RESUMO

The objective of this survey was to investigate the prevalence of Hepatozoon infection in dogs in the rural and urban areas of Uberlândia, Brazil by PCR and molecular characterization. DNA was obtained from blood samples collected from 346 local dogs from both genders and various ages. Seventeen PCR products from positive blood samples of urban dogs and 13 from the rural dogs were sequenced. Partial sequences of the 18S rRNA gene indicated that all 30 dogs were infected with Hepatozoon canis similar in sequence to H. canis from southern Europe. Four local dog sequences were submitted to GenBank (accessions JN835188; KF692038; KF692039; KF692040). This study indicates that H. canis is the cause of canine hepatozoonosis in Uberlândia and that infection is similarly widespread in rural and urban dogs.

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RESUMO

The experiment was carried out with 150 Cobb broiler chickens divided into 3 groups with 50 birds each. The groups of infected chickens orally received 1ml of inoculum containing 3x103 Eimeria acervulina sporulated oocysts at 12 days of age. Group 1 was kept as a positive control with infected non-medicated birds, group 2 was medicated with diclazuril (1%) with a dose of 1mL/4 L of drinking water for 2 successive days, 5 days after infection, while group 3 was kept as negative control with non-infected and non-medicated birds. Oocysts count per gram of feces, score of macroscopic intestinal lesions and weight gain were evaluated. The group treated with diclazuril showed significant and satisfactory improvement in the assessment criteria when compared to the infected non-medicated group. The results revealed more reduction in the total oocyst count and intestinal lesion score in the medicated than in the infected non-treated group. The results confirmed that (1%) liquid diclazuril is effective to control Eimeria infection.

O experimento foi realizado com 150 frangos de corte da linhagem Cobb, divididos em 3 grupos de 50 aves cada. Os frangos de dois grupos (G1 e G2) receberam, por via oral, 1mL de inóculo contendo 3x103 oocistos de Eimeria acervulina aos 12 dias de idade. O grupo 1 foi mantido como grupo controle positivo (infectado não medicado), o grupo 2 foi medicado com diclazuril (1%) na dose de 1mL/4 litros de água potável por dois dias sucessivos, cinco dias após a infecção. Um terceiro grupo (G3) foi mantido como controle negativo não infectado e não medicado. O número de oocistos por grama de fezes (OOPG), o escore de lesões macroscópicas intestinais e o ganho de peso corporal foram avaliados. No G1, o OOPG foi de 3,67x106 e o peso final foi de 1,392kg, valores significativamente diferentes do OOPG de 1,93x106 oocistos e do peso final de 1,613kg do G2 (diclazuril). Os resultados provaram que o diclazuril (1%) em formulação líquida e na dosagem de 1mL/4 litros de água por dois dias sucessivos foi eficaz no controle de Eimeria acervulina.

Assuntos

RESUMO

The experiment was carried out with 150 Cobb broiler chickens divided into 3 groups with 50 birds each. The groups of infected chickens orally received 1ml of inoculum containing 3x103 Eimeria acervulina sporulated oocysts at 12 days of age. Group 1 was kept as a positive control with infected non-medicated birds, group 2 was medicated with diclazuril (1%) with a dose of 1mL/4 L of drinking water for 2 successive days, 5 days after infection, while group 3 was kept as negative control with non-infected and non-medicated birds. Oocysts count per gram of feces, score of macroscopic intestinal lesions and weight gain were evaluated. The group treated with diclazuril showed significant and satisfactory improvement in the assessment criteria when compared to the infected non-medicated group. The results revealed more reduction in the total oocyst count and intestinal lesion score in the medicated than in the infected non-treated group. The results confirmed that (1%) liquid diclazuril is effective to control Eimeria infection.(AU)

O experimento foi realizado com 150 frangos de corte da linhagem Cobb, divididos em 3 grupos de 50 aves cada. Os frangos de dois grupos (G1 e G2) receberam, por via oral, 1mL de inóculo contendo 3x103 oocistos de Eimeria acervulina aos 12 dias de idade. O grupo 1 foi mantido como grupo controle positivo (infectado não medicado), o grupo 2 foi medicado com diclazuril (1%) na dose de 1mL/4 litros de água potável por dois dias sucessivos, cinco dias após a infecção. Um terceiro grupo (G3) foi mantido como controle negativo não infectado e não medicado. O número de oocistos por grama de fezes (OOPG), o escore de lesões macroscópicas intestinais e o ganho de peso corporal foram avaliados. No G1, o OOPG foi de 3,67x106 e o peso final foi de 1,392kg, valores significativamente diferentes do OOPG de 1,93x106 oocistos e do peso final de 1,613kg do G2 (diclazuril). Os resultados provaram que o diclazuril (1%) em formulação líquida e na dosagem de 1mL/4 litros de água por dois dias sucessivos foi eficaz no controle de Eimeria acervulina.(AU)

