News15(1&2)

ACLAD NEWSLETTER Vol. 15, Nos. 1&2

American Committee on Laboratory Animal Diseases

September 1994

Editor:
Stephen S. Morse The Rockefeller University 1230 York Avenue, Box 120 New York, NY 10021-6399 Telephone: (212) 327-7722 FAX: (212) 327-7172 OR 327-7974 E-mail: morse@rockvax.rockefeller.edu * Items for the Newsletter, general comments

Editorial Assistant:
Joan Bailie Section of Comparative Medicine Yale University School of Medicine P.O. Box 208016 New Haven, CT 06520-8016 Telephone: (203) 785-2507 FAX: (203) 785-7499 * Changes of address; general comments; questions about mailing or dues

ACLAD Officers 1993-94: PRESIDENT: Bob Jacoby SECRETARY-TREASURER: Steve Morse COUNCILLORS: Tony Allen, Jim Fox, Pat Manning, Dean Percy, Abbe Smith, Jerry van Hoosier, Pravin Bhatt ACLAD BUSINESS MEETING (PITTSBURGH): SUNDAY, OCTOBER 16, 5 PM, VISTA HOTEL (CRAWFORD ROOM-W) ACLAD SCIENTIFIC PROGRAM (PITTSBURGH): Annual Seminar and Wallace P. Rowe Lecture TUESDAY, OCTOBER 18, 8 A.M. TO NOON, Room S-4, CONVENTION CENTER 8-11 AM: Symposium, "Genetic Basis of Resistance to Infectious Diseases" 11AM-Noon: Wallace P. Rowe Lecture, by Dr. Diane Griffin "The Pathogenesis of Acute Viral Encephalitis"

NEXT ISSUE OF THE NEWSLETTER: MARCH 1995. PLEASE SEND CONTRIBUTION FOR THE NEXT ISSUE BY FEBRUARY 10, 1995. ========================================================== PRESIDENT'S MESSAGE

Fellow Members, The ACLAD Business Meeting will be held from 5-6 PM on Sunday, October 16, in the Crawford/W Room of the Vista Hotel. Among the most important items for discussion will be a critique of proposed revisions to the Constitution and By-Laws. Drafting of the revised Constitutions and By-Laws is still ongoing, and the revisions will be sent out in a separate mailing. We also will be nominating officers for next year's Council. As always, there will be plenty of time for discussion about program initiatives that ACLAD should consider. All members are cordially invited, and encouraged, to attend and participate. Bob Jacoby, President This year's Wallace P. Rowe lecturer is Dr. Diane Griffin. Dr. Griffin, Professor in the Johns Hopkins University School of Medicine, recently assumed the Chair of the Department of Immunology & Infectious Diseases at the Johns Hopkins University School of Hygiene & Public Health. She has long been in the forefront of both immunology and infectious diseases, and is recognized as a leader in research on viral encephalitis and in animal model approaches. Notices for the Business Meeting and the scientific program at Pittsburgh this October are on the next page. Finally, your comments and contributions for this Newsletter are always welcome and encouraged. (For everyone following the evolving story of the hantaviruses, there will be a brief review and updates in the next issue.) (MORE ANNOUNCEMENTS AT END) ==========================================================
ACLAD BUSINESS MEETING AND NOMINATIONS OF COUNCIL DATE: SUNDAY, OCTOBER 16, 1994 TIME: 5 PM PLACE: CRAWFORD/W ROOM, VISTA HOTEL *****************************

ACLAD SCIENTIFIC PROGRAM Annual Seminar and Wallace P. Rowe Lecture Tuesday, October 18 8 A.M.--Noon Room S-4, CONVENTION CENTER 8--11 AM : SEMINAR The Genetic Basis of Resistance to Infectious Diseases: A Dynamic Relationship Between Host and Parasite Moderators: Jim Fox, Dan Ringler JoAnne Flynn: Immune Responses in Experimental Tuberculosis Ann Fortier: Francisella tularensis, a Model of Intracellular Pathogens David G. Brownstein: The Mouse Genome and Resistance to Naturally Occurring Viruses Arthur Like: Parvovirus Induced Autoimmune Diabetes in the Rat 11 A.M.--Noon: WALLACE P. ROWE LECTURE: Diane E. Griffin, M.D., Ph.D. Professor and Chair, Dept. of Immunology & Infectious Diseases Johns Hopkins University School of Hygiene & Public Health, and Professor, Johns Hopkins University School of Medicine, Baltimore "The Pathogenesis of Acute Viral Encephalitis: New Insights from a Murine Model" NOTE: ACLAD will also sponsor an afternoon Forum for trainees and training program directors. Further information is available from Bob Jacoby (or inquire at Pittsburgh). ==========================================================

