Clinical and serological evidence of bovine babesiosis and anaplasmosis in St. Lucia

One hundred fifty-nine Holstein calves were imported into St. Lucia from the U.S.A. An outbreak of babesiosis occurred 17 days post-arrival, and an outbreak of anaplasmosis occurred 5 months after importation. Sera obtained 3, 6 and 12 months post-importation revealed a high prevalence of IFA titres to Babesia bovis and B. bigemina 3 months after arrival and an increase in titres to Anaplasma marginale 6 months after arrival. Sera obtained arrives from native cattle from several places on the island indicated infection rates of 80, 65 and 64% with A. marginale, B. bigemina and B. bovis, respectively. The rapid card test only indicated a 25% prevalence of infection of native cattle by A. marginale. This low prevalence was probably due to deterioration of serological activity during shipment.     

Evaluation of an ELISA for detection of antibodies to Babesia bovis in cattle in Australia and Zimbabwe

An enzyme-linked immunosorbent assay (ELISA) for antibodies to Babesia bovis was evaluated in comparison with the indirect fluorescent antibody test (IFAT) in Australia and Zimbabwe. Positive and negative threshold values for the ELISA were set using sera from cattle of known infection status. Sensitivity and specificity estimates for the ELISA based on 158 positive sera from cattle experimentally infected with Australian isolates of B. bovis and 318 negative sera collected from B. bovis-free herds in Australia were 100% and 99.4%, respectively. The specificity of the assay in Africa, based on 328 sera from B. bovis-free herds in Kenya and South Africa, was 99.7%. The ELISA was compared with the IFAT using sequential sera from 16 calves experiencing primary B. bovis infections, and a total of 777 field sera collected from B. bovis-endemic herds in Australia and Zimbabwe. In primary infections, the ELISA and IFAT detected antibodies at or about the same time. With sera from endemic herds, the performance of the ELISA was at least comparable with that of the IFAT. Two hundred and fourteen of 221 sera that were negative by IFAT, were negative by ELISA, and 428 of 439 sera that were clearly positive by IFAT were positive by ELISA. Of 117 sera that gave equivocal (suspect or weak positive) results in the IFAT, 20 were positive by ELISA, 7 were suspect and 90 were negative. We conclude that the ELISA will be useful for epidemiological studies on B. bovis in Australia and Zimbabwe, and probably elsewhere.     

Identification of an immunodominant 40 kDa merozoite antigen common to the Australian T and Dixie vaccine strains of Babesia bovis and the development of diagnostic tests specific for these strains

Antigenic differences among Australian vaccine and field strains of Babesia bovis were investigated in an attempt to identify strain specific antigens. Immunoblots revealed substantial differences between the current vaccine strains, designated T and Dixie, and previous vaccine strains and field isolates collected on properties where vaccination with the T or Dixie strains had failed to provide complete protection against tick-borne challenge. A major difference was an immunodominant 40 kDa antigen (T40) present in only the T and Dixie strains. The molecular weight and immunodominant nature of this antigen suggest that it may be the equivalent of the major merozoite surface antigen (MSA-1) described by others in North American strains of B. bovis. MSA-1 was shown to be conserved in north American isolates but not in an isolate from Israel or in the Australian S and L isolates. The work presented here suggests that merozoite surface antigen diversity exists among geographically different isolates of B. bovis within Australia.

Monospecific antiserum to T40 was used to develop an indirect fluorescent antibody (IFA) test specific for T and Dixie strain parasites, and a blocking enzyme-linked immunosorbent assay (ELISA) specific for antibody to the T and Dixie strains. In cases of babesiosis in recently vaccinated cattle, the IFA test will be a useful tool for determining whether clinical symptoms are due to a severe vaccine reaction or to a concurrent tick-borne infection. In a preliminary assessment of potential of the ELISA for the serological identification of vaccinated cattle using a total of 160 sera, the test clearly differentiated between animals vaccinated with the T or Dixie strains and non-vaccinated animals, and was not affected by presence of antibodies to other B. bovis strains.     

