RESUMEN
We describe the clinicopathologic features of an arthritis outbreak in sheep induced by small ruminant lentivirus (SRLV), linked to the presence of a new SRLV isolate phylogenetically assigned to caprine arthritis encephalitis virus-like subgroup B2. Thirteen SRLV seropositive Rasa Aragonesa adult ewes were selected from 5 SRLV highly infected flocks (mean seroprevalence, 90.7%) for presenting uni- or bilateral chronic arthritis in the carpal joint. A complete study was performed, including symptomatology, histopathology, immunocytochemistry, immunohistochemistry, in situ hybridization, and microbiology. The carpus was the joint almost exclusively affected, with 10 sheep (76%) showing a moderate increase in carpal joint size (diameter range, 18-20 cm; normal range, 15-16 cm) without signs of locomotion problems and with 3 ewes (23%) showing severe inflammation with marked increase in diameter (21-24 cm), pain at palpation, and abnormal standing position. Grossly, chronic proliferative arthritis was observed in affected joints characterized by an increased thickness of the synovial capsule and synovial membrane proliferation. Microscopically, synovial membrane inflammation and proliferation and hyperplasia of synoviocytes were observed. More positive cases of SLRV infection were detected by immunocytochemistry of articular fluid than of bronchoalveolar lavage fluid. Immunohistochemistry and in situ hybridization also detected positive cells in the subsynovial connective tissue, lung, mediastinal lymph node, mammary gland, and mammary lymph node. All animals were negative for the presence of Mycoplasma or other bacteria in the articular space. The present outbreak likely represents an adaptation of a caprine virus to sheep. Our results underline the importance of the arthritis induced by SRLV in sheep, a clinical form that might be underestimated.
Asunto(s)
Artritis/veterinaria , Infecciones por Lentivirus/veterinaria , Lentivirus/fisiología , Enfermedades de las Ovejas/patología , Animales , Artritis/patología , Artritis/virología , Virus de la Artritis-Encefalitis Caprina/genética , Virus de la Artritis-Encefalitis Caprina/fisiología , Genotipo , Lentivirus/genética , Infecciones por Lentivirus/patología , Infecciones por Lentivirus/virología , Filogenia , Estudios Seroepidemiológicos , Ovinos , Enfermedades de las Ovejas/virología , Especificidad de la Especie , Membrana Sinovial/virologíaRESUMEN
The objective of this study was to assess the effects of Coxiella burnetii shedding or seropositivity on post-partum recovery and subsequent fertility in high-producing dairy cows. Given the difficulty in diagnosing C. burnetii infection at the farm level, an exhaustive series of tests in 43 pregnant animals that delivered at least one live calf were conducted, including blood serology and PCR of milk or colostrum, cotyledons (only at parturition), faeces, vaginal fluid against C. burnetii on gestation Day 171-177, at parturition and on Days 1-7, 8-14, 15-21, 22-28, 29-35 and 90-97 post-partum. During scheduled herd visits, ultrasonography (US) of the genital tract and examination of vaginal fluid were performed on Days 15-21 (V1), 22-28 (V2), 29-35 (V3) and 51-57 (V4) post-partum by the same veterinarian. Logistic regression analysis revealed that the likelihood of suffering endometritis (the presence of echogenic intrauterine fluid (IUF), cervical diameter of ≥4 cm or endometrial thickness ≥0.75 cm) was lower in C. burnetii-seropositive animals (OR = 0.10), compared with C. burnetii-seronegative animals. According to Kaplan-Meier survival analysis, C. burnetii-seronegative and non-shedding cows showed a delayed return to luteal activity and conception was delayed in non-shedding animals, compared with the remaining animals. Overall, the results of our study provide useful insight into the effects of C. burnetii infection on post-partum recovery and subsequent fertility. In particular, animals not infected with Coxiella seem to be susceptible to infection and not protected against the bacterium in dairy herds. The elevated costs of determining an infection at the farm level, make monitoring of cows virtually impossible from a clinical point of view.
