Repositório RCAAP

Conselho Nacional de Desenvolvimento Científico e Tecnológico

Conselho Nacional de Desenvolvimento Científico e Tecnológico

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Canine atopic dermatitis (CAD) is a common dermatosis, defined as a genetic-based disease, which predisposes to cutaneous inflammation and pruritus, mediated by class IgE immunoglobulins directed against specific antigens in most cases. Clinical diagnosis may be later complemented by skin allergic and/or serological tests (ELISA). The aim of these tests is to identify possible allergens in order to enable the clinicians to select candidate antigens for allergen specific immunotherapy. This work aimed to identify the sensitization profile of 58 dogs with atopic dermatitis diagnosis. All animals were submitted to intradermic test (IDT) and screened for the presence of antibodies against different allergens using a serologic test. House dust mites are described as the most frequent allergens in all continents. However, the positivity to C. dactylon is not commonly described and may be characteristic for the region. With this work it was possible to identify the main allergens involved in the immunologic response of atopic dogs residing in Rio Grande do Sul, pointing to the importance to include C. dactylon in screening tests for allergy.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico

Different approaches for the estimation of the uncertainty related to measurement results are found in the literature and in published guidelines. In the present work three approaches were used to estimate the uncertainty of the determination of aflatoxins (AB1, AB2, AG1, AG2) and deoxynivalenol in maize by liquid chromatography coupled to mass spectrometry tandem (LC-MS/MS): the Bottom up approach, adapted from the Guide to the Expression of Uncertainty in Measurement (ISO GUM); the Monte Carlo Method (MCM), which propagates distributions assigned to the input quantities through a numerical simulation; and the Kragten approach wich calculates standard deviations and confidence intervals with a universally applicable spreadsheet technique. A measurement equation was developed for mycotoxins analysis and a cause-and-effect diagram was draft to assist in the identification of the sources of uncertainty associated with the method. Detailed analysis of contributions of the various uncertainty sources was carried out. Measurement uncertainty was determined by the addition of the variances of the individual steps of the test procedure, according to each approach employed. The Bottom up, MCM and Kragten approaches produced very similar estimates to the combined uncertainty and the coefficient of variation (CV) between them was smaller than 1.0%. The main contribution to overall uncertainty is the intermediate precision with contributions over 90,0% for each mycotoxin. The results obtained with this research conclude that the three approaches are adequate for estimating the uncertainty in mycotoxin assays with LC-MS/MS technique. Among them, Bottom up is the most appropriate approach, since it requires that the analyst performs a detailed investigation about dominant components of the measurement uncertainty, allowing for better understanding and improvement of the measurement process. The Monte Carlo and Kragten methods are indicated the data generated by the Bottom up approach.

Infections by protozoa of the Sarcocystidae family have worldwide distribution and are common in ruminants, causing important economic losses. This study evaluated Sarcocystis spp., Toxoplasma gondii and Neospora caninum infections in sheep from Southwest region of Rio Grande do Sul, Brazil. Myocardium samples of 80 sheep raised on extensive system were collected. Tissue cysts were detected by direct examination and the presence of the agents was also confirmed by PCR. Macroscopic evaluation did not reveal changes, but direct microscopic examination showed cysts in 76.2% (61/80) of samples and all cysts were morphologically similar with Sarcocystis tenella or Sarcocystis arieticanis. PCR detected Sarcocystis spp. DNA in 21.2% (17/80) of samples tested and T. gondii DNA in 15% (12/80). In 6.2% (5/80) DNA of both protozoan were detected. Presence of N. caninum nucleic acids was not observed in the samples tested. All PCR-positive samples (23.7% - 19/80) were also positive by direct examination (microscopic cysts). Thus, a high occurrence of microscopic tissue cysts in sheep from the Southwest region of the state of Rio Grande do Sul was detected. Although PCR did not show a good sensitivity to identify the causative agents of these cysts, was possible to verify the presence of Sarcocystis spp. and T. gondii in cardiac muscle samples of the ovine. This may be a risk factor for animal and human contamination, not only through consumption, but also through handling the carcasses of these animals. Equine protozoal myeloencephalitis is a neurologic disease of horses, most often caused by the Sarcocystis neurona. However, the role of horses in the life cycle this parasite is not completely understood. This study attempts to elucidate the role of horses as intermediate hosts in S. neurona cycle, through occurrence of cysts in these animals and determine the antibodies frequency for Sarcocystis spp., Neospora spp. and T. gondii in slaughtered horses. Were collected 197 serum and heart samples of equines. None of the myocardium samples were detected tissue cysts, nucleic acids or histopathological changes associated to Sarcocystis spp. In antibodies detection, 146 (74.1%) serum samples were positive for studied protozoa. Antibodies against Sarcocystis spp. were detected in 36% (71/197), to Neospora spp. in 39.1% (77/197) and to T. gondii in 47,2% (93/197). Thus, the failure in detect tissue cysts, associated with antibodies anti-Sarcocystis spp. detection, increases the role of horses as accidental hosts in the cycle this protozoan, declining your participation in the epidemiology of Sarcocystis infection.

