In this experiment, the effects of (1) lipid type (soy bean oil, tallow and fish oil), (2) the period the fish oil was provided and (3) a dietary supplement containing conjugated linoleic acid (CLA), selenium (Se), vitamin E and vitamin C on pig performance and pork quality were studied. Forty-eight female pigs (PIC hybrids, with a mean live weight of 16.19 kg ± 1.56 SD) were obtained from a single commercial operation in the North Island of New Zealand. The pigs were rank ordered by weight and assigned to one of six dietary treatment groups. The diet base was either a combination of animal and plant feedstuffs (AT and PTS), plant feedstuffs only (PO, POS) or plant feedstuffs combined with fish oil (PFSe and PFSl). The diets also differed depending on the presence or absence of the nutritional supplement SanoviteTM and a vitamin C supplement. SanoviteTM is a trademarked dietary supplement containing CLA (BASF, Auckland, New Zealand), organic Selenium (Alltech Inc., Nicholasville, KY) and vitamin E (Morel et al., 2008). Diets POS, PTS, PFSe, PFSl contained SanoviteTM and a vitamin C supplement. Diets PO and POS were used to establish the effect of the supplementation with SanoviteTM and the vitamin C supplement. Pigs fed diet PFSe received plant feedstuffs and fish oil with supplement between days 1 and 35 and then diet POS up to day 84. Pigs fed diet PFSl received diet POS between days 1 and 35; plant feedstuffs and fish oil with supplement between days 36 and 56 and then diet POS up to day 84. Pigs in group PFSe and PFSl both received the same total amount of fish oil per pig (2.52 l/ 2.31 kg). Between days 1 and 56 of the experiment grower diets were fed, and finisher diets were fed between days 57 and 84 of the experiment. The pigs were kept in pens of six, but fed individually twice daily (at approximately 8 am and 3.30 pm) according to a fixed feeding schedule. Water was available at all times. Individual feed intakes were measured daily and live weight recorded weekly. Faeces were collected once a day during two days in week five of the trail for digestibility determination. Carcass quality characteristics determined at the abattoir included carcass weight and back fat thickness as measured at the end of the slaughter line. Meat quality assessments were performed on the Semimembranosus muscle (SM) from one of the topside cuts of each pig. Measurements of fatty acid profile (loin and backfat) and the Se content (lean meat) were conducted in Singapore by Mrs. J. Leong (MSc). In this study plant or animal feedstuffs, lipid type, SanoviteTM and vitamin C supplementation had no significant effects on growth performance and carcass quality. There were no differences in apparent faecal digestibility characteristics for dry matter (DDM) and organic matter (DOM) in the un-supplemented animal (AT) and plant based (PO) diets. Lipid type had a significant effect on the digestibility of ash (DA), and an increased ratio of unsaturated fatty acid to saturated fatty acid resulted in increases in DDM and DOM. DDM and DOM increased when soybean and linseed oil (POS) were used instead of tallow (PTS) or fish oil (PFS). The main differences in DDM, DOM and DA were observed between diets PO and POS. A positive effect of selenium, vitamin E and CLA supplementation is suggested. Increased cooking temperatures reduced tenderness (higher mean, peak force, yield force and peak force – yield force) and increased cooking loss. There was a significant negative relationship between ultimate pH and relative lightness (L*). There were highly significant positive correlations between all three measurements of expressed juice, and there was a significant positive correlation between cooking loss at 60 and 70oC (P<0.01) but correlations between expressed juice values and cooking loss were not significant. The P-values for the contrasts for cooking loss at 70 0C were significant for all contrasts except for AT vs PO and PFSe vs PFSl. For all other contrasts, the P-values for cooking loss at 70 0C were significantly higher in group POS than for groups PO, PTS and PFSe+PFSl. The P-value for (Peak force – Yield force) at 70°C was significantly higher in samples from group PFSe than for samples from group PFSl. The P-value for the myofibrillar fragmentation index (MFI) was significantly higher for group AT in contrast to group PO. Group POS had a significantly higher P-value for MFI in contrast to groups PO, PTS and PFSe and PFSl. Group POS had a lower P-value for sarcomere length in contrast to groups PFSe and PFSl. Group PFSe had a significantly higher