A review of tuberculosis and parasitic disease co-infection in ungulates, with regard to the potential threat to European bison (Bison bonasus).

Bovine tuberculosis (BTB) is a dangerous zoonosis which presents a serious problem for endangered species such as European bison ( Bison bonasus). Little is known about the influence of parasitic co-infections on the course and diagnosis of tuberculosis in animals. The best known co-infection in cattle is Fasciola hepatica and Mycobacterium bovis. The aim of this study was to review the most recent literature regarding tuberculosis and parasite co-infection in ungulates and relate the results to European bison. Our findings indicate that any comprehensive diagnosis of BTB should include parasitological monitoring, and the possible impact of such invasions on cellular response-based tuberculosis tests should be taken into account. The diagnosis of BTB is complex, as is its pathogenesis, and parasitic infestations can have a significant impact on both. This should be taken into account during further research and monitoring of tuberculosis in European bison.

Suspicion of bovine tuberculosis in sheep in the Malopolskie Voivodeship (southern Poland).

Bovine tuberculosis (BTB) in sheep (Ovis aries) is caused by Mycobacterium bovis and Mycobacterium caprae. Even though sheep have been considered less sensitive to BTB than other ruminants, they have been subject to increasing numbers of tuberculosis cases and it has been suggested that they may act as a disease reservoir in some regions. Aim of the study: Following a report of tuberculous-like gross lesions (repeated cases of purulent or caseous lymphadenitis and a single case of serosal tubercles on the peritoneum) from veterinarians working in a slaughterhouse in the Malopolskie Voivodeship, southern Poland, the aim of this study was to conduct ante-mortem BTB diagnostics in three flocks with suspected BTB. The animals for testing were selected randomly from the flocks; a blood sample for interferon-γ release assay (IGRA) and a single tuberculin skin test (TST) was performed on each sheep. All TST results were negative. The IGRA result was positive in two ewes from the same flock (four and five years of age); these two sheep were tested microbiologically using Stonebrink and Löwenstein-Jensen media. No gross lesions suggesting BTB were observed, and the culture results were negative. Based on the positive IGRA results, together with its high specificity in sheep, and the potential risk to humans posed by consuming local unpasteurized dairy products, we recommend introducing serological BTB screening in sheep from this area, and subjecting the positive results to confirmation by culture.

The effect of heterotypic infections of older horses with equine influenza virus type-2 on some clinical and immunological parameters.

Twelve horses, all of them 10 years old, were vaccinated intramuscularly on 0 and 28 days of the experiment with inactivated vaccine containing only antigens of A-equi-2/Miami/63. Another three unvaccinated horses, each at the age of 10 years, were the negative control group. One, ten-year-old horse was vaccinated with commercial inactivated vaccine containing both antigens of A-equi-2/Miami/63 as well as A-equi-1/Praha/56 as positive control. Three horses were challenged intranasally with homotypic strain of Miami/63, while six other were challenged with heterotypic strains--three with Suffolk/89 and three with Kentucky/86. Three horses vaccinated with vaccine containing only strain A-equi-2/Miami/63 were not challenged. In the group of three unvaccinated horses, each one was challenged intranasally with different strains studied in this experiment. The horse vaccinated with commercial vaccine was not challenged. Replication of each strain was done in chick embryos. During the experiment blood from horses was collected for hematological and immunological examinations (antigen-specific and antigen-nonspecific lymphocyte transformation tests, lymphocyte immunophenotyping, antigen-specific leukocyte migration inhibition test and hemagglutination inhibition test). The statistical analysis showed that the dynamics of lymphocyte immunological reactivity in horses vaccinated with inactivated vaccine containing antigens of A-equi-2/Miami/63 in response to further antigen stimulation (in vitro) was different comparing the homotypic or nearly homotypic challenging with Miami/63 and Suffolk/89 respectively, to the more heterotypic one with the strain Kentucky/86. In horses challenged with classical homotypic strain of Miami/63 no clinical signs were observed. These results confirm that the vaccine shall consist of the strains currently circulating in the horse population.

Some morphological, biochemical and immunological blood parameters in European bison with or without lesions in the digestive, respiratory, urinary and reproductive tracts.

Ninety-seven bison (39 males and 58 females) were culled out during three successive winter seasons. No infection with BHV-1 and BHV-5 or other viruses was detected by PCR and SN methods. Advanced pathological lesions observed in males exhibiting symptoms of posthitis/balanoposthitis are probably due to secondary infection by microorganisms present in the prepuce (Corynebacterium spp., Bacillus spp., Fusobacterium necrophorum, Staphylococcus spp. (coagulase negative) and Staphylococcus hyicus). Histopathological examinations of sections from prepuce and lips of pudendum showed the presence of Onchocercinae nematode (Onchocerca sp.). A comparison of biochemical and morphological blood parameters in the bison with or without lesions in the digestive (presence of parasites), respiratory, urinary and reproductive tracts revealed that the most variable parameter during reproductive organ disorder is the leukocyte count, while disorders in the urinary and digestive tracts coincided with the age of animals. In cases of digestive disorder most changes were observed in the number of neutrophils and concentration of urea. The number of red blood cells, monocytes, the level of alkaline phosphatase (AP) and hematocrit negatively correlated with age. Negative correlation was also observed between the number of neutrophils, level of creatinine and lymphocyte proliferation index. However, positive correlation was seen between erythrocyte sedimentation rate (Spearman's correlation coefficient), level of alanine aminotransferase (ALT) and protein, and age of the bison. Positive correlation was also observed between the lymphocyte count and lymphocyte proliferation index. The attempt to determine the reference values for blood biochemical and morphological parameters showed similarities to those of cattle.

