RESUMEN
Leptospirosis is a global neglected zoonosis, responsible for at least 1 million cases per year and almost 60 thousand deaths. The disease is caused by pathogenic and virulent bacteria of the genus Leptospira, either by direct contact with the bacteria or indirectly by exposure to contaminated water or soil. Domestic and wild animals act as reservoir hosts of infection, shedding leptospires from colonized renal tubules of the kidney, via urine, into the environment. The generation of mutant strains of Leptospira is critical to evaluate and understand pathogenic mechanisms of infection. CRISPR interference (CRISPRi) has proven to be a straightforward, affordable, and specific tool for gene silencing in pathogenic Leptospira. Therefore, the methodological details of obtaining the plasmid constructs containing both dCas9 and guide RNA, delivery of plasmids to Leptospira by conjugation with the E. coli strain ß2163, and transconjugant recovery and evaluation, will be described. In addition, the recently described Hornsby-Alt-Nally (HAN) media allows for the relatively rapid isolation and selection of mutant colonies on agar plates.
Asunto(s)
Leptospira , Leptospirosis , Animales , Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas , Escherichia coli , Silenciador del Gen , Leptospira/genética , Leptospirosis/genéticaRESUMEN
Total thyroxine (T4) concentrations are lower in healthy greyhounds compared to most other non-sighthound breeds. In humans, variations in the structure or concentration of the major thyroid hormone binding proteins are responsible for most reported differences between total T4 concentrations in healthy individuals from different ethnic groups or other subpopulations. The aim of this study was to determine if such variations are also responsible for the lower total T4 concentrations in greyhounds. The predicted protein sequences of thyroxine-binding globulin (TBG), transthyretin and albumin were determined in liver tissue from a euthyroid greyhound with decreased T4 concentration and a Jack Russell terrier using reverse-transcriptase PCR. Sequences were compared to each other and online reference sequences. Serum proteins from 21 greyhounds and 21 non-sighthound dogs were separated by denaturing electrophoresis and immunoblots probed with polyclonal antibodies to human TBG and transthyretin. Reactive bands were quantified by densitrometry, expressed relative to the mean of reference samples included in each gel. Serum albumin concentrations were measured using a commercially-available assay. Several SNPs were identified but none was thought likely to explain the lower total T4 concentrations in greyhounds. There was no significant difference between the quantity of any of the binding proteins in serum from greyhounds and non-sighthound dogs. However, total T4 and transthyretin concentrations were highly correlated in the greyhound group (r = 0.73, P = 0.0002). Variation in the sequence of thyroid hormone binding proteins is not responsible for low greyhound total T4 concentrations. Further evaluation of the role of transthyretin is warranted.
Asunto(s)
Hormonas Tiroideas , Tiroxina , Animales , Anticuerpos , PerrosRESUMEN
Leptospirosis is a neglected, widespread zoonosis caused by pathogenic species of the genus Leptospira, and is responsible for 60,000 deaths per year. Pathogenic mechanisms of leptospirosis remain poorly understood mainly because targeted mutations or gene silencing in pathogenic Leptospira continues to be inherently inefficient, laborious, costly and difficult to implement. In addition, pathogenic leptospires are highly fastidious and the selection of mutants on solid agar media can take up to 6 weeks. The catalytically inactive Cas9 (dCas9) is an RNA-guided DNA-binding protein from the Streptococcus pyogenes CRISPR/Cas system and can be used for gene silencing, in a strategy termed CRISPR interference (CRISPRi). Here, this technique was employed to silence genes encoding major outer membrane proteins of pathogenic L. interrogans. Conjugation protocols were optimized using the newly described HAN media modified for rapid mutant recovery at 37 °C in 3% CO2 within 8 days. Complete silencing of LipL32 and concomitant and complete silencing of both LigA and LigB outer membrane proteins were achieved, revealing for the first time that Lig proteins are involved in pathogenic Leptospira serum resistance. Gene silencing in pathogenic leptospires and rapid mutant recovery will facilitate novel studies to further evaluate and understand pathogenic mechanisms of leptospirosis.
