Effects of vaccination with Brucella melitensis, strains Rev 1 ΔeryCD and Rev 1, on the reproductive system of young male goats.

The present study evaluates the effects of vaccination with Brucella melitensis strains Rev 1 ΔeryCD and Rev 1 on the reproductive system of male goats. Three groups, each of them consisting of 15 six-month-old brucellosis-free male goats, were studied. The first group was vaccinated with the Rev 1 ΔeryCD strain, the second group received Rev 1 and the third group was inoculated with sterile physiological saline solution. The dose of both strains was of 1×109CFU/ml. Over the course of the five months of this study, three males from each group were euthanized every month. Their reproductive tracts, spleens, and lymph nodes were collected to analyze serology, bacteriology PCR, histology, and immunohistochemistry. Results show that vaccination with B. melitensis strains Rev 1 ΔeryCD and Rev 1 does not harm the reproductive system of male goats. Strain B. melitensis Rev 1 ΔeryCD displayed a lower capacity to colonize the reproductive tract than strain Rev 1, which was attributed to its limited catabolic action toward erythritol.

Presence of co-infection between bovine leukemia virus and bovine herpesvirus 1 in herds vaccinated against bovine respiratory complex.

The aim of this study was molecular identification of bovine leukemia virus and possible co-infection with bovine respiratory disease complex (BRDC) viral agents in Mexican dairy herds. We collected 533 blood samples from cattle vaccinated against the BRDC virus in 9 states across Mexico. Peripheral blood leukocytes were removed and genetic material was extracted to detect bovine leukemia virus (BLV), bovine herpesvirus 1 (BoHV-1), bovine viral diarrhea virus (BVDV), bovine parainfluenza virus 3 (BPIV-3), and bovine respiratory syncytial virus (BRSV) infection using polymerase chain reaction. We identified high BLV infection rates in 270 cattle (50.65%). One hundred and thirty-three cows (24.95%) tested positive for BoHV-1, of which 65 samples were positive for both viruses (BoHV-1 and BLV) and 68 were only positive for BoHV-1. Only 4 samples tested positive for BPIV-3 and no sample was positive for BVDV or BRSV. Relative risk and odds ratio analyses did not identify that the presence of BLV infection favors BoHV-1 co-infection in vaccinated herds.

Le but de cette étude était l'identification moléculaire du virus de la leucémie bovine et une éventuelle co-infection par des agents viraux du complexe des maladies respiratoires bovines (BRDC) dans des troupeaux laitiers mexicains. Nous avons recueilli 533 échantillons de sang de bovins vaccinés contre le virus BRDC dans neuf états du Mexique. Les leucocytes du sang périphérique ont été prélevés et le matériel génétique a été extrait pour détecter le virus de la leucémie bovine (BLV), le virus de l'herpès bovin 1 (BoHV-1), le virus de la diarrhée virale bovine (BVDV), le virus parainfluenza bovin 3 (BPIV-3), et le virus respiratoire syncytial bovin (BRSV) par réaction d'amplification en chaîne par la polymérase. Nous avons identifié des taux élevés d'infection par le BLV chez 270 bovins (50,65 %). Cent trente-trois bovins (24,95 %) ont été testés positifs pour le BoHV-1, desquels 65 échantillons étaient positifs pour les deux virus (BoHV-1 et BLV) et 68 étaient uniquement positifs pour le BoHV-1. Seuls quatre échantillons ont été testés positifs pour le BPIV-3 et aucun échantillon n'a été positif pour le BVDV ou le BRSV. Les analyses du risque relatif et des rapports de cotes n'ont pas identifié que la présence d'une infection par le BLV favorise la co-infection par le BoHV-1 dans les troupeaux vaccinés.(Traduit par les auteurs).

Study of the Genetic Expression of Antiretroviral Restriction Factors and Acute Phase Proteins in Cattle Infected with Bovine Leukemia Virus.

The goal of this study was to analyze the genetic expression of antiretroviral restriction factors (ARF) and acute phase proteins (APP), as well as their correlation with proviral and viral loads in cattle with aleukemic (AL) and persistent lymphocytosis (PL). Complete blood samples were collected from a herd of dairy cows, and we extracted genetic material from peripheral blood leukocytes. Absolute quantification of the expression of ARF (APOBEC-Z1, Z2, and Z3; HEXIM-1, HEXIM-2, and BST2) and APP (haptoglobin (HP), and serum amyloid A (SAA)) was performed by qPCR. Statistical significance was observed in the expression of APOBEC-Z3 in BLV-infected animals. We only found positive correlations with a strong expression of the ARF genes in the AL group. The participation of APOBEC (Z1 and Z3), HEXIM-1, and HEXIM-2 was more frequently identified in BLV-infected animals. HEXIM-2 showed active gene expression in the AL group. Although the expression of ARF in early stages of infection (AL) maintains an important participation, in late stages (PL) it seems to have little relevance.

