Adrenomedullin induces pulmonary vasodilation but does not attenuate pulmonary hypertension in a sheep model of acute pulmonary embolism

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Supplementation with protected fat to manage gastro-intestinal nematode infection in Santa Ines sheep

This experiment aimed to determine the influence of supplementation with protected fat on nematode infections in periparturient sheep or sheep in their final stage of pregnancy. Fifty Santa Ines ewes received 200 g of concentrate/animal/day, mineral salt and water and were grouped according to their weight, hematocrit and the number of eggs per gram of feces (EPG). Two diets were used: a control treatment and a treatment consisting of supplementation with 30 g/animal/day of protected fat in the concentrate. Three monthly evaluations were performed over a total of 84 days of testing. The variables analyzed were weight, body condition, EPG and coprocultures; blood tests were performed for the determination of packed cell volume, hemoglobin concentration and total plasma protein, and leukograms and eosinophil counts were performed. For weight and hemoglobin concentrations, interactions were observed between diet and collection date (P<0.05); body condition, total plasma protein, packed cell volume and total leukocytes did not differ statistically among treatments (P> 0. 05) but did differ by collection day (P <0.05). The EPG and eosinophil counts did not differ statistically by either diet or collection date (P>0.05). The genus Haemonchus was predominant, followed by the genera Cooperia, Trichostrongylus and Oesophagostomum. The protected fat did not decrease EPG and did not improve the blood parameters of infected sheep.

Subclinical diagnosis of Caseous lymphadenitis based on Elisa in Sheep from Brazil

Caseous lymphadenitis (CLA), caused by Corynebacterium pseudotuberculosis, is a chronic contagious disease that affects small ruminants and still remains an important problem for many lamb-producing countries. Animals are considered clinically infected when occurs abscesses in superficial lymph nodes. Visceral or internal form can coexist which no apparent clinical signs of infection are seen. The best procedure to avoid spread of the disease is elimination of infected animals. However, as the chronic and subclinical nature of the infection of CLA alternative methods are required for detection and screening. In this study, we described the performance of indirect Enzyme-Linked Immunosorbent Assay (ELISA) for diagnosis of CLA in asymptomatics sheep. Also, test culture and biochemical identification were achieved to confirm CLA infection. The serological diagnostic was performed in sheep symptomatics (n=50) and asymptomatics (n=374) from nine flocks. Analysis reported high positivity of 71% for ELISA in 85% of asymptomatic animal for CLA with a sensitivity of 88% and specificity of 31%. Results from ELISA test in asymptomatic animals against culture for caseous lymphadenitis were more specific (97%) and permitted to exclude healthy animals without symptoms. This study concluded that C. pseudotuberculosis infection could be widely disseminated in sheep flocks in Northwestern region of the state of São Paulo, Brazil and only one screening test is not enough. The association with indirect ELISA test and culture could better indicate the real problem of CLA in sheep flocks.

Detection of serum antibodies to parainfluenza type 3 virus, respiratory syncytial virus, bovine viral diarrhea virus, and herpes virus type 1 in sheep in the region of Botucatu, São Paulo - Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

EC34-48 Farm Sheep Facts

The farm flock and the fattening of western feeder lambs are the only methods of sheep production of importance in Nebraska. The farm flock is not a major enterprise on Nebraska farms. It serves as a side line on farms having a well-drained place which sheep may call their own. This 1934 extension circular contains information on: Nebraska Sheep-industry facts; market, ewe, ram, and lamb facts; shearing and wool facts; two parasites and one pest; grading and marketing wool; and scouring and carding wool for home use.