Asunto(s)
Antígenos Bacterianos/genética , Proteínas de la Membrana Bacteriana Externa/genética , Sistemas CRISPR-Cas/genética , Silenciador del Gen , Leptospira interrogans/genética , Lipoproteínas/genética , Humanos , Leptospirosis/microbiología , Fenotipo , ARN Guía de Kinetoplastida/genéticaRESUMEN
Leptospirosis is a global neglected zoonosis, responsible for at least 1 million cases per year and almost 60 thousand deaths. The disease is caused by pathogenic and virulent bacteria of the genus Leptospira, either by direct contact with the bacteria or indirectly by exposure to contaminated water or soil. Domestic and wild animals act as reservoir hosts of infection, shedding leptospires from colonized renal tubules of the kidney, via urine, into the environment. The generation of mutant strains of Leptospira is critical to evaluate and understand pathogenic mechanisms of infection. CRISPR interference (CRISPRi) has proven to be a straightforward, affordable, and specific tool for gene silencing in pathogenic Leptospira. Therefore, the methodological details of obtaining the plasmid constructs containing both dCas9 and guide RNA, delivery of plasmids to Leptospira by conjugation with the E. coli strain β2163, and transconjugant recovery and evaluation, will be described. In addition, the recently described Hornsby-Alt-Nally (HAN) media allows for the relatively rapid isolation and selection of mutant colonies on agar plates.
RESUMEN
Leptospirosis is a neglected, widespread zoonosis caused by pathogenic species of the genus Leptospira, and is responsible for 60,000 deaths per year. Pathogenic mechanisms of leptospirosis remain poorly understood mainly because targeted mutations or gene silencing in pathogenic Leptospira continues to be inherently inefficient, laborious, costly and difficult to implement. In addition, pathogenic leptospires are highly fastidious and the selection of mutants on solid agar media can take up to 6 weeks. The catalytically inactive Cas9 (dCas9) is an RNA-guided DNA-binding protein from the Streptococcus pyogenes CRISPR/Cas system and can be used for gene silencing, in a strategy termed CRISPR interference (CRISPRi). Here, this technique was employed to silence genes encoding major outer membrane proteins of pathogenic L. interrogans. Conjugation protocols were optimized using the newly described HAN media modified for rapid mutant recovery at 37 °C in 3% CO2 within 8 days. Complete silencing of LipL32 and concomitant and complete silencing of both LigA and LigB outer membrane proteins were achieved, revealing for the first time that Lig proteins are involved in pathogenic Leptospira serum resistance. Gene silencing in pathogenic leptospires and rapid mutant recovery will facilitate novel studies to further evaluate and understand pathogenic mechanisms of leptospirosis.
RESUMEN
MIP and CPAF from Chlamydia have been shown to be effective in inducing immune responses important in clearing chlamydial infections. This study evaluates the protection conferred by MIP and CPAF as novel vaccines in pregnant C. abortus challenged ewes. Fifty C. abortus sero-negative sheep were randomly allocated into 5 groups of 10 according to the treatment they were to receive (1) 100⯵g of MBP-MIP (2) 100⯵g CPAF (3) 50⯵g MBP-MIP and 50⯵g CPAF (4) Tris-buffer (negative control) (5) Enzovax (positive control). Booster inoculations were administered 3â¯weeks after primary inoculations. Blood samples were taken pre-vaccination and weekly for 5â¯weeks. Five months after vaccination the ewes were mated. Pregnant ewes were then challenged on day 90 of gestation. Blood samples taken at four time-points post challenge were analysed for IFNγ levels, TNFα and IL-10 expression and anti-chlamydial antibody levels. Vaginal swabs, placental and foetal tissue and bacterial shedding were analysed using qPCR to quantify levels of C. abortus. Enzovax was 100% effective with no abortions occurring. The MIP/CPAF combined vaccine offered the greatest protection of the novel vaccines with 67% of ewes giving birth to one or more live lambs equating to a 50% vaccine efficacy rate. MIP and CPAF administered singly did not confer protection. Enzovax and MIP/CPAF vaccinated ewes had longer gestations and lambs with higher birth weights than negative control ewes. Aborting ewes shed higher numbers of C. abortus than ewes that had live lambs, all vaccinated ewes demonstrated lower levels of bacterial shedding than negative control ewes with Enzovax ewes shedding significantly fewer bacteria. Ewes that went on to abort had significantly higher levels of IFNγ and IL-10 at day 35 post challenge and significantly higher levels of anti-chlamydial antibodies at 24â¯h post lambing compared to ewes that had live lambs.