Presence of Chlamydia abortus in colostrum, milk, and vaginal discharge samples of sheep

Background: The main transmission route of Chlamydia abortus is by ingesting the microorganism that has been eliminated in vaginal secretions, placental membranes or abortions that contaminate the environment and, possibly, through milk and colostrum. Elimination through vaginal secretions is well documented. However, there are no reports about isolation and identification of C. abortus in the colostrum or milk of infected sheep, so it is important to determine whether or not C. abortus may be present in these secretions, which are the only food of lambs. Objective: To detect C. abortus in colostrum, milk, and vaginal secretions of sheep with a history of reproductive disorders. Methods: Colostrum, milk, and vaginal exudates were collected from 66 sheep. The samples were inoculated in mouse fibroblast cell cultures and the presence of C. abortus determined by direct immunofluorescence. Results: 19 out of 66 colostrum samples (28.7%), 14 out of 66 milk samples (21.2%) and 17 out of 66 vaginal swabs (25.7%) were positive for C. abortus. The 50 samples positive for isolation and detected by immunofluorescence, together with 42 negative samples were subjected to qPCR to amplify a fragment of the ompA gene from C. abortus. Thirty-eight of the 92 samples processed by this technique were positive for C. abortus. Conclusion: The results demonstrated the presence of C. abortus in a high proportion in colostrum, milk and vaginal secretions of infected sheep. To the best of our knowledge, this is the first field study confirming the presence of C. abortus in colostrum, which shows that excretion of Chlamydia by lactogenesis could occur in the first hours after birth.

Antecedentes: La principal vía de transmisión de C. abortus es la ingestión del microorganismo que ha sido eliminado en las secreciones vaginales, membranas placentarias, abortos y, posiblemente, a través de la leche y el calostro. La eliminación a través de secreciones vaginales está bien documentada. Sin embargo, no existen reportes del aislamiento e identificación de C. abortus en el calostro o la leche de ovejas infectadas, por lo que es importante determinar si la bacteria puede o no estar presente en estas secreciones, que son el único alimento de los corderos. Objetivo: Detectar la presencia de C. abortus in calostro, leche y secreciones vaginales de ovejas con antecedentes de problemas reproductivos. Método: Con el propósito de aislar e identificar C. abortus en estas secreciones, se recolectó calostro, leche y exudado vaginal de 66 ovejas. Las muestras fueron inoculadas en cultivos celulares de fibroblastos de ratón y se determinó la presencia de la bacteria por inmunofluorescencia directa. Resultados: Fueron positivas 19 de 66 muestras de calostro (28,7%), 14 de 66 muestras de leche (21,2%) y 17 de 66 hisopos vaginales (25,7%). Las 50 muestras positivas al aislamiento y detectadas por inmunofluorescencia, junto con 42 negativas se sometieron a qPCR para amplificar un fragmento del gen ompA de C. abortus; 38 de las 92 muestras procesadas por esta técnica fueron positivas para C. abortus. Conclusión: Los resultados del presente estudio demostraron la presencia de C. abortus en una alta proporción en el calostro, la leche y las secreciones vaginales de ovejas infectadas. Este es el primer estudio de campo que confirma la presencia de C. abortus en calostro, lo que demuestra que la excreción de clamidia por lactogénesis podría ocurrir en las primeras horas después del nacimiento.

Antecedentes: A principal via de transmissão da Chlamydia abortus é a ingestão do microrganismo que foi eliminado nas secreções vaginais, membranas placentárias ou abortos que contaminam o meio ambiente e, possivelmente, através do leite e colostro. A eliminação pelas secreções vaginais está bem documentada. No entanto, não há relatos de isolamento e identificação de C. Abortus no colostro ou leite de ovelhas infectadas, por isso é importante verificar se a bactéria pode estar ou não presente nessas secreções, único alimento dos cordeiros. Objetivo: Detectar a presença de C. Abortus no colostro, leite e secreções vaginais de ovelhas com histórico de distúrbios reprodutivos Métodos: Para isolar e identificar C. Abortus nessas secreções, foram coletados colostro, leite e exsudato vaginal de 66 ovelhas. As amostras foram inoculadas em cultura de células de fibroblastos de camundongo e a presença da bactéria determinada por imunofluorescência direta. Resultados: 19 de 66 amostras de colostro (28,7%), 14 de 66 amostras de leite (21,2%) e 17 de 66 esfregaços vaginais (25,7%) sendo positivos. As 50 amostras positivas para isolamento e detectadas por imunofluorescência, juntamente com as 42 negativas, foram submetidas a qPCR para amplificar um fragmento do gene ompA de C. Abortus. Trinta e oito das 92 amostras processadas por esta técnica foram positivas para C. Abortus. Conclusão: Os resultados do presente estudo demonstraram a presença de C. Abortus em alta proporção no colostro, leite e secreções vaginais de ovelhas infectadas. Este trabalho é o primeiro estudo de campo na literatura científica confirmando a presença de C. Abortus no colostro, o que mostra que a excreção da clamídia por lactogênese pode ocorrer nas primeiras horas após o nascimento.

Genetic analysis of the pX region of bovine leukemia virus genotype 1 in Holstein Friesian cattle with different stages of infection.

