Western Blot no imunodiagnóstico de lentivírus de pequenos ruminantes.

Background: Small ruminant lentivirus (SRLV) belong to genus Lentivirus, family Retroviridae. These viruses cause caprine arthritis encephalitis (CAE) and maedi visna (MV), infectious diseases that cause economic, production, and reproductive losses. There are no effective treatments or vaccines for these diseases. Thus, early detection via serology has great importance for control of SRLV. Therefore, the objective of this review is to demonstrate the potential of the western blot (WB) test as an immunodiagnostic test for SRLV. Review: In general, immunodiagnosis of SRLV is performed via agar gel immunodiffusion (AGID) and indirect enzyme linked immunosorbent assay (ELISA), which can detect antibodies in several different biological samples but is used pref- erably with serum and blood plasma. However, WB has demonstrated efficacy in the early diagnosis of immunoglobulins against SRLV, presenting higher sensitivity and specificity than the serological tests usually used, because this technique can detect antibodies at a dilution as much as 256 times greater than that of AGID and 32 times greater than that of ELISA. SRLV infection and consequent immunological activation result in the induction of cellular and humoral responses. Ad-ditionally, around the third week, production of antibodies directed mainly toward viral capsid proteins (p25 and p28) occurs. After the fifth week, production of immunoglobulins directed toward other viral proteins occurs. Because of the persistence of SRLV infection, serology is considered to be the most practical means to diagnosis. Each serological test has a percentage specificity and distinct sensitivity, as well as advantages and disadvantages in its applicability. It should be noted that there is no gold standard test for diagnosis of SRLV infection. Moreover, SRLV are characterized by escape mechanisms such as genetic diversity, mutagenic potential, viral intermittence, and the process of compartmentalization, which make immunodiagnosis more difficult. In addition, positive animals tend to present unstable levels of antibodies over weeks, months, and even years. In this context, WB, with early antibody detection, has been proven to be a refined and more accurate technique than other immunodiagnostic tests for SRLV. WB allows the simultaneous resolution of several immunogenic antigens present in a sample, and this feature provides it with greater reliability, differentiates it from other immunological methods, and accredits it as a test of wide applicability. Epidemiological and immunological dynamics studies often use WB in the immunodynamic diagnosis of SRLV. Serum, blood plasma, and seminal plasma are typical biological materials used in the serological diagnosis of SRLV with WB, expanding its potential as an immunodiagnosis method. Conclusion: WB is the most accurate serological technique for SRLV. It is more capable of accurate diagnosis because the genetic diversity that characterizes such lentiviruses and their various immune system escape mechanisms routinely hinder traditional diagnosis. Additionally, this test has been used widely in studies of SRLV for various purposes, but mainly in studies of epidemiological and immunological dynamics, using serum, blood plasma, or seminal plasma. However, independently of the biological sample tested, WB maintains high sensitivity and precision in immunodiagnosis, making it a refined and valid technique for SRLV control programs

Bone grafts and substitutes in small animal orthopaedic reconstrutive surgery: a breaf review

