Protective effect of ß-glucan and glutamine on intestinal and immunological damage in mice induced by cytarabine (Ara-C) / Efeito protetor do ß-glucano e glutamina em lesões intestinais e imunológicas induzidas por ciratabina (Ara-C) em camundongos

Recently, glutamine and ß-glucan have been demonstrated to play an important role in modulation of the immune system and in promoting intestinal health benefits. The aim of this study was to investigate the effect of this intervention on inflammatory responses and intestinal health in mice orally pretreated with soluble Saccharomyces cerevisiae derived 1,3/1,6-ß-glucan (80mg/kg) with or without glutamine (150mg/kg) and then challenged with cytarabine (Ara-C) (15mg/kg). Improvements in villi and crypts were not observed in the ß-glucan group. The intestinal morphometry in the glutamine group showed the best results. ß-glucan in combination with glutamine presented the highest values of IL-1ß and IL-10 and lowest values for leukocytes and INF-γ. Based on these results, combined ß-glucan and glutamine pretreatment reduced intestinal inflammation and improved the immune response after Ara-C challenge.(AU)

Recentemente, glutamina e ß-glucano têm demonstrado desempenhar um papel importante na modulação do sistema imune e na promoção de benefícios para a saúde intestinal. O objetivo deste estudo foi investigar o efeito dessa intervenção sobre as respostas inflamatórias e saúde intestinal de camundongos pré- tratados por via oral com 1,3/1,6-ß-glucano (80mg/kg) derivado de Saccharomyces cerevisiae com ou sem glutamina (150mg/kg) e posteriormente desafiados com citarabina (Ara-C) (15mg/kg). Melhoras em vilosidades e criptas não foram observadas no grupo de tratamento com ß-glucano. A morfometria intestinal no grupo de tratamento com glutamina apresentou os melhores resultados. O grupo em que foi utilizado ß-glucano em combinação com glutamina apresentou os maiores valores de IL-1ß e IL -10 e valores mais baixos para os leucócitos e INF-γ. Com base nestes resultados, o pré-tratamento de ß-glucano combinado com glutamina reduziu a inflamação intestinal e melhorou a resposta imune após o desafio com Ara-C.(AU)

Nickel allergy: blood and periodontal evaluation after orthodontic treatment.

The aim of this study was to assess periodontal status and blood parameters in orthodontic patients with nickel allergy one month after removal of brackets. Ninety-six randomly selected patients were initially evaluated. Allergy to nickel was diagnosed using a patch test. After determining the prevalence of subjects allergic to nickel, two groups were formed: 16 allergic (experimental) and 16 non-allergic (control) patients. Their periodontal status was determined regularly by a single, blinded, duly calibrated examiner using the Löe Index (GI) and their blood was tested (complete blood test, including nickel and IgE levels) after nine months of orthodontic treatment and again one month after removing the orthodontic appliances. Statistical analyses included paired and non-paired t-tests, Mann-Whitney, Wilcoxon, McNemar and linear trend chi-square tests (p≤0.05). Comparison of the values recorded during orthodontic treatment and one month after removing the appliances showed that in the allergic group there was significant increase in eosinophils (p=0.046), basophils (p=0.001) and monocytes (p=0.002), and decrease in number of bands (p=0.000), while in the control group, there was increase in lymphocytes (p=0.039) and decrease in segmented neutrophils (p=0.030) and IgE levels (p=0.001). In both groups, plasma nickel levels increased (p=0.010; p=0.039) and GI scores decreased. One month after removing the brackets, blood and periodontal parameters from patients with and without nickel allergy were similar.