Asunto(s)
Derrame de Bacterias/fisiología , Enfermedades de los Bovinos/microbiología , Coxiella burnetii/aislamiento & purificación , Fiebre Q/veterinaria , Animales , Anticuerpos Antibacterianos/sangre , Bovinos , Femenino , Periodo Periparto , Embarazo , Modelos de Riesgos Proporcionales , Fiebre Q/microbiologíaRESUMEN
AIMS: Copper and manganese levels are altered in mice both lacking PrPc and prion-infected brains. The aim of this study was to analyse the effects of manganese and copper imbalance on neuronal apoptosis in a scrapie-infected Tga20 mouse model. METHODS: Immunoreactivities for the apoptotic proteins Bax and active caspase-3 were evaluated in nine regions of the brain of scrapie-infected and control Tga20 mice treated with one of several diets: depleted cooper (-Cu), loaded manganese (+Mn), depleted copper/loaded manganese (-Cu+Mn) and regular diet. Immunohistochemical determination of NeuN was used to detect possible neuronal loss. RESULTS: Intracellular Bax detection was significantly decreased in animals fed with modified diets, particularly in those treated with copper-depleted diets. A decrease in active caspase-3 was primarily observed in animals fed with enhanced manganese diets. Our results show that the -Cu, -Cu+Mn and +Mn diets protected against apoptosis in scrapie-infected mice. However, NeuN immunolabelling quantification revealed that no diet was sufficient to arrest neuronal death. CONCLUSIONS: With regard to apoptosis induction, the response of Tga20 mice to prion infection was similar to that reported for other mice models. Our results demonstrate the neuroprotective effects of -Cu, -Cu+Mn and +Mn diets in a murine model of scrapie. However, neuronal death induced by infection with prions seems to be independent of apoptosis marker signalling. Moreover, copper-modified diets were neuroprotective against the possible toxicity of the prion transgene in Tga20 control and infected mice even though manganese supplementation could not counteract this toxicity.
Asunto(s)
Apoptosis/fisiología , Encéfalo/metabolismo , Cobre/metabolismo , Manganeso/metabolismo , Neuronas/metabolismo , Scrapie/metabolismo , Animales , Caspasa 3 , Cobre/administración & dosificación , Cobre/deficiencia , Proteínas de Unión al ADN , Dieta , Modelos Animales de Enfermedad , Manganeso/administración & dosificación , Manganeso/deficiencia , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Proteínas del Tejido Nervioso/metabolismo , Proteínas Nucleares/metabolismo , Proteínas Gestacionales/genética , Proteínas Gestacionales/metabolismo , Scrapie/dietoterapia , Proteína X Asociada a bcl-2/metabolismoRESUMEN
Scrapie is the archetype of prion diseases, fatal neurodegenerative disorders that affect humans and animals. Gene expression analysis of normal and infected sheep may provide clues to clarify the molecular mechanisms involved in the neuropathology of these diseases. Real time quantitative PCR has become a powerful and accurate technique for examination of transcription patterns in different biological conditions. One of the critical steps in the comparison of transcription profiles is the selection of stable genes for normalization of expression data. In this work, we have investigated the effect of scrapie on the stability of eight commonly used housekeeping genes in the central nervous system of sheep. We found that their stability decreased in scrapie-infected tissues, with the effect of the disease most evident in the medulla oblongata, a highly affected area of the brain stem. The risk of choosing inappropriate housekeeping genes for expression analysis was evaluated. Although the stability of each reference gene was suitable, a wide variation in expression of target genes (BAX and BCL2) was observed when only one or two housekeeping genes were used to normalize. However, reliable results were obtained with a normalization factor based on three reference genes, regardless of their position in a stability ranking.
Asunto(s)
Genes Esenciales/genética , Inestabilidad Genómica/genética , Scrapie/genética , Animales , Cerebelo/metabolismo , Diencéfalo/metabolismo , Femenino , Expresión Génica/genética , Bulbo Raquídeo/metabolismo , Datos de Secuencia Molecular , Corteza Prefrontal/metabolismo , ARN , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Scrapie/metabolismo , Ovinos/genéticaRESUMEN
Susceptibility/resistance to scrapie in sheep and goats is influenced by host prion protein gene (PRNP) genotype. In this study, we report the analysis of prion protein gene polymorphisms in 137 goats of two Moroccan populations: D'man and Chaouni. We found seven previously described amino acid polymorphisms at codons 37, 127, 137, 142, 154, 222 and 240, as well as three known silent mutations. In addition, we identified three new allelic variants: 101R and 139S in D'man goats and 145D in D'man and Chaouni individuals. The high frequency of the resistant allele 154H could offer genetic protection against the disease to the analysed animals. A total of 12 haplotypes and 28 genotypes were found, the distribution of which shows significant differences between both groups. Moreover, haplotype frequencies were compared with bibliographic data showing that the haplotype distribution of PRNP in Moroccan populations is genetically similar to Southern Italian and Greek goats.
