DNA amplification for direct detection of HIV-1 in DNA of peripheral blood mononuclear cells.

By means of a selective DNA amplification technique called polymerase chain reaction, proviral sequences of the human immunodeficiency virus (HIV-1) were identified directly in DNA isolated from peripheral blood mononuclear cells (PBMCs) of persons seropositive but not in DNA isolated from PBMCs of persons seronegative for the virus. Primer pairs from multiple regions of the HIV-1 genome were used to achieve maximum sensitivity of provirus detection. HIV-1 sequences were detected in 100% of DNA specimens from seropositive, homosexual men from whom the virus was isolated by coculture, but in none of the DNA specimens from a control group of seronegative, virus culture-negative persons. However, HIV-1 sequences were detected in 64% of DNA specimens from seropositive, virus culture-negative homosexual men. This method of DNA amplification made it possible to obtain results within 3 days, whereas virus isolation takes up to 3 to 4 weeks. The method may therefore be used to complement or replace virus isolation as a routine means of determining HIV-1 infection.

Animal-based national surveillance for zoonotic disease: quality, limitations, and implications of a model system for monitoring rabies.

Surveillance for zoonotic diseases among wildlife is a research and public health challenge. The inherent limitations posed by the requisite human-animal interactions are often undefined and underappreciated. The national surveillance system for animal rabies in the United States was examined as a model system; reporting of animal rabies is legally mandated, each case of rabies is laboratory confirmed, and data have been consistently collected for more than 50 years. Factors influencing the monthly counts of animal rabies tests reported during 1992-2001 were assessed by univariate and multivariable regression methods. The suitability of passively collected surveillance data for determining the presence or absence of the raccoon-associated variant of rabies within states and within individual counties was assessed by determining critical threshold values from the regression analyses. The size of the human population and total expenditures within a county accounted for 72% and 67%, respectively, of the variance in testing. The annual median number of rabies tests performed was seven for counties without rabies, 22 for counties with non-raccoon rabies, and 34 for counties with raccoon rabies. Active surveillance may be required in locales with sparse human populations when a high degree of confidence in the status of rabies is required.

Human monocytic ehrlichiosis and human granulocytic anaplasmosis in the United States, 2001-2002.

The epidemiologic features are described of cases of human monocytic ehrlichiosis and human granulocytic anaplasmosis in the United States.

Persistence of elevated rabies prevention costs following post-epizootic declines in rates of rabies among raccoons (Procyon lotor).

Determining the benefits to cost relationships among different approaches to rabies control and prevention has been hindered by the inherent temporal variability in the dynamics of disease among wildlife reservoir hosts and a tangible and objective measure of the cost of rabies prevention. A major and unavoidable component of rabies prevention programs involves diagnostic testing of animals and the subsequent initiation of appropriate public health responses. The unit cost per negative and positive diagnostic test outcome can be reasonably estimated. This metric when linked to methodologies subdividing the epizootic process into distinct temporal stages provided the requisite detail to estimate benefits derived from rabies control strategies. Oral rabies vaccine (ORV), for prevention of the raccoon-associated variant of rabies, has been distributed in Ohio and adjoining states in an effort to develop an immune barrier to the westward spread of epizootic raccoon rabies. The costs of ORV delivery have been quantified. Herein, the cost structures required to assess the benefits accrued by prevention were developed. A regression model was developed effectively predicting (r2=0.70) the total number of rabies diagnostic tests performed by 53 counties in five northeastern (NE) states from 1992 to 2001. Five temporal stages sufficed to capture the range of variability in the raccoon rabies epizootic process. Unit costs, dollars per diagnostic test outcome, were calculated for negative and positive results from published reports. Ohio counties were matched to NE counties based on similar socioeconomic characters. A "pseudo-epizootic" of raccoon rabies was introduced into Ohio and the costs savings from ORV were derived as the excess costs imposed by epizootic spread throughout the state. At 46 km/year (range modeled, 30-60 km/year), the pseudo epizootic spread, and reached the enzootic stage, in all Ohio counties by year 13 (range modeled, 11-17 years). Cumulative excess costs for Ohio ranged between $11 and $21 million; counties of low socioeconomic status experienced the greatest relative excess costs. The costs for rabies prevention activities reached apices during the epizootic stage of raccoon rabies (2.7-10.8 times baseline) an unforeseen finding indicated elevated costs persisted (1.7-7.2 times baseline) into the enzootic stage.

HIV-1 and HIV-2 infection associated with AIDS in Abidjan, Côte d'Ivoire.