O objetivo do presente estudo foi avaliar a condição periodontal e os parâmetros sanguíneos em pacientes alérgicos ao níquel, um mês após remoção dos aparelhos. Noventa e seis pacientes selecionados aleatoriamente foram inicialmente avaliadas. Alergia ao níquel foi diagnosticada usando um teste de contato. Após a determinação da prevalência de alergia ao níquel, formaram-se dois grupos: 16 pacientes alérgicos (experimental) e 16 não alérgicos (controle). Condição periodontal foi diagnosticada através do Índice de Löe (IG). Parâmetros sanguíneos foram determinados por meio de um exame de sangue completo, incluindo a quantificação de níquel e níveis de IgE. Avaliações do estado periodontal foram realizados por um único examinador de forma cega, devidamente calibrado e amostras de sangue foram tomadas depois de nove meses de tratamento e um mês após a remoção dos aparelhos ortodônticos. Análise estatística utilizada foi testes t pareado e não pareado, Mann-Whitney, Wilcoxon, McNemar e qui-quadrado de tendência linear (p≤0,05). Em comparação com os valores observados durante o tratamento, o número de eosinófilos (p=0,046), basófilos (p=0,001) e monócitos (p=0,002) aumentou significativamente depois da remoção dos aparelhos ortodônticos, ao passo que o número de bastões (p=0,000) diminuiu entre os períodos no grupo alérgico. O número de linfócitos (p=0,039) aumentou no grupo controle e o número de segmentados (p=0,030) diminuiu. A diminuição dos níveis de IgE (p=0,001) entre os períodos ocorreu no grupo de controle. Níveis de níquel no plasma aumentou após a remoção de aparelhos ortodônticos em ambos os grupos (p=0,010; p=0,039). O IG diminuiu em ambos os grupos. Parâmetros periodontais e sanguíneos de pacientes com alergia ao níquel foram semelhantes aos não alérgicos um mês após a remoção dos aparelhos.

Nickel allergy: blood and periodontal evaluation after orthodontic treatment / Alergia ao níquel: avaliação periodontal e sanguínea após o tratamento ortodôntico

The aim of this study was to assess periodontal status and bloodparameters in orthodontic patients with nickel allergy one monthafter removal of brackets. Ninety-six randomly selected patientswere initially evaluated. Allergy to nickel was diagnosed using apatch test. After determining the prevalence of subjects allergic tonickel, two groups were formed: 16 allergic (experimental) and 16non-allergic (control) patients. Their periodontal status wasdetermined regularly by a single, blinded, duly calibrated examinerusing the Lõe Index (GI) and their blood was tested (complete bloodtest, including nickel and IgE levels) after nine months oforthodontic treatment and again one month after removing theorthodontic appliances. Statistical analyses included paired andnon-paired t-tests, Mann-Whitney, Wilcoxon, McNemar and lineartrend chi-square tests (p≤0.05). Comparison of the values recordedduring orthodontic treatment and one month after removing theappliances showed that in the allergic group there was significantincrease in eosinophils (p=0.046), basophils (p=0.001) andmonocytes (p=0.002), and decrease in number of bands (p=0.000),while in the control group, there was increase in lymphocytes(p=0.039) and decrease in segmented neutrophils (p=0.030) andIgE levels (p=0.001). In both groups, plasma nickel levels increased(p=0.010; p=0.039) and GI scores decreased. One month afterremoving the brackets, blood and periodontal parameters frompatients with and without nickel allergy were similar.