Comparison of Endocrine Response to Stress Between Captive-Raised and Wild-Caught Bighorn Sheep

Stress hormones in Rocky Mountain bighorn sheep (Ovis canadensis canadensis), produced in response to environmental changes, road development, or high population density, may impact their immune systems to a threshold level that predisposes them to periodic, large-scale mortality. We compared the stress response to a novel environmental situation and repeated handling between bighorn sheep born and raised in captivity (CR) and bighorn sheep born in the wild (WC) and brought into captivity. We measured plasma epinephrine, norepinephrine, cortisol, and fecal glucocorticoid metabolites (FGM). Three weeks after each group’s arrival we used a one-time drop-net event to elicit an acute stress response, and we collected blood samples from each sheep over 35 minutes, as well as one fecal sample. We collected blood and fecal samples from both groups on 7 other occasions over the subsequent 6 months. We also collected fecal samples from the pen at approximately 24-hour intervals for 3 days following every handling event to monitor the stress response to handling. We found that CR sheep had a stronger autonomic nervous system response than WC sheep, as measured by epinephrine and norepinephrine levels, but we found a very similar hypothalamic–pituitary–adrenal axis (HPA) response, measured by cortisol levels, to the acute stress event of a drop-net restraint. We also found that once the WC sheep had acclimated, as indicated by the return to the initial baseline FGM levels within 12 weeks, the CR and WC groups’ HPA responses to sampling events were not significantly different from one another. Fecal samples can provide a noninvasive mechanism for managers to monitor baseline FGM for a given herd. Using long-term monitoring of FGM rather than values from a single point in time may allow managers to correlate these levels to outside influences on the herd and better understand the impacts of management changes, population density, or increased human developments on the health of the sheep population.

Intake, nutrient apparent digestibility and ruminal constituents of sheep fed diets with canola, sunflower or castor oils

The objective in this experiment was to determine the effects of feeding diets with canola, sunflower or castor oils on intake, nutrient apparent digestibility and ruminal constituents of crossbred Dorper x Santa Ines sheep. Four rumen-cannulated animals of 90.2 +/- 11.4 kg average body weight were assigned to a 4 x 4 latin square. Animals remained individually in cages for the metabolism assay and were fed diets containing roughage at 500 g/kg and concentrate based on ground corn and soybean meal also at 500 g/kg. No oil was added to the control diet, whereas the others had canola, sunflower or castor oils at 30 g/kg (DM basis). There was no difference for the intake of DM and nutrients, except for ether extract, which was greater when animals received oil. The digestibility coefficients of dry matter, organic matter, crude protein, non-fiber carbohydrates and neutral detergent fiber were not changed; however, the addition of oil increased the ether extract digestibility. The values of total digestible nutrients (TDN, g/kg of DM), digestible energy (DE, Mcal/kg of DM), TDN intake and DE intake also did not change with the addition of lipids. Concerning the ruminal constituents, the addition of vegetable oils reduced the concentrations of acetate, butyrate and total short-chain fatty acids. Adding canola, sunflower or castor oils at 30 g/kg in diets with 500 g roughage/kg and 500 g concentrate/kg does not impair the intake or digestibility of nutrients in sheep, although it reduces the concentration of short-chain fatty acids in the rumen.

Hollow-fiber liquid-phase microextraction of amphetamine-type stimulants in human hair samples

A fast method was optimized and validated in order to quantify amphetamine-type stimulants (amphetamine, AMP; methamphetamine, MAMP; fenproporex, FPX; 3,4-methylenedioxymethamphetamine, MDMA; and 3,4-methylenedioxyamphetamine, MDA) in human hair samples. The method was based in an initial procedure of decontamination of hair samples (50 mg) with dichloromethane, followed by alkaline hydrolysis and extraction of the amphetamines using hollow-fiber liquid-phase micro extraction (HF-LPME) in the three-phase mode. Gas chromatography-mass spectrometry (GC-MS) was used for identification and quantification of the analytes. The LoQs obtained for all amphetamines (around 0.05 ng/mg) were below the cut-off value (0.2 ng/mg) established by the Society of Hair Testing (SoHT). The method showed to be simple and precise. The intra-day and inter-day precisions were within 10.6% and 11.4%, respectively, with the use of only two deuteratecl internal standards (AMP-d5 and MDMA-d5). By using the weighted least squares linear regression (1/x(2)), the accuracy of the method was satisfied in the lower concentration levels (accuracy values better than 87%). Hair samples collected from six volunteers who reported regular use of amphetamines were submitted to the developed method. Drug detection was observed in all samples of the volunteers. (c) 2012 Elsevier B.V. All rights reserved.