Asunto(s)
Cabras/genética , Haplotipos , Priones/genética , Alelos , Animales , Cruzamientos Genéticos , Frecuencia de los Genes , Marruecos , Filogenia , Polimorfismo GenéticoRESUMEN
The nitrergic system produces nitric oxide as an atypical neurotransmitter in the nervous system. Nitric oxide is produced from l-arginine through specific enzymes known as nitric oxide synthases. Of these, the more abundant form in neurons is the constitutive neuronal nitric oxide synthase, although the inducible isoform can be expressed as well, especially following stress or other injuries. The excessive formation of nitric oxide results in protein nitration, particularly at tyrosine residues, thus the presence of nitrotyrosine can be used as a marker of nitric oxide production. In previous studies we have shown the distribution of the components of the nitrergic system in the cerebellum of rodents, where neuronal nitric oxide synthase immunoreactivity was present in stellate and basket cells, and occasionally in granule cells. Here, we present evidence that in the sheep, as a model of larger mammals, most cerebellar neurons display an intense immunostaining for neuronal nitric oxide synthase, including unipolar brush cells, and Lugaro and Golgi neurons, which are not immunoreactive in rodents. In addition, weak immunoreactivity for inducible nitric oxide synthase and nitrotyrosine was found in particular cell types, indicating a basal expression for these markers. Our results suggest a larger dependence on the nitrergic system for the cerebella of larger mammals. Since this increase happens in both activating and inhibitory neurons of the cerebellar circuitry, we propose that in these animals there is a higher steady-state regulation of the cerebellum based on nitric oxide.
Asunto(s)
Cerebelo/metabolismo , Neuronas/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Óxido Nítrico Sintasa de Tipo I/metabolismo , Tirosina/análogos & derivados , Animales , Inmunohistoquímica , Masculino , Óxido Nítrico/metabolismo , Ovinos , Tirosina/metabolismoRESUMEN
Small ruminant lentiviruses (SRLV) are widely spread in many countries, including Spain. However, little is known about the genetic characteristics of Spanish goat and sheep SRLV. In this study, segments from three genomic regions (pol, gag-p25 and LTR) were amplified using DNA isolated from three Spanish autochthonous sheep (one) and goats (two). Animals (one per flock) belonged to distantly located, single-species flocks (goat or sheep). Sequence analysis showed conservation of regions that are putatively relevant to viral survival. Sequences of Spanish goat and sheep SRLV were allocated into phylogenetic trees (phylograms) with known SRLV groups. The phylograms corresponding to the pol, gag-p25 and LTR regions analyzed presented a compatible topology. This showed that Spanish caprine and ovine SRLV sequences belonged to the A or D phylogenetic groups and were closer to sheep SRLV prototypes (A1 group) than to goat SRLV prototypes (B or C groups), according to the current classification [Shah, C., Boni, J., Huder, J.B., Vogt, H.R., Muhlherr, J., Zanoni, R., Miserez, R., Lutz, H., Schupbach, J., 2004a. Phylogenetic analysis and reclassification of caprine and ovine lentiviruses based on 104 new isolates: evidence for regular sheep-to-goat transmission and worldwide propagation through livestock trade. Virology 319 (1), 12-26]. It was not possible to amplify in the three genetic regions the expected fragment in additional Spanish caprine and ovine SRLV proviral DNA sequences with the PCR primers used. This suggests that there is heterogeneity at the primer binding site among Spanish SRLV sequences. It also illustrates the need to develop diagnostic tests that are sensitive in local breeds.