P-value for sarcomere length in contrast to group PFSl. Group PFSe had a significantly higher P-value for expressed juice percentage loss in weight in contrast to group PFSl. Supplementing with SanoviteTM increased the Se content (P = 0.002) in lean meat as analysed by J. Leong (2010, personal communication). In general it was concluded that an increase in the ratio of unsaturated fatty acid to saturated fatty acid (U/S) in the diet resulted in higher levels of unsaturated fatty acids in loin and backfat. The fatty acid profile in the diet reflected the fatty acid profile of pork. Backfat of pigs fed diets including soybean and linseed oil contained higher levels of linoleic and α-linolenic acids. Diets PO and POS were used to establish the effect of the supplementation of CLA. The backfat of pigs fed diet POS contained higher levels of CLA (C18:2-trans-10, cis-12) and α-linolenic acid than pigs fed diet PO. The loin of pigs fed diet POS contained higher levels of palmitoleic and linoleic acid and CLA (C18:2-cis-9, trans-11) and lower levels of oleic acid than pigs fed diet PO. The use of fish-oil as a lipid type resulted in the highest levels of eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA) in loin and backfat. The loin and backfat of pigs fed fish-oil in the second part of the grower phase (PFSl) contained higher levels of EPA, DPA and DHA than pigs fed fish-oil in the first part of the grower phase (PFSe). By enriching the swine diet with long-chain omega-3 polyunsaturated fatty acids (LC n-3 PUFA) it was possible to increase the EPA, DPA and DHA content of pork. Enriching pork with LC n-3 PUFA will contribute to achieving standards for adequate intake (AI), but might not be suitable to reach suggested dietary targets (SDT). In conclusion, it was possible to change the pork composition by dietary manipulation without compromising pig performance and meat quality. There were a few significant effects from treatments on meat quality characteristics, but differences reported in this study were small and relatively unimportant. A negative influence of the dietary regime on palatability and meat processing was expected, but these issues are beyond the scope of this experiment.

A aferição da pressão intraocular (PIO) é essencial para o exame oftálmico. Objetivou-se estabelecer os valores da curva diária da PIO de porquinhos da índia de diferentes faixas etárias com tonômetro de rebote. A PIO foi aferida às 6, 12, 18 e 24 horas utilizando o tonômetro de rebote (Tonovet®). Os porquinhos da índia foram subdivididos em dois grupos com 10 animais no grupo I e 4 animais no grupo II, considerando as idades, designados por GI (animais entre 2 a 3 meses de idade) e GII (animais entre 2 a 3 anos de idade). Previamente, foram realizados teste da lágrima de Schirmer, prova da fluoresceína, biomicroscopia com lâmpada de fenda e oftalmoscopia indireta em todos animais. O valor médio da pressão intraocular foi de 12,0 ± 1,83 mmHg. Foram encontradas diferenças significativas entres as idades, sendo que os valores médios para o grupo I foram 11,63 ± 0,29 mmHg, e para o grupo II foram 12,82 ± 0,45mmHg (P= 0,0295). Não foram encontradas diferenças entre ambos os olhos (P= 0,7454). Não foram encontradas diferenças significativas nos valores da PIO relacionadas ao sexo (P= 0,1858). Os valores da PIO em porquinhos da índia tiveram alteração nas horas avaliadas. Com base nos resultados obtidos foi possível concluir que os valores da PIO em porquinhos da índia da sofrem alterações ao longo do dia. Os menores valores de pressão intraocular ocorreram às 18 horas. / The measuring of intraocular pressure (IOP) it is essential for the ophthalmic evaluation; the aim of this study was establish the values of the IOP diurnal curve of guinea pigs with different ages using the rebound tonometer (Tonovet®). The animals were divided according to the age in two groups, Group I (GI) constituted with 10 guinea pigs aged between 2 -3 moths, and Group II (GII), 4 animals with 2 -3 years of old. Previously the Schirmer tear test, fluorescein test, biomicroscopy with slit lamp, and direct optalmoscopy were performed on all animals. The men value of IOP was 12.0±0.34 mmHg. Statistical difference were found between ages (mean GI 11.64 ± 0.29 mmHg, and GII 12.83 ± 0.45mmHg), did not have statistical difference related with gender (P= 0.1858) or between eyes (P= 0.7454). In conclusion the guinea pigs had alteration of IOP true the day, the values of IOP are higher in the morning period, decrease true the day and at the night period haves an elevation again.

