Grão de soja moído e níveis de amido na terminação de tourinhos Nelore confinados

Pós-graduação em Zootecnia - FCAV

Desempenho de bovinos Nelore e ½ Angus/Nelore adaptados em confinamento por 9 ou 14 dias

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

Produção de oócitos e embriões bubalinos: efeitos da época do ano e da adição de óleo essencial de Lippia origanoides na maturação in vitro

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Comparison of Gross Pathology, Histopathology, and Mycobacterial Culture for the Diagnosis of Tuberculosis in Elk (Cervus elaphus)

Using the isolation of Mycobacterium bovis as the reference standard, this study evaluated the sensitivity, specificity and kappa statistic of gross pathology (abattoir postmortem inspection), histopathology, and parallel or series combinations of the two for the diagnosis of tuberculosis in 430 elk and red deer. Two histopathology interpretations were evaluated: histopathology I, where the presence of lesions compatible with tuberculosis was considered positive, and histopathology II, where lesions compatible with tuberculosis or a select group of additional possible diagnoses were considered positive. In the 73 animals from which M. bovis was isolated, gross lesions of tuberculosis were most often in the lung (48), the retropharyngeal lymph nodes (36), the mesenteric lymph nodes (35), and the mediastinal lymph nodes (16). Other mycobacterial isolates included: 11 M. paratuberculosis, 11 M. avium, and 28 rapidly growing species or M. terrae complex. The sensitivity estimates of gross pathology and histopathology I were 93% (95% confidence limits [CL] 84,97%) and 88% [CL 77,94%], respectively, and the specificity of both was 89% [CL 85,92%]). The sensitivity and specificity of histopathology II were 89% (CL 79,95%) and 77% (CL 72,81%), respectively. The highest sensitivity estimates (93- 95% [CL 84,98%]) were obtained by interpreting gross pathology and histopathology in parallel (where an animal had to be positive on at least one of the two, to be classified as combination positive). The highest specificity estimates (94-95% [CL 91-97%]) were generated when the two tests were interpreted in series (an animal had to be positive on both tests to be classified as combination positive). The presence of gross or microscopic lesions showed moderate to good agreement with the isolation of M. bovis (Kappa = 65-69%). The results show that post-mortem inspection, histopathology and culture do not necessarily recognize the same infected animals and that the spectra of animals identified by the tests overlaps.

Effect of melatonin on DNA damage of bovine cumulus cells during in vitro maturation (IVM) and on in vitro embryo development

The effect of melatonin during in vitro maturation (IVM) on DNA damage of cumulus cells (CCs) from bovine cumulus-oocyte complexes (COCs) and embryo development was evaluated. COCs from abattoir ovaries were cultured in maturation medium (MM) with 0.5 mu g/ml FSH and 5.0 mu g/ml LH (FSH-LH); 10(-9) M melatonin (MEL) or FSH-LH + MEL (FSH-LH-MEL). After 24 h of in vitro maturation, the CCs surrounding the oocyte were subjected to DNA analysis by Comet assay. After in vitro fertilization and in vitro embryo culture, the embryo development rates were evaluated on day 2 post insemination (cleavage) and days 7-8 (blastocyst). The percentage of CCs with no DNA damage was significantly superior in MEL group (37.6 +/- 2.4) than in FSH-LH-MEL (28.0 +/- 2.4) and FSH-LH (17.8 +/- 2.41) groups. Cleavage and blastocysts rates were similar among groups. Melatonin during IVM protects the CCs from DNA damage but this effect did not influence embryo development in vitro. (C) 2010 Elsevier Ltd. All rights reserved.

