tsushima, Japan. Jpn J Cancer Res 1991; 82: 1-3.
Cytomegalovirus and myoblast transplantation SiR,-Identification of the gene implicated in Duchenne dystrophy (DMD) offers new possibilities for finding a treatment. One possibility is to inject cloned myoblasts from histocompatible donors into the limbs of DMD patients, and human myoblast transplantations have been initiated by research groups in Montreal,l Memphis,2 San Francisco,3 and Quebec.’ Cytomegalovirus (CMV) is an important cause of morbidity and mortality in transplant recipients.5 Ethical problems may arise when myoblasts from seropositive donors are to be transplanted in seronegative recipients, especially if recipients are given immunosuppressive medication. We have evaluated the capacity of cultivated myoblasts to be productively or latently infected with
1. Karpati G. The principle and practice of myoblast transfer. J Neurol Sci 1990; S98: 33. 2. Law PK, Bertorini TE, Goodwin TG, et al. Dystrophin production induced by myoblast transfer therapy in Duchenne muscular dystrophy. Lancet 1990; 336: 114-15. 3. Miller RG, Blau H, Almada A, Steinman L. Myoblast implantation in Duchenne muscular dystrophy. J Neurol Sci 1990; S98: 32. 4. Huard J, Bouchard JP, Roy R, et al. Myoblast transplantation produces dystrophin positive muscle fibers in a 16 year old Duchenne patient. Clin Sci (in press). 5. Rubin RH. Impact of cytomegalovirus infection on organ transplant recipients. Rev Infect Dis 1990; 12: S754-66. 6. Roy R, Dansereau G, Tremblay JP, et al. Expression of major histocompatibility complex antigens on human myoblasts. Transpl Proc 1991; 23: 799-801. 7. Gleaves CA, Smith TF, Shuster EA, Pearson GR. Rapid detection of cytomegalovirus in MRC-5 cells inoculated with urine specimens by using low-speed centrifugation and monoclonal antibody to an early antigen. Clin Microbiol 1984; 19: 917-19. J 8. Saiki RK, Gelfand DH, Stoffel S, et al. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 1988; 239: 487-91. 9. Olive DM, Simsek M, Al-Mufti S. Polymerase chain reaction assay for detection of human cytomegalovirus. J Clin Microbiol 1989; 27: 1238-42. 10. Myerson D, Hackman RC, Nelson JA, Ward DC, McDougall JK. Widespread presence of histologically occult cytomegalovirus. Hum Pathol 1984; 15: 430-39.
Humanised monoclonal
antibody to respiratory syncytial virus
muscular
CMV.
Myoblast clones were obtained from the donor’s muscle biopsy samples by the method of Roy et al.6 Permissivity of three cloned myoblast monolayers on coverslips was tested with an inoculum of the AD-169 strain of human CMV and with a clinical specimen known to be positive for CMV (throat specimen from a 44-year-old man). Infected cells were sought by the shell vial assay with a monoclonal antibody against the 72 kD CMV early antigen .7 Coverslips were stained with a fluorescein-conjugated immunoglobulin. Fluorescent foci were seen after 2-3 days of incubation, whereas uninoculated myoblasts showed no sign of infection. Evidence of secondary infection foci after 7 days of incubation was obtained for one infected clone tested. These results suggest that myoblasts are permissive cells for CMV.