The pX genetic region of bovine leukemia virus (BLV) includes four genes with overlapping reading frames that code for the Tax, Rex, R3, and G4 proteins. These proteins are involved in the regulation of transcriptional and post-transcriptional viral expression, as well as having oncogenic potential. Our goal was to investigate the pathogenicity of the pX region of BLV genotype 1 in terms of lymphocytosis, lymphomas, and proviral DNA load. We screened 724 serological samples from mixed-age Holstein Friesian cattle from six states in Mexico. Peripheral blood leukocytes (PBLs) were isolated from whole blood with anticoagulant, and genomic DNA was extracted from the PBLs using a commercial kit. Then, a set of primers that hybridize in conserved regions of the BLV pX region were used, which allowed for PCR standardization to detect proviral DNA in infected cells. Positive amplicons were sequenced using the Sanger method, resulting in 1156-nucleotide-long final sequences that included the four pX region genes. The experimental group consisted of 30 animals. Twelve of these had lymphocytosis, six had lymphoma, and 12 were apparently healthy cattle without any signs of lymphocytosis or lymphoma. The presence of lymphoma was detected in six bovine tumor tissues using histopathology, and the presence of BLV was detected by in situ hybridization. Phylogenetic analysis demonstrated that the 30 sequences were associated with genotype 1, and the genetic distance between the sequences ranged from 0.2% to 2.09%. We identified two sequences in the G4 gene: one with a three-nucleotide deletion resulting in the loss of a leucine (AGU_7488L, in a cow with lymphocytosis), and one with a nine-nucleotide deletion resulting in the loss of leucine, proline, and leucine (AGU_18A, in a cow without lymphocytosis). Analysis of the PX region indicated that positive selection had occurred in the G4, rex, and R3 genes, and we found no difference in proviral DNA load between the studied groups. We were unable to establish an association between variations in the pX region and the development of lymphocytosis, lymphoma, asymptomatic status, or proviral DNA load in BLV-infected cattle.

Serotyping versus genotyping in infected sheep and goats with small ruminant lentiviruses.

Despite SRLV infection being endemic in Mexico, there is little information regarding which genotypes are present. We compared serotyping and PCR-sequencing results from sheep and goats infected with SRLV. We separated plasma and peripheral blood leukocytes (PBL) from 1940 blood samples from sheep and goats from 12 states across Mexico. To detect SRLV infection, we tested plasma samples using two commercial ELISA kits (VMRD and Eradikit SRLV Screening). Then, we serotyped the infecting virus (A/ B) using Eradikit SRLV Genotyping. PBL DNA was used to detect the proviral genome via PCR. Positive amplicons were sequenced to identify viral genotypes using a phylogenetic analysis. Also, we analysed for residues differences in the sequences of a capsid epitope between genotypes. The serological results indicated a higher detection of seropositive animals using the VMRD ELISA compared to Eradikit, with 21 % and 15.3 % more in sheep and goats respectively. Only 25.7 % of the ELISA serotyping results matched those from PCR-sequencing. PCR-sequencing was able to identify genotype A, B and coinfections in animals classified as indeterminate by the ELISA test. This lack of sensitivity may be related to the lack of epitopes from the matrix and transmembrane peptides used by ELISA screening. Sequences analysis revealed that SRLVs found in sheep cluster with genetic subtypes A2 and B1, while those in goats cluster with subtypes A1 and B1. Serotyping did not prove to be an adequate method for predicting the viral genotype (A and / or B) in infections caused by SRLV.

First evidence of bovine immunodeficiency virus infection in Mexican cattle.

This study set out to identify the presence of bovine immunodeficiency virus (BIV) in animals geographically located in Mexico. BIV was first discovered in the United States in a dairy cow with persistent lymphocytosis, lymphoid hyperplasia and lymphocytic encephalitis. Many studies indicate that BIV infection is globally distributed, but its presence in Mexico remains unknown. We collected 1,168 heparinized blood samples from cattle in ten states across the Mexican Republic, then separated plasma using centrifugation and tested for antibodies against BIV. We used an indirect ELISA based on the use of a synthetic peptide derived from transmembrane glycoprotein (gp45/TM). In order to identify the viral genome, we designed a synthetic gene as a PCR control, as well as a pair of oligonucleotides for amplifying a 519 bp product of the env gene which encodes the surface protein. Positive amplicons were purified and subjected to nucleotide sequencing. A total of 189 (28.94%) tested plasma samples suggest the presence of specific anti-BIV antibodies in all states studied except for Chiapas. Additionally, PCR results identified six positive cows in the states of Puebla and Coahuila. BIV in these cows was confirmed via nucleotide sequencing and in silico analysis of these samples. This is the first report of the presence of BIV in Mexican cattle.

Signature patterns in region V4 of small ruminant lentivirus surface protein in sheep and goats.

