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The epidemiological, clinic and morphological (pathological and ultrastructural) aspects of four outbreaks of copper deficiency affecting 21- to 90-day-old pigs in the Northeast region of Brazil are reported. Clinical signs began with paraparesis and ataxia and progressed to flaccid or spastic paralysis of the pelvic and thoracic limbs, followed by sternal and/or lateral recumbence. In addition, some animals showed dog-sitting position and intention tremors. The clinical manifestation period was 5-20 days. Significant gross lesions were not observed; however, microscopically, symmetrical degeneration of the white matter with ballooned myelin sheaths containing occasional macrophages was observed, mainly in the spinal cord. Two pigs presented with necrosis ad loss of Purkinje cells and ectopic Purkinje cells in the granular layer and cerebellar white matter. A ultrastructural analysis showed different degrees of damage of myelinated axons in the spinal segments, including an absence of the axoplasm structures with only axonal residues remaining. The myelin sheaths were degenerated and often collapsed into the space previously occupied by the axon. These results suggest that myelin degeneration is secondary to the axonal lesion. Finally, the concentration of copper in the liver was determined using atomic absorption spectrophotometry and was found to be low (ranging from 2.2 to 10.8 ppm). In conclusion, in the Brazilian semiarid region, Cu deficiency occurs in 21 to 90-day-old pigs that ingested different types of waste in their food. RESUMO: São relatados os achados epidemiológicos, clínicos e morfológicos (patológicos e ultraestruturais) de quatro surtos de deficiência de cobre em suínos afetados entre 21 e 90 dias de idade na região Nordeste do Brasil. Os sinais clínicos iniciaram com paraparesia e ataxia, que progrediu a paralisia flácida ou espástica dos membros pélvicos e torácicos, seguido de decúbito esternal e/ou lateral. Além disso, alguns animais apresentaram posição de cão sentado e tremores de intenção. O período de manifestação clínica variou de 5-20 dias. Não foram observadas lesões macroscópicas significativas; no entanto, microscopicamente, foi observada degeneração simétrica da substância branca com fragmentação das bainhas de mielina, contendo ocasionais macrófagos, principalmente na medula espinal. Dois suínos apresentaram necrose e perda de células de Purkinje e células de Purkinje ectópicos na camada granular da substância branca cerebelar. A análise ultraestrutural mostrou diferentes graus de lesões em axônios mielinizados em segmentos da medula espinhal, incluindo o desaparecimento de estruturas do axoplasma, restando apenas restos axonais. A bainha de mielina encontrava-se degenerada e muitas vezes, colapsada dentro do espaço previamente ocupado pelo axônio. Esses resultados sugerem que a degeneração da mielina é secundária à lesão axonal. Finalmente, a concentração do cobre no fígado foi determinada usando espectrometria de absorção atômica e revelou baixos valores (variando de 2,2-10,8ppm). Conclui-se que na região semiárida do Brasil ocorre deficiência de cobre em suínos de 21 a 90 dias de idade alimentados com diferentes tipos de resíduos.

Nos anos de 2001 e 2002, 46 cabritos (CAB) e 35 cordeiros (COR) de uma propriedade do agreste do Estado de Pernambuco foram acometidos por ataxia enzoótica (AE) de forma tardia. Houve aumento da incidência do 1o ano (46,3% - CAB; 24,2% COR) para o ano subseqüente (100% - CAB e COR). Somente no ano de 2001, houve maior incidência em CAB (P