Influence of different shrinking temperatures and vacuum conditions on the ability of psychrotrophic Clostridium to cause 'blown pack' spoilage in chilled vacuum-packaged beef

This study determined the ability of psychrotrophic Clostridium strains isolated from vacuum-packaged beefs and abattoir environments to cause 'blown-pack' spoilage of vacuum-packaged beef stored at 2 and 15 degrees C. The influence of shrinking temperatures (83, 84 and 87 degrees C) and vacuum pressure (6 and 9 mbar) on the occurrence of such spoilage as well as the effects of simulated transportation (500 km) on the integrity of packages was determined. At 15 degrees C and 2 degrees C, twelve and six strains caused 'blown-pack' spoilage, respectively. The combination of vacuum pressure (9 mbar) combined with shrinking temperature (87 degrees C) retarded the occurrence of spoilage. The simulated transportation under the experimental conditions did not affect the integrity of packages. More studies that assess the factors that may contribute for the occurrence of 'blown-pack' spoilage should be performed to avoid the occurrence of such spoilage during its shelf-life. (C) 2012 Elsevier Ltd. All rights reserved.

Gas-producing and spoilage potential of Enterobacteriaceae and lactic acid bacteria isolated from chilled vacuum-packaged beef

This study aimed to enumerate and identify lactic acid bacteria and Enterobacteriaceae from spoiled and nonspoiled chilled vacuum-packaged beef and determine their potential to cause blown pack spoilage. These microbial groups were also enumerated in nonspoiled samples and detected in abattoir samples. The potential of isolates to cause blown pack spoilage of vacuum-packaged beef stored at chilled temperature (4 degrees C) and abuse temperature (15 degrees C) was investigated. Populations of lactic acid bacteria in exudate of spoiled and nonspoiled samples were not significantly different (P > 0.05), whereas the number of lactic acid bacteria on the surface was significantly higher (P < 0.05) in spoiled samples as compared to nonspoiled samples. The population of Enterobacteriaceae species in exudate and on the surface of samples were significantly higher (P < 0.05) in spoiled packs in comparison with nonspoiled packs. Results of the deterioration potential showed that blown pack spoilage was noticeable after 7 days at 15 degrees C and after 6 weeks at 4 degrees C for samples inoculated with Hafnia alvei.

L'epatite E nei suidi: epidemiologia, diagnosi e filogenesi

Hepatitis E is an infectious viral disease with clinical and morphological features of acute hepatitis. The aetiological agent is the Hepatitis E virus (HEV). The disease represents an important Public Health problem in developing countries where is frequently epidemic and primarily transmitted by fecal-oral route. In the last few years, a certain number of sporadic cases have been also described in industrialized countries, Italy included. A swine HEV was first identified in 1997 and is now considered an ubiquitous virus. Human and swine strains from the same geographical region have shown to have a high level of nucleotidic omology and in experimental infections, the possibility of interspecific transmission of swine strains to humans and of human strains to non-human primates has been demonstrated. Furthermore, some seroepidemiological studies have demonstrated that people working in contact with swine have a higher risk to get infected than normal blood donors. Recently, cases of HEV hepatitis have been directly associated to the ingestion of uncooked tissues from pigs, wild boar or deer and today the disease is considered an emerging zoonosis. The aims of this thesis were: evaluate HEV prevalence in Italian swine herds (both in fattening and in breeding animals); investigate the possibility of finding HEV in livers used for human consumption; investigate if there is any correlation between HEV infection and the presence of macroscopical lesions; investigate HEV prevalence in a demographic managed wild boar population; phylogenetically analyse viral strains identified. During an internship period at Veterinary Laboratories Agency (Weybridge, UK), furthermore, swine samples at different stages of production and slurry lagoons have been analysed. Six swine herds located in North Italy have been sampled at different stage of production. The overall prevalence resulted 42%, and both breeding and fattening animals were positive for HEV infection. A longitudinal study has been conducted in a herd across all stages of production until the slaughtering age. Livers have been collected from the animals at the abattoir and 11.8% of them were positive for HEV infection. No correlations have been identified between HEV infection and macroscopical lesions in pigs affected by different pathological conditions. Of 86 wild boars tested 22 (25%) were positive for HEV. Of the swine tested in UK 21,5 % and 2 of the 9 slurry lagoons (22,2%) were positive for HEV infection. All the strains identified belonged to genotype 3 and showed high percentages of nucleotidic identity with humans and swine strains identified in Europe. The high prevalence detected in these studies confirms the widespread diffusion of HEV in swine populations in Italy and in UK. Phylogenetical analysis of identified strains, similar to those identified in autochthonous human hepatitis E cases of the same geographical area, confirm the hypothesis that pigs can be a font of zoonotical infection. The finding that a fraction of the livers inserted in the food chain are positive for HEV infection it’s of some concern for Public Health. The finding of a high HEV prevalence in all examined farms, together with the observation that infection may be sub-clinical and affect animals at slaughtering age, raise concern because of the possible risk of transmission of HEV to humans by either direct contact with infected pigs, indirect contact with environment and working instruments contaminated with pig feces, or ingestion of contaminated undercooked meat.