Long-term study of incidence and financial loss due to cattle babesiosis in an Argentinian dairy farm

The incidence and direct financial loss caused by babesiosis were evaluated in 121 Holstein Friesian female cattle that formed eight cohorts (1981–1988) of a dairy farm located approximately 24° 55′S 65° 29′W in Salta, Argentina. Female calves born in 1986 and 1987 (n=32) were vaccinated with a live Babesia vaccine at six months of age.

No cases of babesiosis occurred in the vaccinated cattle. The incidence of babesiosis in the six non-vaccinated cohorts was 23.6% (21/89). Eighteen of the cases were the result of Babesia bovis, one to Babesia bigemina and two to a mixed infection. Two cattle died of B. bovis infection in spite of drug treatment (diaminazene, 3.5 mg kg⁻¹). No disease occurred in cattle younger than seven months or older than 24 months. The number of cases according to age of cattle was: 7–9 months, 5; 10–12 months, 6; 13–24 months, 10.

Financial loss for the six cohorts that suffered clinical cases amounted to US $ 1624.6 (prices in October 1990)—62% were the result of physical losses and 38% to costs of control. A benefit-cost analysis of vaccination was carried out assuming that 95% of the mortality and morbidity losses of the six non-vaccinated cohorts was prevented after a single inculation of a live vaccine (cost of a dose plus administration was US $ 4.2. The benefit-cost ratio was 4:1 for each US dollar expended.     

Management factors associated with Babesia bovis seroprevalence in cattle from eastern Yucatán, Mexico.

The effect of management on the seroprevalence of Babesia bovis was studied in 399 Bos indicus cattle 1–2 years old from 92 farms in the eastern Yucatán, México. The management factors studied were: farm-type, production system, herd size, farm size, stocking density, vector control, dipping interval, type of dipping, type of acaricide and cattle introduction to the farm. A cross-sectional study was carried out (2-stage cluster sampling). The number of serum samples was proportionally distributed according to the number of farms in the nine locations of eastern Yucatán, México (399 animals from 92 farms). Antibody activity to B. bovis was tested using an indirect ELISA. The farms with a seroprevalence ≤75% were considered as cases and those with seroprevalence >75% were considered as controls. The variables with p ≤ 0.20 were included in fixed effects logistic regression. The seroprevalence of the zone was 73.8% (66.3–81.3%). The following risk factors were found: Stocking density (<1 head/ha, OR = 4.04, CI (OR) = 1.20–13.62) and dipping interval (>60 days, OR = 5.07 CI (OR) = 1.26–20.48).     

Effect of agro-ecological zone and grazing system on incidence of East Coast Fever in calves in Mbale and Sironko Districts of Eastern Uganda

Between May 2002 and February 2003 a longitudinal survey was carried out in Mbale and Sironko Districts of Eastern Uganda to determine the influence of agro-ecological zones (AEZ) and grazing systems on tick infestation patterns and incidence of East Coast Fever (ECF) in bovine calves. The study area was stratified into AEZ (lowland, midland and upland) and grazing systems {zero grazing (ZG), restricted-outdoor grazing (ROG) and communal grazing (CG)}, whose strata had previously been shown to influence the prevalence of ECF, babesiosis and anaplasmosis. One hundred and eighty-five smallholder dairy farms with a total of 198 calves of both sexes, between the ages of 1 day and 6 weeks, were purposively selected from the AEZ–grazing system strata. Nine dynamic cohorts (11–51 calves in each) of these calves were examined and sampled monthly. Ticks infesting the calves were counted from one side of the animal body and categorized into the different species, sex and feeding status. Sera were collected at recruitment and monthly thereafter and antibodies against Theileria parva, T. mutans, Babesia bigemina, B. bovis and Anaplasma marginale were measured using ELISA. Tick challenge (total and specific) varied with AEZ and grazing system. The risk of infection with T. parva was higher in the lowland zone compared to the upland zone (hazard ratio (HR) = 2.59; 95% CI: 1.00–6.34). The risk of infection with T. parva was higher in the CG system than the ZG system (HR = 10.00; 95% CI: 3.61–27.92). The incidence risk for sero-conversion, over the 10 months study period, was 62, 16 and 9% in the lowland, midland and upland zones, respectively. Ninety-eight percent of the calves in lowland-CG stratum sero-converted by the age of 6 months, while 56 and 8% did so in the lowland-ROG and the lowland-ZG stratum, respectively. The results of this study show the need to consider farm circumstances and the variation in ECF risk, both spatially and temporally when designing control strategies for ECF.     