O objetivo do presente estudo foi avaliar a condição periodontal eos parâmetros sanguíneos em pacientes alérgicos ao níquel, ummês após remoção dos aparelhos. Noventa e seis pacientesselecionados aleatoriamente foram inicialmente avaliadas. Alergiaao níquel foi diagnosticada usando um teste de contato. Após adeterminação da prevalência de alergia ao níquel, formaram-sedois grupos: 16 pacientes alérgicos (experimental) e 16 nãoalérgicos (controle). Condição periodontal foi diagnosticadaatravés do Índice de Lõe (IG). Parâmetros sanguíneos foramdeterminados por meio de um exame de sangue completo,incluindo a quantificação de níquel e níveis de IgE. Avaliações doestado periodontal foram realizados por um único examinador deforma cega, devidamente calibrado e amostras de sangue foramtomadas depois de nove meses de tratamento e um mês após aremoção dos aparelhos ortodônticos. Análise estatística utilizadafoi testes t pareado e não pareado, Mann-Whitney, Wilcoxon,McNemar e qui-quadrado de tendência linear (p≤0,05). Emcomparação com os valores observados durante o tratamento, onúmero de eosinófilos (p=0,046), basófilos (p=0,001) e monócitos(p=0,002) aumentou significativamente depois da remoção dosaparelhos ortodônticos, ao passo que o número de bastões(p=0,000) diminuiu entre os períodos no grupo alérgico. O númerode linfócitos (p=0,039) aumentou no grupo controle e o número desegmentados (p=0,030) diminuiu. A diminuição dos níveis de IgE(p=0,001) entre os períodos ocorreu no grupo de controle. Níveisde níquel no plasma aumentou após a remoção de aparelhosortodônticos em ambos os grupos (p=0,010; p=0,039). O IGdiminuiu em ambos os grupos. Parâmetros periodontais esanguíneos de pacientes com alergia ao níquel foram semelhantesaos não alérgicos um mês após a remoção dos aparelhos.

Babesia bovis: expression of adhesion molecules in bovine umbilical endothelial cells stimulated with plasma from infected cattle / Babesia bovis: expressão de moléculas de adesão em células endoteliais de cordão umbilical de bovinos estimuladas com plasma de bovinos infectados

Ten male, 12-month-old Jersey with intact spleens, serologically and parasitologically free from Babesia were housed individually in an arthropod-free isolation system from birth and throughout entire experiment. The animals were randomly divided into two groups. Five animals (group A) were intravenously inoculated with 6.6 X10(7) red blood cells parasitized with pathogenic sample of Babesia bovis (passage 7 BboUFV-1), for the subsequent "ex vivo" determination of the expression of adhesion molecules. Five non-inoculated animals (group B) were used as the negative control. The expression of the adhesion molecules ICAM-1, VCAM, PECAM-1 E-selectin and thrombospondin (TSP) was measured in bovine umbilical vein endothelial cells (BUVECs). The endothelial cells stimulated with a pool of plasma from animals infected with the BboUFV-1 7th passage sample had a much more intense immunostaining of ICAM-1, VCAM, PECAM-1 E-selectin and TSP, compared to the cells which did not received the stimulus. The results suggest that proinflammatory cytokines released in the acute phase of babesiosis may be involved in the expression of adhesion molecules thereby implicating them in the pathophysiology of babesiosis caused by B. bovis.

Dez bezerros machos, da raça Jersey, com 1 ano de idade com baços "in situ", sorológica e parasitologicamente livres de Babesia, foram mantidos em baias individuais no isolamento a prova de artrópodes do Depto de Veterinária desde o nascimento e ao longo de toda a experimentação. Os animais foram divididos aleatoriamente em dois grupos. Cinco animais (grupo A) foram inoculados por via intravenosa com 6,6 x10(7) hemácias parasitados com amostra patogênica de Babesia bovis (BboUFV - 1 7ª passagem) , para a determinação subseqüente "ex vivo" da expressão de moléculas de adesão . Cinco animais não inoculados (Grupo B ) foram utilizados como controlo negativo . A expressão de moléculas de adesão ICAM - 1, VCAM , PECAM - 1, E - selectina e trombospondina ( TSP ) foi medida em células endoteliais da veia umbilical de bovinos (BUVECs). As células endoteliais estimuladas com um pool de plasma proveniente de animais infectados com BboUFV - 1 7ª passagem tinham uma imunocoloração muito mais intensa de ICAM - 1 , VCAM , PECAM - 1 de E - selectina e de TSP , em comparação com as células que não receberam o estímulo . Os resultados sugerem que as citocinas pró-inflamatórias liberados na fase aguda da babesiose pode estar envolvida na expressão de moléculas de adesão , implicando , assim, elas na fisiopatologia da babesiose causada por B. bovis.