MINUTES OF THE ACLAD ANNUAL BUSINESS MEETING, 1993 Nashville, Tennessee, Sunday, November 14, 1993

Councillors attending: Jacoby (presiding), Fox, Morse, Percy, Ringler, Smith, Van Hoosier. The meeting was called to order at approximately 5:00 P.M. by President Jacoby, who began by briefly reviewing the history of ACLAD programs. ACLAD's first program was the Wallace P. Rowe Lectureship. This tradition has continued, and over the years the program has expanded to become an entire morning session. Recent and contemplated changes in ACLAD Bylaws will provide a structure with increased membership participation, and continuity provided by 2-year terms for officers. At the same time, AALAS is undergoing major governance changes, and the attendance and focus at national AALAS meetings have changed over the years. The floor was then opened to general discussion. Dr. Prattis and several other members suggested that ACLAD could include other animal disease questions in addition to its traditional emphasis on infectious (especially viral) diseases. Dr. Collins suggested that a future role for ACLAD was as an information resource, for example maintaining directories of experts who could be consulted when there were specialized questions concerning new or unfamiliar disease problems. Dr. Van Hoosier noted that the increased use of transgenic technology (perhaps, Dr. Jacoby suggested, even including transgenic primates in the future), will lead to many problems in detecting diseases in these unconventional backgrounds, and a forum is needed to discuss these problems and make available necessary specialized expertise.
Dr. Jacoby led a discussion of possible formats for the ACLAD Annual Scientific Session at AALAS. Several suggestions were considered. An open session, for informal discussion and impromptu presentations, had been a feature of early ACLAD meetings but had been dropped in recent years. Many of the members at the Business Meeting thought it worth reviving in some form, the major question being at what hour it should be scheduled. It was suggested that the Trainee's Roundtable, introduced this year as a luncheon meeting, might fulfill this role. Suggestions for the Newsletter were also solicited. Dr. Cruise suggested a "Disease of the Month" featuring a brief review of an important disease, such as MHV, in each issue. A "Transgenic Corner" (a correspondence section on people's problems with transgenic animals) was also suggested. Dr. Rahija suggested publishing an ACLAD membership directory as an annual supplement to the Newsletter. The financial statement (prepared by Ms. Joan Bailie) was read by the Secretary-Treasurer. As of November 1, 1993, total deposits for 1993 were $2,300 (most from 1993 dues payments) and debits were $243.67. The opening balance in January 1993 was $6,212.23, leading to an account blance of $8,299.56 as of November 1. It is expected that most of this balance will be required to pay speakers' expenses and other costs of the annual meeting. Membership renewals were discussed. Dues, currently $20 per year, will continue to be due on a calendar year basis as in the past. In addition to the regular annual dues mailing (usually in the spring), each issue of the Newsletter will contain a membership application/renewal form. It was suggested that year of expiration be printed on the mailing label so members will be reminded when their renewals are due.

In order to plan for the future, several committees were formed at the meeting (additional volunteers are welcome):

* Programs and Planning: J. Fox (Chair); A. Allen; R. Rahija; D. Ringler. Purpose: To develop the programs for future ACLAD meetings (including the 1994 and 1995 meetings), and to make recommendations, as appropriate, concerning additional future activities.

* Publications: S. Morse (Chair); J. Hilliard; D. Percy; F. Quimby; A. Smith. Purpose: To examine the Newsletter and recommend improvements in content and format, and to assist the Editor in implementing these recommendations by identifying sources for new material; as appropriate, to consider future needs for ACLAD publications. [This committee met on Monday afternoon, November 15, and discussed plans for new regular features.]