Asunto(s)
Infecciones por Chlamydia/inmunología , Infecciones por Chlamydia/prevención & control , Chlamydia/inmunología , Chlamydia/patogenicidad , Endopeptidasas/inmunología , Vacunación/métodos , Aborto Veterinario/prevención & control , Animales , Vacunas Bacterianas/inmunología , Vacunas Bacterianas/uso terapéutico , Endopeptidasas/metabolismo , Femenino , Embarazo , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/metabolismo , Ovinos , Enfermedades de las Ovejas/inmunología , Enfermedades de las Ovejas/prevención & controlRESUMEN
Leptospirosis is a neglected zoonosis of global importance with a complex epidemiology that affects humans, domestic and wild mammals. However, due to the diversity of clinical signs and difficulties of establishing a confirmatory laboratory diagnosis, the disease remains poorly investigated, particularly in the developing world. In Morocco, a descriptive study of the seroprevalence of Leptospira infection in animals has never been undertaken. To fill this gap, the current study was conducted on a subset of animals in north-west Morocco as a preliminary step towards understanding the epidemiological patterns of animal leptospirosis in the country. The study was conducted on 289 serum samples collected between January and April 2012 from dogs, cattle, sheep, goats and donkeys in the areas of Rabat-Temara, Sidi Kacem and Oulmes. All serum samples were tested by the MAT with 14 reference strains of the most prevalent pathogenic serovars of Leptospira and two serovars of non-pathogenic Leptospira. The overall seroprevalence of Leptospira in cattle, sheep, goats, dogs and donkeys was 15%, 18%, 20%, 21% and 20%, respectively. The most prevalent serogroups found in each species were Ballum, Sejroe, and Australis in cattle, Ballum, Australis and Sejroe in sheep, Australis and Ballum in goats, Javanica and Australis in donkey and Australis, Ballum and Canicola in dogs. Of all the serogroups tested in this study, Icterohaemorrhagiae, the only serogroup which has been previously reported in humans in Morocco, was rarely reactive. The majority of reactive sera were collected from low land areas. A large number of sera samples classified as seronegative when tested against pathogenic leptospires were positive when tested against non-pathogenic leptospires; this is suggestive of possible novel, as yet unclassified, Leptospira serovars in Morocco. Eleven of thirteen sheep urine samples were positive by real-time PCR confirming their role as Leptospira carriers in Morocco.
Asunto(s)
Animales Domésticos/microbiología , Leptospira/inmunología , Leptospirosis/epidemiología , Animales , Anticuerpos Antibacterianos/sangre , Bovinos , Perros , Equidae , Cabras , Leptospira/clasificación , Leptospira/aislamiento & purificación , Leptospirosis/microbiología , Leptospirosis/veterinaria , Marruecos/epidemiología , Prevalencia , Reacción en Cadena en Tiempo Real de la Polimerasa , Estudios Seroepidemiológicos , Serogrupo , OvinosRESUMEN
A total of 474 serum samples from client owned Irish dogs were tested for the presence of antibodies to serovars Canicola, Icterohaemorrhagiae, Bratislava, Autumnalis, Pomona, Altodouro, Grippotyphosa, Mozdok, Hardjobovis and Ballum. Six per cent of dogs presented to veterinary practitioners for problems unrelated to leptospirosis showed evidence of prior exposure to leptospiral serovars belonging to the serogropus Ballum, Australis, Pomona and Sejroe. One unvaccinated dog suspected to have leptospirosis showed seroconversion to serogroup Icterohaemorrhagiae. Based on these results the authors conclude that canine exposure to serogroup Ballum should be monitored because dogs may serve as sentinels for this serovar in the environment. Vaccination with multivalent vaccines containing serovar Bratislava in addition to serogroups Icterohaemorrhagiae and Canicola is advisable.