O trabalho objetivou conhecer os teores de Cu, Mo, Fe e Zn em soro e fígado de ovinos e caprinos criados na região semiárida do estado de Pernambuco e abatidos nas épocas da chuva e seca, e estabelecer se a carência de Cu é causada por deficiência primária de Cu ou secundária à ingestão de quantidades excessivas de Fe ou Mo. Amostras de soro e fígado de 141 ovinos e 141 caprinos foram submetidas à digestão úmida em ácido nítrico-perclórico e, posteriormente, analisados em espectrofotômetro de absorção atômica indutivamente acoplado (ICPOES). A concentração sérica de Cu em caprinos teve uma média de 9,85±2,71µmol/L e em caprinos de 11,37±2,57µmol/L, enquanto que a concentração hepática média foi de 158,45±83,05mg/kg para ovinos e 152,46±79,58mg/kg para caprinos. Os teores séricos de Fe foram de 35,58± 14,89µmol/L em ovinos e de 25,06±8,10µmol/L em caprinos e as concentrações no fígado foram de 156,10±55,99mg/ kg em ovinos e 210,53±121,99mg/kg em caprinos. As concentrações médias séricas de Mo foram de 0,28±0,11µmol/ L em caprinos e 0,31±0,16µmol/L em ovinos, enquanto que no fígado sua concentração foi de 6,53±4,13mg/kg e 8,10± 4,01mg/kg, respectivamente. A concentração sérica de Zn foi de 11,9±6,07µmol/L em ovinos e 11,79±7,42µmol/ L em caprinos e a concentração no fígado foi de 126,43 ±51,50mg/kg e 132,91±55,28mg/kg em ovinos e caprinos, respectivamente. Verificou-se variação na concentração destes minerais considerando os fatores de variação, como o período sazonal, espécie e sexo. Baseado nos valores observados, considerados marginais, e na ocorrência de surtos de ataxia enzoótica em caprinos e ovinos na região, recomenda-se a suplementação com Cu em animais a campo, tanto na seca quanto na chuva. Considerando as concentrações séricas e hepáticas de Fe e Mo, sugerese que as concentrações marginais de Cu não estejam diretamente relacionadas com o excesso destes minerais. Não foram encontradas diferenças significantes nas concentrações séricas e hepáticas de Cu e Mo e nas concentrações séricas de Fe entre as amostras coletadas na época da chuva a as coletadas na época seca. As concentrações hepáticas de Fe e as concentrações séricas e hepáticas de Zn foram significativamente maiores na época da chuva. Considerando que os teores de Zn sérico encontram-se abaixo dos valores considerados como marginais e os hepáticos dentro do limite de normalidade, embora próximos ao limite inferior da referência, recomenda-se a suplementação com Zn, principalmente durante o período da seca. The aim of this research was to determine serum and liver concentrations of Cu, Mo, Fe and Zn in sheep and goats slaughtered in the semiarid region of the state of Pernambuco, northeastern Brazil, during the rainy and dry seasons, and to establish if Cu deficiency which occurs in the region is primary or secondary to high levels of Mo and/or Fe. Serum and liver samples from 141 goats and 141 sheep were submitted to wet digestion in nitric-perchloric acid and analyzed by coupled plasma atomic absorption spectrophotometry (ICPOES). The mean serum concentrations of Cu were 9.85±2.71µmol/L in sheep and 11.37±2.57µmol/L in goats. Mean Cu liver concentrations were 158.45±83.05mg/kg in sheep and 152.46±79.58mg/ kg in goats. Mean serum concentrations of Fe were 35.58±14.89µmol/L in sheep and 25.06±8.10µmol/L in goats. Mean liver concentrations of Fe were 156.10±55.99mg/kg in sheep and 210.53±121.99mg/kg in goats. Mean serum concentrations of Mo were 0.28±0.11µmol/L in goats and 0.31±0.16µmol/L in sheep. Mean liver concentrations of Mo were 6.53±4.13mg/kg in goats and 8.10±4.01mg/kg in sheep. Mean serum concentrations of Zn were 11.9±6.07µmol/L in sheep and 11.79±7.42µmol/L in goats. Mean liver concentrations of Zn were 126.43±51.50mg/kg in sheep and 132.91±55.28mg/kg in goats. Based on Cu serum and liver concentrations and on the sporadic occurrence of enzootic ataxia, Cu supplementation is recommended for grazing sheep and goats during the dry and the rainy seasons. Considering that serum and hepatic concentrations of Fe and Mo are within normal ranges or are marginal, it is suggested that the low Cu concentrations are not related with high Fe or Mo ingestion. Non significant differences were observed between serum and liver concentrations of Cu and Mo, and between serum concentrations of Fe in samples collected during the rainy season and the dry season. Liver concentrations of Fe and serum and liver concentrations of Zn were significantly higher during the rainy season. Taking into account that serum and liver Zn concentrations in sheep and goats are considered marginal, supplementation with Zn is suggested for grazing sheep and goats, mainly during the dry season.