Asunto(s)
Virus de la Artritis-Encefalitis Caprina/genética , Infecciones por Lentivirus/epidemiología , Virus Visna-Maedi/genética , Visna/epidemiología , Secuencia de Aminoácidos , Animales , Productos del Gen gag/genética , Genes pol/genética , Genoma Viral , Cabras , Datos de Secuencia Molecular , Filogenia , Alineación de Secuencia , Ovinos , España/epidemiología , Secuencias Repetidas Terminales/genéticaRESUMEN
A three year long experimental study was carried out to investigate horizontal MVV-infection by PCR and ELISA, in 191 one year-old latxa dairy-sheep raised in two separate groups under low and high MVV-infection pressure, respectively. Sheep originated from a previous MVV-transmission study in lambs and seroprevalence among one year-old sheep in both groups was 15% approximately. The high infection-pressure group (H-group) consisted of 147 replacement ewes that joined a milk-producing, housed dairy-flock with 42-66% MVV-seroprevalence and the low infection-pressure group (L-group) were castrated males raised in a separate shed. In contrast to results obtained when infection was investigated in lambs, the overall degree of agreement between ELISA and PCR results was very good and there was some indication that it increased further as sheep became older. MVV-prevalence did not change in the L-group and increased to 57% in three year-old sheep in the H-group (p<0.001). Random effects logistic regression confirmed seroconversion was significantly higher in the H-group compared to the L-group and was highest during the year after the sheep were introduced in the dairy flock and did not increase with age as in previous studies using less sensitive antibody assays. The evidence that horizontal transmission can be very low in spite of prolonged close contact between infected and non-infected sheep is valuable for MVV-control purposes. Furthermore it highlights the need to investigate virus excretion dynamics in infected animals and animal to animal transmission to improve our overall understanding of horizontal MVV transmission in MVV endemic populations.
Asunto(s)
Transmisión de Enfermedad Infecciosa/veterinaria , Neumonía Intersticial Progresiva de los Ovinos/transmisión , Virus Visna-Maedi/aislamiento & purificación , Envejecimiento , Animales , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Masculino , Oportunidad Relativa , Neumonía Intersticial Progresiva de los Ovinos/inmunología , Neumonía Intersticial Progresiva de los Ovinos/virología , Reacción en Cadena de la Polimerasa/veterinaria , Ovinos , Virus Visna-Maedi/genética , Virus Visna-Maedi/inmunologíaRESUMEN
A recent large-scale experimental study showed that bottle-feeding ovine colostrum from seropositive ewes results in high MVV-seroconversion in lambs. In contrast, relatively few lambs that naturally suckled colostrum from seropositive dams seroconverted as a result of it. Furthermore, lambs fed uninfected bovine colostrum readily seroconverted when mixed with ovine-colostrum lambs indicating that horizontal MVV transmission between lambs was efficient. MVV-infection was further investigated in the same samples using two PCR tests targeting sequences in the long-terminal repeats (LTR) and POL MVV genes. PCR-tests confirmed previous serological findings. However, the LTR-PCR was more sensitive and allowed detecting infection earlier than the other tests, including 5-8% of new-born lambs from seropositive dams, providing more evidence that prenatal MVV-infection may be more important than considered. The degree of agreement between PCR and antibody tests in individual samples was low up to 6 months of age and moderate at 10 months-old. Nine percent of lambs were always PCR-negative but seroconverted and 19% of lambs were PCR-positive at least once and did not seroconvert. However, seroconversion was associated with increasing number of times lambs were PCR-positive and ovine colostrum-fed lambs were more frequently PCR-positive than other lambs. The significance of these findings in terms of MVV-infection, epidemiology and control is discussed.
Asunto(s)
Anticuerpos Antivirales/sangre , Calostro/virología , Transmisión Vertical de Enfermedad Infecciosa/veterinaria , Neumonía Intersticial Progresiva de los Ovinos/inmunología , Neumonía Intersticial Progresiva de los Ovinos/transmisión , Virus Visna-Maedi/genética , Virus Visna-Maedi/aislamiento & purificación , Envejecimiento , Animales , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Masculino , Oportunidad Relativa , Neumonía Intersticial Progresiva de los Ovinos/sangre , Neumonía Intersticial Progresiva de los Ovinos/virología , Reacción en Cadena de la Polimerasa/veterinaria , OvinosRESUMEN
Comparative studies evaluating the performance of Transmissible Spongiform Encephalopathies (TSE) rapid tests (validated for Bovine Spongiform Encephalopathy samples) on Scrapie samples have not been reported widely, particularly those dealing with lymphoreticular system tissues to a much lesser extent. The main objective of this study was to compare the ability of two current rapid tests (Western blot and Luminiscence Immunoassay Prionics-Check; WB and LIA, respectively) to detect PrPsc using central nervous system as well as lymphoreticular system samples corresponding to naturally infected animals. Thirty-four Scrapie-affected sheep, 26 with clinical signs of the disease, were included in the study. Tonsil, retropharyngeal lymph node and medulla oblongata were assessed by three tests: immunohistochemistry (confirmatory test), WB and LIA (rapid tests). The conclusion which can be drawn from this study is the fact that all animals involved in the study, including those at a preclinical stage, could be diagnosed regardless of the test used (with immunohistochemistry consistently showing higher sensitivity) only when the analyses of both the central nervous system and the lymphoreticular system were considered. The choice of these tissues for routine diagnosis is, therefore, proposed as a valuable tool to highly reduce the number of undetected positive cases.