Effects of the two production programs 'Naturafarm' and 'conventional' on the prevalence of non-perforating abomasal lesions in Swiss veal calves at slaughter

Non-perforating abomasal lesions are a considerable problem affecting more than half the population of veal calves. The objective of the present study was to assess the prevalence of pyloric and fundic abomasal lesions in Swiss veal calves at slaughter and to compare the occurrence of non-perforating abomasal lesions between two different production programs ('Naturafarm' and 'conventional'). 'Conventional'-production settings met the minimal standards as defined by the Swiss animal welfare legislation, whereas 'Naturafarm' production complied with increased animal welfare requirements. In order to identify risk factors for the development of abomasal lesions, information on management, housing, and feeding was obtained by a questionnaire. A total of 125 abomasa were randomly selected in one large abattoir. They were examined macroscopically, and the occurence of lesions in either the fundic or pyloric region of the abomasum was recorded separately. Animals raised in the 'conventional'-production setting revealed a significantly higher prevalence of lesions in the fundic part. Factors significantly affecting the prevalence of non-perforating lesions in the fundic part were the 'conventional'-production environment, including missing access to an outside pen, missing access to water and straw as the only roughage, feeding by bucket and the liquid milk by-product Protofit in combination with the powder Sprayfit.

Diagnostic significance of Neospora caninum DNA detected by PCR in cattle serum

A nested PCR that successfully detected Neospora caninum DNA in serum of cattle was used for investigation of selected abortion cases and in a study of healthy pregnant cows at an abattoir. N. caninum DNA was not detected in serum from antibody positive dams that aborted due to N. caninum, but was present in serum of some antibody negative dams that aborted due to other causes. N. caninum DNA was also found in the serum of about half of the animals that aborted of undetermined cause, but was not detected in cow sera from two beef cattle herds in Western Australia with no recent history of abortion. In the abattoir study of 79 dams and their foetuses N. caninum DNA was found in serum of 3 dams and in material from 11 foetuses. The majority of the cows and all foetuses were antibody negative. Our findings suggest that there is no obvious relationship between the presence or absence of N. caninum DNA in serum and the presence of antibodies to N. caninum in dams, the presence of N. caninum DNA in foetuses or abortion due to N. caninum. This is the first report of the detection of N. caninum DNA in serum of cattle rather than the white blood cell fraction. It indicates the presence of free tachyzoites and/or parasite DNA in circulation. The results suggest that persistent infection in the absence of antibodies is a possible outcome of N. caninum infection. Infection of foetuses in the absence of antibodies supports the possibility of persistent infection due to immunotolerance to an early in utero infection. It is therefore important to test for N. caninum DNA as well as antibodies for the detection of exposed and/or infected animals. However, the presence or absence of N. caninum antibodies or DNA did not support nor exclude N. caninum as the cause of abortion. Additional criteria are required for a positive diagnosis of abortion caused by N. caninum.