Asunto(s)
Anticuerpos Antibacterianos/sangre , Enfermedades de los Perros/epidemiología , Leptospira/inmunología , Leptospirosis/veterinaria , Pruebas de Aglutinación/veterinaria , Animales , Vacunas Bacterianas/inmunología , Perros , Irlanda/epidemiología , Leptospirosis/epidemiología , Prevalencia , Estudios SeroepidemiológicosRESUMEN
Leptospirosis is a zoonotic disease with a worldwide distribution affecting most mammalian species. Clinical leptospirosis is common in dogs but appears to be rare in cats. Both dogs and cats, however, can shed leptospires in the urine. This is problematic as it can lead to exposure of humans. The control of leptospirosis, therefore, is important not only from an animal but also from a public health perspective. The aim of this consensus statement is to raise awareness of leptospirosis and to outline the current knowledge on the epidemiology, clinical features, diagnostic tools, prevention and treatment measures relevant to canine and feline leptospirosis in Europe.
Asunto(s)
Enfermedades de los Gatos/prevención & control , Enfermedades de los Perros/prevención & control , Leptospirosis/veterinaria , Animales , Antibacterianos/uso terapéutico , Enfermedades de los Gatos/diagnóstico , Enfermedades de los Gatos/tratamiento farmacológico , Enfermedades de los Gatos/microbiología , Enfermedades de los Gatos/transmisión , Gatos/microbiología , Enfermedades de los Perros/diagnóstico , Enfermedades de los Perros/tratamiento farmacológico , Enfermedades de los Perros/microbiología , Enfermedades de los Perros/transmisión , Perros/microbiología , Humanos , Leptospira , Leptospirosis/diagnóstico , Leptospirosis/tratamiento farmacológico , Leptospirosis/microbiología , Leptospirosis/prevención & control , Leptospirosis/transmisión , Zoonosis/microbiología , Zoonosis/prevención & control , Zoonosis/transmisiónRESUMEN
There is limited evidence and guidance on best practice regarding preoperative dental assessments of patients awaiting cardiac operations. To analyse current practice in oral and maxillofacial units across the United Kingdom, we did a telephone survey to find out how these patients are managed. All maxillofacial departments in the United Kingdom were contacted, with a response rate of 77%. Those attached to dental schools were excluded. Most departments did extractions only, and in many, patients were assessed and treated within 7 days of being referred, although this varied. Most units did not offer antibiotic prophylaxis to patients at high risk of infective endocarditis, but some did after advice had been sought from a cardiologist. Respondents in 76% thought that preoperative dental assessments for cardiac patients were an appropriate use of resources. We propose a more integrated pathway for the management of these patients (particularly considering the pressure on time for treatment) that focuses primary and secondary resources accordingly.
