Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This doctoral thesis involved the study of diseases in pigs, including the description of a case of granulocytic sarcoma and outbreaks of eosinophilic salt poisoning. The study of the characteristics and frequency of disease in swine population in the region covered by the Laboratório de Patologia Veterinária (LPV), Universidade Federal de Santa Maria (UFSM) was conducted through a retrospective study of all autopsy diagnoses. Five hundre and sity four with conclusive diagnoses were set apart. The pig population in the region covered by the study is predominantly family and diagnosed diseases reflect this reality. The infectious and parasitic diseases were the most prevalent [384 (68.1%)], followed, in descending order, nutritional and metabolic diseases [64 (11.,3%)], poisoning and toxi-infections [33(5,8%)] and developmental disorders [17(3.1%)]. Other changes to various etiologies, especially trauma, were cause of death in just over 11% of the protocols examined. Bacterial diseases were responsible for more than half of the causes of death or reason for euthanasia of animals studied, pointing to the influence of management, environmental and nutritional regional husbundry . The main disease diagnosed in pigs in the region covered by the study is the edema disease and along with other forms of infection by Escherichia coli is responsible for 23% of deaths of viral diseases and cancer are not major causes of death in pigs in the region studied. Nutritional hepatose and aflatoxicosis are important diseases in the region and and there cuasation are mainly linked to nutritional management. For description of eosinophilic granulocytic sarcoma, a set of techniques (cytology, histology, histochemistry and immunohistochemistry)were used and which proved to be essential for the confirmation of macroscopic tentive diagnosis. Salt poisoning is a condition recognized in pigs as far back as the first half of the last century and still remains as an important cause of mortality; the large amount of salt accumulated in the body in cases of poisoning can be demonstrated by determination of sodium ion in the liver, muscle, CSF, serum and aqueous humor, and these are acillary data to confirm the diagnosis. Pigs affected by salt poisoning have eosinopenia due to severe meningoencephalitis and infiltration of eosinophils in tissues, acute neuronal necrosis which develops into deeplaminar cortical necrosis as the clinical progresses. / Esta tese envolveu o estudo de doenças de suínos, incluindo a descrição de um sarcoma granulocítico eosinofílico e surtos de intoxicação por sal. O estudo das características e frequência das doenças na população suína na região de abrangência do Laboratório de Patologia Veterinária (LPV) da Universidade Federal de Santa Maria (UFSM) foi realizado através de um estudo retrospectivo de todos os diagnósticos de necropsias. Foram separados 564 casos com diagnósticos conclusivos. A população suína na região de abrangência do estudo é predominantemente familiar e as doenças diagnosticadas, refletem esta realidade. As doenças infecciosas e parasitárias foram as mais prevalentes [384 (68,1%)], seguidas, em ordem decrescente, das doenças metabólicas e nutricionais [64 (11,3%)], intoxicações e toxiinfecções [33 (5,8%)], e distúrbios do desenvolvimento [17 (3,1%)]. Outras alterações de diversas etiologias, sobretudo de trauma, foram causa de morte em pouco mais de 11% dos protocolos examinados. Doenças bacterianas são responsáveis por mais da metade das causas de morte ou razão para eutanásia dos suínos estudados, revelando a influência de fatores de manejo, ambientais e nutricionais do modo de vida local. A principal doença diagnosticada em suínos na região de abrangência do estudo é a doença do edema e juntamente com outras formas de infecção por Escherichia coli é responsável por 23% das mortes Doenças de etiologia viral e neoplasias não são importantes causas de morte em suínos na região estudada. Hepatose nutricional e aflatoxicose são importantes doenças da região e se devem, sobretudo, a fatores de manejo nutricional. Para descrição do sarcoma granulocítico eosinofílico, foi utilizado um conjunto de técnicas (citologia, histologia, histoquímica e imuno-histoquímica), indispensáveis para a confirmação da suspeita macroscópica. Intoxicação por sal é uma condição reconhecida em suínos desde a primeira metade do século passado e continua uma importante causa de mortalidade e a grande quantidade de sal acumulada em casos de intoxicação pode ser comprovada pelas dosagens do íon sódio no fígado, músculo, líquor, soro e humor aquoso, e esses dados servem como forma de diagnóstico. Suínos intoxicados por sal apresentam eosinopenia devido a grave infiltração de eosinófilos e a meningoencefalite eosinofílica evolui para uma necrose neuronal laminar, à medida que se desenvolve o curso clínico da doença.

