Antibody isotype profiles in serum and circulating antibody-secreting cells following mucosal and peripheral immunisations of sheep

The isotype-specific antibody responses of sheep immunised with keyhole limpet hemocyanin by a peripheral route (intramuscular (i.m.) injection) were compared to those induced by immunisation via different mucosal routes: (1) intra-nasal spray; (2) rectal deposition with cholera toxin; (3) injection into the mucosa of the small intestine or rectum. Antigen-specific IgG1 antibodies were induced in the i.m., intra-intestinal and intra-rectal injection groups and in a proportion of the cholera toxin immunised sheep, but not in the intra-nasal immunisation group. IgA was the only antibody isotype detected in serum collected from the intra-nasal immunisation group. No significant differences in serum IgA levels were detected in any of the mucosal immunisation groups as compared to the i.m. injection group. In contrast, analysis of the in vitro antibody profiles secreted by circulating antibody-secreting cells (ASC) revealed significantly higher IgA responses in the supernatants from all mucosal immunisation groups. This suggests that the measurement of antibodies secreted by circulating ASCs may be a better correlate of local mucosal responses in ruminants, as has been previously demonstrated in human studies. In addition to IgG1 and IgA responses, immunisation by direct injection of antigen formulations into the intestinal and rectal mucosa were the only groups to induce consistently high IgG2 antibodies in serum and ASC cultures.     

Association of genes encoding beta2 toxin and a collagen binding protein in Clostridium perfringens isolates of porcine origin

Clostridium perfringens is a cause of economically significant enteritis in livestock. Beta2 toxin, encoded by one of two cpb2 alleles, is implicated as a virulence factor in this disease. Previous studies determined that the consensus cpb2 allele is preferentially associated with C. perfringens isolated from pigs. In C. perfringens strain 13, the consensus cpb2 allele is found on the plasmid pCP13, which also carries cna, encoding a putative collagen binding protein, CpCna. This protein was shown to be a bona fide collagen adhesin, as recombinant, HIS-tagged CpCna bound collagen type I as determined by Far Western blotting. Genomic DNA from C. perfringens isolated from a variety of host species were subjected to PCR to determine the prevalence of cna in these strains and correlate its carriage with the presence and type of cpb2 allele. The cna gene was found in 55.8% of isolates from all host species (n=208) and 68.1% of porcine isolates (n=119). In cpb2+ isolates, cna was present in 69.9% of isolates from all hosts (n=153), but was found in 98.7% of porcine isolates (n=75). Furthermore in porcine isolates, the consensus cpb2 allele and cna were absolutely correlated with the presence of pcp12, a pCP13-encoded gene, and pcp12 was never found in any isolate that lacks either cpb2 allele. The finding that CpCna binds collagen and that the cna gene is associated with the consensus cpb2 allele implicates CpCna as a potential virulence factor in porcine enteritis caused by C. perfringens.     

Pemphigus foliaceus in 91 dogs

A retrospective study of 91 dogs with pemphigus foliaceus was performed. Clinical signs of the disease included crusts (n=79), pustules (n=36), and alopecia (n=33). Lesions were most common on the trunk (n=53), inner pinnae (n=46), face (n=37), and foot pads (n=32). Cytological evaluation revealed acantholytic keratinocytes in 37 of 48 dogs. Results of combination treatment with prednisolone and azathioprine were comparable to results with prednisolone therapy alone. More than half of the dogs achieved remission with appropriate therapy, and another 25% significantly improved.     

Polar constituents from the aerial parts of Origanum vulgare L. Ssp. hirtum growing wild in Greece

From the polar extracts of Origanum vulgare L. ssp. hirtum 19 compounds have been isolated. The structures and relative stereochemistry have been elucidated by spectroscopic analysis and determined as apigenin, luteolin, chrysoeriol, diosmetin, quercetin, eriodictyol, cosmoside, vicenin-2, caffeic acid, p-menth-3-ene-1,2-diol 1-O-beta-glucopyranoside, thymoquinol 2-O-beta-glucopyranoside, thymoquinol 5-O-beta-glucopyranoside, thymoquinol 2,5-O-beta-diglucopyranoside, 12-hydroxyjasmonic acid, 12-hydroxyjasmonic acid 12-O-beta-glucopyranoside, lithospermic acid B, rosmarinic acid, 10-epi-lithospermic acid, and epi-lithospermic acid B. The three latter products display unusual stereochemistry of the 3,4-hydroxyphenyllactic acid unit(s), which to the authors' best knowledge has never been reported before in similar compounds. Moreover, lithospermic acid B (and its stereoisomers), p-menth-3-ene-1,2-diol 1-O-beta-glucopyranoside, 12-hydroxyjasmonic acid, and 12-hydroxyjasmonic acid 12-O-beta-glucopyranoside were isolated for the first time from Origanum species.     

Serological status of Canadian cattle for brucellosis,anaplasmosis, and bluetongue in 2007–2008

A national bovine serological survey was conducted to confirm that the prevalence of brucellosis, bluetongue, and anaplasmosis does not exceed 0.02% (95% confidence) in live cattle in Canada. Sampling consisted of a systematic random sample of 15 482 adult cattle slaughtered in federally inspected abattoirs, stratified by province. Samples were tested to detect antibodies for brucellosis, bluetongue, and anaplasmosis. All samples were negative for brucellosis. Three samples were seroreactors to bluetongue, 2 of which originated from the Okanagan Valley in British Columbia and 1 from Ontario, which after follow-up, was considered an atypical result. A total of 244 samples were seroreactors to Anaplasma and follow-up identified infection in Saskatchewan, Manitoba, and Quebec. In conclusion, the Canadian cattle population remains free of brucellosis and free of bluetongue outside the Okanagan Valley. Canada is no longer free of anaplasmosis and will be unable to claim freedom until eradication measures are completed.     

Compatibility between <Emphasis Type="Italic">Calpurnia aurea</Emphasis> leaf extract,attraction aggregation,and attachment pheromone and entomopathogenic fungus <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> on viability,growth, and virulence of the pathogen

Metarhizium anisopliae sensu stricto (ss) (Metsch.) Sorok. isolate ICIPE 07 is being developed as biopesticide for the control of ticks. In addition, leaf extracts of Calpurnia aurea Benth, and the attraction aggregation and attachment pheromone (AAAP) are being used as ticks’ attractant. The three agents are being considered for use in combination in an autodissemination approach, whereby ticks that are attracted to semiochemicals are infected with the inoculum. Experiments were therefore conducted to evaluate in vitro the compatibility between C. aurea, AAAP, and the M. anisopliae on vegetative growth, conidial production, and spore viability. Calpurnia aurea leaf extract was compatible with the fungus at all the concentrations tested, whereas AAAP inhibited all the fungal growth parameters. The virulence of M. anisopliae formulated in emulsifiable extracts of C. aurea was also tested against different developmental stages of Rhipicephalus appendiculatus in laboratory bioassays. No significant differences in virulence were observed between M. anisopliae applied alone and M. anisopliae formulated in different concentrations of C. aurea leaf extracts. These results suggest that C. aurea leaf extracts is compatible with M. anisopliae and could be mixed together for “spot-spray” treatments as low-cost and environmental-friendly technology to control ticks in grazing field, while AAAP should be used separately.