The env gene in Small Ruminant Lentiviruses (SRLV) encodes the surface glycoprotein (SU) that divides into conserved (C1-C4) and variable regions (V1-V5). SRLV region V4 has been found to be homologous to the V3 region of human lentivirus (HIV). HIV V3 is responsible for tropism and the development of nervous clinical patterns when there is a tendency to conserve amino acids in specific "signature pattern" positions. The goal of this study was to identify signature patterns in the V4 region of the SU, which is encoded by the SRLV env gene. Secondarily, to understand how these signature patterns are associated with different clinical status in naturally infected sheep and goats. Starting with 244 samples from seropositive animals from nine Mexican states, we amplified the V4 region using nested PCR and obtained 49 SRLV sequences from peripheral blood leukocytes. Based on phylogenetic analysis results, we identified three groups: asymptomatic genotypes A (Ssx GA) and B (Ssx GB), as well as animals with arthritic presentation, genotype B (A GB). Similarity levels between group sequences ranged from 67.9%-86.7%, with a genetic diversity ranging from 12.7%-29.5% and a dN / dS ratio that indicated negative selection. Analyses using Vespa and Entropy programs identified four residues at positions 54, 78, 79 and 82 in SU region V4 as possible signature patterns, although with variable statistical significance. However, position 54 residues "N" (p = 0.017), "T" (p = 0.001) and "G" (p = 0.024) in groups A GB, Ssx GA and Ssx GB respectively, best characterized the signature patterns. The results obtained identified a signature pattern related to different genotypes and clinical status by SRLV in sheep and goats.

Lack of association between amino acid sequences of the bovine leukemia virus envelope and varying stages of infection in dairy cattle.

We collected 724 blood samples from dairy cattle from six Mexican states, and tested them for the presence of antibodies against BLV using a commercial ELISA test. Our study groups consisted of 32 samples: 12 asymptomatic cows, 12 cows with lymphocytosis and 8 samples of tumor tissue of the abomasum and heart of cattle with lymphoma. We designed three pairs of primers to amplify the complete BLV env gene, and obtained a fragment of 1548 nucleotides in length with the sequenced products. According to the phylogenetic tree we constructed to identify the viral genotype, 96.87 % of the sequences grouped into genotype 1, while a single sample from a cow with lymphocytosis (3.13 %) was associated with genotype 3 sequences. The similarity between the Mexican BLV sequences ranged from 0.985-1.00. In addition, the proportion of non-synonymous and synonymous mutations indicated negative selection. We did not identify any conserved residues in the viral protein sequences that could be related to BLV infection stage in cattle. Proviral quantification was performed using quantitative polymerase chain reaction, and we used Mood´s median test as statistical analysis. We found no significant association between proviral load and phase of infection. The sequences showed high similarity without any association between BLV surface glycoprotein and the different infection stages, nor differences in the proviral load. BLV genotype 1 was identified as prevalent in the studied samples, and for the first time in Mexico, we identified BLV genotype 3 in cattle.

Distribution of lymphocytes, immunoglobulin-containing cells, macrophages, and dendritic cells in the accessory sex glands of rams experimentally infected with actinobacillus seminis / Distribuição de linfócitos, células carreadoras de imunoglobulinas, macrófagos e células dendríticas em glândulas sexuais acessórias de carneiros infectados experimentalmente com actinobacillus seminis

The distribution of cells involved in the immune response in accessory sex glands of rams experimentally infected with Actinobacillus seminis was studied. Twelve one-year old rams were experimentally infected by intraurethral (IU) (n=4) and intraepididymal (IE) (n=4) route, and four control (CON) animals were used. The animals were slaughtered 35 days post-inoculation, samples were taken from accessory sex glands, and bacteriology and histopathology tests were performed. The presence of CD4, CD8 and TCRγδ (WC1) lymphocytes, CD45RO cells, macrophages (CD14), dendritic cells (CD1b), IgA-, IgG- and IgM-containing cells (IgCC) was determined. Animals of the IE group developed clinical epididymitis. No lesions were seen in rams of the IU group; two of the intraepididymal inoculated CON developed small lesions in the epididymis. A. seminis isolates were achieved from 6:16 (37.5%) accessory sex glands in the IE group, but not in the IU and CON groups. In the CON group, IgA- and IgM- containing cells predominated in the bulbourethral glands and the disseminated prostate, and they were scarce or null in the vesicles and ampullae. A significant increase of IgA-, IgG- and IgM- containing cells was confirmed in the seminal vesicles, the ampullae and the bulbourethral glands in the IE group. In the IE and IU groups, an increase in CD4, CD8, WC1, CD45RO and CD14 was evidenced in the vesicles and ampullae. CD1b dendritic cells were present in the ampullae and vesicles with inflammatory processes. A. seminis triggered a local immune response in the IE and IU groups. These results indicate a different pattern of infiltrating immune cells in the accessory sex glands of infected A. seminis rams.

