Specificity of serological test for detection of tuberculosis in cattle, goats, sheep and pigs under different epidemiological situations.

BACKGROUND: Serum antibody detection has potential as a complementary diagnostic tool in animal tuberculosis (TB) control, particularly in multi-host systems. The objective of the present study was to assess the specificity (Sp) of an enzyme-linked immunosorbent assay (ELISA) based on the new multiprotein complex P22 for the detection of specific antibodies against the Mycobacterium tuberculosis complex (MTC) in the four most relevant domestic animals acting as MTC hosts: cattle, goat, sheep and pig. We used sera from an officially TB-free (OTF) country, Norway, and from a non-OTF one, Spain. The samples included sera from goats that had been vaccinated against M. avium subsp. paratuberculosis (MAP) and sheep from a herd in which Corynebacterium pseudotuberculosis had been isolated. RESULTS: In cattle, the Sp ranged from 92.5 (IC95% 90.7-94) to 99.4% (IC95% 98.3-99.8) depending on the cut-off used and the origin of the samples (Spain or Norway). Sp in cattle (cut-off point 100) was significantly higher (P < 0.05) for Norwegian samples. By contrast, Sp in goats was consistently low at the 100 cut-off [30.9 (CI95%23.4-39.5)-78% (CI95% 68.9-85)]. A higher cut-off of 150 improved Sp in Norwegian goats [97% (CI95% 91.6-99)], but still yielded a poor Sp of 56.1% (CI95% 47.3-64.6) in Spanish goats. In Norway at the 100 cut-off the Sp was 58.3 (CI95% 42.2-72.9) and 90.6% (CI95% 81-95.6) in MAP vaccinated and non-vaccinated goats, respectively, indicating interference due to MAP vaccination. Sp in sheep was between 94.4 (CI95% 91.7-96.3) and 100% (CI95% 96.3-100) depending on the cut-off and country, and no diagnostic interference due to infection with C. pseudotuberculosis was recorded. Sp in pigs was 100%, regardless the cut-off point applied, and no significant differences were observed between pigs from Norway and from Spain. CONCLUSIONS: Due to its excellent Sp in pigs and acceptable Sp in cattle and sheep, this ELISA may constitute a suitable option for TB screening at herd level, particularly in OTF-countries.

Enteropathogens and risk factors for diarrhea in Norwegian dairy calves.

The aims of the current study were to estimate the prevalence of enteropathogens in calves in Norwegian dairy herds, evaluate the clinical consequences of protozoal infections, and identify risk factors for diarrhea. The 135 participating herds were randomly selected from those in The Norwegian Dairy Herd Recording System that had at least 15 cow-years. Each herd was followed for 1 yr. Fecal samples from calves with (n = 68) or without (n = 691) diarrhea were analyzed for the presence of Cryptosporidium, Giardia, and Eimeria species. Diarrheic samples (n = 191) were assayed for rotavirus group A, bovine coronavirus (BCoV), Cryptosporidium, and Escherichia coli F5 by antigen ELISA. Blood samples (n = 1,348) were analyzed for antibodies against BCoV and rotavirus. Potential risk factors for diarrhea were analyzed by using Cox regression analysis adjusted for herd frailty effect. Rotavirus and Cryptosporidium were the most commonly detected enteropathogens in diarrheic samples. A high level of Cryptosporidium shedding or BCoV seropositive calves in a herd was associated with an increased risk of diarrhea. Other factors found to increase the risk of diarrhea were use of slatted concrete floor in group pens versus other floor types [hazard ratio (HR) = 8.9], housing of calves in free-stalls compared with tie-stalls (HR = 3.7), purchasing of calves into the herd versus not purchasing calves (HR = 4.1), and calves being born during winter compared with other seasons of the year (HR = 1.5).

Respiratory infections in Norwegian dairy calves.