Seleção, caracterização e clonagem dos genes fljB e groEL agonistas dos receptores de reconhecimento de padrão do sistema imune inato das aves / Selection, characterization and cloning of the fljB and groEL genes agonists of the innate immune system pattern recognition receptors of birds

The recombinant production of innate immune system pattern recognition receptor agonists has provided a new tool for the production of immunostimulants for animals. The molecular pattern associated with the pathogen (PAMP), flagellin, coded by the fljB gene from Salmonella Typhimirium, and the molecular pattern associated to the damage (DAMP), HSP60, coded by the groEL gene from S. Typhimurium and S. Enteritidis, are recognized by pattern recognition receptors (PRRs) of the innate immune system of birds. In the present study, we performed the cloning of genetic fragments of the genes fljB, from S. Typhimurium, and groEL from S. Typhimurium and S. Enteritidis inserted in expression vector pET100/D-TOPO and transformed in E. coli TO10 cells. The clones were evaluated by colony PCR, plasmidial DNA PCR and genome sequencing in order to confirm the presence of these genes. In the colony PCR, we identified the presence of genes groEL (S. Enteritidis), groEL (S. Typhimurium) and fljB (S. Typhimurium) in 80%, 60% and 80% of the transformed colonies, respectively. The cloning system adopted allowed the production of HSP60 genetic fragment clones and flagellin of Salmonella strains, allowing the posterior use of these clones in gene expression trials, with the future potential of being used as non-specific immunostimulants for birds.

A produção recombinante de agonistas dos receptores do reconhecimento de padrão do sistema imune inato tem fornecido uma nova ferramenta para a produção de imunoestimulantes para animais. O padrão molecular associado ao patógeno (PAMP), flagelina, codificado pelo gene fljB de Salmonella Typhimurium e o padrão molecular associado ao dano (DAMP) HSP60, codificado pelo gene groEL da S. Typhimurium e S. Enteritidis, são reconhecidos por receptores de reconhecimento de padrões (RRPs) do sistema imune inato das aves. No presente estudo, foi feita a clonagem de fragmentos genéticos dos genes fljB de S. Typhimurium e groEL de S. Typhimurium e S. Enteritidis inseridos no vetor de expressão pET100/D-TOPO e transformados em células de E. coli TOP10. Os clones foram avaliados pela PCR de colônia, PCR de DNA plasmidial e sequenciamento genômico para a confirmação da presença desses genes. Na PCR de colônia, foram identificadas em 80%, 60% e 80% das colônias transformadas, a presença dos genes groEL (S. Enteritidis), groEL (S. Typhimurium) e fljB (S. Typhimurium) respectivamente. O sistema de clonagem adotado possibilitou a produção de clones dos fragmentos genéticos da HSP60 e flagelina das cepas de Salmonella, permitindo a utilização posterior desses clones em ensaios de expressão gênica, com potencial futuro de serem utilizados como imunoestimulante inespecífico das aves.

Abortos por Neosporacaninum em bovinos do sul de Minas Gerais / Neosporacaninum abortions in cattle in southern Minas Gerais, Brazil

Este estudo avaliou a participação de Neospora caninum em casos de abortos em bovinos provenientes de propriedades rurais da região sul de Minas Gerais por meio de análises histopatológicas, imuno-histoquímicas (IHQ) e pela reação em Cadeia de Polimerase (PCR). O material utilizado foi obtido de um estudo retrospectivo de casos de aborto recebidos pelo Setor de Patologia Veterinária da Universidade Federal de Lavras e de fetos necropsiados durante os anos de 2011 a 2013. De 60 fetos estudados, 30 (50%) tinham lesões microscópicas. Destes, 19 (63%) apresentaram lesões compatíveis com aborto por N. caninum, caracterizadas principalmente por encefalite não supurativa multifocal, necrose e gliose multifocal, assim como, miocardite e miosite não supurativa. Em 14 fetos chegou-se ao diagnóstico definitivo. Destes, cinco tiveram sua confirmação somente pela marcação IHQ e cinco foram positivos somente na PCR. Quatro fetos foram positivos tanto na IHQ quanto na PCR. Cinco fetos, provenientes do estudo retrospectivo apresentaram lesões compatíveis com N. caninum, mas a presença do protozoário não foi confirmada pela marcação IHQ. Os achados demonstram que o N. caninum é um importante agente associado ao aborto em bovinos na região sul de Minas Gerais. Para tanto, além das lesões microscópicas a associação entre a IHQ e a técnica de PCR foi essencial para a confirmação do diagnóstico.