Asunto(s)
Western Blotting/veterinaria , Sistema Nervioso Central/química , Encefalopatía Espongiforme Bovina/diagnóstico , Proteínas PrPSc/análisis , Scrapie , Animales , Western Blotting/métodos , Bovinos , Encefalopatía Espongiforme Bovina/epidemiología , Encefalopatía Espongiforme Bovina/virología , Inmunohistoquímica , Scrapie/diagnóstico , Scrapie/epidemiologíaRESUMEN
Maedi-visna virus (MVV) seroprevalence associated with consumption of colostrum from seropositive ewes was investigated in 276 housed lambs from birth to 300 days-old. At birth, lambs were allocated to five experimental groups according to the maternal MVV-serological status, source and mode of feeding colostrum (bovine or ovine and bottle fed or suckled from the dam) and type of horizontal MVV-exposure (raised with the dam or separately with other lambs). The risk of being seropositive at 300 days-old was associated with feeding ovine colostrum from seropositive ewes and increased with intake of bottle-fed ovine colostrum and was higher in lambs separated from their dams and raised with other experimental lambs compared to lambs raised with their dams. Approximately 75-87% of ELISA-positive results in lambs that had ovine colostrum was attributable to colostrum itself. However, approximately only 16% of naturally raised and 29-61% of bottle-fed ovine colostrum lambs were ELISA-positive as a result feeding ovine colostrum. These results confirm that ovine colostrum from seropositive ewes can be a major source of MVV but its overall contribution to seroprevalence in natural conditions is relatively low, and shows that horizontal MVV transmission can be an important source of infection in new-born lambs.
Asunto(s)
Calostro/virología , Transmisión Vertical de Enfermedad Infecciosa/veterinaria , Neumonía Intersticial Progresiva de los Ovinos/transmisión , Enfermedades de las Ovejas/transmisión , Virus Visna-Maedi , Envejecimiento , Animales , Anticuerpos Antivirales/sangre , Femenino , Estudios Longitudinales , Riesgo , Estudios Seroepidemiológicos , Ovinos , Virus Visna-Maedi/inmunologíaRESUMEN
Clinical and pathological studies in European badgers (Meles meles) are limited. Badgers play a significant role in the epidemiology of bovine tuberculosis (TB) in some countries and an accurate diagnosis is needed for this infection. However, the lesions of bovine TB are similar to those associated with other pathogens, making pathological diagnosis difficult. In the present study, Streptococcus halichoeri was isolated from a European badger with pyogranulomatous pleuropneumonia and suspected of having tuberculosis. TB and other pathogens able to induce similar lesions were ruled out. Comparative 16S rRNA and rpoB gene sequencing studies showed an identity of 99.51% and 98.28%, respectively, with S. halichoeri. This report represents the third description of this bacterium and the first in an animal species other than the grey seal (Halichoerus grypus). It also shows that S. halichoeri can be associated with a pathological process characterized by granulomatous inflammation and resembling tuberculosis.
Asunto(s)
Mustelidae/microbiología , Filogenia , Pleuroneumonía/veterinaria , Infecciones Estreptocócicas/veterinaria , Streptococcus/genética , Animales , Femenino , Granuloma/microbiología , Granuloma/patología , Granuloma/veterinaria , Pleuroneumonía/microbiología , Pleuroneumonía/patología , ARN Ribosómico 16S/análisis , Infecciones Estreptocócicas/patología , Streptococcus/aislamiento & purificaciónRESUMEN
Small ruminant lentiviruses include viruses with diverse genotypes that frequently cross the species barrier between sheep and goats and that display a great genetic variability. These characteristics stress the need to consider the whole host range and to perform local surveillance of the viruses to opt for optimum diagnostic tests, in order to establish control programmes. In the absence of effective vaccines, a comprehensive knowledge of the epidemiology of these infections is of major importance to limit their spread. This article intends to cover these aspects and to summarise information related to characteristics of the viruses, pathogenesis of the infection and description of the various syndromes produced, as well as the diagnostic tools available, the mechanisms involved in transmission of the pathogens and, finally, the control strategies that have been designed until now, with remarks on the drawbacks and the advantages of each one. We conclude that there are many variables influencing the expected cost and benefits of control programs that must be evaluated, in order to put into practice measures that might lead to control of these infections.