* Bylaws: R. Jacoby (Chair); P. Bhatt; P. Manning. Purpose: To revise the ACLAD Bylaws to reflect current needs; to examine the governance of ACLAD and recommend necessary changes to better serve the membership. A Fundraising Committee was also suggested for the future. After some additional discussion, the meeting was adjourned at approximately 6:15 P.M.

Respectfully submitted, Stephen S. Morse Secretary-Treasurer ==========================================================
RESEARCH UPDATES AND FORUM

Experimental Sendai virus infection in aged BALB/c mice

Robert O. Jacoby, Pravin N. Bhatt, and Stephen W. Barthold

Sendai virus (SV) infection was compared in 2 month-old (young) and 24 month-old (aged) BALB/c mice after intranasal inoculation. Mean virus titers in lung 6 days after inoculation were significantly higher in aged mice. At day 10, aged mice had infectious virus in lung, whereas young mice did not. No virus was detected in either group by day 20. Aged mice with pulmonary tumors had higher virus titers than mice that did not have tumors. The prevalence of SV antigen in lung paralleled the virological results. At days 10 and 20, serum antibody titers measured by enzyme immunoassay were significantly lower in aged mice. Bronchopneumonia and alveolitis developed in both age groups. In the young mice, necrosis and inflammation were prominent on day 10, but were largely resolved by day 20. In the aged mice, necrosis was severe on day 10, but inflammation was less advanced than in young mice. Inflammation was still prevalent in aged mice at day 20 and attempts at repair included prominent epithelialization and squamous metaplasia. These results indicate that aged mice are less efficient than young mice at responding to SV infection and that immunosenescence may be a factor in this deficiency. Future studies will characterize responses of aged mice in more detail and evaluate SV infection as a natural model of viral pneumonia in the aged host, especially as it may contrast with the murine model of adapted influenza infection. These studies will also include further evaluation of a potential correlation between lung neoplasms and susceptibility to viral pneumonia.
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SUMMARIES OF RECENT ARTICLES OF NOTE

by Dean H. Percy
Ontario Veterinary College, Guelph, ONT N1G 2W1

1. Boot, R., Thuis, H.C., Veenema, J.L., Bakker, R.H.G. & Walwoort, H.C.
Colonization andantibody response in mice and rats experimentally infected with Pasteurellaceae from different rodent species. Laboratory Animals 28: 130-137, 1994. In this study, mice and rats were inoculated intranasally with isolates of Pasteurellaceae acquired from mice, rats, hamsters, or gerbils. Strains of P. pneumotropica were the most common isolates studied. Mice and rats were readily colonized with strains isolated from mice and rats, respectively, and to a lesser extent, isolates from gerbils and hamsters. Interspecies colonization of isolates from mice and rats occurred, usually with seroconversion. The organism was frequently recovered from both respiratory tract and cecum. Once infected with mouse strains, rats readily transmitted the organism to cage mates. This study provides additional evidence that interspecies transmission of rodent strains of Pasteurellaceae may occur.

2. Cushion, M., Kaselis, M., Stringer, S.L., & Stringer, J.R. Genetic stability and diversity of Pneumocystis carinii infecting rat colonies. Infect. Immun. 61: 4801-4813, 1993. Based on molecular and antigenic studies, there is evidence that isolates of Pneumocystis carinii from humans, rats and ferrets are different species. In this study, 10 isolates of P. carinii from different colonies of immunodeficient rats were evaluated for diversity at the chromosomal level. Based on differences in gene sequences, electrophoretic karyotypes, and hybridization profiles, it was concluded that these rats were infected with genetically distinct strains of the organism, although the genome of the organism within the host is relatively stable over time.

3. Schoeb, T.R., Dybvig, K., Davidson, M.K., & Davis, J.K. Cultivation of cilia-associated respiratory bacillus in artificial medium and determination of 16S rRNA gene sequence. J. Clin. Microbiol. 31: 2751-2757, 1993. The cilia-associated respiratory (CAR) bacillus is an unclassified gliding bacterium associated with respiratory tract disease in rats, mice and rabbits. In this study, the researchers report the successful cultivation of a reference strain and isolates from a naturally-infected rabbit and rats in Dulbecco's minimal essential medium. Based on morphology and DNA restriction fragment patterns, there were significant differences between the rabbit and rat isolates. Of particular note was the isolation and identification of Mycoplasma spp, including M.pulmonis, from isolates of the CAR bacillus from rats. This study again serves to emphasize the difficulties in defining the roles and possible interactions of these organisms in chronic murine respiratory disease.
Some Other Articles of Note -- Selected References by Dean H. Percy

1. Artwohl, J.E., Cera, L.M., Wright, M.F., Medina, L.V., & Kim, L.J. The efficacy of a dirty bedding sentinel system for detecting Sendai virus infection in mice: A comparison of clinical signs and seroconversion. Lab. Anim. Sci. 44:73-75, 1994.