Fil: Trulls, Horacio Eduardo. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina. Fil: Brem, J. J. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina. Fil: Sánchez Negrette, Marcial. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina. Fil: Pochon, Daniel Osvaldo. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina. Fil: Ortíz, M. L. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina. La acción tóxica del molibdeno, por sí mismo o por su interacción con el metabolismo del cobre, podría ocasionar trastornos sobre diversos tejidos. Con el fin de provocar una deficiencia secundaria de cobre, se procedió al agregado de tetratiomolibdato de amonio (TTMo) en la dieta de ratas, para evaluar sus efectos sobre la función reproductiva. En rumiantes, la toxicidad del Mo es provocada por la formación de tetratiomolibdatos por la microflora ruminal. Con este producto sintetizado in vitro, se pudo reproducir la enfermedad en ratas que, como el resto de los monogástricos, son particularmente resistentes a la intoxicación natural del Mo por carecer del ambiente digestivo adecuado para dicha transformación. Para lograr el objetivo propuesto, se valoró la respuesta de la testosterona tras el estímulo de gonadorrelinas (GnRH) y mediante estudios anatomohistopatológicos se determinó la integridad estructural del tejido testicular. Se utilizaron 40 ratas de sexo macho de la cepa Wistar, de 3 meses de edad; los grupos tratados recibieron durante 120 días dosis de 3 mg de TTMo/100 ml de agua de bebida. En este ensayo de molibdenosis experimental se logró una sintomatología clínica y bioquímica compatible con una deficiencia secundaria de cobre. Las diferencias entre grupos tratados y testigos en la respuesta de testosterona después de la inyección de GnRH, fueron significativas, indicando una función endocrina comprometida. Los ejemplares tratados revelaron disminución del tamaño testicular, túbulos seminíferos con fenómenos degenerativos del epitelio germinal, necrosis y desaparición de espermátides y espermatozoides. Algunos testículos presentaban únicamente espermatogonias y células de Sertoli con fenómenos de vacuolización citoplasmática, picnosis nuclear ycélulas gigantes fagocíticas multinucleadas. Las lesiones en el tejido intersticial se caracterizaron por fibrosis leve y depleción de células intersticiales de Leydig. Molybdenum toxicity may cause disturbances on several tissues either by itself or through its interaction with copper metabolism. To develop a secondary copper deficiency, tetrathiomolybdenum (TTMo) was supplemented to male rats to evaluate its effects on reproductive performance. In ruminants, Mo toxic action is due to the production of tetrathiomolybdates by ruminal microflora. With this in vitro synthesized metabolite, it was possible to reproduce this ruminant disease syndrome in rats, a monogastric species that is particularly resistant to the natural intoxication of Mo, considering the inappropriate digestive environment for such transformation. The objective of this work was to measure testosterone response after gonadorreline (GnRH) injections, as well as to perform anatomopathologic studies to determine the structural integrity of testicular tissues. Forty male 3 months–old Wistar rats, were used. Treated group received daily doses of 3 mg of TTMo/100ml in drinking water during 120 days. In this experimental molybdenosis assay, clinical and biochemical symptomatologies were correlated with a secondary deficiency of copper. The differences between treated and control groups regarding to testosterone response after GnRH injection were significant, showing an abnormal endocrine function. Morphological studies of other two lots showed a decrease in testicular size of treated animals, and seminiferous tubules with degenerative phenomena of the germinal epithelium, necrosis and spermatozoid absence. Some testicles presented only spermatogonia and/or Sertoli cells with citoplasmatic vacuolation, nuclear picnosis and multinucleated phagocytic gigant cells. Lesions of the interstitial tissue were mild fibrosis and depletion of Leydig interstitial cells.

An outbreak of sudden death was observed in cattle in southern Brazil. No significant gross or microscopic lesions were found in 10 cattle at post-mortem examination. To test if the sudden deaths were caused by a toxic plant, 13 weeds were collected and given to rabbits at a total of 440 to 600 g per kg body weight within 7 days. Results were negative. Mean copper levels of the liver in 10 cattle were very low (3.6±1.6 ppm-dry matter) suggesting Cu deficiency as the cause of the disease. Five samples of pasture collected during the outbreak had normal levels of Cu (8.4±0.8 ppm-d.m.) and S (0.2%±0.03-d.m.), but high levels of Fe (522±122 ppm-d.m.). One group of 10 heifers was supplemented with Cu subcutaneously. This group and a similar control group were maintained in the same area where the outbreak occurred. Serum levels of Cu, S, Fe, Mo and ceruloplasmin were determined bimonthly in both groups during one year. Mean serum levels of Cu (1.76±1.06 to 10.34±3.1 μmol/l for the control group and 3.86±1.53 to 10.61±1.34 μmol/l for the treated group) and ceruloplasmin (6.59±3.93 to 18.61±4.14 mg/l for the control group and 10.31±5.48 to 32.49±6.05 mg/l for the treated group) were significantly higher in the supplemented group (P=0.0046 for Cu and P=0.0001 for ceruloplasmin), but they were below normal levels in most samples of both groups. Serum levels of Cu and ceruloplasmin were correlated (r=0.67; P=00.5). In both groups serum