Asunto(s)
Procedimientos Quirúrgicos Cardíacos/estadística & datos numéricos , Servicio Odontológico Hospitalario/estadística & datos numéricos , Cuidados Preoperatorios/estadística & datos numéricos , Cirugía Bucal , Profilaxis Antibiótica/estadística & datos numéricos , Restauración Dental Permanente/estadística & datos numéricos , Personal de Odontología en Hospital/estadística & datos numéricos , Endocarditis Bacteriana/prevención & control , Asignación de Recursos para la Atención de Salud/estadística & datos numéricos , Humanos , Guías de Práctica Clínica como Asunto , Derivación y Consulta/estadística & datos numéricos , Factores de Tiempo , Extracción Dental/estadística & datos numéricos , Reino UnidoRESUMEN
Animal production and health (APH) is an important sector in the world economy, representing a large proportion of the budget of all member states in the European Union and in other continents. APH is a highly competitive sector with a strong emphasis on innovation and, albeit with country to country variations, on scientific research. Proteomics (the study of all proteins present in a given tissue or fluid - i.e. the proteome) has an enormous potential when applied to APH. Nevertheless, for a variety of reasons and in contrast to disciplines such as plant sciences or human biomedicine, such potential is only now being tapped. To counter such limited usage, 6 years ago we created a consortium dedicated to the applications of Proteomics to APH, specifically in the form of a Cooperation in Science and Technology (COST) Action, termed FA1002--Proteomics in Farm Animals: www.cost-faproteomics.org. In 4 years, the consortium quickly enlarged to a total of 31 countries in Europe, as well as Israel, Argentina, Australia and New Zealand. This article has a triple purpose. First, we aim to provide clear examples on the applications and benefits of the use of proteomics in all aspects related to APH. Second, we provide insights and possibilities on the new trends and objectives for APH proteomics applications and technologies for the years to come. Finally, we provide an overview and balance of the major activities and accomplishments of the COST Action on Farm Animal Proteomics. These include activities such as the organization of seminars, workshops and major scientific conferences, organization of summer schools, financing Short-Term Scientific Missions (STSMs) and the generation of scientific literature. Overall, the Action has attained all of the proposed objectives and has made considerable difference by putting proteomics on the global map for animal and veterinary researchers in general and by contributing significantly to reduce the East-West and North-South gaps existing in the European farm animal research. Future activities of significance in the field of scientific research, involving members of the action, as well as others, will likely be established in the future.
Asunto(s)
Crianza de Animales Domésticos , Tecnología de Alimentos , Proteoma , Proteómica , Crianza de Animales Domésticos/tendencias , Fenómenos Fisiológicos Nutricionales de los Animales , Bienestar del Animal , Animales , Animales Domésticos , Acuicultura , Argentina , Australia , Productos Lácteos , Europa (Continente) , Unión Europea , Tecnología de Alimentos/tendencias , Israel , Carne , Nueva Zelanda , Proteómica/tendenciasRESUMEN
Triclosan is a biocidal active agent commonly used in domestic and industrial formulations. Currently, there is limited understanding of the mechanisms involved in triclosan tolerance in Escherichia coli O157. The aim of this study was to identify the differences between a triclosan susceptible E. coli O157:H19 isolate (minimum inhibitory concentration; MIC 6.25 µg/ml) and its triclosan tolerant mutant (MIC>8000 µg/ml) at a proteomic and phenotypic level. Two dimensional DIGE was used to identify differences in protein expression between the reference strain and triclosan tolerant mutant in the presence and absence of triclosan. DIGE analysis indicates the proteome of the reference E. coli O157:H19 was significantly different to its triclosan tolerant mutant. Significant changes in protein expression levels in the triclosan tolerant mutant included the known triclosan target FabI which encodes enoyl reductase, outer membrane proteins and the filament structural protein of flagella, FliC. Phenotypic studies showed that the triclosan tolerant mutant MIC decreased in the presence of efflux inhibitor phenyl-arginine-ß-naphthylamide and biofilm formation was increased in the mutant strain. The data generated indicates that enhanced triclosan tolerance is a result of multiple mechanisms which act together to achieve high-level resistance, rather than mutation of FabI alone.