2. Barthold, S.W., Smith, A.L., & Bhatt, P.N. Infectivity, disease patterns, and serologic profiles of reovirus serotypes 1, 2, and 3 in infant and weanling mice. Lab. Anim. Sci. 43:425-430, 1993.

3. Bray, M.V., Barthold, S.W., Sidman, C.L., Roths, J., & Smith, A.L. Exacerbation of Pneumocystis carinii pneumonia in immunodeficient (scid) mice by concurrent infection with a pneumovirus. Infect. Immun. 61:1586-1588, 1993.

4. Duncan, A.J., Carman, R.J., Olsen, G.J., & Wilson, K.H. Assignment of the agent of Tyzzer's disease to Clostridium piliforme comb. nov. on the basis of 16S rRNA sequence analysis. Internat. J. Systematic Bacteriol. 43: 314-318, 1993.

5. Frith, C.H., Ward, J.M., & Chandra, M. The morphologic, immunohistochemistry, and incidence of hematopoietic neoplasms in mice and rats. Toxicologic Pathology 21:206-218, 1993.

6. Froberg, M.K., Fitzgerald, T.J., Hamilton, T.R., Hamilton, B., & Zarabi, M. Pathology of congenital syphilis in rabbits. Infect. Immun. 61:4743-4749, 1994.

7. Hunskaar, S., & Fosse, R.T. Allergy to laboratory mice and rats: a review of its prevention, management, and treatment. Laboratory Animals 27:206-221, 1993.

8. Lai, W.C., Linton, G., Bennett, M., & Pakes, S.P. Genetic control of resistance to Mycoplasma pulmonis infection in mice. Infect. Immun. 61:4615-4621, 1993.

9. Pohlmeyer, G., & Deerberg, G. Nude rats as a model of natural Pneumocystis carinii pneumonia: Sequential morphological study of lung lesions. J. Comp. Pathol. 109: 217-230, 1993.

10. Robins-Brown, R.M., Tokhi, A.M., Adams, L.M., Bennett-Wood, V., Moisidis, A.V., Krejany, E.O., & O'Gorman, L.E. Adherence characteristics of attaching and effacing strains of Escherichia coli from rabbits. Infect. Immun. 62: 1584-1592, 1994.

11. Rozengurt, N., & Sanchez, S. A spontaneous outbreak of Theiler's encephalomyelitis in a colony of severe combined immunodeficient mice in the UK. Laboratory Animals 27:229-234, 1993.

12. Thurman, J.D., Bucci, T.J., Hart, R.W., Turturro, A. Survival, body weight, and spontaneous neoplasms in ad libidum-fed and food-restricted Fischer-344 rats. Toxicologic Pathology 22:1-9, 1994.
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THE QUALITY ASSURANCE PROGRAM FOR LABORATORY ANIMAL DIAGNOSTIC FACILITIES

by Pamela B. Moore
The Rockefeller University, New York, NY 10021

A quality assurance program for laboratory animal diagnostic facilities was established in 1981 by Dr. Dennis Stark, then Director of the Laboratory Animal Research Center (LARC) at The Rockefeller University, to fill a need seen by several microbiologists for quality assurance in the area of animal diagnostics (1). The program comprises both bacterial cultures and rodent serology testing and is patterned somewhat after the program administered by the Centers for Disease Control for human diagnostic facilities (2,3). Since no standardization of reagents and techniques among participants is made, consensus among labs as to identity only is possible.

Charter members of the program, facilities that have participated in the program since its inception in 1981 and are still participating today, include Anmed Biosafe, Inc. (Rockville, MD), G.D. Searle & Co. (Skokie, IL), The Rockefeller University (New York, NY), University of Miami (Miami, FL), University of Missouri at Columbia, and University of Wisconsin (Madison, WI). At present, there are 17 members in the United States and 3 in Europe that receive samples directly. Several additional labs here and in Europe have used the program but are not included in the compilations. For Europe, Dr. Werner Nicklas (Heidelberg) receives the QAP samples and distributes them to 17 additional European laboratories.