Jatropha curcas pericarp toxicity in sheep

Physic nut (Jatropha curcas) is a plant cultivated for biofuel production. Pericarp is a potential livestock food source by-product. However, its use may be limited due to the presence of toxic compounds, mainly phorbol esters. Thus, this study aimed to evaluate pericarp toxicity. Twenty sheep were divided in four groups, one control group which did not receive the plant and three experimental groups which received pericarp in 15% (G15), 30% (G30) and 45% (G45) concentrations for 23 days. After 10 days of treatment, pericarp ingestion produced food intake decrease, diarrhea, dehydration and loss of body condition. All treated groups showed decrease in alkaline phosphatase activity. G30 animals presented reductions in urea and total protein concentrations, and increase in potassium and sodium levels. G45 animals showed increase in serum aspartate aminotransferase activity and in albumin, creatinin, total and indirect bilirubin levels. Anatomohistopathologic findings included ascites, hydropericardium, congestion of the gastintestinal tract and lungs, pulmonary edema and adhesions in the thoracic cavity, renal tubular cells and centrilobular cytoplasmic vacuolation and lymphohistiocytic pneumonia and lymphoplasmacytic and histiocytic enteritis. On the physiochemical analysis 0.3845mg of phorbol esters/g of pericarp were detected. It is concluded that J. curcas pericarp is toxic and is not recommended for sheep feeding.

Biomechanical in vitro evaluation of three stable internal fixation techniques used in sagittal osteotomy of the mandibular ramus: a study in sheep mandibles

Among the osteotomies performed in orthognathic surgery, the sagittal osteotomy of the mandibular ramus (SOMR) is the most common, allowing a great range of movements and stable internal fixation (SIF), therefore eliminating the need of maxillomandibular block in the postoperative period. Objectives: The purpose of this study was to evaluate the biomechanical resistance of three national systems used for SIF in SOMR in sheep mandibles. Material and methods: The study was performed in 30 sheep hemi-mandibles randomly divided into 3 experimental groups, each containing 10 hemi-mandibles. The samples were measured to avoid discrepancies and then subjected to SOMR with 5-mm advancement. In group I, 2.0x12 mm screws were used for fixation, inserted in an inverted "L" pattern (inverted "L" group). In group II, fixation was performed with two 2.0x12 mm screws, positioned in a linear pattern and a 4-hole straight miniplate and four 2.0x6.0 mm monocortical screws (hybrid group). In group III, fixation was performed with two-hole straight miniplates and eight 2.0x6.0 mm monocortical screws (mini plate group). All materials used for SIF were supplied by Osteosin - SIN. The hemimandibles were subjected to vertical linear load test by Kratos K2000MP mechanical testing unit for loading registration and displacement. Results: All groups showed similar resistance during mechanical test for loading and displacement, with no statistically significant differences between groups according to analysis of variance. Conclusion: These results indicate that the three techniques of fixation are equally effective for clinical fixation of SOMR.

Genotyping of polymorphisms in the prnp gene in Santa Ines sheep in the State of Sao Paulo, Brazil