Prevalence of bovine haemoparasites in Aceh province of Northern Sumatra: Implications for imported cattle

A limited seroepidemiological investigation was conducted in Aceh Province to determine the seroprevalence of Anaplasma marginale and Trypanosoma evansi in indigenous cattle and in groups of Sahiwal cross cattle imported from New Zealand in 1985. The results obtained suggest that these parasites are endemic in the areas surveyed; in local cattle, overall prevalence rates of 82% for A. marginale and 61% for T. evansi were obtained.

Thin blood smears were prepared from 42 anaemic adult cattle and 19 clinically normal calves. Babesia bigemina was detected in 5 blood smears and Theileria orientalis in 51. Sahiwal cattle imported from New Zealand suffered high mortalities during their first year in Aceh and it is suggested that imported naive cattle rapidly become infected with blood parasites.     

Study on some molecular characterization of Babesia orientalis

The study on buffalo babesiosis indicated that its pathogen was different from other Babesia on many aspects such as morphology, transmission and pathogenicity. Therefore, it was named as a new species—Babesia orientalis. In order to prove the validity of this taxon, molecular taxonomic study on the pathogen was done in this experiment. The complete 18S rRNA gene sequence of B. orientalis was determined by PCR. It was sequenced and blasted. The results indicated that the classification of the parasite belonged to the genus Babesia. The 1700 bp complete sequence was compared with 15 other Babesia sp. available in GenBank. The data were analyzed and a phylogenetic tree was established. The results indicated that the hereditary distance of the parasite was close to that of Babesia sp. from South Africa and Babesia ovis, and the hereditary distance was far from Babesia bigemina and B. bovis.     

A study of an enzootic focus of sheep babesiosis (Babesia ovis, Babes, 1892)

Morbidity and mortality due to Babesia ovis in sheep flocks grazing in an enzootic area of Israel occur yearly, about 2 weeks after detection of adult Rhipicephalus bursa ticks on the animals. Disease incidence peaks in May, but lasts throughout the active period of the adult ticks in the spring-summer months of April–July. No clinical cases of babesiosis have been registered during the active period of the preimaginal stages of R. bursa, from October to February. Incidence of parasitaemia during the spring-summer months was variable, ranging between 2 and 25%. However, in the winter months the incidence of parasitaemia in hoggets increased considerably, reaching 4–60% of the animals.

A positive serological response to B. ovis was found in 84.5% of the hoggets and 88.9% of the ewes. In ewes, the prevalence of the serological response showed no marked seasonal variations. Colostral sera of 67.5% and 75% of the ewes and hoggets, respectively, were serologically positive for B. ovis. No antibodies were detected in the sera of lambs less than 3–4 months of age. The epizootiology of sheep babesiosis appears to differ from that of bovine babesiosis.     

Serological reactivity to Mycobacterium bovis protein antigens in cattle.

The serological response to 12 purified Mycobacterium bovis antigens were examined in an ELISA assay. These antigens included the majority of M. bovis protein antigens described to date and in most cases they were very similar to the M. tuberculosis antigens of the same molecular mass.

The purified antigens were tested against sera from M. bovis infected cattle, M. bovis culture-negative cattle from infected herds and animals infected with related microorganisms, mainly other mycobacterial species. All the antigens gave strong reactions with at least some sera from the M. bovis infected group and showed cross-reactivity with some of the sera from the other two groups. The antigen with the highest specificity reacted strongly with only 60% of the M. bovis infected sera. Antigens that reacted with most or all of the M. bovis infected sera also gave the highest cross-reactivity with sera from the other two groups. These results indicate that a serological test based on any one or a combination of these antigens, without removal of the cross-reacting epitopes, would be unsatisfactory.     