We present here an overview of the scientific literature concerning the clinical application of autografts and allografts of the skeletal tissues and of the natural and synthetic bone graft substitutes available in small animal orthopedic surgery for resolution of congenital, traumatic and tumoral etiology situations or for reconstructive artroplastic surgeries. This article focuses the terminology, definitions and characteristics of the bone grafts and bone graft substitutes based in their functions and composition, the process of incorporation of the bone grafts and bone graft substitutes, the collection of autogenous cancellous or corticocancellous bone for grafting, the available commercial forms of bone graft or bone graft substitutes and the biologic/synthetic composite grafts, other accessible options in small animal orthopedic surgery with osteogenic and/ or osteoindutive characteristics (ex. collection and use of the autogenous bone marrow or platelet rich plasma), the principal clinical indications and surgical techniques in which are recommended the use of bone graft and/or bone graft substitutes and how to select the best option for each specific clinical case. At last, the future perspectives in reconstructive orthopedic surgery are also pointed, namely the tissue engineering of bone and cartilage tissues and the medical three dimensional biomodulation.Neste artigo pretendeu realizar-se uma revisão da literatura científica da informação disponível sobre os vários tipos de auto- e aloenxertos do sistema musculo-esquelético e substitutos ósseos naturais e sintéticos do tecido ósseo para utilização em cirurgia ortopédica dos animais de companhia com vista à resolução cirúrgica de situações clínicas do foro da traumatologia e ortopedia de etiologia congénita, traumática, tumoral ou em implantações artroplásticas. Abordam-se assim os conceitos gerais sobre a terminologia, definições e classificação dos enxertos e substitutos ósseos com base nas suas funções e composição, o processo de incorporação dos enxertos e substitutos ósseos, os principais locais de obtenção e a correta técnica de transplante de enxertos ósseos autógenos, os principais tipos de enxertos e substitutos ósseos comercializados e os enxertos compostos, outras abordagens autógenas acessíveis em animais de companhia com potencial osteogénico e/ou osteoindutor (ex. obtenção e utilização de medula óssea autógena ou plasma enriquecido em plaquetas), as principais situações clínicas e técnicas cirúrgicas em que se recomenda a utilização de enxertos e/ou substitutos ósseos e como selecionar em cada caso clínico a melhor opção disponível. Por fim, são também abordadas as perspetivas futuras em cirurgia ortopédica reconstrutiva, nomeadamente a engenharia do tecido ósseo e cartilagem e a biomodelação tridimencional médica

Agronomic descriptors and ornamental potential of passion fruit species.

Abstract: Brazil has great variability, being one of the centers of origin of the genus Passiflora. Passion fruit has a multiple and diversified uses, being little explored as an ornamental plant in the country. The objective of this work was to characterize four passion fruit genotypes (two hybrid materials from the experimental field of passion fruit breeding program of the University of Brasília, and two passion fruit cultivars - BRS Sertão Forte and BRS Mel do Cerrado, both of them from Brazil), based on official and validated morphoagronomic descriptors for the identification of morphological aspects and ornamental potential, under field conditions, in the Distrito Federal, Brazil. The experiment consisted of a randomized block design, with four treatments, four replications, and six plants per plot. Plants were analyzed weekly regarding the characteristics of the branches, leaf blade, petiole, and flowers, with 24 measurements of each structure in the four treatments, which were recorded using a digital camera. Flowers presented pronounced size and color, colored branches, and leaves of varying sizes and shapes. All materials studied showed commercial ornamental potential to be explored as new options for ornamental climbing plants

Diversidade genética entre acessos do banco ativo de gemoplasma de batata-doce mantido na Embrapa Hortaliças.

Resumo

On the static solutions in gravity with massive scalar field in three dimensions

We investigate circularly symmetric static solutions in three-dimensional gravity with a minimally coupled massive scalar field. We integrate numerically the field equations assuming asymptotic flatness, where black holes do not exist and a naked singularity is present. We also give a brief review on the massless cases with cosmological constant.Comment: 11 pages, LaTeX, 1 Postscript figure. Some changes were don

Immune response dynamics of recent and chronic small ruminant lentivirus infection in the male reproductive system.

