Profiles of serological reactivity against cytosoluble proteins of brucella melitensis in infected and vaccinated sheep

Sera from vaccinated lambs with Brucella melitensis Rev. 1 and naturally infected sheep with Brucella melitensis biovar 3 as well as Brucella free sheep, were examined using standard serological tests.No great difference in standard serological tests results between vaccinated and infected animals. These sera were also analyzed for their serological reactivity against whole killed cell [WKC] and cytosoluble protein extract [CPE] antigens using ELISA.The present study showed that ELISA with WKC was unable to differentiate infected sheep from those vaccinated with Rev. I while ELISA with -cytosoluble proteins antigen may able to differentiate antibody response of infected animals from vaccinated ones.Using immunoblot technique, sera from naturally infected animals showed strong antibody reactivity to 28.48KDa and variable reactions to 58.80, 49.70, 39.60, 32.47 and 18.00 KDa. However sera from Rev.1 vaccinated animals showed less intense antibody reactivity which only observed against proteins with molecular masses of 49.70 and 39.60 KDa. It is likely that cytosoluble proteins may provide useful serological reagent for differentiation between infected and vaccinated sheep

Using of brucella cell envelop as an antigen in diagnosis of ovine brucellosis

In this study serum samples from 35 Brucella culture positive and 80 Brucella culture negative sheep were used to evaluate the use Whole Cell Sonicate [WCS] and Cell Envelop [CE] antigens prepared from Brueclla melitensis type 3 [field strain] for ELISA. These samples were also tested using Rose Bengal test [RBT], Buffered acidified plate test [BAPT], Rivanol test [Riv. T.] and Tube agglutination test [TAT]. The results showed that 35 [100%] of the examined sera from the culture positive group were positive in BAPA, RBT and TAT tests while 34 [97.14%] were positive for Riv. T. However, from 80 animal from the culture negative group, 7 [8.75%], 8 [10%], 5 [6.25%], and 8 [10%] were positive for RBT, BAPA, Riv. T and TAT respectively. The serum samples of these animals were then tested by indirect ELISA using WCS and CE antigens. From the 35 culture positive animals 35 [100%] showed positive results, while, from the 80 culture negative animals 5 [6.25%] and 2 [2.5%] were positive in WCS and CE ELISA respectively. The sensitivity of ELISA with both WCS and CE antigens was 100%. It is clear that WCS and CE detect all culture positive sheep. The specificity of ELISA for WSC was 93.75% and for CE was 97.5%. In conclusion CE seems to be effective for diagnosis of Brucella melitensis infection in sheep by ELISA

Serological tests and biochemical profiles in camels infected with brucellosis

Blood samples were collected from 80 camels kept in closed farm [Camel production unit Animal production Institute and others], another 72 blood samples were collected from camels kept in close contact with cattle and other small ruminant from different areas in Gize governorate. As well as 94 blood samples were collected from imported camel from Sudan in camel market. Our data showed that the highest percent of positive reactors was observed in the imported camels [Sudanese camel] in large herd [8.50 to 11.70% [9.50%, 10.60%. 9.50%, 8.51%. 9.57% and 11.70 to 8.50% for RBPT, BAPA, Riv, SAT, MET and DI/ respectively], Camels in contact with other animals [6.94 to 11.10% [8.30%, 9.40%, 8.30%, 6.94%, 8.33% and 8.30 to 11.10% for RBPT, BAPA, Riv, SAT, MET and DIA, respectively] and Camels in closed farms [0.00 to 2.50% [1.25%, 2.50%, 0.00%, 1.25%, 0.00% and 1.25 to 5.00% for RBPT. BAPA, Riv, SAT, MET and DIA, respectively]. The results of sensitivity and specificity of DIA revealed that DIA using n-lauroylsarcosin extract is more specific than DIA whole bacterial antigen. The sera of infected camels with brucclla [either camels contact with animals or imported camels] showed elevated levels in each of the GGT, LDH, ALP, AST, ALT, total protein, albumin, glucose, urea, uric acid and creatinine. The sera of imported camels infected with brucellosis were characterized by increased levels of protein bands with molecular weights 29.83 -30.11, 45.95-46.27 kDa, with increase of 34.64, 35.29, 74.67, 87.74, 98.96, 99.75, 104.62, 110.57, 115.54, 132.63, 134.12, 138.69, 140.25 kDa protein bands in both camels contact with animals and imported camels infected with brucellosis. Protein bands 181.3,1-183.34 and 214.36 KDa were apparent in camels contact with animal's sera infected with brucellosis and protein bands 189.59 and 231.79 were present in imported camel's sera infected with brucellosis especially in 1/320 antibody sera. The LDH and ALP iso-enzymes had a characteristic profile in brucellosis. Our conclusions that imported camels infected brucellosis followed by camels contact with animals infected with brucellosis had more serious biochemical discordance. The results give us an index to diagnosis of brucellosis in the imported camel. The incidence percent of brucellosis in camels in closed farm in dedicate importance, good prognosis mean and diagnosis tools to detect and eliminate the infected camels, and aid in epidemiological controls of the disease

Monitoring of immune response against brucella melitensis Rev- 1 vaccine following administration of some antibiotics [with 4 tables and 4 figures]

The humoral and cellular immune response in guinea pigs vaccinated with Rev-1 vaccine and simultaneously treated with antibiotics [oxytetracycline, streptomycin or enrofloxacin] or 4 weeks post-vaccination were evaluated in the present study. Groups vaccinated and simultaneously treated with oxytetracycline or 4 weeks post-vaccination showed delayed or abrupt reduction respectively in antibody production, lymphocyte transformation, phagocytic percent and intra-cellular killing one week post administration of this antibiotic while groups vaccinated and simultaneously treated with streptomycin or 4 weeks postúvaccination showed abrupt reduction in antibody titer and moderate reduction in phagocytic percent, intra-cellular killing and lymphocyte transformation. Treatment by enrofloxacin simultaneously with Brucella melitensis Rev-1 vaccine or at peak of vaccination [4 weeks postúvaccination] cause non-significant variation in both humoral and cellular immune response. Therefore oxytetracycline and streptomycin should not be recommended for animal treatment during exposure to Brucella melitensis Rev-1 vaccine. In contrast enrofloxacin exhibited a satisfactory margin of safety upon the humoral and cellular immune responses to Brucella melitensis Rev-1 vaccine

Characterization of campylobacter isolates from sheep with particular reference to enterotoxin production

C. jejuni subsp. jejuni, C. coli and C. laridis were isolated from apparently normal and diarrhoeic sheep with an incidence of 10% and 20%, respectively. C jejuni subsp. jejuni was the most predominant species with an overall incidence of 10.7%. All Campylobacter strains isolated from diarrhoeic animals were invasive to Hela cells and their cytotoxin produce degree [++] and [+++] of changes in Vero cells and titer of toxins were high [64-128]. C jejuni enterotoxin isolated from diarrhoeic sheep gave maximum secretion in rat ligated ileal loops. While enterotoxin of other strains evoked less fluid secretion in rat ligated ileal loops. SDS PAGE of heat labile enterotoxin extracted from isolated strains indicated protein profiles ranging from 49.3 to 59.3 KD. In immunized-challenged rats the fluid secretion was reduced by 70 to 80% less than the value for similar challenged unimmunized control rats. Antibody titers measured by ELISA showed four folds higher level than that in control animals