levels of Fe (40.09±5.25 to 78.48±28.23 μmol/l) were higher than normal levels. Samples of forage were collected bimonthly for determination of Cu, Mo, S, Fe and protein in 7 points of the paddock where the outbreak occurred. Levels of Cu (1.36±0.56 to 4.76±1.15 ppm-d.m.) were below the normal ranges. The concentration of Mo (0.17±0.06 to 0.96±0.47 ppm-d.m.) was within normal ranges. Levels of S (0.21±0.04% to 0.5±0.17%) and Fe (172.92±62.64 to 437.24±205.44 ppm-d.m.) were occasionally within toxic level. Levels of protein varied from 7.77±2.6% to 13.16±3.02%. Six samples of water and six of submersed pastures were collected at the end of the experiment when the paddock was flooded. High levels of iron (169.23±83.49 ppm-d.m. ) and S (0.06±0.03%) were found in the water. Levels of Fe and S in the submerse pasture were of 469.3±218.28 ppm and 0.19±0.05%, respectively. The results obtained suggest that the sudden deaths are due to Cu deficiency. Three factors seem to be responsible for the Cu deficiency: 1) high levels of iron in pastures and water; 2) occasionally low levels of copper in the pastures; and 3) the ingestion of S above the requirements due to the levels occasionally high in pastures and high in water. The outbreak reported is similar to other outbreaks of sudden deaths associated with low liver copper previously reported during winter in the same region of southern Brazil. This area is characterized by acid sandy soils with frequent flooding during this season. In periodically flooded acid soils iron is solubilized and absorbed by the plants, decreasing the Cu content and increasing the Fe content of forage. This seems to be the main reason for Cu deficiency in the region., Facultad de Ciencias Veterinarias

Fil: Brem, J. J. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina. Fil: Trulls, Horacio Eduardo. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina. Fil: Sánchez Negrette, Marcial. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina. Fil: Ortíz, M. L. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina. El tetratiomolibdato (TTMo) se sintetiza en los pre–estómagos de los rumiantes por reaccióndel molibdeno con los sulfuros producidos por reducción bacteriana de los elementosazufrados de la dieta. TTMo sería el responsable tanto del bloqueo de la absorción del cobrecomo de su posterior utilización tisular, provocando una deficiencia secundaria del mismo.El objetivo del trabajo fue verificar el efecto del TTMo de amonio sobre la histología dehuesos largos en este modelo experimental con el fin de aportar información acerca de la fragilidadósea que presentan los rumiantes con molibdenosis. Se utilizaron 30 ratas Wistar desexo macho, de 80 a 90 días de edad, agrupadas en forma aleatoria en lotes testigos y tratados,conformados con 15 animales cada uno. Estos últimos recibieron 54 mg/l de TTMo conel agua de bebida durante 100 días y, teniendo en cuenta el contenido de Cu y Mo de la dietay la cantidad de agua y alimento consumido, el lote tratado fue sometido a un desafío de 40ppm de Mo. Los animales mostraron sintomatología clínica claramente compatible con unadeficiencia secundaria de Cu por molibdenosis. Las lesiones más importantes observadas enel tejido óseo de los animales tratados consistieron en engrosamiento del cartílago de crecimiento,con masas de condrocitos hipertróficos y falta de distribución ordenada de células enforma columnar. El tejido subperióstico mostró un incremento de espesor y hemorragias entoda la extensión de diáfisis y metáfisis. También se observó falta de reemplazo del cartílagoy hueso inmaduro por tejido óseo maduro, acompañado por abundante producción de tejidofibroso, alterándose así la estructura ósea normal. Tales lesiones pudieron ser provocadaspor reacciones tisulares de expansión transversal metafisiaria por una mayor deformabilidadósea frente al desafío de cargas que el hueso debió soportar. Ammonium tetrathiomolybdate (TTMo) is synthesized by ruminants in their pre–stomach by means of the reaction of molybdate with sulfides which are produced by bacterial reduction from the sulfurous elements from the diet. TTMo would be responsible either for the blockage of copper absorption as well as for its later tissular use, leading to a secondary copper deficiency. The aim of this work was to demonstrate the effect of TTMo on the histology of osseous tissue from large bones in this experimental model to gather information regarding bone fragility of ruminants with molybdenosis. Thirty male Wistar rats 80–90 days of age, grouped at random in control and treated lots, 15 animals each, were used. The latter group received 54 mg/l of TTMo with drinking water during 100 days. Considering the content of Cu and Mo from the diet and the intake of water and food, the treated group was submitted to a challenge dose of 40 ppm of Mo. Animals showed clinical symptomatology compatible with a secondary copper deficiency. The most important injuries observed in osseous tissue of treated animals consisted in thickening of the growth cartilage, hipertrophic condrocites and disorder in column cells. The subperiostic tissue showed an increase of thickness and hemorrhagic areas in diafisys and metafisys as well. A lack in the replacement of cartilage and immature bone by mature bone tissue, together with a high production of fibrous tissue were observed. This indicates an alteration of the normal structure of bones. These morphologic lesions could be caused by tissue reactions of transverse metaphyseal expansion as a consequence of high deforming forces on bones.