A distribuição das células envolvidas na resposta imune em glândulas sexuais acessórias de carneiros experimentalmente infectados com Actinobacillus seminis foi estudada. Doze carneiros de um ano de idade foram experimentalmente infectados via intrauretral (IU) (n=4) e via intraepididimal (IE) (n=4) e quatro animais controles (CON) foram utilizados. Os animais foram abatidos 35 dias após a inoculação, amostras foram retiradas das glândulas sexuais acessórias e testes bacteriológicos e histopatológicos foram realizados. A presença de linfócitos CD4, CD8 e TCRγδ (WC1), células CD45RO, macrófagos (CD14), células dendríticas (CD1b) e células contendo IgA, IgG and IgM (IgCC) foi determinada. Os animais do grupo IE desenvolveram epididimite clínica. Não foram visualizadas lesões nos carneiros do grupo IU, dois dos CON inoculados intraepididimalmente desenvolveram pequenas lesões no epidídimo. Isolados de A. seminis foram obtidos de 6:16 (37,5%) nas glândulas sexuais acessórias no grupo IE mas não nos grupos IU e CON. No grupo CON células contendo IgA and IgM predominaram nas glândulas bulbouretrais e na próstata e foram escassas ou ausentes nas vesículas e na ampola. Um incremento significativo de células contendo IgA, IgG and IgM foi confirmado nas vesículas seminais, na ampola e nas glândulas bulbouretrais no grupo IE. Nos grupos IE e IU foi evidenciado um aumento em CD4, CD8, WC1, CD45RO e CD14 nas vesículas e ampola. As células dendríticas CD1b estavam presentes na ampola e nas vesículas com processo inflamatório. A. seminis induziu uma resposta imune local nos grupos IE e IU. Estes resultados indicam um padrão diferente de células imunes infiltrantes nas glândulas sexuais acessórias de carneiros infectados por A. seminis.

Oral immunization against porcine pleuropneumonia using the cubic phase of monoolein and purified toxins of Actinobacillus pleuropneumoniae.

The main goal of this work was to obtain an orally administered immunogen that would protect against infections by Actinobacillus pleuropneumoniae. The Apx I, II and III toxins were obtained from the supernatants of cultures of serotypes 1 and 3 of A. pleuropneumoniae. The capacity of monoolein gel to trap and protect the Apx toxins, and the effect of their incorporation on the stability of the cubic phase were evaluated. The gel was capable of trapping a 400-µg/ml concentration of the antigen with no effects on its structure. Approximately 60% of the protein molecules were released from the gel within 4h. Four experimental groups were formed, each one with four pigs. All challenges were conducted in a nebulization chamber. Group A: Control (-) not vaccinated and not challenged; Group B: Control (+) not vaccinated but challenged; Group C: vaccinated twice intramuscularly with ToxCom (a commercial toxoid) at an interval of 15 days and then challenged; and Group D: vaccinated orally twice a week for 4 weeks with ToxOral (an oral toxoid) and challenged on day 28 of the experiment with a same dose of 2.0 × 10(4) UFC of A. pleuropneumoniae serotypes 1 and 3. The lesions found in group B covered 27.7-43.1% of the lungs; the pigs in group C had lesions over 12.3-28%; and those in group D over 15.4-32.3%. No lesions were found in the Group A pigs. A. pleuropneumoniae induced macroscopic lesions characteristic of infection by and lesions microscopic detected by histopathology. The etiologic agent was recovered from the infected lungs, tonsils and spleen. The serotypes identified were 1 and 3. An indirect ELISA test identified the antibodies against the Apx toxins in the serum of the animals immunized orally.

Distribution of immune response cells in the pelvic urethra and the prepuce of rams / Distribuição de células da resposta imune na uretra pélvica e em prepúcio de carneiros

The pathogens of the reproductive system in the male can penetrate and establish by ascending route, from to the prepuce to the urethra, accessory glands, epididymis and testicles. The aim of this paper is determine the distribution and number of cells involved in the immune response in prepuce and pelvic urethra of rams, without apparent clinical alterations in testicle, epididymis and prepuce. [...] Significant differences were found in the total number of CD4, CD45RO, and WC1 lymphocytes, in CD14 macrophages, and CD1b dendritic cells, with mean values being greater in the fornix than in the urethra (p<0.05) in all cases. Only dendritic cells were found in the prepuce. No differences were found in the number of CD8 lymphocytes between both organs. The ratio between each cell type in the connective and the intraepithelial tissues and between organs was 10/1 for CD4 in the fornix (p<0.05), against 7/1 in the urethra (p<0.05), while CD8 had a 1/1 distribution in both mucosae. The WC1 ratio was 5/1 in both mucosae (p<0.05). CD45RO labeling was 19/1 in the prepuce (p<0.05) and 1/1 in the urethra. IgA-containing cells did not show differences in the total number of cells in both tissues. In the urethra, no IgG-containing cells were observed and IgM-containing cells were scarce; in contrast, both cell types were present in the prepuce, in amounts greater than in the urethra (p<0.05). IgA-, IgG-, and IgM-containing cells were located in both organs in the mucosal connective tissue. The presence of antigen-presenting cells, macrophages, and dendritic cells, as well as of lymphocytes CD4, CD8 TCR γδ (WC1), IgA-, IgG and IgM positive cells, and CD45RO cells suggests that both mucosae may behave as inductive and effector sites for the mucosal immune response.