Le phosphore est un élément crucial pour la vie sur Terre, de par son implication dans les processus bioénergétiques, le stockage et le traitement de l'information génétique. C'est également l'un des nutriments limitants en agriculture, aux côtés de l'azote et du potassium. Depuis la révolution verte au milieu du 20ième siècle, le monde agricole est dépendant des engrais phosphorés à bas coûts, fabriqués à partir d'une ressource fossile et nécessaires à l'amélioration des rendements des cultures à même de répondre aux besoins en nourriture d'une population en forte croissance. Cependant cette ressource, la roche phosphatée, s'épuise progressivement. De plus, son utilisation est très peu efficiente : moins de 20% du phosphore extrait se retrouve effectivement dans la nourriture consommée. L'une des raisons de cette faible efficience est la spécialisation de régions entières dans des productions agricoles spécifiques. Ainsi, les régions spécialisées dans les cultures à hauts rendements ont besoin de grandes quantités d'engrais minéraux alors que les régions d'élevage intensif ont des excédents de lisier sans terres agricoles suffisamment grandes et proches pour servir de zones d'épandage. L'épandage excessif de lisier en Bretagne est la cause première d'eutrophisation des cours d'eau. Le phosphore contenu dans le lisier porcin pourrait être recyclé sous forme de struvite (MgNH4PO4,6H2O), un engrais phosphaté à dissolution lente, très concentré et facilement transportable vers les régions de cultures végétales nécessitant une fertilisation phosphatée importante. Le phosphore du lisier porcin étant initialement présent sous une forme minérale solide, il est nécessaire de le dissoudre avant de le précipiter en struvite. Parce-que la dissolution par acidification chimique est trop chère et implique un mauvais bilan environnemental, le procédé développé lors de cette thèse utilise l'acidogénèse, un procédé biologique au cours duquel la matière organique est convertie en acides organiques en absence d'oxygène, acidifiant naturellement le lisier porcin. Différents déchets organiques ont été testés en tant que co-substrats dans du lisier porcin brut ou digéré, provoquant une fermentation de type lactique lorsque le co-substrat possédait une forte teneur en glucides facilement biodégradables, et une fermentation avec de nombreux acides organiques produits lorsque la teneur en glucides facilement biodégradables était faible. Il a pu être démontré que la fermentation lactique était le fait de bactéries appartenant au genre Lactobacillus, alors que divers Clostridiales dominaient lors des autres fermentations avec la production d'acétate, propionate, butyrate et valérate. Un réacteur en semi continu alimenté d'un mélange de lisier brut de petit pois et de carottes a permis la dissolution de 50% du phosphore total soit 750 mg-P/L. Après centrifugation, 3.4 g d'hydroxyde de magnésium par litre de surnageant a été ajouté afin d'élever le pH à 8 et ainsi précipiter la struvite. 99% du phosphore dissous a alors été abattu. Le solide obtenu contenait 70% de struvite, un léger excès de phosphore et de magnésium, ainsi que de la matière organique. L'acidogénèse permet l'hydrolyse de la matière organique complexe et la formation d'acides organiques. De ce fait, ce procédé de recyclage du phosphore contenu dans le lisier porcin pourrait être implémenté dans les nombreuses unités de méthanisation présentes en Bretagne et qui traitent des effluents animaux ainsi que des déchets organiques d'origine agricole, industrielle et municipale. La struvite obtenue pourrait être vendue dans les régions ayant besoin de fertilisation phosphatée alors que la matière organique du digestat pourrait être maintenue en Bretagne. Un tel procédé réduirait significativement l'eutrophisation due à l'épandage excessif du lisier tout en diminuant les besoins en fertilisants minéraux fossiles grâce à une source alternative aux performances fertilisantes équivalentes. / Phosphorus is a crucial nutrient for life, implicated in cellular bioenergetics as well as storage and processing of genetic information. It is also one of the limiting nutrients in agriculture with nitrogen and potassium. Since the green revolution in the middle of the 20th century, agriculture has relied on increasing amounts of cheap mineral P-fertilizers produced from a fossil resource to improve crop yields and sustain population growth. However, the resource is depleting and its use efficiency is poor: less than 20% of extracted P is actually consumed in food. One of the reasons for this is the specialization of entire regions into on type of agricultural production or another. Thus, regions focusing on high yield crops require large applications of fossil mineral fertilizers while intensive livestock breeding areas cannot find an output for their P-rich manure due to the distance with crop fields in need of P fertilization. Over application of animal manure in Brittany is the main cause of eutrophication in the region. Phosphorus could be recovered from pig manure as struvite, a concentrated, slow-release mineral fertilizer easily transported to crop-oriented regions in need of P