Epidemiology of theilerioses in the Trans-Mara division, Kenya: Husbandry and disease background and preliminary investigations on theilerioses in calves

All the calves born (116) into 3 Maasai cattle herds in the Trans-Mara Division of Kenya, between August 1978 and October 1979, were recruited into a monthly health study which concentrated on theileriosis. Twenty-two of the calves died before they were 6 months of age, but the mortality only increased to 25% by the time the calves reached 18 months of age. The mean birth weight of calves was 17.5 kg while at 190 days post-birth the mean weight was 53.4 kg. The main causes of mortality were starvation (7.8%), neonatal diarrhoea (2.6%), chronic indigestion (2.6%) and theileriosis (2.6%) due to Theileria parva and T. mutans infections. The calves were infected with ticks from birth (mostly Rhipicephalus appendiculatus and Amblyomma spp.) and the first Theileria schizonts were detected on Day 17 post birth and reached a maximum of 18.4% of calves in the 11th week post-birth. Seasonal peaks of macroschizont incidence occurred in February and July. All calves had patent Theileria piroplasm infections by the time they were 5 months old and 44% had shown patent Theileria macroschizont infections by 6–7 months of age. Generally low parasitosis of Theileria piroplasms and schizonts occurred. Serology using the indirect fluorescent antibody test showed a high proportion of calves received antibodies against T. mutans and T. parva from their dams by way of colostrum. The majority of calves also had active antibody responses against T. mutans and T. parva by the time they were 6 months of age. There was a correlation between the pre-patent period of piroplasms and active antibody responses to T. mutans and between the prepatent period of schizonts and an antibody response to T. parva. Eighteen older calves developed T. velifera infections. “Turning sickness” due to Theileria infection in the brain was detected in older cattle. Other blood parasites such as Trypanosoma vivax, T. congolense, Anaplasma marginale and Babesia bigemina occurred at patent levels at a lower incidence than Theileria spp. and did not cause disease problems in the calves. The calf population was highly resistant to theileriosis since they had a 100% morbidity, but only 2.6% mortality. Theileria infections would appear to have an important effect on the growth of calves but this and many aspects of the epidemiology of theileriosis in the area required more intensive sampling.     

How to substantiate eradication of bovine brucellosis when aspecific serological reactions occur in the course of brucellosis testing

Collaborative work was financed by the EU to develop and assess new diagnostic tools that can differentiate between bovine brucellosis and bovine infections due to Yersinia enterocolitica O:9 either in conjunction with, or as an alternative to, the classical serological, bacteriological or allergic skin tests. Sixteen heifers were experimentally infected with Brucella abortus biovar 1 (five heifers), Brucella suis biovar 2 (two heifers), Y. enterocolitica O:9 (six heifers) and Y. enterocolitica O:3 (three heifers). Four heifers, naturally infected with Y. enterocolitica O:9 that presented aspecific brucellosis serological reactions were also included in the experiment. A self-limited infection was induced in cattle by B. suis biovar 2. All the brucellosis serological tests used, i.e. the slow agglutination test (SAW), the Rose Bengal test (RB), the complement fixation test (CFT), indirect and competitive ELISA’s, lacked specificity when used to analyze sera from Y. enterocolitica O:9 infected animals. A Yersinia outer membrane proteins (YOPs)-ELISA was also used and although the test is able to detect a Yersinia group infection, it provided no evidence of whether or not there is a possible brucellosis infection when dual infections are present. The brucellergen IFN-γ test showed a lack of specificity also. The only test that was proven to be specific is the brucellergen skin test. All brucellosis serological tests, except the indirect ELISA, were limited in their ability to detect B. abortus persistently infected animals.

Based on these experimental studies, a strategy was implemented as part of the year 2001 Belgian Brucellosis Eradication Program to substantiate the eradication of bovine brucellosis. Epidemiological inquiries have identified risk factors associated with aspecific serological reactions, possible transmission and infection of cattle by B. suis biovar 2 from infected wild boars; and both legal and administrative measures taken by the veterinary services. No cases of bovine brucellosis have been confirmed in Belgium since March 2000.     