NonViral Gene Delivery of Growth and Differentiation Factor 6 (GDF6) to whole bovine Intervertebral Disc

Question: Low back pain is an increasing global health problem, which is associated with intervertebral disc (IVD) damage and de- generation. Major changes occur in the nucleus pulposus (NP), with the degradation of the extracellular matrix (ECM) [1]. Further studies showed that growth factors from the transforming growth factor (TGF) and bone morphogenic proteins (BMP) family may induce chondrogenic differentiation of mesenchymal stem cells (MSC) [2]. Focusing on non-viral gene therapies and their possible translation into the clinics, we investigated if GDF6 (syn. BMP13 or CDMP2) can induce regeneration of degraded NP. We hypothesized that IVD transfected with plasmid over-expressing GDF6 also up-regulates other NP- and chondrogenic cell markers and enhances ECM deposition. Methods: Bovine IVD cells were isolated by pronase/collagenase II overnight digestion. After monolayer expansion up to passage 3, cells were transfected with the plasmid pGDF6 (RG211366, Origene, SF) or with green fluorescence protein (GFP) control using the NeonÒ transfection system (Invitrogen, Basel), both equipped with a Cy- tomegalovirus (CMV) promotor to induce over-expression. We tested a range of yet unpublished parameters for each of the primary disc cells to optimize efficiency. To test a non-viral gene therapy applied directly to 3D whole organ culture, bovine IVDs were harvested from fresh tails obtained from the abattoir within 5 h post-mortem [3]. Discs were then pre-incubated for 24 h in high glucose Dulbecco’s Modified Eagle Medium and 5 % fetal calf serum. Each disc was transfected by injection of 5 lg of plasmid GDF6 (Origene, RG211366) into the center by 25G needle and using Hamilton sy- ringe. Electroporation was performed using 2-needle array electrode or tweezertrodes; 8 pulses at 200mv/cm with an interval of 10 ms were applied using ECM830 Square Wave Electroporation System (Harvard Apparatus, MA) (Fig. 1). After transfection discs were cultured for 72 h to allow expression of GFP or GDF6. Discs were then fixed, cryosectioned and analysed by immunofluorescence against GDF6. Results: We successfully transfected bovine NP and AF cells in monolayer culture with the two plasmids using a 1,400 V, 20 ms and 2 pulses with a *25 % efficiency using 0.15 M cells and 3 lg DNA (Fig. 1). Organ IVD culture transfection revealed GFP6 positive staining in the centre of the disc using 2-needle array electrode. Results from tweezertrodes did not show any GFP posi- tive cells. Conclusions: We identified novel parameters to successfully transfect primary bovine IVD cells. For transfection of whole IVD explants electroporation parameters need to be further optimized. Acknowledgments: This study was supported by the Lindenhof Foundation ‘‘Forschung und Lehre’’ (Project no. 13-02-F). References 1. Roughly PJ (2004) Spine (Phila) 29:2691–2699 2. 3. Clarke LE, McConell JC, Sherratt MJ, Derby B, Richardson SM, Hoyland JA (2014) Arthritis Res Ther 16:R67 Chan SC, Gantenbein-Ritter B (2012) J Vis Exp 60(60):e3490

Findings in uteri and ovaries from Eringer cows, slaughtered due to fertility problems

Eringer cows are often slaughtered due to fertility problems which result from inflammatory and degenerative changes of the uterus or hormonal imbalances. Twenty-one genital tracts from Eringer cows suffering from fertility problems were collected in the abattoir. The purpose of the study was the macroscopic evaluation of the ovaries and the uterus followed by a histological and microbiological analysis of the uterus. Data from inseminations and calvings were provided by the Eringer breeding association and through the internet portal www.agate.ch. Median age of the cows was 6.9 years, number of calves per cow was 2.5 and median period between last calving and slaughter was 1.5 years. In 13 from 21 of the urogenital tracts examined, macroscopic abnormalities of the ovaries and/or histologic or microbiologic findings in the uterus could explain fertility-associated slaughter.