The aims of this study were to estimate the seroprevalence of respiratory agents in Norwegian dairy calves and to identify risk factors for respiratory disease. The participating 135 herds were randomly selected from those in The Norwegian Dairy Herd Recording System with at least 15 cow years. Each herd was followed for 1 yr. Blood samples from calves of >150 d of age (n = 1,348) were analyzed for antibodies against parainfluenza virus 3, bovine coronavirus (BCoV), bovine respiratory syncytial virus (BRSV), and Mycoplasma bovis. Calves reported to have been on pasture (n = 139) were tested for antibodies against Dictyocaulus viviparus. Seroprevalences for parainfluenza virus 3, BCoV, BRSV, and D. viviparus at the calf level were 50.2, 39.3, 31.2, and 4.3%, respectively. No calves were antibody positive for M. bovis. Calves in herds with BCoV-seropositive calves had an increased risk of respiratory disease compared with herds in which BCoV antibodies were not detected [hazard ratio (HR) = 3.9], as had calves in herds in which the majority (>54%) of the sampled calves were seropositive for BRSV (HR = 2.7). Other factors found to increase the risk of respiratory disease in calves were shared housing with cows during the first week of life compared with separate housing (HR = 16.7), a larger herd size (>50 cow years) compared with smaller herds (HR = 8.2), more than an 8-wk age difference between calves housed together in the same group pen compared with having pen mates of a more similar age (HR = 3.9), previous recordings of diarrhea compared with no recorded diarrhea (HR = 3.9), and leaving calves with dams for >24 h after birth compared with earlier separation (HR = 3.5).

Acute toxoplasmosis in three wild arctic foxes (Alopex lagopus) from Svalbard; one with co-infections of Salmonella Enteritidis PT1 and Yersinia pseudotuberculosis serotype 2b.

Acute disseminated toxoplasmosis was diagnosed in three wild arctic foxes (Alopex lagopus) that were found dead in the same locality on Svalbard (Norway). The animals included one adult female and two 4-months-old pups. The adult fox was severely jaundiced. Necropsy revealed multifocal, acute, necrotizing hepatitis, acute interstitial pneumonia, and scattered foci of brain gliosis, often associated with Toxoplasma tachyzoites. One pup also had Toxoplasma-associated meningitis. In addition, the latter animal was infected with Yersinia pseudotuberculosis serotype 2b and Salmonella Enteritidis phage type 1 (PT1), which may have contributed to the severity of the Toxoplasma infection in this animal. The diagnosis of toxoplasmosis was confirmed by positive immunohistochemistry and detection of anti-Toxoplasma gondii antibodies in serum of all foxes. The animals were negative for Neospora caninum, canine distemper virus, canine adenovirus, and rabies virus on immunolabelling of tissue sections and smears.

Bacteriological investigation of infectious keratoconjunctivitis in Norwegian sheep.

Contagious keratoconjunctivitis is a rather common disease in Norwegian sheep. Since the knowledge of its aetiology is limited, the present study was performed to determine the microorganisms involved. Local veterinarians throughout the country collected conjunctival swabs from both sick (n = 43) and healthy (n = 42) sheep on 15 farms with outbreaks of ovine keratoconjunctivitis, and further from healthy sheep (n = 50) on 17 farms not showing any signs of conjunctival disease. All samples were cultivated for bacteria and mycoplasma. Listeria monocytogenes was isolated from 3 cases (1%) in one single herd. Staphylococcus aureus (5%), Corynebacterium spp. (2%) and Escherichia coli (4%) were isolated only in herds with keratoconjunctivitis, but from both sick and healthy animals. Moraxella (Branhamella) ovis was isolated from 28% of sampled animals in affected herds and from 10% of sampled animals in healthy herds. The corresponding numbers for Moraxella spp. were 9%/12%, for Pseudomonas spp. 7%/8%, for Staphylococcus spp. 22//22%, for Bacillus spp. 12%/14%, for Micrococcus spp. 6%/2% and for Streptococcus/Enterococcus spp. 2%/2%. Mycoplasma conjunctivae was isolated from 16 animals with keratoconjunctivitis (37%) and from 3 animals without clinical signs (7%) in farms with keratoconjunctivitis. In farms without clinical signs of keratoconjunctivitis, M. conjunctivae was isolated in 4 animals (8%). To our knowledge, this is the first time M. conjunctivae has been isolated in Norway. Other predisposing agents found were Moraxella (Branhamella) ovis and Listeria monocytogenes. The etiological importance of different microorganisms in ovine keratoconjunctivitis seems to vary; some are probably only present as secondary invaders. Other possible causes of ovine keratoconjunctivitis in Norway, such as Chlamydia psittaci, remain to be investigated.