Os patógenos do aparelho reprodutor do macho podem penetrar e se estabelecer por via ascendente, a partir do prepúcio à uretra, glándulas anexas, epidídimo e testículos. Neste trabalho foi quantificada a distribuição de algumas das células envolvidas na resposta imune, em nível de prepúcio e uretra pélvica, em quatro carneiros de um ano de idade, sem lesões aparentes no testículo, no epidídimo e no prepúcio.[...] Não foram encontradas diferenças significativas no número de linfocitos CD8 entre ambos os orgãos. A relação entre cada tipo celular no tecido conectivo e intra-epitelial e entre os diferentes órgãos, resultou para CD4 10/1 no prepúcio (p<0.05), contra 7/1 na uretra (p<0.05), entretanto os CD8 se distribuíram 1/1 em ambas as mucosas, não sendo significativa as diferenças. Os WC1 foram observados na relação 5/1 em ambas as mucosas (p<0.05). A célula CD45RO, no prepucio, foi observada de 19/1(p<0.05) e na uretra de 1/1, não sendo um resultado significativo. As CC-IgA não mostraram diferença significativa no total de células em ambos os tecidos. Na uretra não foram observadas as CC-IgG, e as CC-IgM foram escassas; em contrapartida, ambos os tipos celulares foram observadas no prepucio, em quantidades menores que na uretra (p<0.05). As CC-IgA, IgG e IgM foram observadas em ambos os tecidos conectivos da mucosa. A presença de células apresentadoras de antígenos, macrófagoss e células dendríticas, assim como de linfócitos CD4, CD8. TCR γδ (WC1), CC-IgA, IgG e IgM e células CD45RO, determinam que ambas as mucosas podem se comportar como locais de indução e promoção da resposta imune das mucosas.

Detección de anticuerpos contra lentivirus de pequeños rumiantes en fetos ovinos y caprinos / Detection of antibodies against small ruminant lentiviruses in ovine and caprine fetuses

The presence of antibodies against Maedi-Visna (MV) and caprine arthritis encephalitis (CAE) was evaluated in ovine and caprine fetuses obtained from pregnant females slaughtered in abattoirs. Sera from 54 caprine fetuses and 65 ovine fetuses were collected and evaluated by the indirect ELISA technique. Antibodies were detected in five caprine fetuses of 80 days of gestation and in four ovine fetuses of 90 to 100 days of gestation; three caprine fetuses and two ovine fetuses of 60 to 65 days of gestation were suspicious. The possibility of uterine infection with these retroviruses is confirmed, and its consequences in current disease control and eradication strategies are discussed.

Se evaluó la presencia de anticuerpos contra los virus del Maedi-Visna (MV) y la artritis encefalitis caprina (AEC) en fetos ovinos y caprinos obtenidos de hembras gestantes sacrificadas para consumo en México. Se recolectaron sueros de 54 fetos caprinos y 65 fetos ovinos que se evaluaron mediante ELISA indirecto. Se detectaron anticuerpos en cinco fetos caprinos de 80 días de gestación y en cuatro fetos ovinos de 90 a 100 días; tres fetos caprinos y dos ovinos de 60 a 65 días de gestación resultaron sospechosos. Se confirma la posibilidad de la infección uterina por estos retrovirus y se discuten sus consecuencias en las actuales estrategias de control y erradicación de las enfermedades.

Evaluación biológica de dos sistemas de apareamiento en ovinos de raza Columbia en producción intensiva / Biological evaluation of two breeding systems of intensive production in the Columbia breed sheep

The biological efficiency of two breeding systems for sheep in the Mexican high plateau was evaluated. A total of 300 adult and 60 replacement sheep of the Columbia breed were randomly distributed into two groups of 150 adults and 30 young ones (mean age to first birth of 18 months). Mates of 36 days with natural mount every eight months (March, November and July) were done in the first group (intensive system). In the second group (annual system), a 45-day mating was done starting on November every year. Marker males were introduced 15 days before mating in all cases, which were replaced on the first day of mating for stallions at a one male for every 20 female ratios. The physical condition of all females was evaluated every month. The animals grazed 8 to 9 hours in grasslands of alfalfa (Medicago sativa) and Orchard (Dactylis glomerata) and Rye (Lolium perenne) grasses irrigated by aspersion. Only the sheep in the intensive system received supplement during lactation and re-mating. Breeding was achieved in both systems in all seasons. The mean fertility of the three breeding of the intensive system was 83.9%; while the mean fertility of the two annual breeding was 88.3% (P < 0.05). Prolificacy was 1.12 vs 1.32 (P < 0.05) for the intensive and annual breeding, respectively; mortality at weaning was 5.3% vs 6.4% (P < 0.05), weaning rate was 0.89 vs 1.09 (P < 0.05), the number of weaned lambs per ewe in two years was 2.5 vs 2.0 and lamb weight was 55.9 vs 44.4 kg (P < 0.05), respectively. Adult ewes were superior to the young ones in all the evaluated parameters, regardless of the kind of mating (P < 0.05). Results in this study allowed to compare the biological efficiency between an intensive breeding system every eight months and an annual breeding system. They also show the possibility of mating sheep of the Columbia breed in an intensive way.

