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1.
Schweiz Arch Tierheilkd ; 164(12): 851-859, 2022 Dec.
Artículo en Alemán | MEDLINE | ID: mdl-36454014

RESUMEN

INTRODUCTION: Contagious ovine digital dermatitis (CODD) is an emerging infectious foot disease in sheep. To date, CODD has been described in Great Britain, Ireland, Sweden and Germany and now in Switzerland for the first time. Unlike foot rot, the CODD lesions do not spread from the interdigital space, but usually begin at the dorsal/abaxial coronary band. The changes can spread to the hoof wall and the sole and finally can lead to exungulation, similar to foot rot. Treponema spp. are often found in CODD lesions analogous to digital dermatitis (Mortellaro's disease) in cattle. Involvement of Dichelobacter nodosus (D. nodosus) is considered a risk factor, but the presence of the bacterium is not mandatory. In February 2022, ulcerative lesions in the dorso-axial coronary band area were noticed on both claws of the left forelimb in an ewe. Histology of the biopsy showed hyperkeratosis and erosion with exocytosis and crust formation. Treponema spp. PCR and fluorescence in situ hybridization (FISH) were positive for Treponema phylotype 1 (PT1). In addition, D. nodosus and Porphyromonas levii could be detected in the biopsy using PCR. A single local application of chlortetracycline spray led to clinical healing within two weeks, no recurrence was seen within the following two months. Three control sheep, which were kept together with the diseased sheep, did not show any clinical signs of CODD. Treponema spp could not be found in interdigital and coronary band biopsies by PCR or FISH. This is the first description of CODD in Switzerland and aims to sensitize veterinarians to CODD as a differential diagnosis for foot diseases in sheep.


INTRODUCTION: La dermatite digitale contagieuse ovine (contagious ovine digital dermatitis; CODD) est une maladie infectieuse des onglons des moutons d'importance croissante. À ce jour, la CODD a été décrite en Grande-Bretagne, Irlande, Suède et Allemagne, et maintenant pour la première fois également en Suisse. Au contraire du piétain, les lésions de CODD ne s'étendent pas à partir de l'espace interdigité, mais elles commencent en général au bord coronaire dorsal/abaxial. De là, les lésions peuvent s'étendre à la corne de la paroi et à la sole, ce qui peut finalement conduire à une perte complète de la boite cornée de l'onglon, comme en cas de piétain. En analogie à la dermatite digitale (maladie de Mortellaro) chez les bovins, des tréponèmes sont souvent mis en évidence dans les lésions de CODD. La présence de Dichelobacter nodosus (D. nodosus) est considérée comme un facteur de risque, mais elle n'est pas indispensable au développement de la CODD. Des lésions ulcératives dans la région du bord coronaire dorso-axial des deux onglons antérieurs d'une brebis ont été remarqués en février 2022. L'examen histologique de la biopsie de la lésion de CODD a montré une hyperkératose ainsi que des érosions avec de l'exocytose et la formation de croûtes. Aussi bien la PCR pour les Treponema spp. que l'hybridisation in-situ à fluorescence (FISH) étaient positives pour Treponema Phylotype 1 (PT1). De plus, D. nodosus et Porphyromonas levii ont été mis en évidence dans la biopsie. Une application locale unique de spray à la tétracycline après le prélèvement de la biopsie a conduit à une guérison clinique en deux semaines, et aucune récidive n'a été observée dans le deux mois suivants. Trois moutons de boucherie qui étaient détenus avec la brebis malade mais ne présentaient pas de lésions de CODD ont servi de contrôles négatifs. Des Treponema spp. n'ont été mis en évidence chez ces animaux, ni dans des biopsies du bord coronaire ni dans celles de l'espace interdigité. Cette étude représente la première description de la CODD en Suisse et est destinée à sensibiliser la profession vétérinaire à la CODD comme diagnostic différentel en cas de maladies des onglons chez les moutons.