The present study evaluates the participation of Neospora caninum in bovine abortions from farms in the southern region of Minas Gerais state, Brazil, through histopathological, immunohistochemical (IHQ) analysis and DNA detection by polymerase chain reaction (PCR). The material used in this study was obtained through retrospective study of bovine abortion cases received by the Setor de Patologia Veterinária da Universidade Federal de Lavras and by necropsies performed in fetuses from 2011 to 2013. From 60 fetuses studied, 30 (50%) had microscopic lesions; 19 (63%) of them had histologic lesions compatible with N. caninum abortion, characterized by multifocal non-suppurative encephalitis, necrosis and gliosis in the central nervous system and multifocal nonsuppurative myocarditis and myositis. From the 19 fetuses with protozoa lesions in 14 the final diagnosis was performed; five were confirmated only by IHQ staining and five were positive only by PCR. Four fetuses were positive in IHC and in PCR. In five fetuses from the retrospective study lesions compatible with N. caninum were seen, but the presence of the protozoan was not confirmed by IHQ labeling. The findings demonstrate that N. caninum is an important agent associated with abortion in cattle from Minas Gerais. Therefore, beyond the microscopic lesions association between IHC and PCR was essential to confirm the diagnosis.

El ejercicio físico como coadyuvante inmunológico: una revisión / Physical exercise as immune adjuvant: review

El uso de coadyuvantes para aumentar el efecto de vacunas ha sido el foco de varios estudios de investigación en el campo de la salud. Sin embargo, algunas sustancias con potencial coadyuvante pueden tener efectos secundarios con una alta toxicidad por lo que son necesarios métodos menos invasivos y más eficaces para realizar esta función. Varios estudios, investigando los efectos del ejercicio físico sobre diversos parámetros inmunológicos, han mostrado que la práctica de ejercicio físico de ciertas intensidades provoca mejoras en la respuesta inmune y que incluso puede actuar como coadyuvante exógeno inmunitario, por lo que puede ser una herramienta importante para aumentar la eficacia de la respuesta inmune después de la vacunación. El objetivo de este trabajo es revisar los estudios que demuestran cómo diferentes tipos, intensidades, y duración de ejercicios físicos pueden actuar como coadyuvantes inmunitarios. Para ello se seleccionaron estudios que investigaron los efectos de diferentes protocolos o modalidades de ejercicio sobre la eficacia de la inmunización después de la vacunación, tanto en humanos como en modelos animales. En cuanto a la intensidad del ejercicio, hay un consenso de que las actividades con intensidad moderada aumentan la eficacia de las vacunas, mientras que las intensidades ligeras no tienen tal efecto. Entrenamientos únicos de ejercicio de fuerza de los miembros superiores han demostrado ser eficaces para la inmunización de jóvenes, mientras que para las personas mayores, tal eficacia se observó con la práctica regular de ejercicios aeróbicos a una intensidad moderada. La literatura estudiada permite afirmar que hay evidencia de que el ejercicio físico se puede utilizar como un complemento de la vacunación, especialmente en la vacunación en ancianos (AU)