Fox encephalitozoonosis: isolation of the agent from an outbreak in farmed blue foxes (Alopex lagopus) in Finland and some hitherto unreported pathologic lesions.

The farmed blue fox (Alopex lagopus) is particularly susceptible to congenital infections of the microsporidian species Encephalitozoon cuniculi. This report is based on an outbreak of the disease in Finland with high mortality. Five pups (four males and one female) with prolonged disease were examined. The pups had moderate pathological alterations in the kidneys and mild lesions were found in the brains, hearts, salivary and prostatic glands. Diagnosis of E. cuniculi infection was made from serological tests (ELISA, CIA, IFAT), and by in vitro isolation of the parasite from the brain of all five pups investigated. The identity was confirmed by molecular means as E. cuniculi strain II ('mouse strain'). Novel histopathological lesions not described as yet in fox encephalitozoonosis are presented. These include cerebral infarction and necrotizing inflammation of the renal pelvis. The sources and mechanisms of spreading of E. cuniculi to blue foxes are discussed.

An indirect ELISA for detection of Encephalitozoon cuniculi infection in farmed blue foxes (Alopex lagopus).

Infection with the intracellular microsporidium Encephalitozoon cuniculi can cause serious disease, encephalitozoonosis, in the blue fox (Alopex lagopus). The disease diagnosis is based on clinical signs and pathological findings, and detection of E. cuniculi or circulating antibodies directed against the parasite. Indirect immunofluorescence (IFAT) and carbon immunoassay (CIA) are the most commonly used serological methods for diagnosis in this species. In the present study, an indirect ELISA (enzyme linked immunosorbent assay) was established and evaluated against IFAT by testing of 205 field samples from blue foxes. There was high agreement between the results of the ELISA and CIA (kappa=0.99), and the ELISA and IFAT (kappa=0.958). There was no significant statistical difference between the tests (p>0.05). It was concluded that the ELISA could be used to identify seropositive farmed blue foxes. The advantage of the ELISA lies in the potential of screening large numbers of animals with the goal of eradicating E. cuniculi infection in the farms.

Isolates of Encephalitozoon cuniculi from farmed blue foxes (Alopex lagopus) from Norway differ from isolates from Swiss domestic rabbits (Oryctolagus cuniculus).

Encephalitozoon cuniculi has a wide host range among mammals, but whether it represents a homogeneous species is a subject of controversy. We have isolated, cultivated (in human MRC-5 cells) and, for the first time, characterized by immunological and molecular biological methods four isolates of E. cuniculi from Norwegian blue foxes with a history of encephalitozoonosis. The isolates were compared with nine isolates from domestic rabbits from Switzerland. Two E. cuniculi subtypes were identified according to their host species. A 5'-GTTT-3' tetranucleotide repeat was present twice in the rDNA intergenic spacer in all isolates from foxes as opposed to three times in all isolates from rabbits. Furthermore, random amplified polymorphic DNA analysis showed one polymorphic band among the subtypes, and Western-blot analysis using serum from an infected fox discriminated between the two subtypes on the basis of their banding patterns in the ranges of 31-33 and 38-40 kDa. The 5'-GTTT-3' tetranucleotide repeat is a valuable genetic marker for these two subtypes of E. cuniculi and will be of use in continued studies on the molecular epidemiology of this parasite.

Malassezia pachydermatis with special reference to canine skin disease.

A review of the diseases caused by Malassezia pachydermatis has led to the conclusion that the yeast is an opportunistic pathogen that depends on predisposing host factors and different immune suppressive mechanisms for clinical manifestation. Until recently, the role of M. pachydermatis in seborrhoeic dermatitis and otitis externa in dogs has been largely unrecognized. The clinical manifestation, aetiology, diagnosis and treatment of disease conditions in dogs caused by M. pachydermatis are reviewed. Human Malassezia furfur infections are briefly described.