Asunto(s)
Dichelobacter nodosus , Dermatitis Digital , Panadizo Interdigital , Enfermedades de las Ovejas , Animales , Femenino , Dermatitis Digital/diagnóstico , Dermatitis Digital/tratamiento farmacológico , Panadizo Interdigital/diagnóstico , Panadizo Interdigital/tratamiento farmacológico , Hibridación Fluorescente in Situ/veterinaria , Ovinos , Enfermedades de las Ovejas/diagnóstico , Enfermedades de las Ovejas/tratamiento farmacológico , Suiza , Treponema/genética
2.
Artículo en Inglés | MEDLINE | ID: mdl-36343421

RESUMEN

In many countries, sheep lameness is a cause of economic concern and a contributing factor to a declining economy. This study aimed to investigate changes in procalcitonin (PCT), acute phase proteins (APPs), and cytokines (CYTs) in response to interdigital dermatitis and footrot in sheep under field conditions, to emphasize their role in the disease pathogenesis, diagnosis, as well as monitoring treatment response. Fifty-three sheep with foot diseases (26 clinical cases with interdigital dermatitis and 27 clinical cases with footrot) and 20 clinically healthy naemi sheep were used in this study. Real time PCR for detection of Fusobacterium necrophorum (F. necrophorum) and Dichelobacter nodosus (D. nodosus) revealed that, all samples collected from lame sheep (N = 53) were positive for D. nodosus (100 %), whereas F. necrophorum was detected in 19 out of 53 samples (35.84 %). The virulent D. nodosus was detected in 48 lameness cases where non-virulent D. nodosus were identified in 5 cases (in concurrent with F. necrophorum). The mean serum levels of PCT, C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin (HP), fibrinogen (Fg) and CYTs (IL1-ß, IL-1α, IFN-γ, IL-6 and TNF-α) in sheep with clinical interdigital dermatitis and footrot were remarkably higher than those detected in control healthy sheep. The serum levels of PCT, CRP, SAA, HP, Fg, and CYTs markers in lame sheep pre- and post-treatment were measured. A substantial decline was detected in serum levels of tested biomarkers of lame sheep after 14 days of treatment. The ROC curves were created. The AUC was assessed to evaluate the accuracy of each variable in distinguishing diseased and healthy sheep. Based on the ROC curves and AUCs; PCT, CRP, SAA, HP, and CYTs were highly diagnostic and predictive for the treatment response of sheep with clinical interdigital dermatitis and footrot. Moreover, all tested biomarkers had a noteworthy role in disease immuno-pathogenesis. Nevertheless, PCT and CRP are better than other tested APPs and CYTs as diagnostic markers for interdigital dermatitis and footrot. However, PCT only has the ability to differentiate sheep with different lameness score.


Asunto(s)
Dermatitis , Dichelobacter nodosus , Panadizo Interdigital , Enfermedades de las Ovejas , Ovinos , Animales , Polipéptido alfa Relacionado con Calcitonina , Cojera Animal/microbiología , Proteínas de Fase Aguda , Citocinas , Enfermedades de las Ovejas/microbiología , Panadizo Interdigital/diagnóstico , Panadizo Interdigital/microbiología , Panadizo Interdigital/patología , Dermatitis/microbiología , Dermatitis/veterinaria
3.
Wilderness Environ Med ; 31(1): 82-86, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-32008950

RESUMEN

Treating skin disorders in wilderness settings is often challenging. In this report we describe common skin conditions affecting the feet of river runners on the Colorado River in Grand Canyon National Park. These conditions are frequently referred to by river runners with a catchall term, "tolio." Several skin disorders have been identified as components of tolio, with the most prevalent currently being pitted keratolysis. We present a case of pitted keratolysis in a river guide occurring during a multiday river trip, where treatment can be difficult. Prevention is often more important.


Asunto(s)
Traumatismos en Atletas/diagnóstico , Traumatismos en Atletas/terapia , Panadizo Interdigital/diagnóstico , Panadizo Interdigital/terapia , Pie de Inmersión/terapia , Deportes Acuáticos , Animales , Arizona , Traumatismos en Atletas/etiología , Panadizo Interdigital/etiología , Humanos , Pie de Inmersión/diagnóstico , Pie de Inmersión/etiología , Masculino , Persona de Mediana Edad , Resultado del Tratamiento
4.
BMC Res Notes ; 12(1): 534, 2019 Aug 22.
Artículo en Inglés | MEDLINE | ID: mdl-31439030

