Comparison of serological tests for the detection of ovine and caprine antibody to Brucella melitensis.

The indirect enzyme-linked immunosorbent assay (IELISA), the competitive enzyme-linked immunosorbent assay (CELISA) and the fluorescence polarisation assay (FPA) were evaluated with sera from sheep experimentally infected with Brucella melitensis and negative Canadian sheep. The sensitivity and specificity of the assays were as follows: IELISA: 91.7% and 97.6%, CELISA: 75.0% and 99.8% and FPA: 91.7% and 89.5%. Sera from the same experimental population were divided according to serological reaction in the rose bengal agglutination test (RBT) and the complement fixation test (CFT). Reactivity relative to the RBT positive and CFT positive sera were as follows: IELISA: 99.7%, CELISA: 93.2% and FPA: 99.1%. Since sera from goats with proven B. melitensis infection were not available, 699 sera from goats judged positive in the buffered antigen plate agglutination test (BPAT) and CFT and 982 BPAT/CFT negative Canadian goats were used. The sensitivity and specificity of the assays relative to the BPAT and CFT positive sera were: IELISA: 99.4% and 98.0%, CELISA: 95.4% and 97.1% and FPA: 92.7% and 99.8%.

Recovery of EDTA and metal precipitation from soil flushing solutions.

This work studies the effectiveness of a process proposed for the recovery of ethylenediaminetetraacetic acid (EDTA) and metal precipitation from soil flushing solutions. Two series of experimental tests were carried out on two samples of a soil artificially contaminated with copper or lead. The metals were extracted from the soil by flushing with a 0.05 M aqueous solution of EDTA sodium salt (E-Na(2)). Cu or Pb extraction efficiencies of about 95 and 98% were observed, respectively. The two extracted solutions were then treated to obtain EDTA recovery and metal precipitation from the aqueous solution. EDTA recovery was achieved in two steps. An initial evaporation treatment lead to reduce the solution volume by about 75%. This was followed by the acidification of the residue solution, which precipitated more than 93% of the used EDTA. The precipitated EDTA was removed by filtration, and was suitable for reuse after adding an alkaline agent. Metal precipitation from the filtered solution was performed using two different methods: an almost total (99.5%) Pb precipitation in alkaline conditions was achieved after complex destabilization through the sequential addition of ferric ions and sodium phosphate, while 93.6% copper precipitation was achieved with ferrous sulfate as a destabilization agent.

Assessment of a monoclonal antibody-based competitive enzyme linked immunosorbent assay (cELISA) for diagnosis of brucellosis in infected and Rev. 1 vaccinated sheep and goats.

In this study a cELISA for the diagnosis of brucellosis due to B. melitensis in sheep and goats was evaluated and its capability of discriminating vaccinated from infected animals was assessed. Information is provided indicating that the cELISA has a diagnostic sensitivity (99.4%) and specificity (98.9%) in sheep and goats comparable to that of many standard indirect ELISA methods. In addition, the test proved able to distinguish between vaccinated and infected animals with an accuracy of up to 90% and results reproducibility of 93%. It was concluded that the cELISA could be useful for differentiation of Rev.1 vaccinated and naturally infected sheep and goats.

Assessment of an indirect ELISA in milk for the diagnosis of ovine Brucellosis.

The possibility of using an ELISA for the diagnosis of ovine brucellosis in milk (M-ELISA) was investigated. The aim of the study was to establish whether the specificity and sensitivity of the M-ELISA would be high enough to detect low levels of Brucella antibodies in ewe milk. The diagnostic performances of the test under study were established by means of reference standards and compared with conventional screening and confirmatory tests under field conditions. The diagnostic specificity of the M-ELISA established on a number of samples from Brucella-free flocks was 100% while relative to RBT and CFT positive reactors the M-ELISA demonstrated sensitivity of 65 and 83% respectively. Its sensitivity relative to culture positive animals was of 92%. The course of Brucella antibodies in milk of positive sheep was evaluated in colostrum and in mature milk for a period of 30 days after delivery and it appeared that concentrations of immunoglobulins in milk tend to sharply decrease soon after parturition while in blood serum these remained constantly high. It was concluded that the M-ELISA for Brucella antibodies in ewe milk can be regarded as a complementary diagnostic tool for individual testing but it would be poorly viable if used as a screening test applied to pooled flock milks.