In September 1987, a seroprevalence study of HIV-1 and HIV-2 infection was conducted among 956 people from different groups in Abidjan, Côte d'Ivoire. Groups examined were hospitalized patients (Internal Medicine and Infectious Disease Departments, Centre Hospitalier Universitaire de Treichville, Abidjan), outpatients at tuberculosis treatment centers, blood donors, women attending an antenatal clinic, and patients attending sexually transmitted disease (STD) clinics. Total HIV infection prevalence ranged from 10% in STD clinic patients and pregnant women to 45% in hospitalized patients on an infectious diseases service. Within groups, HIV-1 infection was 2-6.5 times more prevalent than infection with HIV-2. One-quarter of HIV-seropositive people were serologically reactive to both HIV-1 and HIV-2 on enzyme-linked immunosorbent assay and Western blot. Clinical conditions previously observed in patients with HIV-1 infection were observed in people infected with HIV-2 only, as well as in those with HIV-1 infection and dual serologic reactivity. An isolate of HIV-2 was obtained on culture from a person with wasting disease and chronic fever. The results of this study suggest that infection with HIV-1 and HIV-2 is epidemic in Côte d'Ivoire, and that HIV-2 may be associated with AIDS.

Efficacies of US Food and Drug Administration-licensed HIV-1-screening enzyme immunoassays for detecting antibodies to HIV-2.

To determine the efficacy of enzyme immunoassays (EIAs) for antibodies against HIV-1 in detecting HIV-2-infected blood, we tested 55 HIV-2-positive sera with seven Food and Drug Administration-licensed EIA kits. The percentage detection of HIV-2 sera giving positive reactions with these kits varied between the various manufacturers from 60 to 91%. Observations based on a small number of sera (n = 13), suggest that HIV-2-positive blood collected from apparently healthy people (blood donors, prenatal clinics) are detected with a greater frequency (means = 89%) than blood from AIDS patients or patients (n = 32) hospitalized with other infectious diseases (means = 72%). Based on these results and the low incidence of HIV-2 infection observed in the USA, it was concluded that screening with HIV-2-specific tests would not significantly increase the number of HIV-2-positive people detected by current screening programs. However, due to the poor sensitivity of certain HIV-1 assays for HIV-2 antibodies, HIV-2 sera without cross-reacting antibodies will escape detection. Surveillance for HIV-2 might then be improved by the availability of HIV-1 and HIV-2 combination assays.

Identification of mixed HIV-1/HIV-2 infections in Brazil by polymerase chain reaction.

Analysis of sera from hospitalized Brazilian patients by whole-virus lysate-based enzyme immunoassay and Western blot indicated that 0.4% were reactive to HIV-2 alone while 4% were reactive to both HIV-1 and HIV-2. When these sera were tested for HIV antibody by type-specific peptide enzyme immunoassays, dual seropositivity was confirmed in only 0.4% of patients. To define genetically the HIV strains within the population, we analyzed peripheral blood mononuclear cells from selected seropositive patients for the presence of HIV-1 and HIV-2 proviral DNA using the polymerase chain reaction (PCR). Independent primers/probes sets were used for the amplification and detection of viral sequences from the long terminal repeat (LTR), gag, and protease (prt) gene regions. Our findings confirmed the serologic evidence of HIV-2 in Brazil and determined the extent of mixed HIV-1 and HIV-2 infections. Detailed evaluation of the amplified viral protease sequences by endonuclease restriction analysis and DNA sequencing independently confirmed mixed HIV-1 and HIV-2 infections in the two patients seropositive for HIV-1 and HIV-2. The data further indicated that these isolates are distinct from the HIV laboratory standards. We interpret the combination of culture and PCR findings to demonstrate the presence of both HIV-1 and HIV-2 in Brazil.

A simplified surveillance case definition of AIDS derived from empirical clinical data. The Clinical AIDS Study Group, and the Working Group on AIDS case definition.

