Purification of Cowdria ruminantium by immunoadsorbent affinity chromatography

Immunoselective methods with special reference to immunoadsorbent affinity chromatography as a means for the isolation of Cowdria ruminantium are reviewed. Attention is given to the source of the organism, immunization, purification of antibodies, coupling of antibodies to insoluble matrixes and desorption procedures.     

Serological identification of a new porcine mycoplasma species, M. flocculare

In a preceding paper (Friis 1972), the so-called SAR (M. suipneumoniae antibody resistant) mycoplasma strains were described. As the reported findings indicated that these isolates belonged to a separate species, a representative cloned strain, Ms42, was selected for serological comparison with the known acid-producing mycoplasma species.     

深圳地区动物肉源沙门菌的分离鉴定及耐药性分析

在改进沙门菌分离方法的基础上,分离并鉴定深圳市动物肉源沙门菌,并进行耐药性分析。结果:采集自深圳地区的1 055个动物肉源样品的沙门菌总阳性率为21. 04%,其中:鸡肉(28. 71%)>猪肉(24. 95%)>牛肉(15. 00%)>虾肉(2. 82%)。基于最小抑菌浓度(MIC)测定去除重复的耐药表型,得到370株非重复沙门菌株;其中的75. 2%多重耐药,53株(14. 10%)表现ACSSuT^R耐药表型。经血清型鉴定,得到38种血清型及327株确定血清型的沙门菌,优势血清型包括Derby、1,4,[5],12: i: -、Rissen、Agona和8,20: z23,z4: -。在分离动物肉源沙门菌的过程中,四硫酸盐肉汤(TT)相比于氯化镁孔雀绿肉汤(RV)更有效。本研究表明,深圳地区动物肉源样品中,沙门菌分离率鸡肉和猪肉高于牛肉和虾肉,血清型以Derby、1,4,[5],12: i: -、Rissen、Agona和8,20: z23,z4: -为主,且多重耐药比例较高。     

Serological characterization of Actinobacillus pleuropneumoniae serotype 2 strains by using polyclonal and monoclonal antibodies

The antigenic differences between strains of serotype 2 of both biotypes I and II of Actinobacillus pleuropneumoniae were studied by using several serological techniques. Monoclonal antibodies (MAbs) against A. pleuropneumoniae biotype I serotype 2 were produced by fusion of spleen cells of BALb/c mice immunized with whole-cell (WC) suspension with SP2/O-Ag14 murine myeloma cells. Desirable MAbs were selected by enzyme-linked immunosorbent assay (ELISA) using WC as antigen. MAbs MK-7 and MK-10 identified multiple bands of lipopolysaccharide in Western-blot. Treatment of WC with proteinase K and sodium periodate indicated that both MAb binding sites were carbohydrates in nature. In both ELISA and Western-blot, MAbs MK-7 and MK-10 recognized only biotype I serotype 2 isolates. Neither MAb MK-7 nor MK-10 reacted with reference strains of remaining serotypes of A. pleuropneumoniae and other Gram-negative bacteria tested. The results obtained with various serological tests showed that strains of serotype 2 biotype I shared antigenic determinants with strain N-282 of serotype 2 biotype II, but not with strain N-273 of serotype 1 biotype II. It is suggested that data obtained from this study may be helpful in the development of specific serotyping and serodiagnostic reagents of A. pleuropneumoniae strains.     

Detection of Rhodococcus equi by polymerase chain reaction using species-specific nonproprietary primers.

Species-specific primers for the polymerase chain reaction (PCR) for the detection of Rhodococcus equi were developed. These primers were based on unique DNA fragments produced from R. equi reference strains and field isolates. Following random amplification of polymorphic DNA from R. equi and R. rhodochrous with a set of 40 arbitrary 10-base pair (bp) primers, a pair of species-specific primers was designed to detect a unique 700-bp fragment of R. equi chromosomal DNA. This PCR product was limited to R. equi and was not detectable in other Rhodococcus species or in a panel of additional gram-positive and gram-negative bacteria.     

Identification of monoclonal antibodies that cross-react with cytokines from different animal species

Eleven monoclonal antibodies specific for ovine, bovine and human cytokines were investigated by flow cytometry for cross-reactivities with cytokines produced by peripheral blood mononuclear cells (PBMCs) from sheep, cattle, goat, swine, horse, dog, mink, rabbit and human. Four antibodies specific for IL-4, IL-8, IFN-gamma and TNF-alpha cross-reacted with cytokines from a majority of the species investigated. These antibodies can be applied to flow cytometric studies of cytokine production by PBMCs from several veterinary species. Another five antibodies specific for IL-2, IL-6, GM-CSF and IFN-gamma (two antibodies) cross-reacted weakly and with a variable number of animal species. These antibodies could in certain situations be useful in flow cytometry. In a number of cases the immunological cross-reactivities were confirmed by Western blot analyses. Overall, the results of this study will remedy some of the lack of species-specific anti-cytokine antibodies in veterinary research.     