SIR,-Respiratory syncytial virus (RSV) is a major cause of bronchiolitis and pneumonia in children less than 2 years of age.1 Epidemics of RSV infection result in substantial morbidity and some mortality, especially in those who are immunocompromised or have congenital heart disease. There is no vaccine and treatment for severe RSV infections is mainly supportive care. Ribavirin has been approved for the treatment of infants in hospital but prolonged aerosol administration is required to produce any significant clinical improvement and reduction in virus shedding.2 Animal studies indicate that passively transferred antibody may be useful in the control of RSV infection,3,4 and clinical trials show that human gammaglobulin containing high levels of neutralising antibody to RSV produce significant reductions in virus shedding and clinical improvement.s However, large amounts (2 g/kg body weight of ’Sandglobulin’) were required to reduce virus shedding. Certain murine monoclonal antibodies (mAbs) to the fusion glycoprotein of RSV inhibits virus-induced cell fusion and are highly effective against RSV infections in mice.6 However, their clinical use is limited by the immune response to mouse protein, and the production of human mAbs from human antibody-producing cells has proved difficult. One solution is to transfer the antigenbinding regions (complementarity-determining regions from
a mouse
mAb to
a
or
CDR)
human mAb.7 We have used this CDR
1412
grafting technique to reshape a murine mAb, specific for the fusion protein of RSV, as a human IgG mAb.8 This "humanised" mAb recognised all of 24 clinical isolates of RSV tested (representing subgroup A and B strains), neutralised RSV in vitro, inhibited virus-induced cell fusion, and prevented and cleared RSV infections in mice as effectively as did the original mouse
mAb:
MAb19 Property Concentration giving 50% reduction in 1-7 Ilgjml virus plaques in vitro Concentration giving 50% reduction in 6-3 Ilgjml giant cells in vitro Dose giving > 105 reduction m virus titre in lung 5 mg/kg
Hu 19 0-4
(ig/ml
4-0
Ilg/ml
5
mg/kg
A
single dose of 1-25 mg/kg body weight of humanised mAb prevented infection when given intraperitoneally 24 h before intranasal inoculation of BALB/c mice with 104 plaque-forming units of RSV and eliminated virus from the lungs when given on day 4 of infection. A dose of 5 mg/kg body weight was required to eliminate RSV when the virus challenge was increased 10-fold. These studies raise hopes that this humanised antibody will provide an effective treatment of this serious childhood disease. AFRC Institute for Animal Health, Compton, Newbury RG160NN, UK
G. TAYLOR J. FURZE P. R. TEMPEST P. BREMNER
Scotgen Ltd, Aberdeen
F.J. CARR W. J. HARRIS
1. Stott EJ, Taylor G. In: Dimmock NJ, Minor PD, eds. Immune responses, virus infections and disease. London. IRL Press, 1989: 85-104. 2. Conrad DA, Christenson JC, Waner JL, Marks MI Aerosolised ribavirin treatment of respiratory syncytial virus infection in infants hospitalised during an epidemic. Pediatr Infect Dis 1987; 6: 152-58. 3. Prince GA, Horswood RL, Camargo E, Koenig D, Chanock RM. Immunoprophylaxis and immunotherapy of respiratory syncytial virus infection of cotton rats. Infect Immun 1985; 42: 81-87. 4. Hemming VG, Prince GA, Horswood RL, et al. Studies of passive immunotherapy for infections of respiratory syncytial virus in the respiratory tract of a primate
model. J Infect Dis 1985; 152: 1083-87. 5. Hemming VG, Rodriguez W, Kim HW, et al. Intravenous immunoglobulin treatment of respiratory syncynal virus infections in infants and young children. Antimicrobial Ag Chemother 1987; 31: 1882-86. 6. Taylor G, Stott EJ, Bew M, et al. Monoclonal antibodies protect against respiratory syncytial virus infection in mice. Immunology 1984; 52: 137-42. 7. Jones PT, Dear PH, Foote J, Neuberger M, Winter G. Replacing the complementarity determining regions in a human antibody with those of a mouse. Nature 1986; 321: 522-25. 8. Tempest PR, Bremner P, Lambert M, et al. Reshaping a human monoclonal antibody to inhibit human respiratory syncytial virus infection in vivo. Bio Technol 1991; 9: 266-71.