RESUMEN

OBJECTIVE: Dichelobacter nodosus is the primary aetiological agent of footrot in sheep. Ovine footrot causes considerable economic losses and substantial animal welfare issues in the Australian sheep industry. Current methods for detecting D. nodosus are difficult, laborious and time-consuming. Recently, we developed a robust LAMP assay (VDN LAMP) that was able to identify aprV2 positive D. nodosus in-field. A major advantage of LAMP technology is the ability of the assay to be performed by non-specialists with minimal training. We aimed to assess the performance of the VDN LAMP in-field in comparison to a laboratory-based aprV2/aprB2 rtPCR when used by secondary users after training by the authors. RESULTS: Two animal health officers (termed secondary users) from Department of Primary Industries and Regions, South Australia (PIRSA) were trained in the use of VDN LAMP, before carrying out in-field testing on several locations in South Australia. The performance of VDN LAMP assay by secondary user 1 was shown to successfully detect 73.91% (n = 53) aprV2 positive samples, while secondary user 2 detected 37.93% (n = 30) aprV2 positive samples. Overall, the ability to identify virulent D. nodosus by VDN LAMP by secondary users was mixed for various reasons, however, this could be rectified by additional training and commercial production of the LAMP kits to increase stability. We envisaged in the future VDN LAMP will able to be used by non-specialists to aid control programs.


Asunto(s)
Técnicos de Animales/estadística & datos numéricos , Proteínas Bacterianas/genética , Dichelobacter nodosus/genética , Panadizo Interdigital/diagnóstico , Infecciones por Bacterias Gramnegativas/veterinaria , Técnicas de Amplificación de Ácido Nucleico/métodos , Serina Endopeptidasas/genética , Enfermedades de las Ovejas/diagnóstico , Técnicos de Animales/normas , Animales , Dichelobacter nodosus/fisiología , Panadizo Interdigital/microbiología , Infecciones por Bacterias Gramnegativas/diagnóstico , Infecciones por Bacterias Gramnegativas/microbiología , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Ovinos , Enfermedades de las Ovejas/microbiología , Australia del Sur
5.
Schweiz Arch Tierheilkd ; 161(7): 465-472, 2019 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-31298215

RESUMEN

INTRODUCTION: Ovine footrot caused by Dichelobacter nodosus is a highly contagious and painful disease representing an economic as well as an animal welfare problem. In order to get more information on the infection dynamics, 26 lambs and 4 ewes enrolled in an in-contact infection trial were monitored over two weeks for the presence of D. nodosus-specific DNA. Two D. nodosus-positive ewes were housed together with 13 confirmed negative lambs. The control group consisted of another 13 lamb siblings and two confirmed D. nodosus-negative ewes. Every foot of all sheep was sampled seven times over the two weeks experiment period and subsequently analyzed for the presence of D. nodosus by quantitative real-time PCR. The control group was negative at the beginning and the end of the experiment and showed no clinical symptoms of footrot. The two positive ewes showed a high, but hundred fold differing level of virulent D. nodosus that remained constant over time with one of the ewes being also weakly positive for benign D. nodosus. All lambs of the infection group were positive for virulent D. nodosus at 14 days post infection (dpi). The first positive animals were observed on 3 dpi. The D. nodosus load remained at a low level and only increased in a few lambs at the end of the trial. Five of the contact lambs showed suspicious clincal signs (score 1-2) at 14 dpi corroborating the PCR results and indicating that the disease starts as early as two weeks after contact with positive sheep.


INTRODUCTION: Le piétin causé par Dichelobacter nodosus est une maladie hautement contagieuse et douloureuse qui représente à la fois un problème économique et de bien-être animal. Pour avoir plus informations sur la dynamique de l'infection, 26 agneaux et 4 brebis appartenant à un groupe d'essai d'infection par contact ont été contrôlés pendant deux semaines quant à la présence d'ADN spécifique de D. nodosus. Deux brebis positives pour D. nodosus ont été mises en contact avec 13 agneaux confirmés négatifs. Le groupe témoin était formé de 13 autres agneaux et deux brebis confirmées négatives. Sept échantillons sur écouvillon ont été prélevés sur chaque pied de chaque mouton au cours des deux semaines de la période expérimentale et analysés quant à la présence de D. nodosus par PCR quantitative en temps réel. Le groupe témoin était négatif au début et à la fin de l'expérience et n'a montré aucun symptôme clinique de piétin. Les deux brebis positives ont montré une forte présence de D. nodosus virulent, mais de cent niveaux différents, qui est restée constant dans le temps, l'une des brebis étant aussi faiblement positive pour D. nodosus bénin. Tous les agneaux du groupe infecté étaient positifs pour D. nodosus virulent 14 jours après l'infection (dpi). Les premiers animaux positifs ont été observés à 3 dpi. La charge de D. nodosus est restée faible et n'a augmenté que chez quelques agneaux à la fin de l'expérience. Cinq des agneaux en contact ont présenté des symptômes suspects (score 1-2) à 14 dpi, corroborant les résultats de la PCR et indiquant que l'infection commence dès deux semaines après le contact avec des moutons positifs.