Made available in DSpace on 2016-12-08T16:24:24Z (GMT). No. of bitstreams: 1 PGCV06MA014.pdf: 2536288 bytes, checksum: 2ced4e5eb18884533f8a66a86da4f665 (MD5) Previous issue date: 2006-07-04 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Aujeszky s disease (AD) is an infect-contagious illness that causes serious economical damages to the producer and the swine industry. Aiming to develop mechanisms and to improve technologies that are faster, more sensitive and more specific for diagnosis and for use in free areas or in AD s eradication programs, the sequence codifier of the glycoprotein E (gE) of Aujeszky s disease virus (ADV) was amplified, cloned and expressed. Through genetic engineering the sequence of the gE gene was propagated in an host organism. The gE was amplified by the technique of polimerase chain reaction (PCR), cloned in the vector pGem®-T Easy and transformed in competent cells of Escherichia coli, DH-5a . The clone obtained was sub-cloned in the expression plasmid pFastBac 1, which contains the promoter gene of the polyhedrin. The obtained subclone was analyzed inside for certification of its correct plasmid orientation with the restriction endonucleases BamH I and EcoR I. The sub-clone with the correct orientation had its DNA extracted and used for transposition inside the bacmid (recombinant baculovirus and helper plasmid with competent DH10Bac cell). Colonies with inserted gE were selected by the phenotype of the colony, which expresses white color when cloned. White recombinant colonies had their DNA extracted and used for cotransfection in insect cells Trichoplusia ni (BTI-Tn5B1-4). The recombinant-gE baculovirus was inoculated in cultured cells and expressed the recombinant gE by PCR and Western blotting . The recombinant-gE baculovirus containing only the gE gene of the VDA will be used for antigen and monoclonal antibodies production, which will aid in the development of a more sensitive, specific and safer for the use in VDA free regions / A doença de Aujeszky (DA) é uma enfermidade infecto-contagiosa que causa graves prejuízos econômicos ao produtor e à agroindústria suinícola. Com o objetivo de desenvolver insumos e aprimorar tecnologias que sejam mais rápidas, sensíveis e específicas de diagnóstico para uso em regiões livres ou em erradicação da DA, a seqüência codificadora da glicoproteína E (gE) do vírus da doença de Aujeszky (VDA) foi amplificada, clonada e expressada. Através da engenharia genética a seqüência do gene da gE foi propagada em um organismo hospedeiro. A gE foi amplificada pela técnica da reação em cadeia da polimerase (PCR) , clonada no vetor pGem®-T Easy e transformada em células competentes de Escherichia coli, DH-5a . O clone obtido foi subclonado no plasmídeo de expressão pFastBac 1, o qual possui o sítio promotor do gene da poliedrina. O subclone obtido foi analisado para certificação de sua orientação correta dentro do plasmídeo com as endonucleases de restrição BamH I e EcoR I. O subclone com a orientação correta teve seu DNA extraído e usado para a transposição dentro do bacmid (baculovírus recombinante e plasmídeo helper em célula competente DH10Bac ). As colônias com inserto gE foram selecionadas pelo fenótipo da colônia, a qual expressa cor branca quando clonada. As colônias brancas recombinantes tiveram seu DNA extraído e usado para a co-transfecção em células do inseto Trichoplusia ni (BTITn5B1- 4). O baculovírus gE-recombinante ao ser inoculado em cultivo celular, expressou a gE recombinante, comprovada pela técnica de PCR e Western blotting . O baculovírus gE-recombinante contendo apenas o gene da gE do VDA será utilizado para produção de antígeno e de anticorpos monoclonais, o que auxiliará no desenvolvimento de um teste de diagnóstico mais sensível, específico e mais seguro para uso em áreas livres do VDA

This study was conducted to compare the availability of Zn from two Zn sources, an amino acid chelate and ZnSO₄. In three experiments, 78 Zn depleted and 24 Zn adequate pigs were used. Pigs were depleted of Zn by feeding an isolated soy protein, semi purified diet containing 17 ppm Zn. Of the 78 depleted pigs, 60 pigs were Zn repleted. During Zn repletion in Exp. 1, depleted pigs were fed the low Zn diet supplemented with 5, 15, or 45 ppm Zn either as ZnSO₄ or as Zn amino acid chelate (ZnAAC). In Exp. 2 and 3, low Zn diets were only supplemented with 45 ppm Zn. Zinc adequate pigs, used in Exp. 1 and 2, were fed the 45 ppm supplemental Zn diets. To evaluate differences in site and rate of apparent Zn absorption, chromic oxide was added to the diets of depleted pigs in Exp. 1 and 3. In all experiments, a 24-d period was sufficient to severely deplete the porcine body Zn stores, and to cause parakeratosis and growth retardation. Serum Zn concentrations and serum ALP-activities of depleted pigs dramatically decreased (P < .01) during the first 14 d of Zn depletion. At the end of Zn depletion, Zn contents in liver, kidney, pancreas, brain, and small intestine tissues of pigs fed the low Zn diet were reduced (P < .01) by 10 to 40 % compared with the adequate pigs fed the ZnSO₄ and ZnAAC diets. In Exp. 2, the growth retardation was associated with a low (P < .05) serum mitogenic activity and pituitary RNA content of depleted pigs compared with pair-fed adequate pigs. Moreover, the growth hormone mRNA fraction tended to be reduced (P < .10) for the Zn depleted pigs. In Exp. 1, the apparent absorption of Zn was higher (P < .01) for pigs fed ZnAAC compared with the ZnSO₄ group; however, this was not confirmed in Exp. 3 unless coefficients were corrected for Cr recovery. Furthermore, absorption of Zn occurred primarily within jejunal and distal segments of the small intestine. In the balance of Exp. 3, disappearance rates of Zn, Cu, Fe and DM were higher (P < .01) in depleted pigs fed ZnAAC compared with ZnSO₄. The recovery of Cr also was different (P < .01) between pigs fed the ZnSO₄ (87 %) and ZnAAC (70 %) diets. Moreover, the moisture content of the fecal matter was 11 % higher (P < .01) for the ZnAAC group compared with pigs fed ZnSO₄. In Exp. 1, depleted pigs fed the 15 ppm ZnSO₄ and ZnAAC diets regained their ability to grow, however, replenishment of body fluid and tissue Zn pools did not occur within the 24-d Zn repletion period. Both the 5 ppm ZnSO₄ and ZnAAC groups did not respond to Zn repletion within a 12-d period. In all experiments, the rate and degree of repletion of body fluid and tissue Zn stores was not different between pigs fed the 45 ppm ZnSOq and ZnAAC diets, although a higher (P < .05) serum mitogenic activity was observed for the adequate pigs fed ZnAAC compared with ZnSO4. In conclusion, an amino acid chelate did not improve growth, or rate and degree of replenishment of body fluid and tissue levels of Zn compared with pigs fed ZnSO₄. However, ZnAAC may have influenced intestinal luminal conditions since a higher rate of disappearance of Zn, Cu, Fe, Cr, and DM was measured. / Ph. D.