A clinical AIDS case definition is needed for surveillance in countries where the CDC case definition is not practical. To derive such a definition, we compared 110 HIV-seropositive and 135 randomly selected HIV-seronegative adult medical-ward inpatients in Brazil. Multivariate analysis of clinical signs and symptoms and simple diagnoses resulted in a discriminant function with sensitivity of 89% and specificity of 96% in predicting for AIDS. These data were the empirical basis for a clinical definition of AIDS in adults drafted in a Caracas, Venezuela, workshop sponsored by the Pan American Health Organization. The revised "Caracas" definition presented here requires a positive HIV serology, the absence of cancer or other cause of immunosuppression, plus > or = 10 cumulative points, as follows: Kaposi's sarcoma (10 points); extrapulmonary/noncavitary pulmonary tuberculosis (10); oral candidiasis or hairy leukoplakia (5); cavitary pulmonary/unspecified tuberculosis (5); herpes zoster < 60 years of age (5); CNS dysfunction (5); diarrhea > or = 1 month (2); fever > or = 1 month (2); cachexia or > 10% weight loss (2); asthenia > or = 1 month (2); persistent dermatitis (2); anemia, lymphopenia, or thrombocytopenia (2); persistent cough or any pneumonia except TB (2); and lymphadenopathy > or = 1 cm at > or = 2 noninguinal sites for > or = 1 month (2). This definition has a sensitivity of 95% and a specificity of 100% (91% without HIV serology) when applied to the Brazilian patients in this study. The Caracas definition has been adopted by Brazil, Honduras, and Surinam, and is in validation elsewhere. The use of a reasonably sensitive and specific case definition commensurate with available diagnostic resources should facilitate AIDS surveillance in developing countries.

Mammalian reservoirs and epidemiology of rabies diagnosed in human beings in the United States, 1981-1998.

Between 1981 and 1998, 37 cases of rabies were diagnosed in human beings in the United States. Information directly linking the cause of infection to animal bite was available for only eight of these cases. Indirect incrimination of the vector by analysis of cDNA sequences obtained by reverse transcriptase polymerase chain reaction of samples indicated that for all cases (12/12) believed to have been acquired in foreign countries, variants of the rabies virus (VRVs) associated with dogs (7/12 involved known bite histories) were the cause of the rabies infections. In contrast, VRVs associated with bats (bat-associated VRVs or BAVs) were implicated as the cause of 88% (22/25) of infections believed to have been acquired within the United States (1/22 involved known bite histories). Sequence analyses revealed that a single BAV (Ln/Ps), associated with rabid silver-haired (Lasionycteris noctivagans) and Eastern pipistrelle (Pipistrellus subflavus) bats, was implicated in 73% (16/22) of bat-associated infections. Silver-haired bats are predominantly solitary and migratory. Eastern pipistrelle bats may occur individually or in small clusters. Both species are only infrequently submitted for rabies testing. Unrecognized bites and unique properties of the Ln/Ps BAV may explain its association with the majority of rabies infections in human beings in the United States.

Mastomys (rodentia: muridae) species distinguished by hemoglobin pattern differences.

Hemoglobin electrophoresis patterns were found to be reliable markers for distinguishing two species of Mastomys in Sierra Leone having 32 and 38 chromosomes. All 32-chromosome animals exhibited a single hemoglobin pattern, whereas those with 38-chromosomes had four distinguishable patterns. Both karyotypes were present throughout Sierra Leone. The 38-chromosome species was more prevalent in the Guinea savanna zone to the north, while the 32-chromosome species was most dominant in human-modified high forest areas of the eastern and southern parts of the country. In almost all situations the 32-chromosome species was more common in houses than in bush habitats; the reverse was true for Mastomys having 38 chromosomes. Analysis of hemoglobin patterns thus becomes useful for species identification, and is necessary to understand the roles of the different Mastomys forms as reservoirs of human diseases, such as Lassa fever in West Africa.

Isolation of human immune deficiency virus from African AIDS patients and from persons without AIDS or IgG antibody to human immune deficiency virus.

We previously reported a high incidence of acquired immune deficiency syndrome (AIDS) in Kinshasa, Zaire, as well as a high frequency of antibody to human immunodeficiency virus (HIV), which includes HTLV-III and LAV viruses, in persons without AIDS. In this report we assessed the frequency of HIV virus infection in persons with and without clinical AIDS and the association of virus isolation to presence of antibody. We isolated HIV from 27 (77%) of 35 patients with AIDS, and 5 of 9 patients with AIDS-related complex (ARC). Virus was also isolated from plasma and cerebrospinal fluid of patients in the study. The presence of antibody was a reliable marker for virus infection in African patients with AIDS. HIV was isolated from 5 of 27 control patients without AIDS, 3 of whom had normal T helper to T suppressor ratios and normal numbers of T helper cells. Two of these patients had no detectable antibody to HIV by ELISA or Western blot methods. In a population, such as the general heterosexual population of Kinshasa, with frequent infection by HIV and with few clearly definable risk groups, screening for antibodies to HIV may not be sufficient to identify some virus infected persons.