Asunto(s)
Infecciones por Bacterias Gramnegativas/veterinaria , Animales , Carga Bacteriana , Dichelobacter nodosus/fisiología , Femenino , Panadizo Interdigital/diagnóstico , Panadizo Interdigital/epidemiología , Panadizo Interdigital/microbiología , Panadizo Interdigital/transmisión , Infecciones por Bacterias Gramnegativas/diagnóstico , Infecciones por Bacterias Gramnegativas/epidemiología , Infecciones por Bacterias Gramnegativas/transmisión , Reacción en Cadena de la Polimerasa , Ovinos , Enfermedades de las Ovejas/diagnóstico , Enfermedades de las Ovejas/epidemiología , Enfermedades de las Ovejas/microbiología , Enfermedades de las Ovejas/transmisión , Suiza/epidemiología
6.
PLoS One ; 13(9): e0204310, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-30260992

RESUMEN

Dichelobacter nododus is the causative agent of footrot, a major disease of sheep that creates welfare concerns and large economic loss. The virulence of D. nododus depends on the presence of extracellular proteases, AprV2 and AprB2, which differ by one amino acid. Strains possessing AprV2 can cause clinically virulent disease, while AprB2 may cause clinically benign disease. Current methods for detecting D. nodosus are difficult, laborious and time consuming. New techniques capable of rapidly detecting and typing D. nodosus are needed to aid control programs. Molecular methods, like real-time polymerase chain reaction (rtPCR) can detect aprV2 and aprB2, however, this assay is not field-deployable and cannot support local decision-making during an outbreak. Here we present a field-based molecular assay for detecting aprV2, using loop mediated isothermal amplification (LAMP). The aprV2 LAMP (VDN LAMP) assay was optimised to reliably detect aprV2 from laboratory purified genomic (gDNA) of virulent D. nodosus down to 5x10(-3) ng µL-1, with time to positive (Tp) ≤ 16 minutes, while aprB2 was unreliably detected at 5 ng µL-1 from 16-20 minutes. The use of field collected samples that were rtPCR positive for aprB2 resulted in no amplification, while aprV2 positive field samples by VDN LAMP assay are defined as having Tps' of < 20 minutes and melting temperature between 88.0-88.9°C. When compared to rtPCR, the VDN LAMP was shown to have a diagnostic specificity of 100% and sensitivity of 83.33%. As proof of concept, the VDN LAMP was taken on farm, with all processing occurring in-field. The on farm VDN LAMP successfully detected 91.67% aprV2 positive samples, no aprB2 positive samples (n = 9) or D. nodosus negative (n = 23) samples, with a kappa agreement of 'almost perfect' to rtPCR. This highlights the potential of the assay to inform local treatment decisions for management.


Asunto(s)
Proteínas Bacterianas/genética , Dichelobacter nodosus/patogenicidad , Panadizo Interdigital/diagnóstico , Infecciones por Bacterias Gramnegativas/veterinaria , Serina Endopeptidasas/genética , Enfermedades de las Ovejas/microbiología , Animales , Australia , Dichelobacter nodosus/genética , Diagnóstico Precoz , Panadizo Interdigital/microbiología , Infecciones por Bacterias Gramnegativas/diagnóstico , Reacción en Cadena de la Polimerasa/métodos , Sensibilidad y Especificidad , Ovinos
7.
Artículo en Alemán | MEDLINE | ID: mdl-30142653