Asunto(s)
Infecciones por Lentivirus/veterinaria , Lentivirus/genética , Rumiantes/virología , Enfermedades de las Ovejas/diagnóstico , Animales , Enfermedades de las Cabras/diagnóstico , Enfermedades de las Cabras/etiología , Enfermedades de las Cabras/prevención & control , Cabras , Especificidad del Huésped , Interacciones Huésped-Patógeno , Lentivirus/fisiología , Infecciones por Lentivirus/diagnóstico , Infecciones por Lentivirus/etiología , Infecciones por Lentivirus/prevención & control , Ovinos , Enfermedades de las Ovejas/etiología , Enfermedades de las Ovejas/prevención & control , Oveja DomésticaRESUMEN
This work was aimed at studying the effect of maedi-visna virus (MVV) infection in vitro on the ability of sheep cells to adhere to staphylococci (Staphylococcus aureus and Staphylococcus epidermidis), and phagocytose these bacteria. Adherence was studied in sheep choroid plexus cells (SCPC) using an ELISA test and phagocytosis was studied in pulmonary alveolar macrophages (PAM) by chemiluminescence. A 5- and 7-day of in vitro MVV infection resulted in syncytium formation and a significant increased adherence (P < 0.01) of SCPC to bacteria. SCPC endogenous fibronectin was significantly higher (P < 0.01) on days 5 and 7 than on day 0 of MVV infection. A significantly decreased phagocytosis (P < 0.05) was also observed on days 5 and 7 of MVV infection in PAM when compared to MVV-free controls. Comparatively, phagocytosis was highest for S. aureus non-slime producing strains, followed by S. epidermidis, and S. aureus slime producing strains, in that order. Finally, increased expression of both, class I and class II major histocompatibility antigens was also observed in MVV-infected PAM on days 5 and 7, whereas SCPC only demonstrated upregulation of MHC class I. These results, indicative of an alteration of some cell functions in MVV-infected cells, may help to understand interactions between MVV-infected cells and bacteria in simultaneous infections and may provide clues to the possible in vivo interactions of both pathogens.
Asunto(s)
Adhesión Bacteriana , Fagocitosis , Ovinos/virología , Staphylococcus , Visna/inmunología , Animales , Plexo Coroideo/inmunología , Plexo Coroideo/virología , Ensayo de Inmunoadsorción Enzimática , Antígenos de Histocompatibilidad Clase I/análisis , Antígenos de Histocompatibilidad Clase II/análisis , Cinética , Mediciones Luminiscentes , Macrófagos Alveolares/inmunología , Macrófagos Alveolares/fisiología , Macrófagos Alveolares/virología , Staphylococcus aureus , Staphylococcus epidermidisRESUMEN
The aim of this work was to investigate whether an enzyme-linked immunosorbent assay (ELISA) was useful for early detection of maedi-visna virus (MVV) infection in sheep under field conditions. An ELISA based on p25 recombinant protein and a gp46 synthetic peptide was used. Sequentially obtained serum samples (n = 1,941) were studied for 4 years. ELISA results were compared with those of the agar gel immunodiffusion (AGID) test, and results of both tests were compared with a reference result established using consensus scores for at least 2 of 3 serologic techniques (AGID, ELISA, and western blotting, which was used to resolve result discrepancies between the other 2 techniques). A total of 247 discrepancies were observed between ELISA and AGID. Of these, 131 were due to an earlier detection of 120 sera by the ELISA and 11 sera by AGID. The remaining discrepancies (116) were due to the presence of false reactions in both tests. Fewer false-negative results were found by ELISA than with AGID (6 vs. 69 sera, respectively), whereas the number of false-positive results was virtually the same for ELISA and AGID (21 vs. 20, respectively). In relation to the reference result, ELISA sensitivity and specificity were 97.8% and 98.2%, respectively, whereas values for AGID were 76.3% and 98.3%, respectively. The agreement between ELISA and the reference result was higher than that between AGID and the reference result (K value: 0.96 and 0.77, respectively). A variation in the ELISA signal (based on optical density) was observed during the study period, suggesting different antibody levels throughout the animal's life. The ELISA was useful for detecting MVV-infected sheep in field conditions and has potential for use in control and eradication programs.
