298 Abstracts / Veterinary Immunology and Immunopathology 128 (2009) 211–347 latory effects on innate immune responses, particularly neutrophil recruitment, after saponin inoculation. Further- more, we observed a delay in cell infiltration in the Sv group, which may reflect the immunomodulatory properties of saliva. Even though, edematous reactions have frequently been observed after saponin injection or in combination with WPA, the induction of a strong cellular response, sim- ple formulation, safety, and the low costs, allow its use as alternative adjuvant in veterinary medicine. doi:10.1016/j.vetimm.2008.10.185 Selection of recombinant peptides for canine leishma- niasis detection by phage display Juliana Franco Almeida * , Guilherme Rocha Lino Souza, Fausto Emíllio Capparelli, Carlos Roberto Prudêncio, Ana Paula Peres Freschi, Rone Cardoso, Flávia Figueira Messias, Luiz Ricardo Goulart Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, MG, Brazil Keywords: Canine leishmaniasis; Epitopes; Phage display E-mail address: almeidajf@hotmail.com (J.F. Almeida). Species: Canine Human visceral leishmaniasis (HVL) is a canine zoonose leading to 500,000 new cases/year, in which no vaccine is available, and chemotherapy is highly toxic. Canine leishmaniasis (CL) is a systemic disease caused by differ- ent Species of the genus Leishmania that is transmitted by blood sucking phlebotomine sandflies. Dogs are con- sidered the main domestic reservoir of the parasite, constituting part of the epidemiological cycle of human transmission, accounting for more than 90% of visceral leishmaniasis cases described in the world. Phage dis- play has been utilized for numerous purposes, including mapping protein–ligand interactions, identifying binding antagonists and enzyme inhibitors through the design of mimotopes. This technology was used to identify pep- tides against HVL antibodies expanding its purpose to CL epitopes screening in the perspective of using them as vac- cine or diagnostics’ antigens. IgG was purified from sera of human visceral leishmaniasis patients, and used for screen- ing against a random peptide phage library. The selection was performed in liquid phase through agarose beads (G protein agarose). After four rounds of biopanning, phage clones were isolated, sequenced, translated, and submitted to bioinformatic analyses. Peptide analysis revealed a high degree of sequence similarities between important para- site proteins involved in the disease development, such as gp46, gp63, kinesin K39, DNA topoisomerase II, LACK antigen, among others. Two highly frequent epitopes were identified, and the phagotopes were used in ELISA assays against positive and negative canine sera, showing differ- ential reactivity, with 83% of sensitivity. The highly reactive clones may be useful in diagnostics and therapeutics of canine leishmaniases. Financial support: FINEP, CNPq, UFU, DiaMed and ImunoScan. doi:10.1016/j.vetimm.2008.10.186 Detection of anti-Leishmania chagasi IgG subclass anti- body in infected and vaccinated dogs Trícia Maria Ferreira De Sousa Oliveira * , Melina Bason, Tiago Wilson Patriarca Mineo, Rosangela Zacarias Machado Universidade Estadual Paulista (UNESP), Jaboticabal, SP, Brazil Keywords: Leishmania; Subclass; IgG antibodies; ELISA E-mail address: triciaoliveira@yahoo.com.br (T.M.F. De Sousa Oliveira). Species: Canine Visceral leishmaniasis is a zoonotic disease, caused by Leishmania chagasi and transmitted by the bite of sandflies from Lutzomiya longipalpis species in Brazil. In this coun- try, it is an emerging disease in urban areas, and dogs are the main domestic reservoir. The aim of this study was to analyze the presence of anti-Leishmania IgG subclass anti- bodies by ELISA in 121 sera of seropositive dogs from Belo Horizonte (MG) endemic area, and 40 sera of vaccinated animals with vaccine Leishmune (Fort Dodge ® ). This study also aims to verify the difference of IgG subclass among the groups. The proportion of dogs witch produced detectable levels of anti-Leishmania IgG subclass antibodies from Belo Horizonte was 80,17% (97/121) for IgG1; 0.83% (1/121) for IgG2; 39.67% (48/121) for IgG3 and 55.37% (67/121) for IgG4. The results showed significant higher percentage in IgG1 antibodies (p < 0.01). The proportion of vaccinated dogs which produced detectable levels of anti-Leishmania IgG subclass antibodies was 22% (9/40) for IgG1; 87.5% (35/40) for IgG2; 2.50% (1/40) for IgG3 and 5% (2/40) for IgG4. Result analysis disclosed significant increase in IgG2 antibodies (p < 0.01) in vaccinated dogs, and that the IgG2 response of vaccinated dogs was significantly higher than in infected dog (p < 0.01). These results suggest that there was great variation in levels of IgG1 and IgG2 between the two groups, but it needs more studies to use the markers of resistance or susceptibility of individual dogs (25.4 mm). doi:10.1016/j.vetimm.2008.10.187 Comparison between antigen total ELISA and FML ELISA in diagnosis of visceral canine leishmaniasis Teresinha Cristina Cândido * , Tatiana de Oliveira Gerzoschkwitz, Maria Cecília Rui Luvizotto, Valéria Marc ¸ al Felix de Lima Departamento de Apoio Produc ¸ ão e Saude Animal, Faculdade de Odontologia de Arac ¸ atuba, Universidade Estadual Paulista Júlio de Mesquita Filho, Brazil Keywords: Diagnosis; ELISA; Visceral leishmaniasis Species: Canine Visceral canine leishmaniasis or calazar is Brazilian an endemic antropozoonosis caused by Leishmania L. chagasi.