PIP: This study represented the 1st attempt to isolate human immunodeficiency virus (HIV) from African acquired immunodeficiency syndrome (AIDS) patients and controls. HIV was isolated from 27 (77%) of 35 Zairians with AIDS and from 5 (55%) of 9 patients with AIDS-related complex (ARC). In addition, 5 (19%) of 27 controls admitted to Zaire's Mama Yemo Hospital for causes unrelated to AIDS were found to be positive for antibodies to HIV. Next, an effort was made to isolate the virus from 42 AIDS or ARC patients on whom data were already available on the results of an enzyme-linked immunosorbent assay (ELISA). HIV was isolated from 30 (81%) of 37 patients with positive ELISA tests and from none of the 5 patients with a negative assay. Among controls, antibodies were found in a higher proportion of patients with abnormal helper: suppressor ratios or a low absolute T helper cell count. On the other hand, these abnormalities were not found in 3 of the 5 control patients from whom HIV was isolated, including 2 without HIV antibody. This suggests that neither of these criterion are good indicators of virus infection. The isolation of HIV infection from 5 hospital controls with no clinical signs of infection suggests that either the rate of asymptomatic HIV virus infection is high in Zaire or that common tropical diseases such as malaria or tuberculosis may be associated with HIV infection. The frequency of HIV isolation from AIDS and ARC patients in this study is higher than that in earlier reports from non-Africans, but is comparable to current statistics from the US.

National surveillance for Rocky Mountain spotted fever, 1981-1992: epidemiologic summary and evaluation of risk factors for fatal outcome.

Between 1981 and 1992, the Centers for Disease Control collected and summarized 9,223 cases of Rocky Mountain spotted fever (RMSF) reported from 46 states. Four states (North Carolina, Oklahoma, Tennessee, and South Carolina) accounted for 48% of the reports. The annual incidence per million U.S. population decreased from a high in 1981 of 5.2 to a low in 1992 of 2.0, primarily due to decreased incidence in the southeast. Case report forms were filed on 7,650 patients, of whom 4,217 had laboratory-confirmed RMSF. The age group with the highest incidence was those 5-9 years of age. Most cases (90.0%) occurred between April 1 and September 30 and included a history of tick attachment (59.6%). Reported symptoms included fever (94.0%), headache (86.2%), myalgia (82.5%), and rash (80.2%). The case-fatality ratio was 4.0%. Risk factors associated with death included older age, delay in treatment or no treatment, and treatment with chloramphenicol (compared with tetracycline); however, insufficient data existed to fully assess the confounding effect of severity of illness on antibiotic choice.

Rocky Mountain spotted fever in the United States, 1993-1996.

During 1993 through 1996, 2,313 cases of Rocky Mountain spotted fever (RMSF) were reported to the Centers for Disease Control and Prevention (CDC) by 42 states and the District of Columbia through the National Electronic Telecommunications System for Surveillance (NETSS). During this same interval, 1,752 case report forms (CRFs) were submitted to CDC and 1,253 (70%) of the cases were categorized as confirmed RMSF by laboratory testing. On the basis of analyses performed with NETSS data, the average annual RMSF incidence during 1993-1996 was 2.2 cases per million persons; the incidence rose from 1.8 in 1993 to 3.3 per million persons in 1996. Incidence for confirmed cases was highest among children 5-9 years of age (3.7 per million) and lowest among individuals older than 70 years of age (1.4 per million). The south Atlantic region accounted for the largest proportion of confirmed cases (52%). The case-fatality rate was highest among persons 70 years of age and older (9.0%) and lowest among adults 40-49 years of age (0.6%).

A household-based, case-control study of environmental factors associated with hantavirus pulmonary syndrome in the southwestern United States.

During an outbreak of hantavirus pulmonary syndrome (HPS) in the southwestern United States, trained environmental assessment teams conducted surveys at 17 case-patient homes and matched controls from June through August 1993. Variables related to rodent abundance were quantified and standardized rodent trapping was conducted around and within households. The majority of households were located in pinon-juniper vegetation zones, and there were no significant differences in the type of house in which cases and controls lived. The only environmental factor that distinguished case households from controls was significantly higher small rodent densities (median trap success for case sites = 17.3%, 12.7% for near controls, and 8.3% for far controls). Frequency of hantaviral infection in deer mice (Peromyscus maniculatus) did not vary significantly among households of cases and controls, with a range of 27.5-32.5% antibody-positive. Indices of rodent fecal contamination were slightly higher in case houses. The data indicate that higher rodent densities were associated with households in which HPS cases occurred. Strategies that control rodent numbers and decrease rodent access to dwellings may reduce risk of human infection.

