Alexey Leon Gomel Bogado

Humoral and cellular immune responses in pigs immunized intranasally with crude rhoptry proteins of Toxoplasma gondii plus Quil-A.

We evaluated the humoral and cellular immune responses in pigs immunized intranasally with crude rhoptry proteins of Toxoplasma gondii plus Quil-A. The experiment used 13 mixed-breed pigs divided into the following three groups: G1 (vaccinated-challenged, n=6), which received the rhoptry vaccine (200(g/dose); G2 (adjuvant-challenged, n=4), which received PBS plus Quil-A; and G3 (unvaccinated-challenged, n=3), which was the control group. The treatments were performed intranasally at days 0, 21, and 42. Three pigs from G1 produced IgG and IgM antibody levels above the cut-off in the ELISA on the challenge day. Partial protection was observed in G1 at the chronic phase of infection when compared with G3. The preventable fractions were 41.6% and 6.5%, in G1 and G2, respectively. The results of this study suggest that rhoptry proteins plus Quil-A stimulated humoral, local, and systemic immune responses, which were able to partially protect the brain from cyst formation.

Eimeria tenella: utilization of a nasal vaccine with sporozoite antigens incorporated into Iscom as protection for broiler breeders against a homologous challenge.

The aim of this study was to evaluate a nasal vaccine using antigens derived from sporozoites of Eimeria tenella incorporated into Iscom to protect broiler chicks. Forty-five one-day-old chickens (Cobb), unvaccinated against coccidiosis, were used in this experiment. The birds were maintained in separated battery cages and divided into three groups: G1 (n=15), G2 (n=15), and G3 (n=15). G1 received 50 microg of sporozoites+Iscom vaccine, G2 received Iscom without antigens, and G3 received only PBS. The treatments were administered by nasal route on days 0, 7, and 21 of the experiment. On the 28th day, all birds were challenged with 105 sporulated oocysts of E. tenella. On the challenge day, three birds from each group were euthanized to evaluate lymphocyte proliferation. Lesion scores were obtained from five birds from each group, 7 days after challenge. The remaining animals were euthanized on the 50th day. The mean lymphocyte proliferation responses were significantly different (P=0.03); G1 was 2.3-2.6 times more elevated than G2, and G3 (P<0.001). 83% of the birds from G1 showed an IgY antibody reaction by ELISA at challenge. The means for oocysts shedding were 16,890+/-20,511, 48,080+/-50,047, and 65,020+/-74,461, for G1, G2 and G3 birds, respectively. There was no significant difference (P>0.17) in oocysts shedding between groups. However, the G1 and G2 chicks demonstrated reduction in percentage of oocyst shedding when compared to control birds (G3) by 74.02% and 26.05%, respectively. The average lesion scores were G1=0.4, G2=1, and G3=2. This study demonstrated that the lowest lesion score and oocyst shedding were observed in the birds from the group that received antigens derived from sporozoite with an Iscom adjuvant (G1). These results suggest that this vaccine can induce protection against avian coccidiosis.

Oocyst shedding in cats vaccinated by the nasal and rectal routes with crude rhoptry proteins of Toxoplasma gondii

During this study, cats were immunized by the intranasal and rectal routes with crude rhoptry proteins of Toxoplasma gondii admixed with Quil-A. Twenty-five domestic short hair cats divided into five groups (n=5) were used during this evaluation: G1 and G3 cats received 200 μg of the rhoptry proteins with Quil-A (20 μg) by the intranasal and rectal routes, respectively; G2 and G4 cats received bovine serum albumin (BSA, 200 μg/dose) with Quil-A (20 μg); and G5 animals served as unvaccinated controls. All treatments were performed at days 0, 21, 42, and 63. The challenge was done with 800 cysts of the ME49 of T. gondii strain at day 70 (challenge day). The serum IgG, IgM, IgA, and fecal IgA antibody levels were evaluated by using the indirect enzyme-linked immunosorbent assay (ELISA). Some animals produced antibody levels beyond cut-off; however, two animals from G1 (OD(mean)=0.308, OD(cut-off)=0.200) and three from G3 (OD(mean)=0.254) demonstrated IgG levels on being challenged, with similar results occurring in two cats from G1 to IgM (OD(mean)=0.279, OD(cut-off)=0.200). Fecal IgA levels were detected in all G1 cats (OD(mean)=0.330, OD(cut-off)=0.065), and in one cat from G3 (OD(mean)=0.167). The serum and fecal humoral immune responses did not correlate with oocyst shedding. Oocyst shedding varied from 98.4% (G1), 87.5% (G2), 53.0% (G3), to 58% (G4), and was lower than that of G5 cats. The prepatent period of cats vaccinated intranasally (G1) was reduced from 6-9.6 to 2.8 days, suggesting protection of environmental contamination, considering cats as the primary source of contamination. The intranasally and rectally administered rhoptry vaccines were able to partially protect cats against T. gondii cysts on being challenged; however, the intranasal method of vaccination yielded better results relative to the rectal route.

