Detection of Anti-Neospora caninum Antibodies on Dairy Cattle Farms in Southern Italy.

Neosporosis is recognized as one of the major causes of bovine abortion worldwide. Canids are the main definitive host for this parasite and the presence of dogs in the farm is an important factor for the Neospora caninum infection in bovines. Since, in the province of Lecce, located in the Apulia region of Southern Italy, there are no studies showing the presence of the infection in farm animals, the objective was to perform a serological evaluation for anti-N. caninum antibodiesin serum from 706 dairy cattle and 21 farm dogs located in 40 farms uniformlydistributed over the territory.The presence of N. caninum infection was confirmed in 90.0% (36/40) of the 40 farms examined. The results obtained on all serum samples by an enzyme-linked immunosorbent assay (ID Screen®Neospora caninum competition ELISA kit) for anti-N. caninum antibodies showed a seropositivity rate of 21.1% (149/706) among dairy cows, with a statistically significant higher percentage of positive subjects in the animals over two years old and a positivity rate of 42.9% (9/21) in tested dogs. The obtained data confirmed the presence of neosporosis even in the Lecce area, where it could therefore represent an important cause of abortion and economic losses.

Evaluation of a Real-time PCR in Combination with a Cultivation Method for the Detection of Brucella Abortus in Organs of Infected Cattle in Southern Italy.

Introduction: Brucellosis is a widespread zoonosis of great economic importance for livestock farming in many areas of the world. It is a highly infectious disease which is diagnosed using conventional serological and microbiological methods. The aim of this study was to assess the efficiency of a specific real-time PCR in combination with broth cultivation in detecting Brucella spp. in organs of infected cattle, in order to compare the sensitivity of the two approaches and the time needed in them until a correct diagnosis is made. Material and Methods: We examined 67 organs collected from 10 cattle slaughtered following a brucellosis outbreak which occurred in February 2016 in southern Italy. The research was carried out by enrichment broth cultivations in combination with a real-time PCR every week for six weeks. Results: Brucella strains were isolated by cultivation from 44 enrichment broths of organs. All isolates were later identified as Brucella abortus by real-time PCR. Using this method in combination with cultivation made it possible to identify the same percentage of infected animals faster than by cultivation alone. Moreover, the same diagnostic results were obtained, on average two weeks before they would have been using only cultivation. In almost all cases, Brucella was detected by real-time PCR after the first week of cultivation in pre-enrichment Brucella broth, while the bacterial growth was evident usually after 2 or 3 weeks. Conclusion: Real-time PCR has allowed results to be obtained faster than in the classical microbiological method, reducing the response times to identify positive animals by half.

Serological surveillance of Leptospirosis in Italy: two­year national data (2010­2011).

Nowadays, leptospirosis is a re­emerging widespread infectious disease often underestimate worldwide. The National Reference Centre for Leptospirosis (NRCL), at the Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna, Brescia (Italy), with the cooperation of all the other Istituti Zooprofilattici Sperimentali (IIZZSS), evaluated the distribution of such important zoonosis in Italy. Serological data obtained between 2010­2011 by each laboratory were collected by the NRCL and discussed. Serum samples collected from 43,935 animal specimens were analysed by the Microscopic Agglutination Test (MAT), using a panel of 8 serogroups as antigens (Australis, Ballum, Canicola, Grippotyphosa, Icterohaemorrhagiae, Pomona, Sejroe, Tarassovi). A MAT cut­off of 1:100 was used to identify the serological positivities, 6,279 sera showed positive titers. Bovine (46.9%), swine (27.5%), ovine and goat (7.4%), dog (6.9%), and wild boar (4.5%) samples were delivered to the Laboratories more frequently than equine and other species sera. Data analysis showed that the most common serogroups in Italy are: Australis present in dogs, wild boars, horses, hares, swine, foxes, and rodents; Sejroe detected in cattle, sheep, goats, and buffaloes; Icterohaemorrhagiae present in dogs, goats, and foxes; Pomona detected in swine, cattle, and wild species; Grippotyphosa reported in hares.

Serological diagnosis of bovine brucellosis using B. melitensis strain B115.

Bovine brucellosis is diagnosed by official tests, such as Rose Bengal plate test (RBPT) and Complement Fixation test (CFT). Both tests detect antibodies directed against the lipolysaccharide (LPS) of Brucella cell wall. Despite their good sensitivity, those tests do not discriminate between true positive and false positive serological reactions (FPSR), the latter being generated by animals infected with other Gram negative microorganisms that share components of Brucella LPS. In this study, an antigenic extract from whole Brucella melitensis B115 strain was used to set up an ELISA assay for the serological diagnosis of bovine brucellosis. A total of 148 serum samples from five different groups of animals were tested: Group A: 28 samples from two calves experimentally infected with Yersinia enterocolitica O:9; Group B: 30 samples from bovines infected with Brucella abortus; Group C: 50 samples from brucellosis-free herds; Group D: 20 samples RBPT positive and CFT negative; Group E: 20 samples both RBPT and CFT positive. Group D and Group E serum samples were from brucellosis-free herds. Positive reactions were detected only by RBPT and CFT in calves immunized with Y. enterocolitica O:9. Sera from Group B animals tested positive also in the ELISA assay, whereas sera from the remaining groups were all negative. The results obtained encourage the use of the ELISA assay to implement the serological diagnosis of brucellosis.