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Case Description—A 7-year-old Golden Retriever was examined because of anorexia, lethargy, vomiting, and gradual weight loss. Clinical Findings—Splenomegaly, pancytopenia, high serum calcium concentration, and high alkaline phosphatase activity were detected. Magnetic resonance imaging revealed an enlarged mesenteric lymph node and increased signals from the bone marrow of the ilium and vertebral bodies. Histologic examination and immunophenotyping of biopsy specimens confirmed a stage V (b) T-cell malignant lymphoma. Treatment and Outcome—Clinical remission was attained by use of 2 chemotherapy cycles, followed by an allogeneic hematopoietic cell transplant performed at 18 weeks after diagnosis. A donor was identified by molecular dog leukocyte antigen typing methods. The patient was conditioned with 2 fractions of 4 Gy total body irradiation delivered 3 hours apart at 7 cGy/min, followed by an IV infusion of recombinant canine granulocyte colony-stimulating factor mobilized leukapheresis product and postgrafting immunosuppression with cyclosporine. Chimerism analyses revealed full donor engraftment that has been maintained for at least 58 weeks after transplant. Remission has been confirmed by normal results of serum thymidine kinase assays and the absence of peripheral blood clonal T-cell receptor gene rearrangements. Clinical Relevance—Systemic chemotherapy induces remissions; however, most dogs succumb to disease recurrence because of multidrug resistance. Outcome of allogeneic hematopoietic cell transplantation in dogs can be excellent because of improved donor-recipient selection by use of molecular dog leukocyte antigen typing, compared with early attempts, and better prevention of graft versus host disease, better supportive care, and substitution of peripheral blood mononuclear cells for bone marrow.

Workers engaged in the care and handling of laboratory animals perform tasks in a variety of indoor and outdoor settings on behalf of the civil and uniformed services, academic institutions, hospitals and non-profit entities, contract research organizations, pharmaceutical and biotechnology companies, diagnostic facilities, and animal breeding facilities, among others. The vivarium environment is one where both animals and human workers spend extended time in close proximity, and can involve both terrestrial and aquatic animals of highly diverse species for in-life as well as post-mortem stages of research, teaching, and testing. In these environments, workers may encounter exposures to a wide range and intensity of biological, physical, chemical, and psychosocial health hazards. Managers of laboratory animal facilities therefore need to simultaneously consider the health and well-being of the animals as well as that of the animal handlers and other staff in the facility. In many laboratory animal settings, however, preventive programs for animal care workers have been less developed and comprehensive than those for workers in other industries (McMurry and Key, 1999).

A prospective study was conducted that evaluated duration of virus shedding through acute and experimentally-induced recurrent disease episodes in 12 cats, and tissue distribution of latent infections, following intranasal vaccination with a temperature sensitive (ts) mutant strain of feline herpesvirus 1 (FHV1). Six of these cats were challenged with a virulent field strain of the agent to assess the extent to which vaccination affected subsequent shedding of virus and the establishment of latent infections. Virus isolation (VI) tests were done in parallel with a polymerase chain reaction (PCR) assay to compare the performance of each diagnostic method. The PCR confirmed that all 12 cats shed virus throughout the periods of vaccination, challenge or mock-challenge, and a cyclophosphamide-dexamethasone stress protocol to reactivate latent infections. Shedding to the tsFHV1 was documented by VI for up to 25 days following vaccination and for up to 15 days following challenge, but not after experimental stress. Overall, FHV1 was present in 144 of 300 (48%) cat-days of testing by PCR compared to 32 of 300 (11%) by VI. The frequency and distribution of latent FHV1 detected in neurologic, ophthalmic, and other tissues by PCR were identical for vaccine-only and vaccine-challenge groups, thereby disproving previous hypotheses that tsFHV1 mutants administered by this route protect against latency.