Conselho Nacional de Desenvolvimento Científico e Tecnológico

The navicular disease represents a degenerative disease of the navicular bone and associated structures. The diagnosis is based on history, clinical signs and nerve or nerve or joint blocks in addition to abnormalities seen in the radiographs. In order to evaluate the relationship of the radiographs findings and histology lesions inside the navicular bursa and adjacent structures 62 distal limb specimen were used in this study. The specimens were radiographed (dorso-palmar, lateral and skyline views) and a bursagram was performed. Then the specimens were processed to harvest the navicular bone and adjacent structures for histopathology. The results of this study showed that the positive predictivity was 71%. Considering only lesions seen at the bursagram the positive predictivity was 50%

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The efficiency of superovulation protocols with different levels of progesterone in dairy cows was evaluated on the ovulated follicles rate. Twenty five high producing female were distributed in control group (n = 5), and four treatments with progesterone device (CIDR). Used CIDR for 4 and 5 days, new CIDR for 4 and 5 days (n=5 in each treatment). All cows received a pre synchronization with prostaglandin F2α (PGF) and GnRH. On Day 10 of the estrous cycle started the follicle stimulating hormone (FSH) treatment in nine decreasing doses for 4.5 days. Two applications of PGF were also used along with the 5th and 6th FSH injection. Ovaries were evaluated daily by transrectal ultrasonography to determine the start of the second follicular wave, evaluation of recruitment, follicles growth, also for monitoring the ovulation. Superovulation with used CIDR 4 resulted in a higher ovulation rate between 36-60h after device removal (94.9%, P <0.0001) compared to the control group (49.5%), used CIDR 5 (68.3%), new CIDR 4 (52.8%), or new CIDR 5 (70.6%). Thus, artificial insemination could be performed within 24-36h after removal the used 4 days CIDR. The number of follicles in control cows (38 ± 9.27) was similar (P=0.739) to used CIDR 4 (31.4 ± 6.8), new CIDR 4 (31 ± 4.6), used CIDR 5 (33.4 ± 8.4), or new CIDR 5 (38.6 ± 10.6). Our finds supported the hypothesis that low levels of exogenous progesterone during superovulation were sufficient to allow pulses of luteinizing hormone to enable ovulation induction.

The objective of this study was to evaluate the performance, the microbiological profile, the enzyme profile and the detection of pathogenic strains by multiplex PCR of 40 weaned piglets (females and males). The experimental desing was completely randomized with fours treatments, five replicates and sex as a bloking factor. The treatments consistes of: T1 (control); T2 (Plant Extract (EV) (500 ppm)); T3 (Amoxicilin (A) (20 mg Kg-1) and T4 (A (20 mg kg-1)+EV (500ppm). There was no effect (P>0.01) between treatments for initial and final weight and gain daily weight, but the males in the control group had a mean daily consumption of 1.8% over diet (P<0.01) to the other treatments. The total count control of bacterial colonies was 35.9%, 70.9% and 63.8% higher (P<0.01) to treatment with A, EV+A and EV, respectively. In the midst MacConkey, the treated group A was 88.44%, 91.78% and 56.50%, higher (P<0.01) compared to EV+A, EV and control, respectively. The antibiogram 48 stool samples showed that Amoxicilin disk was 85.7%, 72.7%, 44.5% and 100% resistant in the control treatment, EV, A and EV+A, respectively. The bioflavonoids and ascorbic acid and the interaction with Amoxicilin not change the performance of pigs in the nursery phase, but reduce the colony count bacterianas. By the dosing biochemical CK, AST (U L-1), AST, TBARS and the frequency of fimbriae genes (F41, F6 (R 987), F4 (K88), F18 and F5 (K99) and toxins (LT, STx, STa and STb) of E. coli strains isolated from pigs with diarrhea using multiplex PCR with primers specific to these genes. Of the 64 samples 55 (85.93%) had genes for fimbriae and toxins, 23 (35.93%) had only genes for fimbriae and only two samples had genes for toxins only. The frequency of the detected virulence factors alone or in combination with other factors were: F41 (10.93%), F6 (21.87%), F4 (37.5%), F18 (70.31%), LT (39.6%), Stx (42.18%), STa (60.93%) and STb (75%). In all 64 samples was not found any of virulence factors for F5 (K99). The use of bioflavonoids and ascorbic acid did not change the performance of pigs in the weaned piglets. The use of plant extracts and associated with Amoxicilin reduces the count of bacterial colonies. Showed high resistance of the samples studied to Amoxicilin, the Neomycin and Norfloxacin. The antimicrobial Cefepine, ceftriaxone, Merepenem were more efficient in inhibiting the growth of E. coli strains isolated from pigs supplemented with ascorbic acid and bioflavonoid. The EV supplementation to the diets of weaned piglets does not alter the enzymatic profiles (ALT, AST), muscle (CK) and oxidative (TBARS). Animals receiving IV or its interaction with Amoxicilin presented to fimbriae genes and toxins. EV supplementation to weanling pigs a diet did not cause lack genes for the presence of fimbriae and toxins. The presence of genes for fimbriae and toxins does not determine the occurrence of diarrhea, as they may be present, but are not being expressed.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