Evaluation of colorimetric pH strips for wild animal meat using image analysis

Colorimetric pH strips (MColorpHast™) are very convenient to use, but the decision of pH is based on an individual's color perception and is, therefore, subjective. We developed a pH calculation program for the image of coloring strips on CIE1976 (L*a*b* color space), which slightly underestimated human judgment as the color of pH darker. This image analysis and three individuals' judgments were used for evaluating the strip's features for various qualities of meat from wild animals, and the results were compared with the assessments based on potentiometric pH. In both methods, dipping the strips in distilled water just before use improved the regression coefficient compared with that mentioned in the manual. The image analysis showed higher correlation than human judgments but slightly underestimate pH by a maximum of 0.13 unit from the regression line of the potentiometric pH. In addition, the image analysis revealed meat pigment changed pH higher on the color scale in the lower meat pH region. The strips must be used according to the manual, but dipping is effective when the meat surface is dry, and keeping the strips from touching the meat drip is important in lower pH region because the pigment affects the color of pH.     

Serological diagnosis of Echinococcus granulosus infection in sheep using cyst fluid antigen processed by antibody affinity chromatography

Serum antibody responses in sheep naturally or experimentally infected with Echinococcus granulosus and/or other larval cestodes were examined using an enzyme-linked immunosorbent assay (ELISA) with various antigens prepared from sheep hydatid cyst fluid ( SHCF ). Serum donors included: sheep experimentally infected with E. granulosus and their age-matched non-infected controls; sheep experimentally infected with other helminth parasites; sheep naturally infected with E. granulosus both from Tasmania and the Australian mainland; sheep from Tasmania naturally infected with larval cestodes other than E. granulosus; and naturally reared sheep completely free from infection with larval cestodes. Attempts were made to eliminate serological reactions which were not specific for E. granulosus by using a series of antibody affinity chromatography steps to deplete crude SHCF antigen; these included adsorption with a monoclonal antibody, 3EgH 29-2, removal of host IgG using rabbit anti-sheep IgG antibody, and removal of antigens which bound non-specifically to normal sheep immunoglobulin. The final affinity-depleted antigen product was designated AD SHCF . Specific serological reactivity in infected sheep was very low. Affinity depletion of SHCF using 3EgH 29-2 did not appear to increase the specificity of serological diagnosis of E. granulosus infection when experimentally infected sheep were compared with their non-infected controls provided the latter were age-matched with experimental animals. The other affinity adsorption steps significantly reduced non-specific background binding to antigen by normal sheep serum. Despite this reduction in background in the ELISA, only low levels of antibody could be detected in naturally-infected sheep.(ABSTRACT TRUNCATED AT 250 WORDS)     

Serological classification of Japanese isolates of Haemophilus paragallinarum using two serovar-specific monoclonal antibodies

A serological classification of 106 Japanese isolates of Haemophilus paragalinarum recovered from 1960 to 1984 was performed by dot-blotting and hemagglutination-inhibition (HI) tests using two serovar-specific monoclonal antibodies (MAbs), E5C12D10 and F2E6. By the dot-blotting test, 49 of the isolates were serovar A and 55 isolates were serovar C, and the two remaining isolates did not react with either MAb. These two nontypable strains had no hemagglutinating activity against chicken erythrocytes and were nonpathogenic to chickens. Although 49 serovar A isolates were serotyped by the HI test, only 23 of the 55 serovar C isolates could be serotyped. The remaining 32 isolates could not be serotyped because no or low hemagglutinating activity could be detected. Our results indicate that H. paragallinarum serovars A and C have both been present in Japan since 1960, with serovar A isolates being dominant before 1970 and serovar C isolates more prevalent than serovar A since 1970.     

An immunodiffusion method for the identification of the species of origin of meat samples

SUMMARY The agar gel diffusion method employing suitable antiserums has been used as a convenient method for identifying species of origin of meat samples. The method can detect contamination of one species by another species at a level of between 5 to 20% depending on the sensitivity of the particular antiserum used and the species being tested, and provides a result within 24 hours.     

High-performance liquid chromatography determination of erythrocyte membrane phospholipid composition in several animal species

High-performance liquid chromatography (HPLC) was used to determine the phospholipid (PL) composition of ovine, equine, bovine, porcine, and canine RBC membranes. Procedural modifications of established techniques provided for separation of 7 PL within a 15- to 20-minute sample run. Significant (P less than 0.05) differences were detected in RBC membrane PL composition among the various species. The concern for physiologic properties associated with hemolysis and/or sedimentation rate must include evaluation of differences in the PL bilayer structure.     

Molecular identification of Eimeria species infecting market-age meat chickens in commercial flocks in Ontario

A previously described multiplex PCR was evaluated for the identification and prevalence of Eimeria species in market-age commercial chicken flocks in Ontario. The multiplex PCR based on species-specific RAPD-SCAR markers was able to distinguish six available laboratory strains of Eimeria species (E. tenella, E. maxima, E. necatrix, E. mitis, E. acervulina, and E. brunetti) and E. tenella, E. maxima and E. acervulina in unknown field samples, including multiple infections in single reactions. No backyard (0/77) and 20/360 market-age commercial chickens were oocyst-positive using standard fecal flotation methods. PCR identified E. tenella alone (9/360, 2.5%), E. maxima alone (5/360, 1.38%), E. maxima plus E. tenella (5/360, 1.38%) and E. acervulina alone (1/360, 0.27%) in market-age commercial broilers. This is probably the first time the multiplex PCR has been evaluated in poultry establishments in Canada and illustrates the value of the tool in coccidiosis epidemiology on commercial farms.