Epidemiological aspects of canine babesiosis in the semiarid area of the state of Minas Gerais, Brazil

Epidemiological aspects of Babesia vogeli infection were studied in the canine population of a rural town located in the Brazilian “Drought Polygon” of the state of Minas Gerais, Brazil. The survey was carried out in March 2003, when 505 dogs were identified and their characteristics registered on appropriate forms. Blood samples were collected at this time and again in June, September and December 2003. Serum samples were tested by the indirect fluorescent antibody test (IFAT) to detect antibodies against B. vogeli. The prevalence of anti-B. vogeli antibodies was 18.8%; however, no correlations were found between prevalence of infection and the age or gender of the animals. Cross-bred dogs presented a higher chance of acquiring infection in comparison to pure-bred dogs. Significant differences concerning the incidence of the disease were found during the period April–June in comparison to other months, demonstrating that transmission of B. vogeli is related to seasonal variations of tick infestations. The results indicate that climatic factors within the semiarid area interfere directly in the epidemiology of canine babesiosis.     

Serosurvey of Babesia canis, Babesia gibsoni and Ehrlichia canis in pound dogs in California, USA

The seroprevalence of three canine tick-transmitted parasites, Babesia gibsoni, Babesia canis and Ehrlichia canis, was estimated in selected regions of California. Blood smears and sera were obtained from 971 dogs in seven animal shelters: four in Los Angeles County, one in Yolo County, one in El Dorado County in California and one in Minden, Nevada. Seroprevalence in Los Angeles County shelters were 0–13%, 0–2.6% and 0% for B. canis, B. gibsoni and E. canis, respectively. Seroprevalences of the same three parasites in Yolo County and El Dorado County Shelters were 0% except for a 1% seroprevalence of B. canis in dogs from Yolo County Shelter.

Potential risk factors (breed, age, sex and evidence of ticks on the dogs) for B. canis seropositivity were evaluated. Dogs 3 years of age or older had a significantly higher risk (odds ratio 5.04) of being seropositive to B. canis compared with dogs less than 1 year old. Breed, sex and evidence of ticks were not associated with seropositive reactions to B. canis. Of 29 coyotes captured in Los Angeles County, three (10.3%) were seropositive for B. gibsoni, with titers of 1280 to 2560. This study indicated that dogs in Los Angeles County were at higher risk of being seropositive and potentially infected with canine babesial parasites than dogs in Yolo and El Dorado Counties. Movement of chronically infected dogs from Los Angeles County into other areas could contribute to the spread of these important pathogens.     

Seroepidemiology of canine babesiosis and ehrlichiosis in a hospital population

Canine ehrlichiosis and babesiosis have a worldwide distribution with geographic variation in prevalence and main clinical manifestations. We prospectively determined seroprevalence of canine babesiosis and ehrlichiosis, and risk factors for seropositivity. Three hundred and eighty-one dogs were randomly selected to represent the canine population at a Veterinary Teaching Hospital in south Brazil (latitude 23° S). Dogs were tested with a point-of-care ELISA for Ehrlichia canis antibodies and IFA to confirm previous exposure to Babesia vogeli. Multiple logistic regression analysis was then used to estimate adjusted odds ratio (OR) and their 95% confidence intervals. One hundred and thirty-six (36%) dogs were seropositive for B. vogeli antibodies, whereas 87 (23%) dogs were seropositive to E. canis antibodies. Fifty-four (14%) dogs seroreacted to both agents. Adult dogs previously infested with ticks were more likely to seroreact to B. vogeli or E. canis. Superficial bleeding (OR = 12.4) was more common in dogs exposed to B. vogeli, whereas neurological signs (OR = 7.7) were more common in dogs seropositive to E. canis. Neurological signs (OR = 12.0) and lameness (OR = 12.8) were more prevalent in dogs that seroreacted to both organisms. Owners of dogs with ticks were more likely to have been exposed to ticks themselves (OR = 3.2). Canine babesiosis and ehrlichiosis appear to be highly prevalent in this hospital population. Clinical signs differed from the most common signs in other regions with bleeding occurring more in dogs seropositive to babesiosis, but not ehrlichiosis; neurologic signs in dogs with E. canis antibodies; and lameness in dogs that seroreacted to both organisms.     