Waterborne outbreak of Escherichia coli 0157 SIR,-4 patients with Escherichia coli 0157 infection from the village of Tarves (population 1700) in Grampian presented during the very hot summer of 1990. All 4 cases (3 boys aged 4, 8, and 9 years; 1 woman aged 20 years) had faecal isolates of verotoxinproducing E coli 0157 H7 phage-type 2. In addition, 2 cases (both boys aged 8 and 9 years) also had Campylobacterjejuni 5/23 isolated from their faeces. All cases gave a history of haemorrhagic colitis with cramping abdominal pain followed by bloody diarrhoea; nausea and vomiting were reported by 3 patients, in 2 of whom a low-grade pyrexia developed. A food history pointed to meat from two barbecue parties as possible causes in 2 cases. However, no pathogens were grown from either left-over food or food obtained from retail outlets that sold the same meat. Some children had been "off sick" from school with diarrhoea for a day or two during the same period as 2 cases, but they had not sought medical advice and no faecal specimen had been submitted for culture from these other cases. Analysis of the drinking water supply to patient’s homes revealed heavy contamination with faecal E coli, but E coli 0 157 was not isolated. Tarves obtains its water supply from a reservoir that is located on a hillock in the village. This supply was beginning to become inadequate for local needs because of heavy demand during the hot
dry weather. A subsidiary water supply was opened up from two nearby reservoirs that had not been used for some time. The supply to one of the subsidiary reservoirs that augmented the main reservoir came from a source that both resembled a field-drain system and may have been contaminated by cattle slurry. The subsidiary reservoir has now been cut off permanently. No further cases of haemorrhagic colitis have been reported. The identification of 4 cases of E coli 0157 H7 as phage type 2, and Cjejuni as both Lior-type 5 and Penner-type 23 confirmed the belief that this outbreak came from a single source. Type 2 is one of eight E coli 0157 H7 phage types commonly seen in Grampian. The serotype of Cjejuni is rarely isolated in Grampian (about one isolate per month). If this outbreak was waterbome, it is disappointing, although not surprising, that we failed to isolate either pathogen from the water supply. Such isolation has been achieved previously for Cjejunil but only once for E coli 0157 H7.12 Recovery of these organisms from environmental samples is often technically difficult because of the altered physiological state that bacteria must exist in to survive a hostile environmentMoreover, any contamination of the water supply is likely to have been intermittent. We thank Dr B. Rowe, Division of Enteric Pathogens, Colindale, for further identification of E coli 0157 and Dr D. Jones, Manchester, for
campylobacter typing. Department of Public Health Medicine, Grampian Health Board, Aberdeen AB9 8QP, UK
V.
Department of Environmental Health, Gordon District Council, Ellon, Aberdeenshire
M. MAIN
Department of Microbiology, City Hospital, Aberdeen
I. GOULD
J. DEV
Melby K, Dahl OP, Crisp L, Penner JL. Epidemiology, clinical, and serological manifestations in patients during a waterborne epidemic due to Campylobacter jejum. J Infect 1990; 21: 309-16. 2. McGowan K, Wickersham E, Strockbine N. Escherichia coli O157:H7 from water Lancet 1989; i. 967-68. 3. Roszak DB, Colwell RR. Survival strategies of bacteria in the natural environment. Micobiol Rev 1987; September: 365-79. 1.
Frozen
biopsy specimens and culture rates of Helicobacter pylori
SllR,-Culture of Helicobacter pylori from gastric biopsies is the reference method to show its colonisation of the gastroduodenal tract; this bacterium is thought to be a major cause of gastritis in man.1 Biopsy specimens are routinely frozen at -80°C when immediate innoculation is not possible, but the influence of freezing on subsequent isolation rates has not been evaluated. Over 15 months, we examined prospectively 177 consecutive patients to investigate the influence of freezing. Most patients had either gastroduodenal ulcers or non-ulcer dyspepsia. Three antralmucosal biopsy specimens were taken from each patient during endoscopy; one was fixed for histopathogical examination, the second was placed in a polystyrene tube ("dry" biopsy), and the third was placed in 0-5 ml of BSK-II medium (commonly used for culture of Borrelia burgdorferP’), which we have previously found to be satisfactory as a transport medium ("wet" biopsy). The wet and dry biopsy specimens were sent to the laboratory for bacteriological examination and were randomly either inoculated within four hours after sampling or frozen at - 80°C. 91 (51 %) of 177 paired biopsy samples were frozen at - 80°C. Our results showed that the isolation rates decreased after freezing, irrespective of whether the biopsy was placed (p < 0-001) or not (p < 0-01) in transport medium: Number positive/number tested Frozen Without transport medium With transport medium p
Non-frozen
19/91(2.?%) 36/86 (42%) 34/91 (37%) 54/86 (63%)