Se evaluó la eficacia biológica de dos sistemas de apareamiento en ovinos del altiplano central de México. Un total de 300 ovejas adultas y 60 de reemplazo de la raza Columbia se distribuyeron al azar en dos grupos de 150 adultas y 30 jóvenes (edad promedio al primer parto de 18 meses). En el primer grupo (sistema intensivo) se realizaron empadres de 36 días, con monta natural cada ocho meses (marzo, noviembre y julio), mientras que en el segundo grupo (sistema anual) se llevó a cabo un empadre de 45 días, iniciando en noviembre de cada año. En todos los casos, quince días antes del empadre se introdujeron machos marcadores, que en la fecha de inicio del empadre se sustituían por sementales en proporción de un macho por cada 20 hembras. Mensualmente se evaluó la condición física de todas las hembras. Los animales pastorearon de ocho a nueve horas diarias en praderas de alfalfa (Medicago sativa) y pastos orchard (Dactylis glomerata) y rye grass (Lolium perenne) irrigadas por aspersión. Solamente las ovejas del sistema intensivo recibieron complemento durante la lactancia y el reempadre. En los dos sistemas se lograron apareamientos en todas las épocas, la fertilidad en los tres empadres del sistema intensivo fue de 83.9%, mientras que el promedio de los dos empadres anuales fue de 88.3% (P < 0.05). En el mismo orden, la prolificidad fue de 1.12 vs 1.32 (P < 0.05); la mortalidad al destete, 5.3% vs 6.4% (P > 0.05); la tasa de destete, 0.89 vs 1.09 (P < 0.05); el número de corderos destetados por oveja en dos años fue de 2.5 vs 2.0, y de kg de cordero de 55.9 vs 44.4 kg (P < 0.05). Independientemente del tipo de empadre, las ovejas adultas fueron superiores a las jóvenes en todos los parámetros evaluados (P < 0.05). Los resultados de este estudio permiten comparar la eficacia biológica entre un sistema de apareamiento intensivo cada ocho meses y uno anual, también muestra la posibilidad de aparear ovejas de la raza Columbia en forma intensiva.

Selenio y salud animal Importancia, deficiencia, suplementación y toxicidad / Selenium and animal health: importance, defficiency, supplementation and toxicity / Selênio e saúde animal importância, deficiência, suplementação e toxidez

El selenio (Se) es un mineral esencial en la nutrición animal y se considera su participación en diversos procesos asociados a la producción animal, tan diversos como la fertilidad de la especie y la prevención de enfermedades. Laglutatión-peroxidasa (GSH-Px), fue la primera enzima en que se demostró la presencia activa del selenio y su importancia al evitar el daño oxidativo de las membranas celulares. Con anterioridad se había demostrado que la conocida como “Enfermedad del músculo blanco” era consecuencia de la deficiencia de Se, determinando muerte en animales recién nacidos y ocasionalmente en animales en desarrollo y aún en adultos, en particular en rumiantes. Actualmente ha quedado claro que el Se también es crítico en la estructuración de las enzimas necesarias para la síntesis de la hormona tiroidea y para su activaciónen los tejidos periféricos, pasando de T4 a T3. La deficiencia de Se afecta seriamente la capacidad de respuesta inmune de los animales. El diagnóstico de la deficiencia debe fundamentarse en los niveles del elemento en los suelos, las plantas forrajeras y la condición del animal en sangre y tejidos. Se han instrumentado diversas formas de suplementación del elemento que pueden emplearse dependiendo de las condiciones productivas de los animales y sus niveles previos de Se. La intoxicación por Se (Selenosis) debe tenerse en cuenta en los programas de suplementación, y puede ocurrir en formas crónicas o agudas. Las formas crónicas ocurren en regiones con suelos ricos en el elemento y condiciones que favorecen su absorción por las plantas forrajeras. Las formas agudas generalmente ocurren por excesos del elemento en las dietas o las sales que se administran a los animales o por errores de dosificación en las formulaciones parenterales. La relación metabólica del Se entre la hembra gestante, el feto y el recién nacido, es un área que requiere mayor investigación.

O selênio (Se) é um mineral essencial na nutrição animal e se considera sua participação em diversos processos associados à produção animal, tão diversos como na fertilidade das espécies e prevenção de enfermidades. A glutation peroxidase (GSH-Px), foi a primeira enzima em que se demonstrou a presença ativa do selênio e sua importância para evitar o dano oxidativo nas membranas celulares. Anteriormente sabia-se que a “enfermidade do músculo branco” era conseqüência da deficiência de Se, determinando a morte em animais recém-nascidos e ocasionalmente em animais em desenvolvimento, animais adultos e em particular em ruminantes. Atualmente, está claro que o Se é crítico na estruturação das enzimas necessáriaspara a síntese de hormônios da tireóide e para sua ativação nos tecidos periféricos, passando de T3 a T4. A deficiência de Se afeta seriamente a capacidade da resposta imune dos animais. O diagnóstico da deficiência deve fundamentar-se nos níveis deste elemento no solo, nas plantas forrageiras e nas condições do animal. Indicam-se diversas formas de suplementação deste elemento, que podem ser empregadas dependendo das condições produtivas dos animais e seus níveis prévios de Se. As intoxicações por Se (selenose), devem ser consideradas nos programas de suplementação, que pode ocorrer de forma crônica ou aguda. As formas crônicas ocorrem em regiões com solos ricos no elemento e nas condições que favorecem a absorção pelas plantas forrageiras. As formas agudas geralmente ocorrem por excesso do elemento nas dietas, ou sais administrados aos animais ou por erros na dosagem das formulações parenterais. A relação metabólica do Se na fêmea gestante, o feto e orecém-nascido, é uma área que requer uma maior investigação.