Anti-Neospora caninum antibody detection and vertical transmission rate in pregnant zebu beef cows (Bos indicus): Neospora caninum in pregnant beef cows (Bos indicus)

The aim of the present study was to evaluate anti-Neospora caninum antibodies and the vertical transmission rate in naturally infected pregnant zebu beef cows (Bos indicus) reared on pasture. The present study began with 200 cows from four farms (50 cows from each farm), and these animals were submitted to timed artificial insemination (TAI). After ultrasonography, 76 pregnant cows were selected, 22, 15, 22, and 17, respectively, from farms 1, 2, 3, and 4. Blood samples were taken from cows thrice during the first, second, and third trimester of gestation, and a blood sample was collected from 31 calves before colostrum milking. From 76 cows 23 (30.3%) had anti-N. caninum antibodies detected by indirect ELISA (Idexx), and 53 (69.7%) did not. Sixty-four cows that initiated the experiment were negative to N. caninum and 11 became positive either during the second or third trimester of gestation, this mean an infection incidence of 17.2% (11/64). OD for ELISA was higher (OD=2.08) during the second and third (OD=2.10) trimesters of pregnancy when compared with the first (OD=1.81), however, there were no statistical differences (P=0.45). The vertical transmission was calculated to be 29.0% (9/31), and the risk of vertical transmission of N. caninum in seropositive dams was 26.25 times higher than seronegative animals (OR=26.25, 2.38<OR<289, P=0.007). In conclusion, the rate of vertical transmission of N. caninum in pregnant zebu beef cows was 29%, and the risk was 26.25 higher in seropositive dams relative to than seronegative animals.

Molecular cloning, sequencing, and expression of Eimeria tenella HSP70 partial gene

Members of the Eimeria genus are protozoan parasites of the subphylum Apicomplexa (Eimeriidae family), and belong to the coccidia group. Eimeria tenella is one of the most pathogenic species owing to its ability to penetrate the mucosa, and cause inflammation and damage. It is an obligate intracellular parasite that causes disease by destroying the host cells during multiplication. Heat shock protein 70 (HSP70) is a molecular chaperone that prevents cellular stress. The objective of this study was to clone, sequence, and express E. tenella HSP70 protein. After selecting the region of highest hydrophilicity in the hsp70 gene, we cloned complementary DNA (cDNA) into a pTrcHis2-TOPO vector and transformed it into TOP10 Escherichia coli cells; after induction, the bacteria expressed a 23-kDa protein with insoluble expression levels of approximately 5 mg/L. In summary, the partial hsp70 gene was successfully expressed in E. coli, producing a 23-kDa protein under insoluble conditions, and the antigen characteristics predicted by hydrophilicity analysis suggest the development of a vaccine for use in avian coccidiosis.