Santos C.R., Mori E., Leao D.A. & Maiorka P.C. 2012. [Genotyping of polymorphisms in the prnp gene in Santa Ines sheep in the State of Sao Paulo, Brazil.] Genotipagem de polimorfismos no gene prnp em ovinos Santa Ines no Estado de Sao Paulo. Pesquisa Veterinaria Brasileira 32(3):221-226. Laboratorio de Neuropatologia Experimental e Comparada, Departamento de Patologia, Faculdade de Medicina Veterinaria e Zootecnia, Universidade de Sao Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Cidade Universitaria, Sao Paulo, SP 05508-270, Brazil. E-mail: caio-patologia@usp.br Enzootic paraplexia or scrapie is a fatal neurodegenerative disease affecting mainly sheep and rarely goats. The disease is influenced by polymorphisms at codons 136, 154 and 171 of prnp gene that encodes the prion protein. The animals may be susceptible or resistant to the development of the disease according to the allelic sequences observed in these codons. In Brazil there were only cases of scrapie in imported animals, therefore the country is considered free of the disease. This study performed the genotyping of different polymorphisms associated to the development of scrapie. Then, based on these findings the animals were categorized in resistant and susceptible. A total of 118 samples were sequenced from the Santa Ines sheep raised on properties located in the State of Sao Paulo. From these samples, 6 alleles and 11 genotypes were identified (ARQ / ARQ, ARR / ARQ, ARQ / AHQ, ARQ / VRQ, AHQ / AHQ, ARR / ARR, ARR / AHQ, VRQ / VRQ, ARQ / TRQ, TRR / TRR, TRQ / TRQ), the genotype ARQ / ARQ presented a frequency of 56.7%. It was also detected the presence of tyrosine at codon 136, which may be considered a rare observation, since there is no report regarding Santa Ines breeding presenting this polymorphism. These results showed the great genetic variability in Santa Ines in Sao Paulo and only 1,69% of the genotypes observed are extremely resistant to scrapie. These data demonstrate that the Santa Ines sheep can be considered potentially susceptible to scrapie.

Clinical, haematological and biochemical responses of sheep undergoing autologous blood transfusion

Background: This study aimed to evaluate the clinical, haematological and biochemical responses to autologous blood transfusion and the feasibility of this practice in sheep. Thus, we used eight male, 8 months old sheep, weighing on average 30 kg, from which 15 mL/kg of whole blood was collected and stored in CPDA-1 bags. Blood samples were refrigerated for 8 days and subsequently re-infused. The clinical, haematological and biochemical parameters were evaluated before blood collection and reinfusion, after 10 minutes of collection and reinfusion, after 3, 6, 12, 24, 48, 96 and 192 hours after collection and reinfusion. Results: With respect to clinical parameters, we observed a decrease in heart rate after 24, 48 and 196 hours from reinfusion compared to basal values (p <0.05). Haematological variables including globular volume and erythrocyte counts showed a significant decrease (p <0.01) at all time points after collection and increased (p <0.01) at all time points after reinfusion. There was a significant increase in total protein and calcium at all time points after reinfusion (p <0.05). Conclusion: Autologous transfusion in sheep slightly altered the physiological, biochemical and haematological responses of sheep, indicating that the technique proposed is safe and can be applied in the clinical practice of this species. The 8 d period was not sufficient for complete recovery of the haematological parameters after blood collection.

The effect of sodium fluoride preservative and storage temperature on the stability of cocaine in horse blood, sheep vitreous and deer muscle