The use of adjuvants in order to enhance the effect of vaccines has been the focus of several research studies. However, some substances with the potential of being used as adjuvants may exhibit high toxicity and side effects. In this sense, new methods which are less invasive and more effective to perform this function are being developed. Thus, several studies have investigated the effects of exercise on immune parameters. It is suggested that physical activities can improve the immune response and may even act as exogenous adjuvants, increasing the host’s immune response efficiency after vaccination. Therefore, the objective of this study was to review the literature in order to find out how different types, intensities and duration of exercise can act as adjuvants for immunization. To this end, studies that investigated the effects of different exercise modalities or protocols were selected, all of them investigating the effectiveness of immunization after vaccination, both in humans and in animal models. Regarding exercise intensity, there is a consensus that moderate activities increase the efficacy of vaccines, while light intensities do not cause any effect. Acute sessions of resistance exercise for upper limbs demonstrated to be effective for immunization of young people, while for the elderly, such efficacy was observed with the regular practice of aerobic exercise at a moderate intensity. In conclusion, from what has been observed in the literature, it seems to exist some evidence that physical exercise can be used as a complement to vaccination, especially when it comes to immunization of the elderly (AU)

Antibody kinetics in goats and conceptuses naturally infected with Neospora caninum.

Neospora caninum is a protozoan which can cause abortions in caprines. However, information regarding the humoral immune response and the occurrence of reproductive disorders is scarce. This is the first study in which the kinetics of antibodies is studied in pregnant goats naturally infected by N. caninum, as well as their respective conceptuses. The subclasses of IgG (IgG1 and IgG2) were also evaluated in pregnant goats. Reproductive problems related to neosporosis (abortion and stillbirth) occurred in 15.38% of the goats. There was a statistically significant association between the increased titres of maternal IgG in the second half of the gestational period with the occurrence of endogenous transplacental transmission. The rate of congenital transmission was 77%. During the gestational period of the seropositive goats, there was mainly a predominance of the subclass IgG2, although mixed patterns of IgG2-IgG1 and the IgG1 pattern were also observed. These results indicate that N. caninum is responsible for the occurrence of important alterations in the humoral immune response of naturally infected goats, and is also a potential causative agent for reproductive disorders in goats. The high proportion of infected conceptuses reinforces the suggestion that congenital infection is one of the main routes of parasite transmission in goats.

Congenital neosporosis in goats from the State of Minas Gerais, Brazil.

Congenital Neospora caninum infection was diagnosed in two Saanen goat kids from two distinct herds with a history of abortion and weak newborn goat kids in the Southern region of the State of Minas Gerais, Brazil. The first kid was weak at birth, had difficulty to rise and was unable to nurse. Gross lesions of porencephaly and hydrocephalus ex vacuo were seen. Multifocal necrosis, gliosis and non-supurative encephalitis were observed in the brain. Several parasitic cysts with a thick wall that reacted strongly only with polyclonal antiserum to Neospora caninum were seen in the cerebral cortex, brain stem and cerebellum. The second kid was born from a Neospora caninum seropositive mother that aborted in the last pregnancy. It was born without clinical signs. The diagnosis of neosporosis was based on antibody titer of 1:800 to N. caninum by indirect fluorescence antibody test obtained from blood collected before the goat kid ingested the colostrum and Neospora caninum DNA was detected by polymerase chain reaction and sequenced from placenta. This is the first report of neosporosis in goats in the southeast region of Brazil.

[Immunization with synthetic peptide anti-tick Rhipicephalus (Boophilus) microplus SBm7462 via mucosal routes]. / Imunização com o peptídeo sintético anti carrapato Rhipicephalus (Boophilus) microplus SBM7462® por vias de mucosa.

The mucosal immunization consists on antigen administration in these surfaces it is a not invasive method, inductor of local and systemic immune response. This work evaluated the immune response of the synthetic vaccine anti-tick Rhipicephalus (Boophilus) microplus SBm7462®, via oral and nasal routes. Were used 60 BALB/c mice, divided in three groups of 20 animals each (I: oral immunization; II: nasal immunization; III: animals not immunized). Nine and 15 days after the first and the second immunization was collected, separately for each group, T cells from the immunized animals, cultivating them per 10 days. After the incubation, were determined the percentage of cellular viability and the specific nature of this T cells, which had held as memory T cell. The averages of T cell SBm7462-reagents had been submitted to the analysis of variance and comparison for Tukey test, 5% of probability. Group II presented higher cellular proliferation "in vitro". For ELISA test, were positive only two animals in group I and eight in group II. The oral and nasal routs alternatively viable for immunization with the synthetic peptide SBm7462®, however require greater number of doses to induce responses with high levels of immunoglobulins.