Isolation of the causative agent of hantavirus pulmonary syndrome.

Investigation of a recent outbreak of acute respiratory illness in the southwestern United States resulted in the recognition of a new disease, hantavirus pulmonary syndrome (HPS) with high mortality. Different animals and cell lines were used in attempts to isolate the causative agent. A previously unknown hantavirus was passaged in laboratory-bred deer mice, recovered from lung tissues of a deer mouse, Peromyscus maniculatus, and propagated in the E6 clone of Vero cells. Virus antigen was readily detected in the infected cells by an indirect immunofluorescence assay, using convalescent-phase sera from HPS patients. By electron microscopy, the virus was shown to have the typical morphologic features of members of the genus Hantavirus, family Bunyaviridae. Virus sequences corresponded to those previously detected by a nested reverse transcriptase-polymerase chain reaction assay of hantavirus-infected specimens from rodents and humans. This newly recognized virus, the etiologic agent of HPS, has been tentatively named Muerto Canyon virus.

Rabies epizootics among raccoons vary along a North-South gradient in the Eastern United States.

The characteristics of rabies epizootics among raccoons were investigated in 11 eastern states along a North-South gradient from New York to North Carolina. Epizootics were defined as discrete intervals of time of at least 5 months in duration, when reported cases of raccoon rabies from an individual county exceeded the median value of raccoon rabies cases reported by that county over the entire period rabies was present among raccoons in the county. Over the approximately 20-year study period, 35,000 cases of raccoon rabies were reported, and epizootics were detected from 251 (64.4%) of 390 counties. The median annual incidence was 0.14 epizootics per year. During the first defined epizootic in a county, the median total number of raccoons reported rabid was 47, with a median monthly incidence of rabies in raccoons of 3.1. The median lag time from the first report of a rabid raccoon in a county to the beginning of the first epizootic was 4 months. Significant differences in the annual incidence of epizootics and monthly incidence of rabid raccoons during epizootics were observed among different states. Although human population density and per capita health spending within counties were positively associated with increasing magnitude of epizootics, a significant difference in the characteristics of rabies epizootics in northern and southern states was apparent. We hypothesize that environmental conditions and perhaps human influence resulted in rabies epizootics in southern states that were smaller, less-frequent, and lacking in well-defined temporal structure compared with those in northern states.

Survey of rabies preexposure and postexposure prophylaxis among missionary personnel stationed outside the United States.

BACKGROUND: Of the 36 cases of human rabies that have occurred in the United States since 1980, 12 (33%) were presumed to have been acquired abroad. In the United States, it is recommended that international travelers likely to come in contact with animals in canine rabies-enzootic areas that lack immediate access to appropriate medical care, including vaccine and rabies immune globulin, should be considered for preexposure prophylaxis. In 1992, the death of an American missionary who had contracted rabies while stationed in Bangladesh highlighted this high-risk group. METHODS: To assess their knowledge of rabies risk, rabies exposures, and compliance with preventive recommendations, we asked 695 missionaries and their family members to complete questionnaires about their time stationed abroad. RESULTS: Of the 293 respondents stationed in countries where rabies is endemic, 37% reported prior knowledge of the presence of rabies in their country of service. Only 28% of the personnel stationed in rabies-endemic countries received preexposure prophylaxis. Having preexposure prophylaxis specifically recommended increased the likelihood of actually receiving it (O.R. 15.6, 95%CI 7.4 - 34.9). There were 38 reported exposures (dogs = 66%, another human = 20%), proven or presumed to be rabid. Three of the people exposed received rabies immune globulin and vaccine; 11 received vaccine alone; 8 received only basic first aid, and 16 received no treatment. CONCLUSIONS: Although American missionaries stationed abroad are at an increased risk for exposure to rabies, compliance with established preventive measures was low. Prior to being stationed abroad, an educational rabies-prevention briefing, including encouragement to receive preexposure prophylaxis, could be an effective intervention for missionaries to decrease their risk of rabies.

Surveillance and spatiotemporal associations of rabies in rodents and lagomorphs in the United States, 1985-1994.