RESUMEN

OBJECTIVE: The assessment of pain in sheep is a clinical challenge, because being a prey species, they tend to mask it. Since pain in animals cannot be measured directly, various methods are applicable to detect its effects on physiological, biochemical and ethological processes. Clinicians experienced in behavioural assessment are able to reliably determine the level of pain endured by an animal. The objective of this study was to identify behaviours that reflect acute and chronic pain as well as stress in sheep. METHODS: The behaviour of 36 Meat Merino ewes with contagious footrot (contagious interdigital dermatitis) was analysed during normal activities in the flock and during footrot treatment, which included claw trimming to remove abnormal horn. The behaviour during treatment was compared with that of 12 healthy control sheep of the same breed during routine foot trimming. Grading of the observed behaviours was made using a numerical score. The occurrence of footrot was defined as, and equated with, chronic pain, and treatment was equated with acute pain. For the assessment of stress induced by the manipulative treatment, the data of the healthy sheep were taken as a comparison. RESULTS: The strongest indications of chronic pain were non-weight bearing lameness of an affected limb, shifting of weight and abnormal gait patterns. Additionally, sheep with footrot urinated more frequently than healthy animals. Stress-related bruxism occurred in healthy and ill sheep during foot trimming, but not during normal activities in the flock. Tail wagging and strong defensive movements of all limbs commonly occurred during footrot treatment. Head shaking appeared to be primarily stress-related in association with dorsal recumbency on the tilt table, but was exacerbated by pain elicited by hoof trimming. CONCLUSION: The behaviours selected in this study were useful for the identification of chronic and acute pain in sheep; however, correct interpretation of behavioural changes requires an experienced individual.


Asunto(s)
Dolor Agudo/veterinaria , Dolor Crónico/veterinaria , Panadizo Interdigital/diagnóstico , Enfermedades de las Ovejas/diagnóstico , Dolor Agudo/diagnóstico , Dolor Agudo/etiología , Crianza de Animales Domésticos , Animales , Dolor Crónico/diagnóstico , Dolor Crónico/etiología , Femenino , Panadizo Interdigital/etiología , Ovinos , Enfermedades de las Ovejas/etiología , Oveja Doméstica
8.
J Clin Microbiol ; 56(4)2018 04.
Artículo en Inglés | MEDLINE | ID: mdl-29436426

RESUMEN

Virulent footrot is an economically significant disease in most sheep-rearing countries. The disease can be controlled with vaccine targeting the fimbriae of virulent strains of the essential causative agent, Dichelobacter nodosus However, the bacterium is immunologically heterogeneous, and 10 distinct fimbrial serogroups have been identified. Ideally, in each outbreak the infecting strains would be cultured and serogrouped so that the appropriate serogroup-specific mono- or bivalent vaccine could be administered, because multivalent vaccines lack efficacy due to antigenic competition. If clinical disease expression is suspected to be incomplete, culture-based virulence tests are required to confirm the diagnosis, because control of benign footrot is economically unjustifiable. Both diagnosis and vaccination are conducted at the flock level. The aims of this study were to develop a PCR-based procedure for detecting and serogrouping D. nodosus directly from foot swabs and to determine whether this could be done accurately from the same cultured swab. A total of 269 swabs from the active margins of foot lesions of 261 sheep in 12 Merino sheep flocks in southeastern Australia were evaluated. DNA extracts taken from putative pure cultures of D. nodosus and directly from the swabs were evaluated in PCR assays for the 16S rRNA and fimA genes of D. nodosus Pure cultures were tested also by the slide agglutination test. Direct PCR using extracts from swabs was more sensitive than culture for detecting and serogrouping D. nodosus strains. Using the most sensitive sample collection method of the use of swabs in lysis buffer, D. nodosus was more likely to be detected by PCR in active than in inactive lesions, and in lesions with low levels of fecal contamination, but lesion score was not a significant factor. PCR conducted on extracts from swabs in modified Stuart's transport medium that had already been used to inoculate culture plates had lower sensitivity. Therefore, if culture is required to enable virulence tests to be conducted, it is recommended that duplicate swabs be collected from each foot lesion, one in transport medium for culture and the other in lysis buffer for PCR.