A polymerase chain reaction (PCR) assay was developed and used to detect feline herpesvirus-1 (FHV-1) in conjunctival and oropharyngeal swabs, and in latently infected tissues (trigeminal ganglia, optic nerves, optic chiasma, olfactory bulbs and corneas) collected from 10 experimentally infected cats. There was good agreement between parallel tests of the swab specimens by PCR and virus isolation assay during the phase of acute, latent and recurrent disease episodes (kappa=0.63, P

In a survey of seven species of wild rodents (n = 423) collected between October 1993 and March 1994 from the three principal ecological biomes of North Carolina (USA), we found hantavirus antibodies in seven (2%) of 301 Peromyscus spp. Hantavirus antibodies were detected in P. leucopus and P. maniculatus captured from mountain and coastal island bi-omes. Three mice were positive for Sin Nom-bre virus, while four others had antibodies to Seoul virus or a related agent. Two mice serologically positive for Sin Nombre virus were collected from inside a private mountain domicile. We conclude that the risk of human exposure to hantaviruses in North Carolina resembles that for most other areas of the continental United States.

Epidemiology is defined as the study of the distribution and determinants of disease within populations. In addition to the requirements for disease surveillance, epidemiologic methods have numerous applications in laboratory animal science and can reveal important insights into the multifactoral mechanisms of disease, thereby aiding in the design of optimized intervention strategies. Observational approaches to data collection can be used to quantify the role of causal factors under natural circumstances, complementing the value of experimental studies in this field. The meaning and appropriate use of standard measures of disease frequency and exposure-disease relationships are reviewed, along with explanations of bias and confounding. Recommendations for reporting the methods and findings from this type of work in comparative medicine literature are presented. Aspects of model-based approaches to data analysis are introduced, offering further opportunities for gaining needed information from epidemiologic study of problems in laboratory animal medicine and management.

Results from a 1999 economic survey of laboratory animal veterinarians were used to develop a multivariable linear regression model capable of predicting income in light of a large set of characteristics of veterinarians and their employers. When considered simultaneously, the variables of experience, supervisory responsibility, ACLAM board certification, geographic region, type of employer, size of animal facility, and job title provided the best model fit, in agreement with an earlier statistical model of income for persons in this profession. Aspects of the demographics of faculty appointments and secondary sources of professional income available to some laboratory animal veterinarians also were evaluated.

To determine whether feline herpesvirus 1 (FHV-1) DNA is in the corneas of clinically normal cats and cats with eosinophilic keratitis or corneal sequestration.Corneal biopsy specimens obtained from cats referred for treatment of corneal sequestration or eosinophilic keratitis.Corneal scraping or keratectomy specimens collected from clinically normal cats, cats with eosinophilic keratitis, and cats with corneal sequestration were evaluated for FHV-1 DNA by use of polymerase chain reaction (PCR). DNA was extracted from the tissue, and 1 microgram was assayed for FHV-1 by use of a single-round (40 cycles) PCR assay with primers directed at a 322-bp region of the thymidine kinase gene. Polymerase chain reaction positivity for clinically normal and affected cats of various breeds was compared by chi 2 analysis at alpha = 0.05.The FHV-1 DNA was detected in 5.9% (1/17) of corneas from clinically normal cats, in 55.1% (86/156) of corneal sequestra, and in 76.3% (45/59) of scraping specimens from cats with eosinophilic keratitis. Prevalence was significantly (P0.001) greater for cats with corneal sequestration or eosinophilic keratitis than for clinically normal cats. For cats with corneal sequestration, prevalence of FHV-1 DNA was significantly lower in Persian and Himalayan, compared with domestic shorthair and longhair breeds.Data strongly imply involvement of FHV-1 in the pathogenesis of eosinophilic keratitis and corneal sequestration. In Persian and Himalayan breeds, however, other nonviral factors also appear to be involved.Feline herpesvirus 1 must be considered when treating cats with corneal sequestration or eosinophilic keratitis.

A polymerase chain reaction (PCR) assay was developed and used to detect feline herpesvirus-1 (FHV-1) in conjunctival and oropharyngeal swabs, and in latently infected tissues (trigeminal ganglia, optic nerves, optic chiasma, olfactory bulbs and corneas) collected from 10 experimentally infected cats. There was good agreement between parallel tests of the swab specimens by PCR and virus isolation assay during the phase of acute, latent and recurrent disease episodes (kappa = 0.63, P0.001). The PCR reliably detectedor = 240 copies of FHV-1 template DNA, significantly improving upon previously published PCR assays for the agent.