Analysis and design of a thrombectomy device by using simulation techniques

Purpose: In this work, we present the analysis, design and optimization of one experimental device recently developed in the UK, called the 'GP' Thrombus Aspiration Device (GPTAD). This device has been designed to remove blood clots without the need to make contact with the clot itself thereby potentially reducing the risk of problems such as downstream embolisation. Method: To obtain the minimum pressure necessary to extract the clot and to optimize the device, we have simulated the performance of the GPTAD analysing the resistances, compliances and inertances effects. We model a range of diameters for the GPTAD considering different forces of adhesion of the blood clot to the artery wall, and different lengths of blood clot. In each case we determine the optimum pressure required to extract the blood clot from the artery using the GPTAD, which is attached at its proximal end to a suction pump. Result: We then compare the results of our mathematical modelling to measurements made in laboratory using plastic tube models of arteries of comparable diameter. We use abattoir porcine blood clots that are extracted using the GPTAD. The suction pressures required for such clot extraction in the plastic tube models compare favourably with those predicted by the mathematical modelling. Discussion & Conclusion: We conclude therefore that the mathematical modelling is a useful technique in predicting the performance of the GPTAD and may potentially be used in optimising the design of the device.

Analyse de la distribution d'Escherichia coli, utilisée comme un marqueur microbiologique, dans un réseau de production porcin

La présence d’Escherichia coli pathogènes en élevages porcins entraine des retards de croissance et la mortalité. La transmission des E. coli pathogènes entre les élevages et l'abattoir d’un même réseau de production n'est pas bien décrite. La détection des gènes de virulence des E. coli pathogènes pourrait permettre d’identifier un marqueur de contamination dans le réseau. L’objectif de cette étude a été d’identifier un marqueur de contamination E. coli dans un réseau de production porcine défini afin de décrire certains modes de transmission des E. coli pathogènes. Pour ce faire, une région géographique comprenant 10 fermes d’engraissement, un abattoir et un réseau de transport a été sélectionnée. Trois lots de production consécutifs par ferme ont été suivis pendant 12 mois. Des échantillons environnementaux ont été prélevés à l’intérieur et à l’extérieur des fermes (3 visites d’élevage), dans la cour de l’abattoir (2 visites lors de sorties de lot) et sur le camion de transport. La détection des gènes de virulence (eltB, estA, estB, faeG, stxA, stx2A, eae, cnf, papC, iucD, tsh, fedA) dans les échantillons a été réalisée par PCR multiplexe conventionnelle. La distribution temporelle et spatiale des gènes de virulence a permis d’identifier le marqueur de contamination ETEC/F4 défini par la détection d’au moins un gène d’entérotoxine ETEC (estB, estA et eltB) en combinaison avec le gène de l’adhésine fimbriaire (faeG). La distribution des échantillons positifs ETEC/F4 qualifie la cour de l’abattoir comme un réservoir de contamination fréquenté par les transporteurs, vecteurs de contamination entre les élevages. Ceci suggère le lien microbiologique entre l’élevage, les transporteurs et l’abattoir jouant chacun un rôle dans la dissémination des microorganismes pathogènes et potentiellement zoonotiques en production porcine.

Use of duckweed, bentonite and acid to improve water quality of effluent discharge from abattoirs

Intensive animal industries create large volumes of nutrient rich effluent, which, if untreated, has the potential for substantial environmental degradation. Aquatic plants in aerobic lagoon systems have the potential to achieve inexpensive and efficient remediation of effluent, and to recover valuable nutrients that would otherwise be lost. Members of the family Lemnaceae (duckweeds) are widely used in lagoon systems, but despite their widespread use in the cleansing of sewage, only limited research has been conducted into their growth in highly eutrophic media, and little has been done to systematically distinguish between different types of media. This study examined the growth characteristics of duckweed in abattoir effluent, and explored possible ways of ameliorating the inhibitory factors to growth on this medium. A series of pot trials was conducted to test the tolerance of duckweed to abattoir effluent partially remediated by a sojourn in anaerobic fermentation ponds, both in its unmodified form, and after the addition of acid to manipulate pH, and the addition of bentonite. Unmodified abattoir effluent was highly toxic to duckweed, although duckweed remained viable and grew sub optimally in media with total ammonia nitrogen (TAN) concentrations of up to 100 mg/l. Duckweed also grew vigorously in effluent diluted 1:4 v/v, containing 56 mg TAN/L and also modified by addition of acid to decrease pH to 7 and by adding bentonite (0.5%).