Interested diagnostic facilities receive four shipments during the year. Each shipment contains one to two bacterial cultures, either pure or as mixed cultures, and one serologic sample, mouse or rat. The bacteria are reisolated and identified, and antibiotic sensitivities tested. The sera are tested by the facilities' rodent serology screens. Results are returned to Rockefeller for tabulation.

The bacterial cultures generally are isolates from cultures submitted to LARC for identification, although environmental samples from various animal rooms at LARC are sometimes used. Two years ago, the bacterial samples were supplemented with pure cultures obtained from American Type Culture Collection in an attempt to give this part of the program a firmer basis for identification. This also allows for less frequently encountered organisms to test our skill.

Results from several years have been compiled as an assessment of the success of the program. As can be seen in Table 1, the majority of organisms in the QAP have been gram negative. Agreement as to identity is highest for the most commonly encountered organisms. Those which are infrequently seen in a lab animal colony vary considerably in agreement. Several significant changes in animal serologic diagnostics has occurred over the years. Complement fixation assays are generally no longer performed by any of the recipient institutions. The majority of the rodent viral antibodies are now discerned by ELISA assays, with immunofluorescence (IFA) and hemagglutination inhibition (HAI) primarily used as confirming tests. In some instances, the receiving facilities submit these samples to outside independent testing labs, rather than doing their screening in-house.

Outside testing labs showed about the same degree of variation as was seen between in-house testings. A compilation of serology results for several individual rodent viral antibodies is shown in Table 2. In general, tests positive by ELISA are usually also positive by IFA but about evenly split for HAI.

The program has been useful for individual laboratories to assess their test results against those of other laboratories, both here and in Europe. The major drawback is the increasing inability to obtain specific viral positive rodent sera. Programs to improve health status and quality control over the years have resulted in progressively cleaner animals, especially for the viruses most commonly tested. This should be taken as a positive sign. But, ironically, as suppliers and facilities continue this process of improvement, fewer samples of positive sera from naturally infected animals are available, especially in quantities sufficient for the QAP.

References:

1. Stark, D.M. A quality assurance program for laboratory animal diagnostic facilities. Lab Animal 13:25-31, 1984.

2. Lamotte, L.C. and Griffin, C.W. CDC reports results of lab performance test. Lab. Mgmt. 15:28-31, 1977.

3. Taylor, R.N. and Fulford, K.M. Assessment of laboratory improvement by the Centers for Disease Control diagnostic immunology proficiency testing program. J. Clin. Microbiol. 13:356-368, 1981.

Table 1 QAP: Bacteriology Results*Organism Total Consensus/ % Agreement in QAP Sample Total Received ___________________________________________________________________ Acinetobacter calco.,anitratus 19/22 86 Aeromonas sobria 7/35 20 Aeromonas hydrophila 33/35 94 Bacillus sp 13/22 59 Bacillus sp 9/22 41 Bacillus sp 16/21 76 Bordetella bronchiseptica 16/21 76 Bordetella bronchiseptica 16/16 100 Bordetella bronchiseptica 19/24 79 CDC Group IV, c-2 10/18 56 Citrobacter freundii 28/35 80 Citrobacter freundii 32/37 86 Enterobacter faecalis 22/37 59 Enterobacter sakazakii 25/28 89 Edwardsiella ictaluri 1/32 3 Flavobacter indologenes 11/16 69 Klebsiella pneumoniae 18/22 82 Klebsiella pneumoniae 19/24 79 Morganella morganii 27/33 82 Moraxella sp 11/32 34 Nocardia asteroides 4/35 11 Providencia rettgeri 24/33 73 Pseudomonas fluorescens 7/34 21 Pseudomonas putida 18/34 53 Salmonella sp 16/17 94 Salmonella sp 32/35 91 Salmonella arizonae 33/35 94 Salmonella arizonae 31/35 88 Staphylococcus aureus 17/17 100 Staphylococcus aureus 21/24 88 Vibrio sp 5/20 25 ___________________________________________________________________ *Represents 22 QAP shipments from 1989 to 1994. Duplicate entries mean same organism was sent out more than once, in different QAP shipments. ================================================================== Table 2 QAP: Serology Results* ___________________________________________________________________ Species Antibody Number ELISA IFA HAI Total Assays +/- +/- +/- +/- __________________________________________________________________ Mouse MHV 10 123/15 23/5 3/2 149/22 Sendai 7 81/20 16/5 8/4 105/29 LCM 1 5/2 0/3 0/0 5/5 MVM 3 17/2 1/4 5/2 23/8 PVM 3 28/6 2/4 3/5 33/15 GD7 3 33/2 0/0 1/7 34/9 Rat M pul 11 153/16 15/3 0/0 168/19 Sendai 15 226/18 25/14 16/12 267/44 PVM 12 152/9 9/9 18/9 179/27 KRV 17 200/9 12/10 40/30 252/49 RCV 14 190/18 34/4 7/0 231/22 ___________________________________________________________________ *Represents 22 QAP shipments from 1989 to 1994. "Number Assays": Number of times (QAP shipments) serum positive for agent shown was sent out.========================================================== **ANNOUNCEMENTS**