After an episode of clinical Pasteurella pneumotropica infection was diagnosed in a C57BL/6N mouse, a randomly amplified polymorphic DNA polymerase chain reaction assay (RAPD-PCR) was developed and used to genetically characterize and differentiate 52 field isolates and laboratory reference strains of P. pneumotropica and related bacteria. A survey of rodents in the facility recovered 36 isolates of P. pneumotropica from 30 mice, six isolates from hamsters, and three isolates from rats during the follow-up investigation. Antibiograms and routine bacteriologic evaluations for morphologic and biochemical characteristics on selective media did not substantively aid in the differentiation of these isolates, but the RAPD-PCR revealed four strains of P. pneumotropica in the colony, two of which were confined to rats and hamsters. The RAPD-PCR unambiguously differentiated Heyl and Jawetz biotypes of P. pneumotropica recovered from mice, identified two additional genetic groups for rat and hamster isolates, and clearly distinguished P. pneumotropica from related bacteria. Most field isolates were genetically consistent with the Jawetz biotype of P. pneumotropica. The RAPD-PCR is a fast, sensitive, and efficient method for identifying genetic differences between strains of the P. pneumotropica complex and can contribute substantially in addressing the epidemiology, pathogenesis, and taxonomic classification of this common opportunistic pathogen.

Pasteurella pneumotropica, a gram-negative opportunistic pathogen, can be isolated from the oropharynx, intestinal tract, and reproductive tract of clinically normal mice and has been associated with various clinical syndromes, including conjunctivitis, infections of the reproductive tract, otitis, and subcutaneous abscess formation. Enrofloxacin, a fluoroquinolone bactericidal antimicrobial, has been shown to be effective in eliminating P. multocida from rabbits. We sought to determine whether enrofloxacin would eliminate P. pneumotropica from mice known to be asymptomatically infected with the agent. Pasteurella pneumotropica-positive (culture and immunofluorescence assay) male (n = 55) and female (n = 55) C57BL/6N mice were randomly assigned to one of seven treatment groups or to a control group. These groups were designed to evaluate the efficacy of enrofloxacin administered orally via the drinking water or parenterally at three dosages (8.5, 25.5, and 85.0 mg/kg of body weight per day) over a 14-day treatment period. A tetracycline-treated group (60 mg/kg per day) and an untreated control group were included for comparisons. Repeated oropharyngeal swab and fecal specimens were obtained for culture through posttreatment day 30, and specimens from numerous enteric and reproductive organs collected during necropsy were used to evaluate group differences. Enrofloxacin eliminated evidence of P. pneumotropica from all sites when administered at 25.5 or 85 mg/kg but not at 8.5 mg/kg by either route for at least 30 days after treatment. Tetracycline-treated and control groups remained consistently culture-positive throughout the study. We concluded that the oral route may be a more practical method for treating large numbers of mice. Enrofloxacin may be a practical and inexpensive alternative to cesarian rederivation or embryo transfer for the elimination of P. pneumotropica in mice.

Hantaviruses have emerged as agents of significant morbidity and mortality in human beings. These pathogens are maintained in nature as not clinically apparent, persistent infections in roden reservoirs, varying with each virus, and are shed via rodent feces, urine, and salivary excretions. Human exposure to hantaviruses principally occurs through the respiratory tract route and is focal and discontinuous, paralleling the distribution of virus in reservoir species and the likelihood of human-rodent interactions. Prior to 1993, hantaviruses were established etiologic agents of human febrile nephropathies and hematologic abnormalities on several continents, exclusive of North America. An episode of severe respiratory tract illness developed in association with a group of novel hantaviruses in the United States during 1993-1994, resulting in at least 98 confirmed cases of disease from 21 states and 51 deaths. Current evidence suggests that hantaviruses of zoonotic potential have long existed across widespread areas of the United States but have gone unrecognized.