The broad objective of the study was to determine the economic value of local pigs in marketand subsistence-oriented production systems in communal areas of Southern Africa. Data were collected from 288 households to investigate farmer perceptions, effects on pig production and handling of disease outbreaks such as classical swine fever (CSF) in market- and subsistenceoriented production systems. The utilisation of local pigs in these market- and subsistenceoriented production systems in improving people‟s welfare was evaluated. Climate change was identified by farmers in these production systems as a major constraint to pig production hence an experiment was carried out in the hottest season to determine diurnal heat-related physiological and behavioural responses in Large White (LW) and South African local pigs. The same genotypes were used to determine effects of diurnal heat-related stress on their growth performance. Choice experiment was done to determine farmer preferences for local pig traits and implicit prices for these traits in CSF-affected and unaffected areas that were under subsistence- and market-oriented production systems. In this experiment, the importance of heat tolerance was assessed relative to other productive and climate change adaptation traits. Significantly more pigs were culled in the CSF-affected areas that were market-oriented (8.0 ± 1.76) than subsistence-oriented (4.1 ± 1.00) production system. The risk of parasites and disease challenges was high in subsistence-oriented production system and coastal areas. In both production systems, CSF was perceived as destructive since the culling of pigs affected pork availability and income generation. The high risk of disease outbreaks and threat of climate change caused farmers in subsistence-oriented production system to select local pigs for their adaptive traits while those in the market-oriented production system focused on productive imported pigs. Farmers (83 %) indicated that they wanted pig genotypes that were adapted to climate change effects such as hot conditions. Local pigs were found to have superior heat tolerance over LW pigs (P < 0.05) in terms of lower heart rate and skin surface temperature. Frequency per day and duration for behavioural heat loss activities such as wallowing, sleeping in a prostrate posture and sprawling in slurry were also lower (P < 0.05) for local than LW pigs. The superiority of heat tolerance of local over LW pigs was further confirmed by their uncompromised growth performance under high diurnal temperatures. The Pearson‟s product moment correlation coefficient between temperature and feed conversion ratio for LW pigs was strongly positive (r = 0.50; P < 0.001) unlike the weak and positive correlation for local pigs (r = 0.20; P < 0.05). There was a quadratic relationship between temperature and average daily gain (ADG) for both pig genotypes. The regression coefficients for ADG were higher (P < 0.001) for LW than local pigs. It was concluded that at high ambient temperatures, performance of local pigs was less compromised than for LW pigs. Although local pigs were found to be heat tolerant, results of choice experiment showed that this trait was not selected for relative to other traits. Keeping pigs that required bought-in feeds, fell sick often and produced low pork quality (eating quality based on farmer perceptions) negatively affected farmers‟ livelihoods more in subsistence- than market-oriented production system. Farmers in market-oriented production system derived more benefit from productive traits such as heavier slaughter weights and large litter size than subsistence-oriented farmers. Under the subsistence-oriented production system, farmers in CSF-affected areas placed high prices on adaptive traits than the unaffected areas. Subsistence-oriented farmers who were affected by CSF wanted a total compensation price of R10 944.00 (USD1563.43) for keeping a pig genotype with unfavourable traits when compared to R4235.00 (USD605.00) for their CSF-unaffected counterparts. Implicit prices for traits could not be determined for market-oriented production system. It was concluded that farmers in CSFaffected areas placed high economic values on pig traits than farmers from the CSF-unaffected areas. The findings suggest that adapted local pigs can be promoted in subsistence-oriented production systems while productive imported pigs and their crosses with local pigs can be kept in market-oriented production systems.