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This study aimed to determine the pre-patent period and to evaluate the kinetics of cyst elimination and the systemic humoral (IgA, IgG(1), IgG(2a), IgM, IgE) and intestinal secretory (IgA) immune responses in gerbils (Meriones unguiculatus) experimentally innoculated with different doses of Giardia duodenalis trophozoites. Forty-eight animals aged 6-8 weeks were used, equally distributed among six groups, five groups innoculated with different doses of trophozoites (10(1), 10(2), 10(3), 10(4), 10(5)) and one control (non-infected) group. Coproparasitological examinations were carried out daily up to 91 days after inoculation (d.a.i.) to determine the pre-patent period and the kinetics of cyst elimination. Blood and stool samples were weekly collected for antibody assays. The pre-patent period was observed from the 9 d.a.i. onwards, with intermittent elimination of variable quantities of cysts up to 27 d.a.i.. All infected gerbils, irrespective of the dose received, were able to mount systemic humoral immune responses as evidenced by specific IgM titers from 7 to 28 d.a.i., corresponding to the peak of cyst elimination, followed by high and persistent IgG1 titers. Intestinal secretory responses were also seen with two peaks of fecal IgA titers, corresponding to IgM and IgG1 response peaks, respectively. In conclusion, systemic and intestinal humoral immune responses were related to the control of giardiasis in this experimental model.

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This study aimed at measuring intestinal villi and assessing the intestinal absorptive area in broilers infected with Eimeria acervulina under different treatments to control coccidiosis. The experiment was divided into two stages, carried out in successive housings, raised in the same environment (or aviary). In the first stage, on 25 May 2008, fifty 12-day-old birds were orally inoculated with 3 x 10(3) oocysts of E. acervulina. In the second stage, on July 2008, other 50 birds were allocated on litter contaminated by the feces of birds on the first housing (natural infection by oocysts present in the reused litter). The experiment was arranged in a complete randomized design with five treatments and three replicates of 10 chicks per treatment. Broiler chicks were housed at 1 day of age and autopsies were performed at 21 days of age. Three 2-cm-long segments of the duodenum were excised from each bird and fixed in 10% buffered formalin. A total of 30 slides were prepared for each treatment, totaling 150 evaluated histological sections using H&E staining. Villus morphology was carried out by the HL Image 97 software. The intestinal absorptive area was calculated and macroscopic lesions were classified according to standard lesion scores. Results showed that intestinal villus measurements and absorptive area are directly affected by E. acervulina and that there is direct and positive correlation between the macro and microscopic findings observed in intestinal coccidiosis. E. acervulina causes shortening of villi and reduction in the intestinal absorptive area, affecting broiler growth. The prevention method of litter fermentation during the interval between housings and oral administration of Diclazuril can reduce the severity of intestinal lesions by E. acervulina in broilers impairing oocyst virulence or viability.

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Since it is known that Entamoeba dispar is non-pathogenic and morphologically similar to E. histolytica, there are many targets used in PCR for differentiating these species. However, obtaining high quality DNA from fecal samples is fundamental for PCR. Most methods are laborious or use kits that make diagnosis expensive. In the present work, a new simple, fast and cheap technique of DNA extraction from fecal samples was combined with a PCR for an episomal target in order to identify E. histolytica and E. dispar in feces.

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Two different methods to detect Fasciola hepatica in faeces using kappa index were compared with four tamises technics and sequencial fitration in two tamises using Flukefinder® apparatus. High accordance between the two methods was observed. The values showed high sensibility and specificity.(AU)

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Two different methods to detect Fasciola hepatica in faeces using kappa index were compared with four tamises technics and sequencial fitration in two tamises using Flukefinder® apparatus. High accordance between the two methods was observed. The values showed high sensibility and specificity.

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Infection rates with Giardia duodenalis and Cryptosporidium spp. were compared among dogs living under different conditions. Stool samples (n = 433) collected from dogs of different ages, gender, living conditions and origin were analyzed using three techniques, i.e., centrifugal flotation in zinc sulfate solution, centrifugal flotation in sucrose solution, and methylene blue gram safranin staining. Eighty-nine of the samples were from stay dogs living in shelters run by animal protection societies, 199 were from kennels and 122 from households. A total of 119 (29.0%) had G. duodenalis cysts and six (1.4%) were positive for Cryptosporidium spp. oocysts. Dogs from kennels were most frequently affected by G. duodenalis (49.7%) while those from shelters showed a higher prevalence of Cryptosporidium spp. (2.2%). A significant difference (p < or = 0.05) was observed between immature dogs and adults only with respect to Giardia infection. There was no significant difference between the gender with regard to the presence of either protozoan.

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The susceptibility of dogs to experimental inoculation with trophozoites and cysts of human isolates of Giardia duodenalis and the clinical and laboratory profiles of infection of these animals were studied. Two groups (A and B), each comprising three dogs, were inoculated with G. duodenalis trophozoites and cysts, respectively. A third group of two dogs was not inoculated and remained as control. After inoculation feces were collected daily to determine the pre-patent period, by flotation in 33% zinc sulfate solution. Blood samples (5mL) were collected from animals at 15-day intervals during the 165 days of the experimental period and were used to carry out the hemogram and biochemical evaluation of the levels of total protein, albumin, alanine aminotransferase, gamma glutamyltransferase, urea and creatinine. A prepatent period was observed at 5-6 days post-inoculation (p.i.) in the inoculated dogs, with cysts eliminated for approximately 3 months. No alterations were seen in the clinical parameters evaluated. Anemia was observed at 15 p.i. in the inoculated dogs. The mean eosinophil count of inoculated groups was higher than that of the control (p< or =0.05) but none of the biochemical parameters analyzed presented significant differences. The results of this study show that G. duodenalis from human isolates is able to infect dogs with minimal systemic manifestations without producing clinical signs of giardiasis.

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