Resumo: O objetivo desse estudo foi analisar a resposta imunológica aos lentivírus de pequenos ruminantes (LVPR) com enfoque no sistema reprodutor de machos com infecção recente e crônica. Para isso, foram selecionados 12 reprodutores caprinos, sendo seis soronegativos e seis soropositivos com infecção natural crônica há mais de 18 meses (grupo com infecção crônica). Após seleção dos animais, os seis machos soronegativos foram inoculados com a cepa viral do vírus da artrite encefalite caprina (CAEV)-Co, título 10- 5,6 TCID50/mL, por via intravenosa. A partir da inoculação viral este agrupamento passou a ser denominado de grupo com infecção recente e juntamente com o grupo com infecção crônica foram acompanhados, semanalmente por 180 dias, com análise dos testes de Western Blot (WB) no soro sanguíneo e plasma seminal. Nos animais com infecção crônica para LVPR, 18,94% (50/264) apresentaram anticorpos antiLVPR em pelo menos uma das distintas amostras, e 81,06% (214/264) tiveram resultados negativos. Das amostras de soro sanguíneo do referido grupo, em 27,27% (36/132) detectou-se anticorpos anti-LVPR, enquanto que no plasma seminal 10,60% (14/132) foram reagentes no teste de WB. Nos animais inoculados com o CAEV-Co, ocorreu a soroconversão após a terceira semana da inoculação viral. Nesse grupo, 31,06% (41/132) das amostras de plasma seminal tiveram anticorpos anti-LVPR, sendo que dessas 41, 70,73% (29/41) coincidiram com resultado das amostras de soro sanguíneo. Nos 29,27% (12/41) restante, houve a detecção somente no plasma seminal e eram amostras provenientes de três animais (AIR2, AIR3 e AIR5). Em um dos animais com infecção recente, só foi identificado anticorpos anti-LVPR em amostras de plasma seminal, possivelmente em função da compartimentalização do vírus. Intermitência viral foi observada em ambas as amostras biológicas, independentemente do estágio de infecção. Conclui-se que a resposta imunológica em reprodutores com infecção recente LVPR é mais acentuada do que em animais cronicamente infectados. E, independentemente do estágio da infecção há uma flutuação nos níveis de anticorpos, sendo, portanto, um fator de risco, em virtude da existência de amostras falso-negativo ao realizar o diagnóstico.Título em português: Dinâmica da resposta imunológica da infecção recente e crônica por lentivírus de pequenos ruminantes no sistema reprodutor de machos

Avaliação da toxicidade de solvente de extratos vegetais com ação antiviral em sêmen caprino refrigerado.

[Evaluation of Solvent Toxicity of Plant Extract with Antiviral Action in Refrigerated Goat Semen]. Background: Caprine Arthritis Encephalitis Virus have been detected in sperm of breeding goats causing economic losses. In order to control the virus, researches aiming to identify natural extracts with potential antiviral effects are performed. However, aqueous or ethanolic extracts must be diluted in dimethyl sulfoxide (DMSO), which is a substance with unknown effects in sperm quality when present in diluting media. Therefore, this study aimed to evaluate sperm viability of refrigerated caprine semen diluted in media containing DMSO. This was performed to provide data that aid in researches involving the use of this component with natural extracts that may inactivate the caprine lentivirus in sperm. Materials, Methods & Results: The experiment was performed at the Laboratory of Seminal Technology in Embrapa Goats and Sheep in the city of Sobral, Brazil. Sperm viability was assessed in caprine semen refrigerated in two dilution media with crescent concentrations of DMSO. Sperm samples of five goats seronegative for the caprine lentivirus were pooled and diluted in minimal essential medium (MEM) enriched with glucose at 0.01 M added of crescent concentrations of DMSO (0%, 1.5%, 1.75%, 2.0%, 2.25% and 2.5%). The same breeders provided the pool of sperm to test Tris added 2.5% of egg yolk and the same concentrations of DMSO previously mentioned. Treatments were refrigerated at 7°C and evaluated up until four h after DMSO addition. Individual progressive motility (MIP), sperm vigor (V), percentage of spermatozoa reactive to hypoosmotic test (HO) and morphologically normal (NOR) were evaluated. IPM, vigor and NOR remained within normal standards for the caprine species in all treatments test. Percentage results of spermatozoa reactive to hypoosmotic was higher in Tris yolk with values ranging between 34.66% to 46.33%. Sperm vigor was positively correlated (r = 0.85) with IPM in the MEM diluted pool of sperm. In Tris yolk, vigor and hypoosmotic test correlated moderately (r = 0.63, r = 0.54, respectively) with IPM. Tris yolk medium added DMSO presented the highest percentage of reactivity to hypoosmotic test in all treatments when compared to MEM added DMSO. Discussion: The fact that DMSO is easily homogenized in water, ethylic alcohol and most organic solvents favors its use in diluting natural extracts. These components are a possible source of products that inactivate caprine arthritis encephalitis virus in sperm, which is the key to promoting the safe use of genetic material of infected breeders, in addition to commercial use of germplasm. In this study, there was no interference of DMSO in the analyzed parameters when added in a maximum concentration of 2.5% to MEM and Tris yolk, which is in accordance with standard values for goats. In addition, Tris yolk may promote greater protection to the membrane of sperm cells, which was demonstrated by hypoosmotic test. This medium could be ideal to be used in new methodologies that incorporate DMSO. In conclusion, DMSO added to dilution media Tris yolk and MEM did not interfere with the quality of refrigerated caprine sperm, which maintained viability. These results indicate that this substance did not present harmful effects to the genetic material, promoting the use as solvent of extracts from plant compounds with potential anti-viral effect. The information in this study may aid new research performed in this area.[ALICE ANDRIOLI PINHEIRO]