DEATHS It is with great sadness that we learned of the deaths of two members, Norman Somerson, PhD (Ohio State University) and Jerjang Chang, DVM, PhD (University of North Carolina) in this past year. They will be missed. Our condolences go to their families and colleagues.

NATIONAL CAPITAL AREA BRANCH, AALAS The National Capital Area Branch, AALAS, presents its 22nd Annual Meeting on September 22-24, 1994, at the Turf Valley Hotel and Country Club in Ellicott City, MD. The meeting topic is "The Laboratory Animal Science Team: Quality Animal Care + Quality Animal Management = Quality Science". Interest from ACLAD members is welcome; for further information: Jesse Price, (703) 659-4474.

THANKS As always, I am delighted to acknowledge the excellent help of Ms. Joan Bailie, Editorial Assistant, of Dr. Dean Percy, Ontario Veterinary College, and of the other contributors to this issue. Many thanks for all the indispensble help from our volunteers and contributors!

FUTURE PLANS

1. COUNCIL NOMINATIONS, BYLAWS CHANGES: This is an ELECTION YEAR for ACLAD. Nominations for Council and discussion of new By-Laws will be topics at the Annual Business Meeting this October in Pittsburgh. When they are completed, the suggested By-Law revisions (for consideration by the membership) will be mailed to all members.

2. NEWSLETTER PLANS: Future plans for the NEWSLETTER call for further expansion. The Publications Commitee, which is responsible for overseeing the Newsletter and for developing the content that will appear in the Newsletter, will meet in Pittsburgh (time and place will be set at the Business Meeting). All interested members are invited to participate actively and to join the Committee!

3. MEMBERSHIP DIRECTORY?: We are looking into the feasibility of sending out a Membership Directory in a mailing to all members, and are hopeful that this may be possible in the near future. (The limiting factor is cost).

4. REAGENT DIRECTORY: We are beginning the process of revising the Directory of Reagents that first appeared in 1990. Anyone who has diagnostic reagents to add to the list, please send information to the Editor (those who were listed in the previous edition will receive copies for updating). Suggestions from members for new items to include are invited.

* Items for the Newsletter are always welcome. Please send them to Steve Morse (address below). * Information on ACLAD membership ($20/yr) is available from Steve Morse (Secretary-Treasurer) or Ms. Joan Bailie (ACLAD Editorial Asst.) at the address below. New members are always welcome.

Stephen S. Morse [Newsletter Editor & ACLAD Secretary-Treasurer] The Rockefeller University 1230 York Avenue, Box 120 New York, NY 10021-6399 Telephone: (212) 327-7722 FAX: (212) 327-7172 OR 327-7974 E-mail: morse@rockvax.rockefeller.edu * Items for the Newsletter, general comments, ACLAD business

Editorial Assistant: Joan Bailie Section of Comparative Medicine Yale University School of Medicine P.O. Box 208016 New Haven, CT 06520-8016 Telephone: (203) 785-2507 FAX: (203) 785-7499 * Changes of address; general comments; questions about mailing or dues

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