fertilization. P in pig slurry is mostly under a solid inorganic form, requiring dissolution prior to precipitation as struvite. Because chemical acidification is too expensive and harmful to the environment, the process developed in this PhD relied on acidogenesis, a biological process in which organic matter is converted to organic acids under anaerobic conditions, thus naturally acidifying the swine slurry. Various organic wastes were tested as organic co-substrates on raw and digested pig slurry, leading to lactic acid fermentation when the co-substrate had a high content in easily biodegradable carbohydrates and a fermentation with diverse organic acids produced at low content in easily biodegradable carbohydrates. Lactobacillus was the genus responsible for lactic acid fermentation and various Clostridiales dominated otherwise, producing acetate, propionate, butyrate and valerate. A reactor was operated with semi-continuous feeding of raw swine slurry and carrot/pea, leading to the dissolution of 50% total-phosphorus or 750 mg-P/L. After centrifugation, struvite was precipitated in the supernatant by adding magnesium hydroxide to increase the pH to 8. 99% of dissolved P precipitated. The solid recovered contained 70% of struvite, a slight excess of P and Mg as well as organic matter. Because hydrolysis of organic matter and production of organic acids occurs during acidogenesis, the process could be implemented in the many anaerobic digestion units installed in Brittany treating animal manure and agricultural, industrial and municipal organic waste. The struvite recovered could be sold to regions in need while the digestate impoverished in P and rich in organic matter could be kept locally. Such process would reduce eutrophication due to over application of pig manure and also reduce the reliance on fossil P fertilizer by offering an alternative source with equivalent fertilizing performances.

La réduction des apports alimentaires de phosphore (P) ainsi que l’optimisation de son utilisation par les animaux représentent des enjeux majeurs pour assurer la durabilité des filières avicole et porcine. Le développement de systèmes d’alimentation assurant une utilisation efficace du phosphore est donc crucial en particulier durant la période de finition. Ce travail de thèse a permis de redéfinir le besoin en P des animaux durant cette phase et ce en lien avec les autres constituants de la matrice alimentaire reconnus pour impacter de façon majeure sur l’utilisation globale de P (Ca, phytase microbienne). Ce travail a également permis de quantifier l’impact du Ca et de la phytase microbienne sur les flux digestifs et métaboliques de P. Les connaissances générées contribuent à développer d’une part, des modèles mécanistes simulant le devenir de P à l’échelle de l’animal et d’autre part, de nouveaux systèmes d’alimentation répondant aux enjeux de la durabilité. / The reduction of dietary phosphorus (P) and the optimisation of its utilisation by the animals represent major challenges for the sustainability of poultry and pig production. The development of new feeding strategies ensuring an efficient utilization of P appears crucial particularly during the finishing period. The present work contributed to redefine the requirement of P of the animals during this phase in conjunction with the other constituents of the feed matrix well-known to significantly impact on the overall utilisation of P (Ca, microbial phytase). Moreover, this work quantified the impact of Ca and microbial phytase on the digestive and metabolic flows of P. On one hand, generated knowledge helps developing mechanistic models simulating the fate of P in the scale of the animal and on the other hand new feeding systems to meet the challenges of sustainability.

The aim of the experiment was to evaluate the effect of pig slurry applications in the growing crop of potatoes for industrial processing during the growing season. In 2015, he was at an altitude of 560 m based experiment, where the effect was evaluated by pig slurry at a variety Eurostarch experiment was divided into three variants, each variant had four repetitions, was a variant without application of slurry, 1 times applied manure and 2 times applied manure. For each experiment evaluated the proportion of emerged plants, tuber yield, starch content, average weight 1 tubers and the number of tubers in a bunch. In the reporting year and the effect of the application of manure. From the experimental results it is evident that the manure contributing to higher tuber yield, starch content and increased weight of tubers as the variant with one and two applications of slurry.

The aim of this study was to evaluate weight losses of pork meat during storage, especially during the chilling proces 24 hours after slaughter. In the theoretical part there are described the history of the origin and development of Prestice black-pied pig, the chemical composition of meat, slaughtering and chilling. The practical part dealt with the evaluation of weight losses in pig carcasses after twenty four hours of chilling. Data for this study was obtained from the carcasses of Prestice black-pied pig breed.