Mycoplasma agalactiae em rebanhos leiteiros no estado do Ceará em associação com o vírus da artrite encefalite caprina.

Background: Contagious agalactia is an infectious disease caused by Mycoplasma agalactiae (M. agalactiae) that occurs in small ruminants leading to productive and economic losses. Due to the similarity of clinical signs presented in Caprine Arthritis Encephalitis (CAE), which is a viral disease, a differential diagnosis is important. Therefore, this study aimed to investigate the presence of anti-Mycoplasma agalactiae antibodies in dairy goat flocks in Ceará State and possible correlation with CAE. Materials, Methods & Results: The research was performed in four mesoregions in Ceará State (Metropolitan Region of FortalezaMRF; Northeast Ceará - NeC; North Ceará - NC; Sertões in Ceará - SC), from which 16 productions located in 10 cities with the highest representativeness for goat milk production within the State or mesoregion were sampled. A total of 417 females and 69 males (486 animals) of breeds with dairy production aptitude, pure or crossbreed, maintained in semi-intensive or intensive systems, were tested. Blood serum was obtained by venipuncture of the jugular vein with vacuum pressure syringe followed by centrifugation at 1,500 g for 10min. Antibodies against the caprine arthritis encephalitis virus (CAEV) were detected with micro technique of agarose gel immunodiffusion (AGID) and Western Blot (WB). The anti-Mycoplasma agalactiae antibodies were detected with commercial kit of enzymatic immunoassay (IDEXX Laboratories). Seroprevalence of M. agalactiae in dairy goat flocks in Ceará State was 0.62% (3/486). From the total of 16 visited productions, 18.75% (3/16) had seropositive animals for M. agalactiae located in MRF, NC and SC mesoregions. CAE was diagnosed in 56.25% (9/16) of productions with AGID and in 81.25% (13/16) with WB. In addition, 5.2% (25/486) of animals were seropositive for CAE with AGID and 16.6% (80/486) with WB. Animals that reacted positive for M. agalactiae were all females of pure breed with milk production aptitude in distinct mesoregions submitted to intensive rearing system. None of these animals was positive in neither test (AGID or WB) for CAE. Therefore, no correlation of results obtained in diagnosis of M. agalactiae by ELISA and CAEV by AGID or WB (P < 0.05) was identified. However, two out of three productions that were positive for M. agalactiae presented positive results for CAEV with frequencies of 10% and 20%. Discussion: Seroprevalence of M. agalactiae in Ceará State was low in comparison with other Brazilian states and even other countries. However, the presence of the pathogen in more than one mesoregion indicates that the disease occurs in different locations within the State. Therefore, flocks in Ceará are susceptible to the infection, which may be favored by uncontrolled commerce that occurs with deficient surveillance, associated with the importation of animals to improve flock genetic quality. The presence of the pathogen in dairy goats may contribute to significant losses in the local production. On the other hand, CAE was diagnosed in nearly all productions proving the dissemination of this lentivirus infection among dairy goat flocks in Ceará State. Although an association between these diseases was not identified, the presence of a retrovirus in the organism may favor co-infection with another micro-organism, promoting the deficiency in the immune system of the host. In conclusion, M. agalactiae is present in different mesoregions of the Ceará State and control measures should be adopted in short term to prevent pathogen dissemination and, consequently reduce economic and productive losses in the local dairy goat production. No correlation was identified between the prevalence of infection by CAEV and M. agalactiae in this study. [Mycoplasma agalactiae in dairy goat flocks bred in Ceará State of Ceará in association with Caprine Arthritis Encephalitis Virus].[ALICE ANDRIOLI PINHEIRO]