Asunto(s)
Dichelobacter nodosus/clasificación , Panadizo Interdigital/diagnóstico , Infecciones por Bacterias Gramnegativas/veterinaria , Reacción en Cadena de la Polimerasa/veterinaria , Enfermedades de las Ovejas/diagnóstico , Animales , ADN Bacteriano/aislamiento & purificación , Dichelobacter nodosus/aislamiento & purificación , Brotes de Enfermedades/veterinaria , Panadizo Interdigital/microbiología , Infecciones por Bacterias Gramnegativas/diagnóstico , Pezuñas y Garras/microbiología , Pezuñas y Garras/patología , ARN Ribosómico 16S/aislamiento & purificación , Serotipificación , Ovinos , Enfermedades de las Ovejas/microbiología , Oveja Doméstica , Vacunación
9.
Artículo en Alemán | MEDLINE | ID: mdl-30616280

RESUMEN

Infectious causes of lameness in sheep remain of considerable clinical importance. Because of the availability of newly licensed drugs, important changes in therapy options, particularly for ovine footrot, have occurred. This paper provides an overview of common and rarer infectious causes of lameness in sheep, and presents a detailed review of recent advances in research regarding the aetiology, diagnosis, prevention, treatment and control of ovine foot rot. Despite the recent release of several antibiotics licensed for the treatment of footrot in sheep, the use of footbaths following the cascade of regulations remains a crucial part of many integrated treatment and control programmes. The sustainable control of foot rot requires recent advances in science to be put into practice, with tailor-made control programmes for each individual farm. The article presents various treatment options and potential routes of control and eradication.


Asunto(s)
Antibacterianos/uso terapéutico , Panadizo Interdigital/terapia , Enfermedades de las Ovejas/terapia , Animales , Panadizo Interdigital/diagnóstico , Panadizo Interdigital/tratamiento farmacológico , Panadizo Interdigital/prevención & control , Pezuñas y Garras/fisiopatología , Cojera Animal/diagnóstico , Cojera Animal/tratamiento farmacológico , Cojera Animal/prevención & control , Cojera Animal/terapia , Ovinos , Enfermedades de las Ovejas/diagnóstico , Enfermedades de las Ovejas/tratamiento farmacológico , Enfermedades de las Ovejas/prevención & control
10.
J Vet Diagn Invest ; 30(2): 205-210, 2018 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-29105601

RESUMEN

Virulent ovine foot rot is a contagious foot disease. Given the development and validation of a real-time PCR to detect Dichelobacter nodosus isolates that contain the virulence-associated protease genes aprV2 and aprB2, the diagnosis of foot rot has made considerable progress. We evaluated pooling methods to reduce the number of samples during a foot rot control program. Samples of individual feet were compared to a 4-feet sample of the same sheep. All further analyses based on 4-feet samples (pools-of-5 and pools-of-10 4-feet samples) were compared to samples of individual sheep, and a risk-based herd sampling was evaluated and compared to the whole flock. The sensitivity and specificity of the 4-feet samples for detection of aprV2-positive strains was 93.8% (CI: 87.6-97.5%) and 98.3% (CI: 96.5-99.3%), respectively. The sensitivity and specificity of the pools-of-10 was 86.7% (CI: 78.4-92.7%) and 100.0% (CI: 97.4-100%), respectively. Pools-of-5 were not significantly more sensitive than pools-of-10. The pooling of 4 individual foot samples into one 4-feet sample is an adequate method to reduce the number of samples of individual sheep. The sensitivity of pools-of-5 and pools-of-10 is too imprecise for a control program. Risk-based sampling allowed for a substantial reduction of samples to be tested, had a sensitivity of 95.8% (CI: 78.9-99.9%) and specificity of 100.0% (CI: 88.1-100.0%) when determining the foot rot flock status, and represents an adequate methodology to predict within-flock freedom from infection.