Between 1985 and 1994, 368 cases of rabies in rodents (95% of reports) and lagomorphs (5%) were reported to the Centers for Disease Control and Prevention, Atlanta, Georgia (USA), from 22 states. This was a 354% increase from the period 1971 to 1984. Most reports were cases of rabies in woodchucks (Marmota monax) (n = 317), primarily from the eastern United States, which has been recently experiencing an epizootic of raccoon (Procyon lotor) rabies. Cases of rabies in woodchucks were temporally and spatially associated with reports of raccoon rabies. Antigenic or genetic characterization of variants of rabies viruses from rodents and woodchucks corresponded to the variants associated with the major terrestrial wildlife reservoir within the geographic region of specimen origin. Although rodents and lagomorphs are infrequently infected with rabies and human contact with these animals rarely requires postexposure treatment, appropriate health authorities need to evaluate individual circumstances surrounding potential exposures.

Rabies surveillance in the United States during 1992.

In 1992, 49 states, the District of Columbia, and Puerto Rico reported 8,644 cases of rabies in nonhuman animals and 1 case in a human being to the Centers for Disease Control and Prevention. Almost 92% (7,912 cases) were wild animals, the largest number of wild animals ever reported, whereas 8.5% (732 cases) were domestic species. The total number of reported cases increased 23.9% over that of 1991 (6,975 cases), with most of the increase resulting from continued spread of rabies in raccoons. The 2 epizootics of rabies in raccoons (Northeastern/mid-Atlantic region and Southeastern region) are now approaching convergence in North Carolina (49 reported cases of rabies in 1992). Massachusetts (57 cases), New York City (41 cases), and New Hampshire (10 cases) became new additions to the epizootic in the Northeast, with Maine, Rhode Island, and Vermont the only states in the region without cases associated with the raccoon strain of rabies. The state of New York (including New York City) reported 1,761 cases (79% in raccoons) of rabies, the largest number ever recorded for any state. Increases attributable to epizootics of rabies in other species were reported by Alaska (25 cases in 1992, compared with 12 in 1991, mainly attributable to rabies in foxes) and Kansas (374 cases in 1992, compared with 63 in 1991, mainly attributable to rabies in skunks). Reported cases of rabies in coyotes (75) increased 50% over those for 1991 (50 cases). In the southern portion of Texas (reporting 70 of the 75 cases in coyotes), there was a similar increase (55%) in reported cases of rabies in dogs, whereas nationally, reported cases of rabies in dogs (182) increased 17%. Twenty states, the District of Columbia, and Puerto Rico reported decreases in rabies in animals in 1992, compared with 16 states in 1991. Hawaii was the only state that did not report a case of rabies in 1992.

Rabies surveillance in the United States during 1999.

During 1999, 49 states, the District of Columbia, and Puerto Rico reported 7,067 cases of rabies in nonhuman animals to the Centers for Disease Control and Prevention, a decrease of 11.2% from 7,961 cases in nonhuman animals and 1 case in a human being reported in 1998. More than 91% (6,466 cases) were in wild animals, whereas 8.5% (601 cases) were in domestic species (compared with 92.4% in wild animals and 7.6% in domestic species in 1998). No cases of rabies were reported in human beings in 1999. Decreases were evident in all major species groups, with the exception of cattle, sheep/goats, and swine. The relative contributions of the major groups to the total reported were as follows: raccoons (41.0%; 2,872 cases), skunks (29.4%; 2,076), bats (14.0%; 989), foxes (5.4%; 384), cats (3.9%; 278), cattle (1.9%; 135), and dogs (1.6%; 111). Reported cases (6) associated with the epizootic of rabies in raccoons in Ohio declined from the 26 cases reported in 1998. Fifteen of the 19 states where the raccoon variant of the rabies virus is enzootic reported fewer cases of rabies during 1999. Massachusetts and Rhode Island, states with enzootic rabies in raccoons, each reported more rabid skunks than rabid raccoons for the third consecutive year. In Texas, cases associated with the enzootic canine variants of the rabies virus remained low (10 cases), whereas cases associated with the gray fox variant of the virus increased (66). Cases of rabies in skunks decreased by 8.6%, compared with those reported in 1998. Michigan reported the largest percentage increase in rabid skunks (950.0%; 2 cases in 1998 to 21 in 1999). Cases of rabies in horses and mules declined 21%, from 82 cases in 1998 to 65 in 1999. Cases of rabies reported in bats (989) were similar in number to those reported in 1998 (992) and represented almost 14.0% of the total number of rabid animals reported during 1999. Reported cases of rabies in cats (278) and dogs (111) decreased by 1.4% and 1.8%, respectively, whereas cases in cattle (135) increased by 16.4%, compared with those reported in 1998.