Babesia divergens-like organisms from free-ranging chamois (Rupicapra r. rupicapra) and roe deer (Capreolus c. capreolus) are distinct from B. divergens of cattle origin - an epidemiological and molecular genetic investigation

In 2005 and 2006, three adult female chamois (Rupicapra r. rupicapra) were found dead with signs of acute babesial infection in the eastern Swiss Alps. PCR on DNA extracted from blood or spleen of the carcasses revealed sequence identity of the amplified part of the 18S rRNA gene with GenBank entries attributed to Babesia divergens of cattle origin or B. capreoli of wild ruminant origin which have never been described before in this region. Examination of 424 blood samples from 314 head of cattle from this area by IFAT, microscopy and PCR provided no evidence for babesial infection. Six of 887 ticks collected from cattle were PCR-positive, and sequencing revealed Babesia sp. genotype EU1 in five and B. divergens/B. capreoli in one of them. A Babesia isolate of chamois, two isolates of roe deer from the same region and one isolate of a roe deer from the north-western Swiss Alps were genetically compared with two Swiss B. divergens isolates of cattle origin by analysing the genomic rDNA locus. Whereas the near full length sequences of the 18S rRNA gene were virtually identical among all six isolates (>99.4% identity), distinct differences between the two isolates from cattle on the one hand and the four isolates from free-ranging ruminants on the other hand were observed in the sequences of the internal transcribed spacers 1 and 2 (ITS1, ITS2) and part of the 28S rRNA gene. These results indicate that, albeit genetically very closely related, these babesial organisms from cattle and from free-ranging ruminants indeed are distinguishable organisms with different host specificities, and they support the use of the discrete species name B. capreoli for the B. divergens-like organisms from chamois and roe deer.     

Evaluation of a commercially available enzyme-linked immunosorbent assay for detecting antibodies to Fasciola hepatica and Fasciola gigantica in cattle, sheep and buffaloes in Australia

A commercially available ELISA for detecting antibodies to liver fluke was evaluated for use in Australia. Milk and serum samples from cattle and sheep in which infection with Fasciola hepatica was confirmed by detection of eggs in faeces were used to estimate sensitivity. Similar samples collected from cattle and sheep outside the F. hepatica-endemic area were used to estimate specificity. The ELISA was also evaluated for detecting antibodies to F. hepatica in milk from sheep and antibodies to Fasciola gigantica in sera from cattle and buffaloes, but with small numbers of samples. In cattle, the sensitivity and specificity of the ELISA were 98.2% and 98.3% using serum and 97.7% and 99.3% using milk. In infected herds, 41.4% and 41.5% of animals were positive in the serum and milk ELISAs, respectively, whereas F. hepatica eggs were found in faecal samples from 26.5% of animals. In sheep, the sensitivity of the ELISA was 96.9% and the specificity was 99.4%. In infected flocks, 60.2% of animals were positive in the serum ELISA and F. hepatica eggs were found in faecal samples 52.2% of animals. There was perfect agreement in the ELISA between paired serum and milk samples collected from ewes. The assay detected antibodies in sera from cattle and buffaloes with natural and experimental F. gigantica infections. In the experimentally infected animals, antibodies were detected 2 weeks post-infection. We conclude that the ELISA will be a valuable tool for diagnosing F. hepatica infections in cattle and sheep. The assay may also be useful for diagnosing F. gigantica infections but further studies are required to establish sensitivity and specificity.     

牛传染性胸膜肺炎抗体间接ELISA检测方法的建立

旨在建立一种检测牛传染性胸膜肺炎（CBPP）血清抗体的间接ELISA方法,用于该病的监测。利用生物信息学软件对CBPP的病原丝状支原体丝状亚种国内分离株Ben-1株的全基因组进行分析,选取脂蛋白rP0308作为包被抗原,通过一系列反应条件的优化,建立了基于rP0308蛋白的间接ELISA抗体检测方法,并对其性能进行评价。结果显示该方法的敏感性为92%,特异性为96%,与牛支原体、牛鼻支原体、无乳支原体、口蹄疫、牛病毒性腹泻、牛传染性鼻气管炎和牛结核分枝杆菌等阳性血清均不发生交叉反应,批内变异系数在2.41%~6.03%,批间变异系数在2.94%~6.59%,重复性良好。利用本方法和商品化的cELISA试剂盒分别对实验室保存的1 648份临床样品进行检测,该方法与商品化试剂盒的阴性符合率为89.1%,阳性符合率为79.2%,总符合率为88.7%。以上研究结果表明,本研究建立的间接ELISA方法具有良好的敏感性、特异性和重复性,具有良好的临床应用前景。     