A protein immunoblot (western blot) assay was developed for detection of IgG and IgM antibodies to cytomegalovirus (CMV) in banked serum and was used to study the epidemiology of infection in a breeding population of rhesus macaques. All juveniles and adults were antibody-positive to CMV. Repeated samples from 1990-born infants (n = 28) over a 12-month period allowed retrospective assessment of CMV seroconversion. Most infants (23 of 25 seroconversions) were apparently exposed to CMV during their first year of life. The IgM testing permitted identification of seroconversion time points in eight infants. Risk factors that significantly affected seroconversion probabilities in the study population were not found. Cytomegalovirus-free rhesus macaques are potentially valuable for viral teratology and pathogenesis research projects; however, present epidemiologic findings suggest that establishment of CMV-free colonies will be difficult in conventional settings.

B virus is a zoonotic alpha-herpesvirus enzootic in Asian monkeys of the genus Macaca. At least 25 cases of human disease caused by B virus have occurred to date, leading to death in 16 instances. Advances in the technology available for the diagnosis of B virus infection and in the agents for its treatment are improving the prognosis for cases in human beings. Efforts are under way at several institutions in the United States to establish B virus-free colonies of rhesus macaques for use in biomedical research. Unfortunately, the epidemiology of B virus in group-housed macaques is poorly understood. The elucidation of factors important in the transmission of B virus between monkeys will greatly enhance efforts to eradicate this virus and may help to minimize further human exposure to the agent.

Data on costs associated with episodes of disease and disease prevention, including expenditures for veterinary services, were collected from 57 California beef cow-calf herds during 1988-1989 as part of the National Animal Health Monitoring System. Mean cost associated with episodes of disease was $33.90/cow-year, with $0.78 and $1.37/cow-year being spent for veterinary services and drugs, respectively. The highest cost for veterinary services related to episodes of disease were for dystocia, lameness, and ocular carcinoma. For disease prevention, mean expenditures for veterinary services were $1.67/cow-year, nearly all of which was spent on prevention of reproductive tract conditions. Preventive expenditures for veterinary services related to female infertility (pregnancy examination), vaccination against brucellosis and male infertility (breeding soundness examination) were $0.72, $0.39, and $0.22/cow-year, respectively. Many costs associated with episodes of disease and disease prevention were similar to those reported from Colorado National Animal Health Monitoring System beef herds.

Twenty-nine California dairy herds were studied over a 12-month period from 1988 to 1989 as part of the National Animal Health Monitoring System. Monthly interviews administered to dairy producers were used to measure the costs of all health-related expenditures and disease incidence in these herds. Of the total $1,523,558 reported, $1,355,467 (89%) was attributed to cost of disease events and $168,091 (11%) to cost of disease prevention. Most (78%) of the cost of disease events was attributable to death and culling losses. Veterinary services accounted for only $54,099 (4%) of total costs, 64% of which was used for disease prevention, compared with 36% for disease treatment. Udder disease was the most costly category of diseases reported at an average of $49.85/head at risk annually, followed by reproductive problems at $38.05. Through the use of sampling strategies less biased than those used in other surveys, the National Animal Health Monitoring System is designed to provide statistically-valid estimates of disease incidence and costs across broad geographic areas, potentially benefiting all those interested in the economics of livestock diseases in the United States.

Sixty cow-half herds of more than 50 cows each were randomly selected for a prevalence survey of bovine trichomoniasis in California. Herd size, as judged by the number of bulls, ranged from 1 to 210 bulls (median = 8; mean = 59 +/- 15.8). Preputial smegma was collected from 729 bulls (median = 6 bulls/herd) and cultured for Tritrichomonas foetus. Of 57 herds from which samples were collected, 9 (15.8%) had at least one infected bull. Of the 729 bulls from which samples were cultured, 30 (4.1%) were infected. Correcting for sensitivity of the diagnostic test yielded a prevalence of 5.0%. Infection rates for bulls greater than 3 years old and less than or equal to 3 years old were 6.7% and 2.0%, respectively (P less than 0.025). Median herd sizes were 14 bulls (range, 6 to 114) for infected herds and 7 (range, 1 to 210) for uninfected herds. These findings suggest that trichomoniasis is common in California beef herds. Because several bulls less than 4 years old were infected, we suggest that control measures stressing replacement of older bulls with younger ones should be combined with diagnostic procedures in those younger replacements, to ensure that they are not already infected.