[Molecular analysis of the antigenic sequence (SBm7462®) from Bm86 of the Rhipicephalus (Boophilus) microplus and similarity with Rhipicephalus sanguineus and Hyalomma anatolicum anatolicum]. / Análise molecular da sequência antigênica (SBm7462®) da BM86 doRhipicephalus (Boophilus) microplus E similaridade com Rhipicephalussanguineus E Hyalomma anatolicum anatolicum

The vaccination represents optimum method evaluated with effective cost to prevent economic losses and to increase the duration and quality of life of the production animals. . Diverse vaccines are produced from the intestinal protein Bm86 of the Rhipicephalus. (B.) microplus. The knowledge of the conservation of the gene bm86 is very important to evaluate the vaccine efficiency and the possibility of reaction crossed between different species of ticks. Samples of R. (B.) microplus come from different localities had been sequenced. The analyses of multiple alignments of the sequences had been made through the BioEdit program 7.0.5.3 version and the verification of polymorphism for visual inspection. In this work the alignment of all was become fulfilled sequences using itself BLAST in the search for similarity. Similarity was observed enters the sequenced fragments of R. (B.) microplus with the sequence of the protein Rs86 de Rhipicephalus sanguineus and with protein HA98 of the tick Hyalomma anatolicum anatolicum. The results give molecular support to synthetic the vaccine use based in the gene bm86 (SBm7462®) to be used in different species of ticks.

[Elaboration and evaluation of a candidate to the DNA vaccine using synthetic genes derived from the peptídeo SBm7462 against the carrapato Rhiphicephalus (Boophilus) microplus]. / Elaboração E avaliação de um candidato à vacina de DNA, utilizando-se genes sintéticos derivados do peptídeo SBm7462contra o carrapato Rhiphicephalus (Boophilus) microplus.

Rhiphicephalus (Boophilus) microplus is one of the most important arthropods in veterinary medicine due economic losses and health problems caused in cattle production. The vaccination represents optimum method evaluated with effective cost to prevent economic losses and to increase the duration and quality of life of the production animals. A synthetic peptide, SBm 7462, derived from Bm86, has been shown great results in control of ticks. The construction and synthesis of one nucleotide sequence based on this peptide might be useful for design a DNA vaccine that has many advances than peptide vaccine. A gene, called seq1, was constructed with a three repetition of nucleotide sequence of SBm 7462. It was cloned into a pCIneo vector expression in mammals and injected in BALB/c mouse. When mice were inoculated with the expression cassette they did not response in ELISA. They elevated antibody titles only when vaccinated with the synthetic peptide SBm7462®. And, the best titles of immunoglobulins were seen when the SBm7462® was administered subcutaneously.

Polymorphism of the bm86 gene in South American strains of the cattle tick Boophilus microplus.

Thirty Boophilus microplus strains from various geographic regions of Brazil, Argentina, Uruguay, Venezuela and Colombia were analyzed for the bm86 and bm95 gene. A fragment of cDNA of 794 base pairs of the parasite larvae, included between nucleotides 278-1071s, was amplified and cloned on the pGEM-T vector. Two random clones were sequenced for each population and the nucleotides 278-1071 and predicted amino acid sequences compared with the bm86 and bm95 genes. Variations from 1.76 to 3.65% were detected in the nucleotides sequence when compared with the homologous sequence of the bm86 gene and a 3.4-6.08% in the homologous amino acid sequence of the Bm86 protein. When the sequences obtained were compared with the bm95 gene, variations from 0.50 to 3.15% were detected. Variations from 1.14 to 4.56% were detected for the Bm95 protein homologous sequences in the deduced amino acid sequence. Only five of the 30 strains analyzed presented two different types of alleles expressed and the two alleles of the Alegre population and allele 1 of the Betim population were the most divergent of all those analyzed.