Estimation of sensitivity, specificity and predictive values of two serologic tests for the detection of antibodies against Actinobacillus pleuropneumoniae serotype 2 in the absence of a reference test (gold standard)

Latent-class models were used to determine the sensitivity, specificity and predictive values of a polyclonal blocking enzyme-linked immunosorbent assay (ELISA) and a modified complement-fixation test (CFT) when there was no reference test. The tests were used for detection of antibodies against Actinobacillus pleuropneumoniae serotype 2 in a survey of respiratory diseases in Danish finishing pigs. The estimates were obtained by maximum-likelihood and also by a Bayesian method (implemented with Gibbs sampling). Possible dependence of diagnostic errors was investigated by comparing models where independence was assumed to models allowing for conditional dependence, given the true disease status.

No strong evidence of conditional dependence in either test sensitivity or specificity was found. Assuming independence, maximum-likelihood estimates and 95% confidence intervals of the sensitivity and specificity of the ELISA were 100% and 92.8% (90.1–95.5%) and the corresponding values of the CFT were 90.6% (85.8–95.4%) and 98.6% (98.0–99.3%), respectively. Bayesian estimates and posterior 95% credible intervals of the sensitivity and specificity of the ELISA were 99.7% (98.7–100%) and 92.7% (89.9–95.3%) and of the CFT were 90.6% (86.0–95.3%) and 98.7% (98.0–99.3%). The sensitivity and specificity of a combined test, where the CFT is subsequently applied to the pig sera that test positive in the ELISA, were estimated at 90.2% (85.6–95.0%) and 99.9% (99.8–100%), respectively. The cost of the combined test was less than the cost of the use of the CFT alone, at prevalences <54%. Prevalences and predictive values and their 95% limits were estimated in six sub-samples of data. The estimates of sensitivity and specificity obtained in the present investigation generally validate those reported from other sources.     

Serologic survey of wild cervids for potential disease agents in selected national parks in the United States

A total of 589 serum specimens were collected from mule deer (Odocoileus hemionus) (133) and wapiti (Cervus elaphus) (456) in eight national parks and/or adjacent lands in the western USA. Thirty two percent of the samples were collected from immobilized animals and 68% from hunter-killed animals in or near Glen Canyon National Recreation Area, Bryce Canyon National Park (NP), and Zion NP, Utah; Yosemite NP, California; Rocky Mountain NP, Colorado; Upper Yellowstone NP, Montana, and Grand Teton NP, Wyoming. Serum specimens were tested for the presence of antibodies against selected disease agents. Overall seroprevalences for mule deer were 77/133 (58%) for parainfluenza-3 virus (PI-3), 42/133 (32%) for bovine herpesvirus-1 (BHV-1), 79/133 (59%) for bovine virus diarrhea virus (BVD), 73/133 (55%) for respiratory syncytial virus (RSV), 14/133 (11%) for bluetongue virus (BT), 18/133 (14%) for epizootic hemorrhagic disease vims (EHD), 3/133 (2%) for Borrelia burgdorferi, and 1/133 (1%) for Francisella tularensis. None of the deer sera presented antibodies for Leptospira spp., Brucella abortus and Anaplasma marginale. For wapiti, overall prevalences were 262/456 (57%) for PI-3, 211/456 (46%) for BHV-1, 251/456 (55%) for BVD, 247/456 (54%) for RSV, 1/456 ( < 1%) for BT, 16/456 (4%) for Leptospira pomona, 13/456 (3%) for Leptospira hardjo, and 8/456 (2%) for B. abortus. No antibody titers were detected for EHD, A. marginale, and other Leptospira serotypes. This survey documents seroprevalence of selected park cervid populations to domestic livestock pathogens. Further research on the epidemiology of these potential pathogens in wild ungulates in national parks is recommended.