Asunto(s)
Escherichia coli O157/enzimología , Proteómica/métodos , Triclosán/química , Acil-CoA Deshidrogenasas/química , Adhesión Bacteriana , Biopelículas , Células CACO-2 , Carbocianinas/química , Celulosa/química , Dipéptidos/química , Farmacorresistencia Bacteriana/efectos de los fármacos , Electroforesis en Gel Bidimensional , Escherichia coli O157/efectos de los fármacos , Perfilación de la Expresión Génica , Humanos , Immunoblotting , Espectrometría de Masas , Pruebas de Sensibilidad Microbiana , Mutación , Oxidorreductasas/metabolismo , Fenotipo , ProteomaRESUMEN
This study investigated the effects of cortisol and insulin, hormones that affect both glycaemic status and vascular function, on the in vitro contractility of isolated healthy equine small laminar veins. Small veins (150-500 µm) draining the digital laminae from healthy horses or ponies were investigated by wire myography. Concentration response curves were constructed for noradrenaline (NA), phenylephrine (PE), endothelin-1 (ET-1) and 5-hydroxytryptamine (5-HT) in the presence of either cortisol (10(-6 ) m) or insulin (1000 µIU/mL). Cortisol significantly increased the maximum contractility of laminar veins to the vasoconstrictors NA and 5-HT but decreased the maximal contraction to ET-1. Insulin decreased the contractility of vessels to PE and ET-1. It is possible that short-term cortisol excess could enhance venoconstrictor responses to 5-HT and NA in laminar veins in vivo, thereby predisposing to laminitis. Additionally, a reduction in the ability of insulin to counteract alpha-adrenoreceptor and ET-1-mediated contraction, likely to occur in subjects with insulin resistance, may further exacerbate venoconstriction in animals prone to laminitis. These mechanisms may also predispose horses with disorders such as equine Cushing's disease and equine metabolic syndrome to laminitis.
Asunto(s)
Pezuñas y Garras/irrigación sanguínea , Enfermedades de los Caballos/etiología , Hidrocortisona/farmacología , Insulina/farmacología , Vasoconstricción/efectos de los fármacos , Venas/efectos de los fármacos , Animales , Endotelina-1/farmacología , Pezuñas y Garras/patología , Enfermedades de los Caballos/metabolismo , Caballos , Inflamación/etiología , Inflamación/metabolismo , Inflamación/veterinaria , Norepinefrina/farmacología , Fenilefrina/farmacología , Serotonina/farmacologíaRESUMEN
Concern has been expressed about the overuse of biocides in farm animal production and food industries. Biocide application can create selective pressures that lead to increased tolerance to one or more of these compounds and are concomitant with the emergence of cross-resistance to antibiotics. A triclosan sensitive Salmonella enterica serovar Typhimurium and the isogenic triclosan tolerant mutant were studied at the proteomic level in order to elucidate cellular mechanisms that facilitate biocide tolerance. 2-D differential fluorescent gel electrophoresis (DIGE) compared protein profiles of parent and mutant Salmonella, in the presence and absence of triclosan. Differentially expressed proteins were identified by mass spectrometry and divided into two groups: Group A describes proteins differentially expressed between susceptible and triclosan tolerant Salmonella and includes the known triclosan target FabI which contained a mutation at the triclosan target binding site. Group B identified proteins differentially expressed in response to triclosan exposure and defines a general cell defence network. Only four proteins were common to both groups highlighting the diverse range of pathways employed by Salmonella to counteract biocides. These data suggest that sub-lethal concentrations of triclosan induce discernible changes in the proteome of exposed Salmonella and provide insights into mechanisms of response and tolerance.
Asunto(s)
Proteínas Bacterianas/metabolismo , Desinfectantes/farmacología , Farmacorresistencia Bacteriana/fisiología , Proteoma/metabolismo , Salmonella typhimurium/efectos de los fármacos , Salmonella typhimurium/metabolismo , Triclosán/farmacología , Especificidad de la EspecieRESUMEN
Toxoplasma gondii, an intracellular protozoan parasite, is one of the major causes of infectious abortion in sheep. To further understand the pathogenesis of toxoplasmosis, serum, amniotic and allantoic fluids and foetal stomach contents were collected from experimentally infected pregnant ewes to determine pathogen numbers and other markers of infection. Fifteen pregnant ewes (90 days of gestation) were each orally inoculated with 3000 sporulated oocysts of T. gondii. Serum samples were collected weekly following challenge. Amniotic and allantoic fluids and foetal stomach contents were collected at 21, 25, 28, 33 and 35 days post-infection. Characteristic placental lesions were detected in 1 of 4 challenged ewes at day 25, 3 of 4 challenged ewes at day 28 and in all challenged ewes at days 33 and 35 post-infection. T. gondii was detected only sporadically in amniotic and allantoic fluids before 35 days of infection, by real-time PCR, and only in ewes with placental lesions. At 35 days post-infection, high numbers of parasite were detected in both amniotic and allantoic fluids. An increase in the number of fluids from challenged animals with IgM and IgG was detected over time, except for IgG in allantoic fluid, which was detected in all samples from day 21 post-infection. IgG in amniotic and allantoic fluids was shown to be specific for T. gondii, and reacted with antigens with an apparent molecular mass of approximately 22 kDa and 30 kDa. Results suggest a maternal source of immunoglobulin in the allantoic fluid and a foetal source of immunoglobulin in the amniotic fluid early in infection but that both sources may contribute immunoglobulin to both fluids at a later stage.