Bovine pulmonary lesions found in post-mortem inspection in slaughterhouse in state Paraná

Polyana Carolina Marino, Gercio Luiz Bonesi, Luiz Carlos Negri Filho, Daniella Furlan, Débora Nayara Augusto, Alexey Leon Gomel Bogado, Rogério Anderson Marcasso, Luiz Cesar da Silva, Marcelo Diniz dos Santos, Werner Okano 1 Docente. Departamento de Clínica Médica e Cirúrgica de Grandes Animais. UNINGA, Maringá, PR. pollyveterinaria@gmail.com 2 Docente. Medicina Veterinária. UNOPAR. Arapongas, PR. gercio.bonesi@agricultura.gov.br; rogeriomarcasso@gmail.com 3 Mestrando do Programa de Pós Graduação em Saúde de Produção de Ruminantes. PROSUP/CAPES. UNOPAR. Arapongas, PR. luiz.negri@hotmail.com; furlandanie@hotmail.com; debora_augusto@outlook.com 4 Docentes do Programa de Pós Graduação em Saúde de Produção de Ruminantes. UNOPAR. Arapongas, PR. alexey_leon@hotmail.com; silvaluizcesar@gmail.com; werner.okano@unopar.br 5 Docente do Programa de Pós Graduação em Biociência Animal. UNIC. Cuiabá, MT. marcelo.diniz@kroton.com.br __________________________________________________________________________________

Liver abscess in cattle slaughter coming from 14 counties of north central mesorregion of Parana

The beef liver has important nutritional properties in food, but can transmit zoonoses important to humans. Service inspection of products of animal origin used to give food security in the consumption of these products. Liver abscesses occur by different etiologies in bovine liver. From the record of liver condemnations of cattle slaughter in abatoir under state inspection, 2,256 cattle slaughtered between August 2011 and March 2014, in Rolandia county from 14 counties of north central mesorregion Parana were detected 8,37% of liver abscesses. All 189 livers containing single or multiple abscesses were condemned by the standard inspection service, causing economic losses resulting decreased feed efficiency, and decreased carcass yield beyond the value of the viscera which is no longer marketed. Keys words: slaughterhouse, liver, Animal Standard Meat Inspection _______________________________ 1 . Acadêmico de Medicina Veterinária / UNOPAR Arapongas/PR. Bolsista PIBIC CNPq 2 . Acadêmico de Medicina Veterinária / UNOPAR Arapongas/PR. IC voluntário 3 . Acadêmico de Medicina Veterinária / UNOPAR Arapongas/PR. Bolsista PIBIT CNPq 4 . Mestrando Programa em Saúde e Produção de Ruminantes / UNOPAR UEL 5 . Docente de Medicina Veterinária / UNOPAR/PR – Arapongas 6 . Docente de Medicina Veterinária / UNOPAR Arapongas / PPG Saúde e Produção de Ruminantes UNOPAR – UEL 7 . Docente de Medicina Veterinária UNIC Cuiabá / PPG Biociência Animal Autor para correspondência * vetwerner@gmail.com Recebido em 30/006/2014. Aprovado em 20/09/2014

Nematode resistance to anthelmintics nitroxynil 34% and 1% ivermectin from sheep flock in São João do Ivaí county, Parana

Resistência de nematoides aos anti-helmínticos nitroxinil 34% e ivermectina 1% em rebanho ovino no município de São João do Ivaí, Paraná Nematode resistance to anthelmintics nitroxynil 34% and 1% ivermectin from sheep flock in São João do Ivaí county, Parana Luiz Eduardo Vila Nova 1 , Marcelo Eduardo Costa 1 , Priscilla Gomes Carneiro Ferreira de Melo 2 , Luiz Fernando Coelho Cunha Filho 3 , Flavio Antonio Barca Junior 3 , Luiz César da Silva 3 , Werner Okano 3 , Alexey Leon Gomel Bogado 3

Hematological parameters of Holstein breed bovines: the influence of by the tuberculin comparative test

Hematological parameters of Holstein breed bovines: the influence of by the tuberculin comparative test Luiz Carlos Negri Filho1, Lucas Vinícius de Alcântara Andrade2, Pedro Henrique Newbery Chineze3, Mauricio Zampronio Affonso 4 , Marcelo Diniz dos Santos 5 , Flávio Antônio Barca Junior 6 , Alexey Leon Gomel Bogado 6 , Luiz Cesar da Silva 7 ,Werner Okano 7* _________________________________________________________________________________________________________________