Selenium (Se) is an essential trace element for animal nutrition and plays multiple actions related to animal production, fertility and prevention of diseases. Glutathione peroxidase enzyme (GSH-Px) was the first seleno enzyme to demonstrate the active presence of selenium and its importance to prevent cellular membrane oxidative damage. Formerly, White muscle disease (WMD) was recognized as resulting from Se deficiency, determining new born mortality and adult animals, especially in ruminants. Nowadays, it is clear that Se is critical for thyroid hormone synthesis and its activation in peripheral tissues – from T3 to T4. Lack of Se seriously affects the capacity of immune response by the animals. Se status in the soil, plants, animal blood and tissues can be used as a tool to diagnose Se deficiency. Several forms of Se supplementation are described which can be applied depending of the productive capacity of the animals and their previous levels of Se. Acute and chronic Se intoxication (Selenosis) should be considered in supplementation programs as it may occur in a chronic or acute way. Chronic forms are associated to regional seleniferous soils, with permanent or repeated consumption of seleniferous plants. Acute forms are associated with high ingestion of Se as a result of wrongly designed diets, salt provided to the animals or parenteral injection dosage mistakes. The metabolic relation among Se, the fetus, the newborn and the pregnant dam requires further investigations.

Interacción de cepas verocitotóxicas de Escherichia coli y rotavirus en un brote de diarrea en becerros / Interaction of Escherichia coli verocytotoxin strains and rotavirus in an outbreak of calves diarrhoea

Se estudió un brote de diarrea hemorrágica en becerros de 1 a 20 días de edad, de curso agudo, con edema gastrointestinal, signos nerviosos y sin respuesta al tratamiento con antibióticos. Se realizaron estudios parasitológicos, bacteriológicos e identificación de rotavirus. Se determinó en las cepas de Escherichia coli aisladas la capacidad de producir citotoxinas sobre células vero. El 63.7 por ciento de las cepas aisladas de E.coli resultaron verocitotóxicas (productoras de toxina vero o de toxina shiga, Stx), estas cepas se aislaron del 86.5 por ciento de los animales afectados. En 24 por ciento de estos animales se determinó la presencia de rotavirus mediante electroforesis en PAGE. Se encontraron variaciones en la movilidad electroforética de las bandas. Todos los casos con identificación de rotavirus correspondieron con la presencia de cepas de Escherichia coli productoras de toxina siga (STEC) en el intestino. El porcentaje de riesgo calculado, de presentar infección con rotavirus atribuible a la infección con E.coli STX, fue de 100, no existió, en cambio, riesgo de presentar cepas STX si se presentaba una infección primaria con rotavirus. Los datos sugieren que las cepas citotóxicas son agentes primarios de infección y toxemia neonatal en bovinos; a la vez, se refuerza el probable papel de los bovinos como reservorios de cepas citotóxicas que afectan al humano y determinan el síndrome urémico hemolítico.

Lesiones por parapoxvirus en veterinarios de México / Parapoxvirus infection in veterinary practitioners of México

Se describen tres casos de lesiones en las manos médicos de veterinarios, producidas por parapoxvirus. En dos de los casos, las lesiones pudieron atribuirse al virus del ectima contagioso, tomando en cuenta la historia clínica. En todos los casos el diagnóstico se confirmó mediante la observación por microscopía electrónica de las partículas virales características (Tipo 2 o C)

Evaluación experimental de un inmunógeno de Pasteurella heamolytica en corderos / Experimental evaluation of Pasteurella haemolytica immunogen in lambs

Se evaluó el efecto de la inoculación de cordero con bacterias vivas y citotoxina de Pasteurella heamolytica al desafío. Se utilizaron 40 corderos distribuidos al azar en 4 grupos. l grupo 1 (lote testigo) se le inoculò, por vía subcutánea, sólo medio de cultivo; el grupo 2 fue inmunizado con cultivo vivo de P. Haemolyticaserotipo Al por vía subcutánea; el grupo 3 recibío el mismo inmunógeno por vía intradérmica y el grupo 4 con citotoxina de P. Heamolytica por vía subcutanea más adyuvante completo de Freund. Los animales fueron expuestos al virus de P13 y desafiados una semana después con una cepa de P. Heamolytica Al. fueron sacrificados para la evaluación del daño pulmonar. Los animales del grupo testigo mostraron fiebre después del desafío bacteriano. Los títulos de anticuerpos anticápsula y antitotoxina fueron más altos para los grupos inmunizados. comparados con el grupo testigo (P < 0.05). Las lesiones en pulmón fueron más severas para el grupo testigo (P < 0.05), sin diferncia significativa entre las diversas vías de inmunización. Según los resultados obtenidos, se puede considerar que la protección conferida por la inmunización con P. heamolytica así como con citotoxina, es eficiente para reducir en forma considerable las lesiones pulmonares ocacionadas por esta bacteria experimentalmente. Asimismo, se demostró la inocuidad de los inmunogenos por ausencia de reacciones posvacunales.