Asunto(s)
Ensayo de Inmunoadsorción Enzimática/veterinaria , Neumonía Intersticial Progresiva de los Ovinos/diagnóstico , Virus Visna-Maedi/inmunología , Animales , Western Blotting/veterinaria , ADN Viral/genética , Electroforesis en Gel de Agar/veterinaria , Neumonía Intersticial Progresiva de los Ovinos/inmunología , Reacción en Cadena de la Polimerasa , Sensibilidad y Especificidad , Pruebas Serológicas/veterinaria , Ovinos , Virus Visna-Maedi/patogenicidadRESUMEN
Paratuberculosis lesions in naturally infected sheep were classified histologically. Adult sheep (n = 166) culled for various reasons, from four flocks in which clinical cases of the disease had occurred, were studied. Eight-two sheep (49.4%) showed lesions that could be divided into three main categories. Type 1 lesions, found in 24.1% of these animals, consisted of small granulomata formed by macrophages and were located exclusively in the ileocaecal Peyer's patch. In type 2 lesions, found in 4.8% of the sheep, granulomata were also observed in the mucosa associated with Peyer's patches. Type 3 lesions were characterized by granulomata in areas of the mucosa associated with, and also distinct from, the Peyer's patches. Three subtypes of type 3 lesions were recognized. In subtype 3a, found in 4.2% of the sheep, multifocal granulomata appeared in different areas of the lamina propria; they were not apparently associated with lymphoid tissue, and neither did they modify the morphology of the affected areas. Subtype 3b, found in 13.9% of animals, consisted of large numbers of macrophages, widespread in the lamina propria; in subtype 3c (2.4% of sheep) lymphocytes were the main inflammatory cell, with some macrophages scattered amongst them. In subtypes 3a and b, villi were distended and the mucosa appeared thickened. Mycobacteria could be demonstrated in tissue sections from all the samples with subtype 3b lesions and in almost all of those with type 2 and 3a lesions; these organisms were absent or sparse, however, in type 1 and 3c lesions. Macroscopical lesions were clearly visible only in sheep with type 3b and 3c lesions. Mycobacterium paratuberculosis was cultured from all the animals with type 3 lesions, from 87.5% of those with type 2 lesions, and from 47.5% of those with type 1. Type 3b lesions resembled the "borderline-lepromatous" form of mycobacterial lesions, whereas type 3c lesions resembled the "borderline-tuberculoid" form. The relationship between intestinal lymphoid tissue and paratuberculosis lesions is discussed, and the diagnostic importance of histological examination of the ileocaecal valve emphasized.
Asunto(s)
Granuloma/clasificación , Granuloma/patología , Paratuberculosis/patología , Enfermedades de las Ovejas/clasificación , Enfermedades de las Ovejas/patología , Animales , Granuloma/veterinaria , Mycobacterium avium subsp. paratuberculosis/aislamiento & purificación , Paratuberculosis/microbiología , Ovinos , Enfermedades de las Ovejas/microbiologíaRESUMEN
A study of the pathogenesis of paratuberculosis and the effects of vaccination was carried out on 17 Rasa lambs, allocated to four groups. Vaccination seemed to accelerate the progress of the infection, and led quickly to healing. The only site of early lesions was the interfollicular areas in the Peyer's patches. This would suggest an explanation both for the location of paratuberculous lesions in the ileum in clinical cases, and for the lower susceptibility to infection of adult animals, in which the intestinal organized lymphoid tissue is greatly diminished. Thus, a critical role of the Peyer's patch in the establishment of M. paratuberculosis infection is suspected.
