THE PULMONARY AND METABOLIC EFFECTS OF SUSPENSION BY THE FEET COMPARED WITH LATERAL RECUMBENCY IN IMMOBILIZED BLACK RHINOCEROSES (DICEROS BICORNIS) CAPTURED BY AERIAL DARTING.

Aerial translocation of captured black rhinoceroses (Diceros bicornis) has been accomplished by suspending them by their feet. We expected this posture would compromise respiratory gas exchange more than would lateral recumbency. Because white rhinoceroses (Ceratotherium simum) immobilized with etorphine alone are hypermetabolic, with a high rate of carbon dioxide production (VCO2), we expected immobilized black rhinoceroses would also have a high VCO2. Twelve (nine male, three female; median age 8 yr old [range: 4-25]; median weight 1,137 kg [range: 804-1,234] body weight) wild black rhinoceroses were immobilized by aerial darting with etorphine and azaperone. The animals were in lateral recumbency or suspended by their feet from a crane for approximately 10 min before data were collected. Each rhinoceros received both treatments sequentially, in random order. Six were in lateral recumbency first and six were suspended first. All animals were substantially hypoxemic and hypercapnic in both postures. When suspended by the feet, mean arterial oxygen pressure (PaO2) was 42 mm Hg, 4 mm Hg greater than in lateral recumbency (P=0.030), and arterial carbon dioxide pressure (PaCO2) was 52 mm Hg, 3 mm Hg less than in lateral recumbency (P=0.016). Tidal volume and minute ventilation were similar between postures. The mean VCO2 was 2 mL/kg/min in both postures and was similar to, or marginally greater than, VCO2 predicted allometrically. Suspension by the feet for 10 min did not impair pulmonary function more than did lateral recumbency and apparently augmented gas exchange to a small degree relative to lateral recumbency. The biological importance in these animals of numerically small increments in PaO2 and decrements in PaCO2 with suspension by the feet is unknown. Black rhinoceroses immobilized with etorphine and azaperone were not as hypermetabolic as were white rhinoceroses immobilized with etorphine.

A 3-D airway epithelial cell and macrophage co-culture system to study Rhodococcus equi infection.

We developed a 3-D equine bronchial epithelial cell (BEC) culture that fully differentiates into ciliary beating and mucus producing cells. Using this system, we evaluated how mucus affects the phagocytic activity of macrophages. Adult horse monocyte-derived macrophages were incubated with Rhodococcus equi for 4h either in the mucus layer of in vitro generated airway epithelium or on collagen coated membranes. Using light and electron microscopy, we noted that the number of macrophages with intracellular bacteria, and the number of intracellular bacteria per macrophage were lower in the presence of mucus. TNFα measurements revealed that the presence of BECs promoted TNFα production by R. equi-infected macrophages; a decrease in TLR-2 (involved in R. equi recognition) and an increase in EGF-R (involved in mucin production) mRNA expression were also noted. Interestingly, when foal macrophages were added to foal BECs, we made the opposite observation, i.e. many macrophages were loaded with R. equi. Our in vitro bronchial system shows great potential for the identification of mechanisms how BECs and mucus play a role in phagocyte activation and bacterial clearance. Further studies using this system will show whether the airway environment in the foal responds differently to R. equi infection.

Detection of rubella virus RNA in serum samples by RT-PCR assay / Detecçäo do RNA do vírus da rúbeola no soro através de RT-PCR

Em geral, a reaçäo de polimerizaçäo em cadeia precedida de retrotranscriçäo (RT-PCR) tem demonstrado ser um método rápido e sensível para detectar o RNA do vírus rubéola em uma variedade de materiais clínicos. A reaçäo de RT-PCR foi utilizada para detectar o vírus no soro e confirmar o teste sorológico em indivíduos infectados com vírus da rubéola. Para este estudo, uma fita simples de RNA viral, extraída do soro, foi usada como fita-molde para para a transcriçäo reversa, seguida da amplificaçäo com dois diferentes pares de olinucleotídeos iniciadores e de uma segunda amplificaçäo com outros pares de oligonucleotídeos. O método do RT-PCR foi utilizado para se examinar amostras de soro de nove pacientes com sorologia confirmada para o vírus da rubéola. Quatro soros testados por RT-PCR tiveram como resultado infecçäo pelo vírus da rubéola. Os soros foram primeiramente analisados utilizando-se de 500µl para a extraçäo de RNA. O vírus da rubéola foi também analisados utilizando-se 500µl para a extraçäo de RNA. O vírus da rubéola foi também analisado utilizando-se volumes menores de soro, e somente uma amostra foi amplificada a partir da extraçäo do RNA viral de 300µl. Neste estudo, a estratégia de se utilizar soro no desenvolvimento do teste RT-PCR sugere ser uma alternativa a mais para o diagnóstico da infecçäo pelo vírus da rubéola

Detecçäo de vírus Dengue sorotipos 1 e 2 pela técnica de immunoblotting utilizando proteínas recombinantes / Detection of dengue virus serotypes 1 and 2 by immunoblotting techniques using recombinant proteins

O vírus dengue säo patógenos que vêm afetando milhöes de pessoas durante os dois últimos séculos. No presente trabalho, comparou-se a técnica tradicional de MAC-ELISA e o teste de Immunoblotting, utilizando-se proteínas recombinantes, para pesquisa de anticorpos da classe M, contra os vírus Dengue sorotipos 1 e 2. Foram testadas 100 amostras de soro de pacientes, com suspeita clínica de dengue, por MAC-ELISA e Immunoblotting. Estes soros quando submetidos ao MAC-ELISA resultaram 56 positivos para dengue e 44 negativos. Estas mesmas amostras avaliadas por Immunoblotting erevelaram 56 soros negativos, 36 soros positivos para DEN-1, 6 para DEN-2 e 2 para DEN-1 e 2. Proteínas recombinantes (DEN-1 e DEN-2) utilizadas como antígeno em Immunoblotting possibilitam um diagnóstico diferencial dos sorotipos de dengue circulantes em área, enquanto o teste MAC-ELISA apesar de sensível e rápido näo tipifica o sorotipo de dengue, dado importante durante os períodos de epidemias. (AU)

Dengue viroses (DEN-1 to 4) are human pathogens affeeting millions of people In the last two eenturies. Results of a eomparative study between the traditional MAC-ELISA method and the Immunoblotting teehniques using reeombinant proteins of DEN-l and DEN-2 to deteet vírus speei- fie class M Immunoglobulins, are reported in this paper. One hundred sera samples were studied. Fifty six samples were positive to dengue by MAC-ELISA and 44 were positive by Immunoblotting teehni- que: 36 to DEN-1, 6 to DEN-2 and 2 to DEN-1 and 2 sorotypes. So, the serotype identifieation of the dengue vírus eireulantig in a determined área is possible by using these reeombinant proteins in Immunoblotting teehniques. These data are importante for epidemiologieal surveillanee mainly during the oeeurrenee of an epidemie. (AU)

Growth and maintenance of Aedes Albopictus cell line, clone C6/36, in different media

The sensitivity of Aedes albopicts cell line, clone C6/36, to arbovirus isolation and growth has been being shown by several authors. The present work has verified which, among several media under the same conditions, would be the most efficient one for C6/36 cultivation and viral isolation. The L-15 medium has proved to be the best among others (MEM,DMEM and 199) with the quickest viral isolation(DEN-1). Also, in this medium, the samples could be observed until the 14th day, without significative cellular death. The results reccommend L-15 medium as the most efficient and economic one for the purposes