The aim of the thesis was to capture trends in population of Prestice Black Pied breed. In the literature section there was described formation and regeneration of breed characteristics and typical features of Prestice Black Pied breed, breed performance, the structure of the breeding base and the expected utilization and marketing. In own work there were described changes in the breeding base, which were in western Bohemia and moved to other places in the country. The largest companies that breed Prestice Black Pied breed are the ZD Mladotice and Žihelský statek SpA, located in the Plzeň region and Zemet Ltd. Tečovice, based in Zlin region and Chovservis Inc. Radostov breeding, based in Hradec Kralove region. In 2013 it was bred 348 sows, in 2014 404 sows, 2015 sows and 413 in 2016, 369 sows. Furthermore, the age structure was evaluated in basic herd of sows. The farmers had the most sows in the first and second litter, compared to that of the fifth litter sows had a few breeders. The requirement for longevity therefore not been complied with. The results of performance tests were evaluated reproductive performance of sows of basic herd in collections (the number of all born piglets, the number of live born piglets, the number of surviving piglets and milkiness). The average value of a breeding population did not meet the target. Some companies, however, had excellent results, especially ZD Mladotice and Zemet Ltd. Tečovice. In field tests were evaluated gain, the percentage of lean meat and backfat thickness. Breeding goal was achieved with values backfat thickness and percentage of lean meat. Gilts had lower gain than required by breeding goal. The gain for young boars has been in 2013 and 2016 higher than the required target and breeding in 2014 and 2015 below the required target. There were also evaluated linear track sires in breeding. It was found that, of the 10 extant lines has long represented the biggest line Wiskont and least represented of the Sudet line. The entire work was focused on monitoring results of individual breeders and their comparison with the breeding goal for Prestice Black Pied breed. Furthermore, the evaluation of the quality of semen of boars Prestice Black Pied breed, operating in the artificial insemination center in Kout Na Šumavě. All boars met the minimum parameters.

A aferição da pressão intraocular (PIO) é essencial para o exame oftálmico. Objetivou-se estabelecer os valores da curva diária da PIO de porquinhos da índia de diferentes faixas etárias com tonômetro de rebote. A PIO foi aferida às 6, 12, 18 e 24 horas utilizando o tonômetro de rebote (Tonovet®). Os porquinhos da índia foram subdivididos em dois grupos com 10 animais no grupo I e 4 animais no grupo II, considerando as idades, designados por GI (animais entre 2 a 3 meses de idade) e GII (animais entre 2 a 3 anos de idade). Previamente, foram realizados teste da lágrima de Schirmer, prova da fluoresceína, biomicroscopia com lâmpada de fenda e oftalmoscopia indireta em todos animais. O valor médio da pressão intraocular foi de 12,0 ± 1,83 mmHg. Foram encontradas diferenças significativas entres as idades, sendo que os valores médios para o grupo I foram 11,63 ± 0,29 mmHg, e para o grupo II foram 12,82 ± 0,45mmHg (P= 0,0295). Não foram encontradas diferenças entre ambos os olhos (P= 0,7454). Não foram encontradas diferenças significativas nos valores da PIO relacionadas ao sexo (P= 0,1858). Os valores da PIO em porquinhos da índia tiveram alteração nas horas avaliadas. Com base nos resultados obtidos foi possível concluir que os valores da PIO em porquinhos da índia da sofrem alterações ao longo do dia. Os menores valores de pressão intraocular ocorreram às 18 horas. / The measuring of intraocular pressure (IOP) it is essential for the ophthalmic evaluation; the aim of this study was establish the values of the IOP diurnal curve of guinea pigs with different ages using the rebound tonometer (Tonovet®). The animals were divided according to the age in two groups, Group I (GI) constituted with 10 guinea pigs aged between 2 -3 moths, and Group II (GII), 4 animals with 2 -3 years of old. Previously the Schirmer tear test, fluorescein test, biomicroscopy with slit lamp, and direct optalmoscopy were performed on all animals. The men value of IOP was 12.0±0.34 mmHg. Statistical difference were found between ages (mean GI 11.64 ± 0.29 mmHg, and GII 12.83 ± 0.45mmHg), did not have statistical difference related with gender (P= 0.1858) or between eyes (P= 0.7454). In conclusion the guinea pigs had alteration of IOP true the day, the values of IOP are higher in the morning period, decrease true the day and at the night period haves an elevation again.