Asunto(s)
Vacunas Bacterianas , Paratuberculosis/inmunología , Enfermedades de las Ovejas/inmunología , Enfermedades de las Ovejas/microbiología , Animales , Anticuerpos Antibacterianos/sangre , Ensayo de Inmunoadsorción Enzimática/veterinaria , Heces/microbiología , Granuloma/microbiología , Granuloma/veterinaria , Ganglios Linfáticos Agregados/patología , OvinosRESUMEN
The finding of brain tissue fragments in blood and lungs of cattle stunned in slaughterhouses has raised concerns about food safety in the context of the bovine spongiform encephalopathy epidemic. In the present study, the possible occurrence of brain tissue emboli in animals killed in traditional Spanish bullfighting was investigated. Thorough histological analysis of multiple possible target organs was carried out in 434 bulls. No evidence of brain tissue embolism was obtained, but emboli from diverse sources were detected in pulmonary and hepatic tissue of a significant number of animals. These emboli seem to have been caused by the use of a long sword, which extensively disrupts intra-thoracic and intra-abdominal organs and vascular structures.
Asunto(s)
Embolia/veterinaria , Encefalopatía Espongiforme Bovina/transmisión , Deportes , Animales , Bovinos , Brotes de Enfermedades/prevención & control , Brotes de Enfermedades/veterinaria , Embolia/etiología , Embolia/patología , Encefalopatía Espongiforme Bovina/epidemiología , Masculino , España/epidemiologíaRESUMEN
OBJECTIVE: To investigate the relation between lesions and serologic response in sheep with naturally acquired paratuberculosis and to evaluate the efficacy of agar gel immunodiffusion (AGID) and ELISA as diagnostic tests. ANIMALS: 134 adult sheep, culled by the owner from flocks with history of severe chronic weight loss. PROCEDURE: Before euthanasia, blood samples were collected and serum was obtained. AGID and ELISA were performed, using a protoplasmic antigen of Mycobacterium paratuberculosis. Sensitivity and specificity of these techniques were evaluated, using histopathologic findings as a reference. Histologic studies were carried out focusing on specimens containing intestinal organized lymphoid tissue. RESULTS: Of the sheep examined, 46.3% had lesions that were classified in 5 categories, according to their location, extension, cellular types, and presence of acid-fast bacilli. Regarding serologic test results, 37.1% of sheep with lesions had a positive AGID result, and all sheep without lesions had negative results. However, ELISA positivity was slightly greater--48.4% of sheep with lesions; however, by use of ELISA, 11.1% of sheep without lesions had positive results. CONCLUSION: A close relation between pathologic findings and serologic response was determined, especially between presence of acid-fast bacilli (potentially heavily shedding sheep) in the intestinal lesions and positivity to serologic tests, especially AGID. CLINICAL RELEVANCE: On the basis of these results, together with its high specificity, reliability, and the difficulty of fecal specimen culture in sheep, AGID is proposed as a useful tool for seeking evidence of paratuberculosis in live sheep, and therefore, is suggested as the basis for control programs in areas in which the disease is a serious threat to sheep production.
Asunto(s)
Paratuberculosis/diagnóstico , Animales , Anticuerpos Antibacterianos/sangre , Antígenos Bacterianos/análisis , Ensayo de Inmunoadsorción Enzimática , Inmunodifusión , Tejido Linfoide/microbiología , Tejido Linfoide/patología , Mycobacterium avium subsp. paratuberculosis/aislamiento & purificación , Paratuberculosis/sangre , Paratuberculosis/patología , Sensibilidad y Especificidad , OvinosRESUMEN
Maedi-visna, a multisystemic disease of adult sheep, was first described in Spain in 1984. To get an idea of the seroprevalence of the disease locally and to estimate the number of seropositive animals with lesions, samples of blood, lungs and mammary glands were taken from 124 randomly selected sheep killed in the main slaughterhouse of Zaragoza. In the agar gel immunodiffusion test, 74 (59.7 per cent) of the sheep were positive and 50 were negative. Among the 74 seropositive animals, 19 (25.6 per cent) had no lesions in any organ, 12 (16.2 per cent) had lesions in the lungs only, 15 (20.2 per cent) had lesions in the mammary glands and 28 (37.8 per cent) had lesions in both organs. In the lungs hyperplasia of lymphoid follicles was more evident than an interstitial infiltrate but in the mammary glands this relationship was not observed. Even when the lesions occurred in both organs, they did not show the expected proportion in terms of either type or severity. Among the 50 seronegative sheep, eight (16 per cent) showed maedi-like lesions, formed exclusively by the hyperplasia of lymphoid follicles.