A systematic sampling technique was used in combination with a highly sensitive and specific ELISA to provide unbiased age-specific prevalence estimates of B virus antibody in rhesus monkeys housed in three different outdoor breeding corrals. Among 146 sampled monkeys, 97% of animals 2.5 years and older were seropositive, while only 22% of younger animals were seropositive. Neither gender nor social dominance ranking were predictive of B virus antibody status. The strong age association was not inconsistent with hypothesized venereal transmission of B virus. Improvements in the epidemiologic understanding of B virus are necessary to assist efforts to eradicate this agent from breeding colonies of rhesus monkeys.

A population of free-ranging koalas in southeastern Queensland was examined to determine the prevalence of Chlamydia psittaci infections. Although C. psittaci was isolated from 46 of 65 (71%) koalas studied, only six (9%) of these had clinical signs of disease. Most adult females (82%) had back or pouch young present even though 67% of them were infected. There were no significant correlations between age, sex or site of sampling (urogenital versus conjunctival tissues) and the isolation of C. psittaci. No other important bacterial or fungal pathogens were isolated. The complement fixation test had a sensitivity of 7% and a specificity of 94% in detecting chlamydial infections, suggesting that it is unsuitable for use as a screening test. Chlamydia psittaci infection within this population appeared to represent a generally well-balanced host-parasite relationship and few animals had clinical signs of disease. Only four of 27 (15%) healthy koalas infected with C. psittaci followed for 24 wk after sampling developed eye disease or "dirty tail." Two koalas with keratoconjunctivitis recovered without treatment during the study period. Additional factors, including the stresses imposed by loss of habitat, may act to produce overt disease in koalas with latent C. psittaci infections.

The IDEIA Chlamydia Test, a commercially available antigen-capture enzyme-linked immunosorbent assay (ELISA) test, based on a monoclonal antibody for the detection of chlamydia in clinical specimens, was evaluated in a population of 65 free-ranging koalas in southeastern Queensland determined to be infected with Chlamydia psittaci. Compared to isolation of the organism in tissue culture, the sensitivity of the IDEIA test ranged from 3 to 11%, and the specificity from 90 to 97%. The results indicated that the IDEIA test is unsuitable for use as a diagnostic screening test for C. psittaci in free-ranging koalas.

DNA-slot hybridization and immuno-slot blot analyses were compared for the detection of Chlamydia psittact in crude swab material from free-ranging koalas. Immuno-slot blot analysis detected chlamydiae in 43 out of 68 koalas, with the sensitivity of the assay varying from 52 to 73% depending on the site of infection. Gene probe analysis was also used employing a genus-specific probe pCKO-10 isolated from a koala chlamydial gene library (ocular strain) and a plasmid probe pCKU cloned from a urogenital strain. The sensitivity of these two assays was comparable and they were considerably more efficient than the immuno-slot blot method for the detection of chlamydiae. Comparison of these data with a cell-culture method of detection, previously used with the same samples, demonstrated that gene probe analysis detected more positives than observed with cell culture. However, this appears to reflect more on the condition of the swab material rather than the sensitivity of the method.

The economic impact of management practices designed to limit the introduction of disease into a parent broiler breeder flock (biosecurity) was evaluated using benefit-cost analysis. Equations were developed to quantify the losses resulting from infection with one of four alternative categories of disease representing incremental levels of pathogenicity. Realistic costs and assumed values relating to the probability of infection were used to evaluate the ameliorative effect of three alternative levels of biosecurity. A microcomputer spreadsheet program was used to confirm that expenditure on protective measures can be justified by both the risk of introducing a disease and the magnitude of losses that may occur following infection.

SUMMARY. The economic impact of management practices designed to limit the introduction of disease into a parent broiler breeder flock (biosecurity) was evaluated using benefitcost analysis. Equations were developed to quantify the losses resulting from infection with one of four alternative categories of disease representing incremental levels of pathogenicity. Realistic costs and assumed values relating to the probability of infection were used to evaluate the ameliorative effect of three alternative levels of biosecurity. A microcomputer spreadsheet program was used to confirm that expenditure on protective measures can be justified by both the risk of introducing a disease and the magnitude of losses that may occur following infection.