The in vitro stability of cocaine in horse blood, sheep vitreous humour (VH) and homogenised deer muscle is described. The stability of cocaine in horse blood was of interest because many toxicology laboratories utilise horse blood for the preparation of calibration and check standards and the latter are typically stored during routine use. The storage stability of cocaine in human VH and muscle has not been previously reported. In the absence of blank human VH and muscle, cocaine stability under varying conditions was demonstrated in animal tissues. Blood and VH were stored with and without addition of NaF at room temperature (RT), 4 degrees C and -18 degrees C for 84 days. Muscle homogenates were prepared in water, water/2% NaF, and phosphate buffer (pH 6.0)/2% NaF, and stored for 31 days at RT, 4 degrees C and -18 degrees C. Cocaine stability in human muscle obtained from cocaine positive forensic cases was assessed following storage at -18 degrees C for 13 months. Cocaine and benzoylecgonine (BZE) were extracted using SPE and quantified by GC-MS/MS. Cocaine was stable for 7 days in refrigerated (4 degrees C) horse blood fortified with 1 and 2% NaF. In the absence of NaF, cocaine was not detectable by day 7 in blood stored at RT and 4 degrees C and had declined by 81% following storage at -18 degrees C. At 4 degrees C the rate of cocaine degradation in blood preserved with 2% NaF was significantly slower than with 1% NaF. The stability of cocaine in horse blood appeared to be less than that reported for human blood, probably attributable to the presence of carboxylesterase in horse plasma. Cocaine stored in VH at -18 degrees C was essentially stable for the study period whereas at 4 degrees C concentrations decreased by >50% in preserved and unpreserved VH stored for longer than 14 days. Fluoride did not significantly affect cocaine stability in VH. The stability of cocaine in muscle tissue homogenates significantly exceeded that in blood and VH at every temperature. In preserved and unpreserved samples stored at 4 degrees C and below, cocaine loss did not exceed 2%. The increased stability of cocaine in muscle was attributed to the low initial pH of post-mortem muscle. In tissue from one human case stored for 13 months at -18 degrees C the muscle cocaine concentration declined by only 15% (range: 5-22%). These findings promote the use of human muscle as a toxicological specimen in which cocaine may be detected for longer compared with blood or VH. (C) 2011 Elsevier Ireland Ltd. All rights reserved.

Trypanosoma (Megatrypanum) melophagium in the Sheep Ked Melophagus ovinus from Organic Farms in Croatia: Phylogenetic Inferences Support Restriction to Sheep and Sheep Keds and Close Relationship with Trypanosomes from Other Ruminant Species

Trypanosoma (Megatrypanum) melophagium is a parasite of sheep transmitted by sheep keds, the sheep-restricted ectoparasite Melophagus ovinus (Diptera: Hippoboscidae). Sheep keds were 100% prevalent in sheep from five organic farms in Croatia, Southeastern Europe, whereas trypanosomes morphologically compatible with T. melophagium were 86% prevalent in the guts of the sheep keds. Multilocus phylogenetic analyses using sequences of small subunit rRNA, glycosomal glyceraldehyde-3-phosphate dehydrogenase, spliced leader, and internal transcribed spacer 1 of the rDNA distinguished T. melophagium from all allied trypanosomes from other ruminant species and placed the trypanosome in the subgenus Megatrypanum. Trypanosomes from sheep keds from Croatia and Scotland, the only available isolates for comparison, shared identical sequences. All biologic and phylogenetic inferences support the restriction of T. melophagium to sheep and, especially, to the sheep keds. The comparison of trypanosomes from sheep, cattle, and deer from the same country, which was never achieved before this work, strongly supported the host-restricted specificity of trypanosomes of the subgenus Megatrypanum. Our findings indicate that with the expansion of organic farms, both sheep keds and T. melophagium may re-emerge as parasitic infections of sheep.

Blood gas profile of copper-poisoned in sheep treated with ammonium tetrathiomolybdate

The aim of this study was to evaluate the blood gas profile of experimentally copper-poisoned sheep (in the pre-hemolytic, hemolytic and post-hemolytic phases) that have been treated or not treated with ammonium tetrathiomolybdate. Ten lambs of the Santa Ines breed were divided into two groups: control and ATTM (treated (ammonium tetrathiomolibydate). The animals were submitted to increasing doses of copper sulfate until macroscopic hemoglobinuria was detected. All of the control animals from died within four days of hemolytic crisis, and one sheep from ATTM died during the treatment. There was no difference in blood gas parameters between experimental groups. Higher values of pCO(2) were observed during the hemolytic crisis (HC) in both groups. The control group had higher mean values of hCO(3) in the times HC and 2 days after hemolytic crisis (dA) when compared with the time 15 before hemolytic crises (dB). The sheep that were treated with ATTM presented lower values of hCO(3) at 7dB and higher levels at the HC. The control and ATTM groups exhibited higher values of BE during the HC. Poisoning resulted in disorder in the acid-base equilibrium, characterized by metabolic alkalosis and respiratory acidosis. Treatment with ATTM was able to reverse the changes in acid-base balance in copper poisoning sheep.