Asunto(s)
Líquido Amniótico/química , Anticuerpos Antiprotozoarios/inmunología , Antígenos de Protozoos/inmunología , Complicaciones Parasitarias del Embarazo/parasitología , Enfermedades de las Ovejas/parasitología , Toxoplasmosis Animal/parasitología , Alantoides , Animales , Anticuerpos Antiprotozoarios/sangre , Anticuerpos Antiprotozoarios/química , Antígenos de Protozoos/sangre , Antígenos de Protozoos/química , Chlorocebus aethiops , Femenino , Inmunoglobulina G/sangre , Inmunoglobulina G/química , Inmunoglobulina M/sangre , Inmunoglobulina M/química , Placenta/parasitología , Placenta/patología , Embarazo , Complicaciones Parasitarias del Embarazo/inmunología , Complicaciones Parasitarias del Embarazo/patología , Ovinos , Enfermedades de las Ovejas/inmunología , Toxoplasmosis Animal/inmunología , Toxoplasmosis Animal/patología , Células VeroRESUMEN
Six ovine fetal brains were harvested 33 to 35 days postchallenge from 5 ewes, each of which was given 3000 Toxoplasma gondii oocysts on day 90 of pregnancy. Histopathologic examination of transverse sections taken at 13 levels in the fetal brains revealed the presence of toxoplasmosis-related lesions in all 6 brains. However, lesions were not randomly distributed (P = .007); they were most numerous at the level of the optic tract, the rostral margin of the pons, and 4 mm caudal to the ansate sulcus and were absent in all sections at the level of the caudal cerebellum. Lesion distribution may be due to hemodynamic factors, differences in the expression of endothelial surface receptor molecules at the level of the blood-brain barrier, or the presence of localized permissive/inhibitory factors within the brain. The results have implications for the selection of areas of brain from aborted ovine fetuses to be examined histopathologically for laboratory diagnosis.
Asunto(s)
Encéfalo/patología , Encéfalo/parasitología , Transmisión Vertical de Enfermedad Infecciosa/veterinaria , Enfermedades de las Ovejas/parasitología , Toxoplasmosis Animal/transmisión , Animales , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Técnicas Histológicas/veterinaria , Embarazo , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Ovinos , Enfermedades de las Ovejas/transmisiónRESUMEN
Pregnant ewes were challenged with Chlamydia abortus at 91-98 days of gestation and euthanised at 14, 21 and 28 days post-challenge. IFNγ mRNA labelling appeared to be co-localised with Chlamydial lipopolysaccharide within trophoblast cells in discrete areas lining the primary villi in the limbus and hilar zone of the placentomes from challenged sheep on days 21 and 28 post-infection. The presence of IFNγ was also demonstrated by immunohistochemistry. No labelling was seen in tissues from the non-infected ewes. The presence of IFNγ in trophoblast cells from infected ewes may indicate an attempt to restrict the replication of the organism and be an important trigger for the inflammatory responses that develop on the fetal side of the placenta in enzootic abortion.