Avian toxoplasmosis: experimental infection of chicken and pigeon.

Two groups of 13 new-laying hens each were infected by crop-route with 5000 and 50,000 infective oocysts of T. gondii. Four groups of 5 pigeons each were inoculated by crop-route with 50, 500, 1000 and 5000 infective oocysts. To each group of infected birds suitable controls were added. Hens from the experiment with 5000 infective oocysts were apparently resistant to the infection and they had no clinical signs in the succeeding 40 days p.i. Hens from the experiment with 50,000 infective oocysts showed an egg-drop and mortality in embryonated eggs, especially during the first 2 weeks p.i. Isolation of the parasite was unsuccessfully attempted from 720 embryonated eggs, produced by infected groups, and tested on various days p.i. and at different stages of infection. The parasite was isolated from the brain, heart, liver, spleen and lung of infected birds 7 and 15 days p.i.; 40 days p.i. it was evident only in brain and heart. IgG onset and mean course were monitored by ELISA and high titers were reached by both groups. Pigeons from groups 500, 1000 and 5000 developed rapidly progressive clinical signs as diarrhea, trembling, incoordination, torticollis and death. They had enlargement of liver and spleen and focal necrosis, nodular features in the crop. Pigeons from expt 50 had no clinical signs in spite of the presence of the parasite in their organs for over 45 days p.i. Parasite was isolated from brain, heart, liver, spleen, lung, kidney, crop and muscles from all infected groups. Histopathological and ultrastructural features revealed the presence of multiplying tachizoites even within cells of the crop. Seroconversion, as monitored by ELISA, was recorded in all infected groups although high ELISA-titres were never reached. One of the negative controls from expt 5000 developed specific antibodies but the parasite was not isolated from its organs.

Enzyme-linked immunoassay in the diagnosis of leptospirosis in domestic animals using peroxidase-conjugated protein-A.

The ELISA test for detection of antibodies to Leptospirosis in domestic animals was performed using Staphylococcal protein-A coupled to peroxidase in place of antisera to IgG. Genus- and type-specific antigens were extracted with SDS technique from four pathogenic serotypes and two non-pathogenic ones, and they were identified with the aid of ELISA using specific rabbit antisera. Micro-agglutination (MA) and ELISA were compared using a total of 48 positive swine sera and a 100% agreement was obtained, since with sera from 16 dogs clinically suspected of Leptospirosis the ELISA resulted highly more sensitive and precocious than MA in detecting specific antibodies.

An occurrence of Newcastle disease in pigeons: virological and serological studies on the isolates.

The antigenic and pathogenetic relationship between pigeon Newcastle disease virus (NDV) isolates during outbreaks of 1982 in Italy and reference pathogen and non-pathogen NDV-strains were investigated. The pigeon-isolates were slow-eluters and showed a thermostability at 56 degrees C of over 30 min. They proved to be lentogenic as measured by the mean-death-time in chicken-embryos, and between lentogenic and mesogenic as measured by the Hanson test. They failed to produce plaques in chicken-embryo-fibroblasts and showed high pathogenicity for experimentally infected pigeons, low-pathogenicity for quails and were not pathogenic for chickens. They were antigenically different from the LaSota strain as measured by the cross-HI-test and induced considerable seroconversion in inoculated animals. The existence of a lentogenic neurotropic pigeon-pathogenic strain was considered.

Presence of Aujeszky's disease virus in organs and materials from experimentally infected dogs.

The presence of Aujeszky's disease virus was investigated in various organs and materials from experimentally infected dogs, employing the fluorescent antibody test (FAT), tissue culture and biological tests in rabbits. The FAT appeared less sensitive for virus detection than virus isolation in cell culture and rabbit tests. The virus distribution in the central nervous system was related to the site of inoculation.