The effect of mineral supplementation on pregnancy rate after synchronization with progestagen and fixed time artificial insemination (FTAI) was evaluated in no pregnant or barren Australian Merino sheep (n = 260) in autumn / winter 2010. Ewes were selected based on their body condition score. Average body condition score was 2.7 (range 1-5) with an average weight of 30.0 kg at TAI. All females were synchronized on a random day of the estrous cycle. The day they received an intravaginal pessary containing 60 mg of medroxyprogesterone acetate (MAP; Progespon®, Intervet) and mineral supplement was considered day zero (DO). After 12 days (D12) 250 IU of eCG (Novormon ®, Syntex, Argentina) were given intramuscular and the progesterone pessary was removed. After 55 to 60 hours (D14) fixed time insemination was performed. Ewes were distributed randomly into four groups (G). Ewes (n = 65) of G1 received 0.375 g anhydrous sodium selenite + 20.0 g anhydrous sodium glycerophosphate (Mineral supplement 1), and 15 g calcium lactobionate + 30 g of magnesium lactobionate + 8.0 g of zinc lactobionate (mineral supplement 2), subcutaneous. The ones from group 2 (G2) received Suplement 1, Group 3 (n = 65) was treated with Supplement 2. Ewes that received a single application of 0.9% saline solution (n=65) subcutaneous, without mineral supplementation constituted the control group (G4). Pregnancy diagnosis was performed by rectal ultrasonography 30 days after TAI resulting in 66.6%, 44.6%, 39.3% and 46.6% pregnancy rate in group 1, 2, 3 and control group, respectively. Mineral supplementation with magnesium, calcium, zinc (Mineral supplement 1) and selenium (mineral supplement 2) resulted in a higher rate of pregnancies (P <0.05), suggesting that these minerals may increase sheep production.

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Neosporosis is a parasite disease caused by Neospora caninum protozoan and characterized as a major cause of abortion in cattle. Eventually is possible birth of animals with neurological signs, stillbirths or birth of clinically normal calves, but chronically infected. Although there are many researches related to bovine neosporosis, there are still doubts about the cycle and the pathogenesis of N. caninum infection, so that is suitable for certain control and prophylaxis strategies to disease. Gerbils are considered highly sensitive of the parasite replication. Thus, this study aimed to verify the occurrence of infection with N. caninum by oral inoculation of tachyzoites in gerbils (M. unguiculatus). Seventeen animals were orally inoculated with 4x105 N. caninum tachyzoites. The detection frequency of total DNA samples of N. caninum showed that the protozoan infectivity occurs via this route of inoculation, noting that 81.8 % of the inoculated animal parasite DNA was detected. In relation to tissues, it can be observed that the DNA parasite was found in all organs evaluated (heart, lungs, kidneys, liver, spleen, brain), by variable frequency, demonstrating the wide distribution of this parasite in animals tissues.

In order to assess and establish the association between ultrasonographic and anatomophatological findings, ultrasonographic examinations were performed, post mortem, on the fetlock and associated structures of 37 equine forelimb specimens. All this specimens showed abnormal images or images identified as suspicious on the ultrasound examination. Subsequently, these limbs were dissected and underwent an anatomopathogical study. The association between ultrasonographic and anatomopathological findings was established by comparing data obtained by both methods. Ultrasonographic changes in size, shape, architecture and echogenicity of ligaments, tendons, joint capsule, articular cartilage and bony surfaces of the metacarpophalangeal joint were associated with the anatomopathological findings.

Conselho Nacional de Desenvolvimento Científico e Tecnológico

The Criollo Horse breed is gaining increasing importance in our state, due to competition to economic and cultural aspects. However, few studies were developed with this horse breed. Under seasonal and natural photoperiod influence, in a stud farm at 29º38 South latitude, the length of gestation, foal s heat occurrence and duration of partum to ovulation interval, fall transition, anestrous, vernal transition and estrous were evaluated in 82 Criollo mares, with age between three and 28 years. Palpation per rectum and ultrasonography were performed daily from the beginning of the breeding season in September/2005 until December/2006. The mean and standard deviation (±SD) of gestation length in 70 mares was 335.6±10.5 days, varying from 312 to 364 days. The mean interval from parturition to first ovulation (±SD) of 42 mares that foaled between September and December of 2005 and 2006 was 20.0±14.0 days. The longer period was 71 days. A foal heat ovulation shorter then 20 days was shown on 83.3% (35/42) of the mares and, on them the mean parturition to ovulation interval (±SD) was 14.2±3.0 days. The results of estrus evaluation, during the six days prior to ovulation, showed a daily enlargement of follicle diameter of about three millimeters. The mean diameter varied from 30.1mm at the 6th day pre-ovulation to 45.6mm on the day before corpus luteum detection. The vernal transition was evaluated in 14 mares in two consecutive years. On them, 64.3% finished their transitional period between the second half of September and the first fortnight of October, close to the spring equinox. The fall transition started on May and 75% (6/8) of the mares entered the anestrous period at the second half of that month