Asunto(s)
Infecciones por Chlamydophila/metabolismo , Chlamydophila , Interferón gamma/biosíntesis , Trofoblastos/metabolismo , Aborto Séptico/inmunología , Aborto Séptico/metabolismo , Aborto Séptico/microbiología , Aborto Séptico/veterinaria , Animales , Infecciones por Chlamydophila/inmunología , Infecciones por Chlamydophila/microbiología , Femenino , Interferón gamma/inmunología , Embarazo , ARN Mensajero/biosíntesis , ARN Mensajero/inmunología , Ovinos , Enfermedades de las Ovejas/inmunología , Enfermedades de las Ovejas/microbiología , Trofoblastos/inmunología , Trofoblastos/microbiologíaAsunto(s)
Antibacterianos/uso terapéutico , Enfermedades de los Perros/transmisión , Enfermedades de los Perros/orina , Leptospirosis/veterinaria , Zoonosis , Enfermedad Aguda , Animales , Derrame de Bacterias , Enfermedades de los Perros/tratamiento farmacológico , Perros , Femenino , Leptospirosis/tratamiento farmacológico , Leptospirosis/transmisión , Leptospirosis/orina , Exposición Profesional , Factores de Riesgo , Orina/microbiologíaRESUMEN
Leptospirosis is a global zoonotic disease. Pathogenic Leptospira species, the causative agent of leptospirosis, colonize the renal tubules of chronically infected maintenance hosts such as dogs, rats and cattle. Maintenance hosts typically remain clinically asymptomatic and shed leptospires into the environment via urine. In contrast, accidental hosts such as humans can suffer severe acute forms of the disease. Infection results from direct contact with infected urine or indirectly, through contaminated water sources. In this study, a quantitative real-time PCR specific for lipL32 was designed to detect the urinary shedding of leptospires from dogs. The sensitivity and specificity of the assay was evaluated using both a panel of pathogenic Leptospira species and clinical microbial isolates, and samples of urine collected from experimentally infected rats and non-infected controls. The lower limit of detection was approximately 3 genome equivalents per reaction. The assay was applied to canine urine samples collected from local dog sanctuaries and the University Veterinary Hospital (UVH) at University College Dublin. Of 525 canine urine samples assayed, 37 were positive, indicating a prevalence of urinary shedding of leptospires of 7.05%. These results highlight the need to provide effective canine vaccination strategies and raise public health awareness.
Asunto(s)
Carga Bacteriana/métodos , Enfermedades de los Perros/diagnóstico , Enfermedades de los Perros/microbiología , Leptospira/aislamiento & purificación , Leptospirosis/veterinaria , Orina/microbiología , Animales , Proteínas de la Membrana Bacteriana Externa/genética , Derrame de Bacterias , ADN Bacteriano/genética , Perros , Hospitales Veterinarios , Irlanda , Leptospirosis/diagnóstico , Leptospirosis/microbiología , Lipoproteínas/genética , Reacción en Cadena de la Polimerasa/métodos , Prevalencia , Ratas , Sensibilidad y EspecificidadRESUMEN
Chlamydophila abortus is an intracellular pathogen and the etiological agent of enzootic abortion of ewes (EAE). C. abortus has a biphasic development cycle; extracellular infectious elementary bodies (EB) attach and penetrate host cells, where they give rise to intracellular, metabolically active reticulate bodies (RB). RB divide by binary fission and subsequently mature to EB, which, on rupture of infected cells, are released to infect new host cells. Pregnant ewes were challenged with 2 x 10(6) inclusion forming units (IFU) of C. abortus cultured in yolk sac (comprising both EB and RB). Serum samples were collected at 0, 7, 14, 21, 27, 30, 35, 40, and 43 days postinfection (dpi) and used to identify antigens of C. abortus expressed during disease. Additionally, sera from fetal lambs were collected at 30, 35, 40, and 43 dpi. All serum samples collected from experimentally infected pregnant ewes reacted specifically with several antigens of EB as determined by one-dimensional (1-D) and 2-D gel electrophoresis; reactive antigens identified by mass spectrometry included the major outer membrane protein (MOMP), polymorphic outer membrane protein (POMP), and macrophage infectivity potentiator (MIP) lipoprotein.