Asunto(s)
Dichelobacter nodosus/aislamiento & purificación , Panadizo Interdigital/diagnóstico , Enfermedades de las Ovejas/diagnóstico , Manejo de Especímenes/veterinaria , Animales , Dichelobacter nodosus/genética , Dichelobacter nodosus/patogenicidad , Panadizo Interdigital/microbiología , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Sensibilidad y Especificidad , Ovinos , Enfermedades de las Ovejas/microbiología , Virulencia
11.
Vet Res Commun ; 41(3): 189-193, 2017 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-28343363

RESUMEN

Dichelobacter nodosus is the principal cause of ovine footrot and strain virulence is an important factor in disease severity. Therefore, detection and virulence determination of D. nodosus is important for proper diagnosis of the disease. Today this is possible by real-time PCR analysis. Analysis of large numbers of samples is costly and laborious; therefore, pooling of individual samples is common in surveillance programs. However, pooling can reduce the sensitivity of the method. The aim of this study was to develop a pooling method for real-time PCR analysis that would allow sensitive detection and simultaneous virulence determination of D. nodosus. A total of 225 sheep from 17 flocks were sampled using ESwabs within the Swedish Footrot Control Program in 2014. Samples were first analysed individually and then in pools of five by real-time PCR assays targeting the 16S rRNA and aprV2/B2 genes of D. nodosus. Each pool consisted of four negative and one positive D. nodosus samples with varying amounts of the bacterium. In the individual analysis, 61 (27.1%) samples were positive in the 16S rRNA and the aprV2/B2 PCR assays and 164 (72.9%) samples were negative. All samples positive in the aprV2/B2 PCR-assay were of aprB2 variant. The pooled analysis showed that all 41 pools were also positive for D. nodosus 16S rRNA and the aprB2 variant. The diagnostic sensitivity for pooled and individual samples was therefore similar. Our method includes concentration of the bacteria before DNA-extraction. This may account for the maintenance of diagnostic sensitivity. Diagnostic sensitivity in the real-time PCR assays of the pooled samples were comparable to the sensitivity obtained for individually analysed samples. Even sub-clinical infections were able to be detected in the pooled PCR samples which is important for control of the disease. This method may therefore be implemented in footrot control programs where it can replace analysis of individual samples.


Asunto(s)
Dichelobacter nodosus/patogenicidad , Panadizo Interdigital/diagnóstico , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Enfermedades de las Ovejas/diagnóstico , Animales , Dichelobacter nodosus/genética , Panadizo Interdigital/microbiología , Vigilancia de la Población/métodos , ARN Ribosómico 16S/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Sensibilidad y Especificidad , Ovinos
12.
J Clin Microbiol ; 55(5): 1313-1326, 2017 05.
Artículo en Inglés | MEDLINE | ID: mdl-28202796

RESUMEN

Dichelobacter nodosus is a fastidious, strictly anaerobic bacterium, an obligate parasite of the ruminant hoof, and the essential causative agent of virulent ovine footrot. The clinical disease results from a complex interplay between the pathogen, the environment, and the host. Sheep flocks diagnosed with virulent but not benign footrot in Australia may be quarantined and required to undergo a compulsory eradication program, with costs met by the farmer. Virulence of D. nodosus at least partially depends on the elaboration of a protease encoded by aprV2 and manifests as elastase activity. Laboratory virulence tests are used to assist diagnosis because clinical differentiation of virulent and benign footrot can be challenging during the early stages of disease or when the disease is not fully expressed due to unfavorable pasture conditions. Using samples collected from foot lesions from 960 sheep from 40 flocks in four different geographic regions, we evaluated the analytical characteristics of qPCR tests for the protease gene alleles aprV2 and aprB2, and compared these with results from phenotypic protease (elastase and gelatin gel) tests. There was a low level of agreement between clinical diagnosis and quantitative PCR (qPCR) test outcomes at both the flock and sample levels and poor agreement between qPCR test outcomes and the results of phenotypic virulence tests. The diagnostic specificity of the qPCR test was low at both the flock and individual swab levels (31.3% and 18.8%, respectively). By contrast, agreement between the elastase test and clinical diagnosis was high at both the flock level (diagnostic sensitivity [DSe], 100%; diagnostic specificity [DSp], 78.6%) and the isolate level (DSe, 69.5%; DSp, 80.5%).