The main aim of the present thesis was to compare different types of slaughter lines and their influence on the occurence of meat defects with emphasis on PSE defect. The research has been concentrated mainly on the slaughter lines which use the technology of stunning the animals with electricity and concentrated CO2. The research monitors the rate of PSE defect from the slaughter to the cutting of the pig (approximately 48 hours). For considering the difference in quality of the meat the basic identification indicators of both pH1 and the draining of meat were created. Furthermore, we monitored pH24 and pH48, too. Analyzing of the above mentioned indicators shows that stunning with concentrated CO2 is more suitable with regard to the quality of the meat. The average pH1 value was 0,3 (p < 0,001) higher, pH24 value was 0,243 (p < 0,001) higher and the average value of meat juice drainage was 2,16% lower (p < 0,001). The anylysis of the PSEi and PSE quality difference based on pH1 when stunning with the use of concentrated CO2 shows lower frequency of PSEi (7,843% lower) and PSE (1,961% lower). Based on meat juice drainage, PSEi frequency was 23,810% lower and PSE 4,762 % lower.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / Two vaccinia viruses (VACV) genetically and phenotypically divergent were isolated, in a mixed infection, from a horse lesion during an outbreak of vesicular and exanthematous disease in horses in Southern Brazil and termed Pelotas 1 (P1V) and Pelotas 2 (P2V). This thesis describes studies performed to investigate the pathogenesis of P1V and P2V infection in rabbits and guinea pigs, and to analyze the sequence of genes potentially involved in their phenotype. Chapter 1 investigated the dose-dependent susceptibility of rabbits to P1V and P2V after intranasal (IN) inoculation. Groups of weaning rabbits were inoculated with three doses of each VACV isolate (102.5 TCID50, 104.5 TCID50 e 106.5 TCID50/rabbit). The inoculation resulted in severe respiratory distress and death of most inoculated rabbits regardless the viral strain. Clinical signs started three to six days post-inoculation (pi) and culminated in death or euthanasia at days 5 to 10 pi. Viremia was detected in animals of all groups. All rabbits surviving the infection beyond day 9 pi developed neutralizing antibodies. Interstitial pneumonia, necrossupurative bronchopneumonia and diarrhea were observed in animals which died or were euthanized in extremis. These results demonstrate that P1V and P2V are virulent for rabbits and show no apparent differences in phenotype in this species. Chapter 2 describes the investigation of the susceptibility of rabbits to intradermal (ID) inoculation to VACV, in single or mixed infection. All inoculated animals developed skin lesions characterized by hyperemia, papules, vesicles pustules and ulcers. Infectious virus was detected in cutaneous lesions, lungs and intestine of animals that died during acute infection. These results demonstrate that rabbits develop cutaneous disease and systemic infection after P1V and P2V ID inoculation. Apparently, co-infected animals developed lesions more severe than those submitted to single virus infection. In chapter 3, the susceptibility and the potential of transmission of P1V and P2V by guinea pigs were investigated. For that, guinea pigs were inoculated IN with both P1V and P2V (106 TCID50/ml). The guinea pigs did not showed clinical signs but developed viremia, shed virus in secretions and seroconverted to VACV. Nevertheless, the virus was not transmitted to guinea pig sentinels maintained in close contact or when exposed to food and feces contaminated with VACV. In Chapter 4, four genes involved in virus phenotype/virulence (C7L, K2L, N1L e B1R) were submitted to nucleotide sequencing and analysis. A 15 nucleotide (nt) deletion in K2L gene was identified in P2V. The same pattern of nucleotide deletion was also detected in other genogroup 1 Brazilian VACV isolates. Point mutations were identified in K2L, C7L and N1R genes from P2V isolates when compared to P1V and to a standard VACV strain. The molecular analysis of these genes would not allow the establishment of association between the sequences/genotype and phenotype. However, this analysis indicate that the 15 nt deletion in K2L gene may be used as a molecular marker for genogroup 1 Brazilian VACV