Seroprevalence of selected viral and bacterial pathogens in free-ranging European bison from the Bialowieza Primeval Forest (Poland).

Blood was collected from bison that were selected between 1991 and 2001 for poor body condition, cachexia, lameness and balanoposthitis. Sera were tested for antibodies to bovine herpes virus type 1 (BHV-1), bovine viral diarrhoea virus (BVDV), foot-and-mouth disease virus (FMDV), parainfluenza virus type 3 (PI-3), Brucella abortus, Chlamydophila abortus, Coxiella burnetti, and Leptospira interrogans. The results of serological tests showed a prevalence of low titers of a serological reaction to Chlamydophila abortus (45.1%), bovine viral diarrhoea virus (29.5%), Leptospira interrogans (21.3%) and parainfluenza virus type 3 (13.9%). There were differences in the prevalence of antibodies to Ch. abortus between female and male bison. Futhermore, a relationship between age and antibodies to BVDV was observed in older bison. These results suggest that there is some transmission of bacterial and viral pathogens occurring between domestic and wild ruminants grazing in the same pastures in the peripherial areas of Bialowieza Primeval Forest.

Relationships among ecology, demography and diseases of European bison (Bison bonasus).

The bison population in the Bialowieza Forest in Poland has now grown to approximately 300, while the herds in the Belarusian part of the forest total about 240 bison. The first signs of a health problem in these herds appeared in 1980, when two cases of balanoposthitis were detected in two bulls (2 and 5 years of age). Since 1980 research has been conducted to explain the cause of diseases, particularly balanoposthitis, and to monitor the health of bison in Bialowieza Forest. A total number of 614 bison (294 male and 320 female) of different ages was eliminated between 1980 and 2000. Not all the culled bison were examined (postmortem analysis, histopathological, bacteriological, virological and toxicological examinations, serological tests, molecular research). Based on the increase in numbers, reproduction in this population for the past 21 years is generally considered successful. Among 182 male bison eliminated during 1990-2000, only 85, or 47%, of the animals had balanoposthitis. Thus, the percentage of balanoposthitis cases went from 100% during the 1980s down to 47% in the past decade. It appears that the culling process has been a major factor leading to this decrease. It can be assumed that a set of factors is involved in the appearance of the disease (Corynebacterium spp., Bacillus sp., Pseudomonas aeruginosa, Escherichia coli, Ureoplasma spp, Fusobacterium necrophorum, Streptoccocus spp., Staphyloccocus spp.) while opportunistic infections including nematodes (Onchocerca spp.) are responsible for the occurrence of secondary lesions.

Serologic survey for bovine pathogens in free-ranging European bison from Poland.

From 1980 to 1983, blood was taken from 60 selected European bison (Bison bonasus) in Poland. Serum samples were tested for the presence of antibodies against Brucella abortus, 14 serovars of Leptospira interrogans, Chlamydia spp., Coxiella burnetti; foot and and mouth disease virus, bovine leukemia virus and bovine herpes virus-1. In addition, an attempt was made to isolate bovine herpes virus-1 from the prepuce of selected bulls. Serological tests suggested chlamydial infection in 28 bison, subclinical Q-fever of a 2-yr-old heifer, subclinical bovine leukemia virus infection in a 12-yr-old bull and bovine herpes virus-1 infection in five bulls and three cows. Attempts to isolate bovine herpes virus-1 were not successful. These results suggest the possibility of cross transmission of several of these bovine pathogens between free-ranging bison and domestic cattle in Poland.

Detection of Cryptosporidium parvum oocysts in bovine feces by monoclonal antibody capture enzyme-linked immunosorbent assay.

A monoclonal antibody enzyme-linked immunosorbent assay (ELISA) was developed to detect Cryptosporidium parvum oocysts in bovine feces. Fecal oocysts were concentrated by centrifugation through Formalin-ethyl acetate solution and captured with monoclonal antibody 18.280.2 reactive with C. parvum oocysts. Captured oocysts were detected with goat anti-oocyst serum, following the addition of a peroxidase conjugate of rabbit anti-goat immunoglobulin and O-phenylenediamine substrate. The assay was specific for Cryptosporidium sp. oocysts and did not detect oocysts of Eimeria auburnensis, Eimeria bovis, Eimeria ellipsoidalis, or Eimeria zuernii. Assay sensitivity allowed detection of 3 x 10(5) oocysts per ml of feces, compared with 1 x 10(6) oocysts per ml detected by examination of acid-fast-stained fecal smears and 1 x 10(3) oocysts per ml detected by indirect immunofluorescence. The capture ELISA was suitable for diagnostic analysis of bovine fecal samples and for assessment of oocyst shedding in experimentally infected calves. This assay may also prove useful for diagnostic assessment of humans in which cryptosporidiosis is suspected.