isolates. In summary, the results obtained in these studies demonstrate: i. P1V and P2V produce systemic and cutaneous disease in rabbits but they do not exhibit evident differences in virulence for rabbits; ii. Guinea pigs are susceptible to mixed P1V an P2V infection but apparently do not effectively transmit the virus; iii. P1V and P2V present some sequence differences in virulence genes and that a 15 nt deletion in K2L gene may be used as a molecular marker to distinguish between VACV genogroups. / Duas amostras de vírus vaccínia (VACV) geneticamente e fenotipicamente distintas foram isoladas de um mesmo animal em um surto de doença vesicular e exantemática em equinos no Rio Grande do Sul, e denominados Pelotas 1 (P1V) e Pelotas 2 (P2V). Esta tese descreve estudos realizados para investigar a patogenia dos isolados P1V e P2V em coelhos e cobaias, e analisar a sequência de genes potencialmente envolvidos no fenótipo desses isolados. O Capítulo 1 relata a investigação da susceptibilidade dose-dependente de coelhos ao P1V e P2V. Os animais foram inoculados pela via intranasal (IN) com três doses (102,5 DICC50, 104,5 DICC50 e 106,5DICC50/coelho) de cada um dos isolados. A inoculação resultou em enfermidade respiratória grave e morte na maioria dos coelhos, independente do isolado utilizado. Os sinais clínicos iniciaram nos dias 3 e 6 pós-inoculação (pi) e culminaram com a morte ou eutanásia dos animais, 5 a 10 dias pi. Viremia foi detectada em coelhos de todos os grupos. Anticorpos neutralizantes foram detectados em todos os animais que sobreviveram além do dia 9 pi. Pneumonia intersticial com broncopneumonia necrossupurativa e conteúdo líquido intestinal foram lesões observadas em animais inoculados com o P1V ou P2V que evoluíram para a morte ou foram motivo para a eutanásia in extremis. Esses resultados demonstram que P1V e P2V são virulentos para coelhos e não apresentam diferenças evidentes de patogenia nessa espécie. No Capítulo 2 foi investigada a susceptibilidade de coelhos após inoculação de VACV pela via intradérmica (ID). Para isso, os coelhos foram inoculados com um dos isolados ou com ambos. Todos os coelhos inoculados apresentaram lesões de pele caracterizadas por hiperemia, pápulas, vesículas, pústulas e úlceras. Excreção viral foi detectada nas lesões cutâneas e também em amostras de pulmão e intestino de animais que morreram durante a fase aguda da infecção. Os resultados desta inoculação demonstraram que coelhos desenvolvem doença cutânea e sistêmica após a inoculação ID de P1V e P2V. Algumas evidências indicam que os coelhos co-infectados desenvolveram lesões mais severas do que na infecção simples. No Capítulo 3, investigou-se a susceptibilidade e o potencial de transmissibilidade dos isolados P1V e P2V por cobaias. Para isso, cobaias foram inoculadas pela via intranasal (IN) com uma mistura dos isolados P1V e P2V (106 DICC50/ml). As cobaias não apresentaram sinais clínicos, porém excretaram o vírus nas secreções nasais, desenvolveram viremia e soroconverteram para VACV. Apesar disso, o vírus não foi transmitido a sentinelas por contato direto, indireto (aerossóis) ou por água e alimentos contaminados com fezes deliberadamente infectadas com o vírus. No Capítulo 4, quatro genes (C7L, K2L, N1L e B1R) envolvidos no fenótipo do VACV foram amplificados por PCR, sequenciados e submetidos à análise molecular. Uma deleção de 15 nucleotídeos (nt) no gene K2L foi identificada no P2V. Essa mesma deleção também foi identificada em isolados brasileiros do VACV pertencentes ao genogrupo 1. Mutações pontuais foram identificadas nos genes K2L, C7L e N1L no P2V comparando-se com o P1V e cepas de referência do VACV. A análise molecular desses genes não permite associar essas deleções/mutações presentes no P2V com o fenótipo, mas sugere que a deleção de 15 nt no gene K2L possa ser utilizado como marcador molecular de isolados de VACV do genogrupo 1. Em resumo, os resultados obtidos nesses experimentos demonstram que: i. P1V e P2V produzem doença sistêmica e cutânea em coelhos, mas não diferem fenotipicamente nessas espécies; ii. cobaias são susceptíveis à infecção mista pelo P1V e P2V, mas aparentemente não transmitem o vírus com eficiência; iii. P1V e P2V apresentam algumas diferenças em genes de virulência, sendo que a deleção de 15 nt no gene K2L pode ser utilizada como marcador de genogrupos de VACV.