Asunto(s)
Dichelobacter nodosus/genética , Dichelobacter nodosus/patogenicidad , Panadizo Interdigital/diagnóstico , Reacción en Cadena de la Polimerasa/veterinaria , Enfermedades de las Ovejas/diagnóstico , Animales , Australia , Proteínas Bacterianas/genética , Dichelobacter nodosus/aislamiento & purificación , Panadizo Interdigital/microbiología , Elastasa Pancreática/análisis , Serina Endopeptidasas/genética , Ovinos , Enfermedades de las Ovejas/microbiología
13.
Vet J ; 216: 189-95, 2016 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-27687951

RESUMEN

In sheep, the diagnosis of foot lesions is routinely based on physical examination of the hoof. Correct diagnosis is important for the effective treatment, prevention and control of both infectious and non-infectious causes of lameness. Therefore, the aim of this study was to evaluate the level of inter-observer agreement for clinical examination of ovine foot lesions. Eight observers of varying experience, training and occupation performed foot examinations on a total of 1158 sheep from 38 farms across North England and Wales. On each farm, a group of two to four observers independently examined a sample of 24 to 30 sheep to diagnose the presence or absence of specific foot lesions including white line lesions (WL), contagious ovine digital dermatitis (CODD), footrot (FR), inter-digital dermatitis (ID) and toe granuloma (TG). The inter-observer agreement of foot lesion assessments was examined using Fleiss kappa (κ), and Cohen's κ examined the paired agreement between the test standard observer (TSO) and each observer. Scoring differences with the TSO were examined as the percentage of scoring errors and assessed for evidence of systematic scoring bias. With the exception of WL (maximum error rate 33.3%), few scoring differences with the TSO occurred (maximum error rate 3.3%). This suggests that observers can achieve good levels of reliability when diagnosing most of the commonly observed foot conditions associated with lameness in sheep.


Asunto(s)
Dermatitis Digital/diagnóstico , Enfermedades del Pie/veterinaria , Panadizo Interdigital/diagnóstico , Enfermedades de las Ovejas/diagnóstico , Animales , Dermatitis Digital/epidemiología , Inglaterra/epidemiología , Pie/patología , Enfermedades del Pie/diagnóstico , Enfermedades del Pie/epidemiología , Panadizo Interdigital/epidemiología , Pezuñas y Garras/patología , Variaciones Dependientes del Observador , Examen Físico , Prevalencia , Reproducibilidad de los Resultados , Factores de Riesgo , Ovinos , Enfermedades de las Ovejas/epidemiología , Gales/epidemiología
14.
Vet Rec ; 176(24): 626, 2015 Jun 13.
Artículo en Inglés | MEDLINE | ID: mdl-25977404

RESUMEN

Five UK goat farms with high levels of lameness (prevalence 14-67 per cent) were investigated. On two farms (farms 1 and 2), the animals presented with typical footrot lesions. The remaining three farms (farms 3, 4 and 5) presented with infected lesions on the foot that did not resemble footrot. These lesions were observed to start from the white line or sole but the interdigital space was rarely affected. Swabs were processed by PCR to assess the presence of Dichelobacter nodosus and three specific treponeme groups (group 1: Treponema medium/Treponema vincentii-like, group 2: Treponema phagedenis-like and group 3: Treponema denticola/Treponema putidum-like) that are reported to be associated with bovine digital dermatitis and contagious ovine digital dermatitis. On farms 1 and 2, 85.7 per cent of samples were found to be positive for D nodosus while only 9.5 per cent were positive for treponeme groups 1, 2 and 3. In contrast, 5.3 per cent of samples from farms 3, 4 and 5 were positive for D nodosus, while 34.2, 68.4 and 36.8 per cent of samples from these farms tested positive for treponeme groups 1, 2 and 3, respectively. T medium/T vincentii-like, T phagedenis-like and T denticola/T putidum-like treponemes were detected on foot lesions of lame goats suggesting that they have a role in the aetiology of this lameness, which has not previously been described in dairy goats.


Asunto(s)
Úlcera del Pie/veterinaria , Enfermedades de las Cabras/microbiología , Cojera Animal/microbiología , Treponema/aislamiento & purificación , Infecciones por Treponema/veterinaria , Animales , Femenino , Panadizo Interdigital/diagnóstico , Úlcera del Pie/microbiología , Enfermedades de las Cabras/epidemiología , Cabras , Cojera Animal/epidemiología , Masculino , Treponema/clasificación , Infecciones por Treponema/microbiología , Reino Unido/epidemiología
15.
Vet J ; 201(3): 265-8, 2014 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-24973004

RESUMEN

The novel sheep disease, contagious ovine digital dermatitis (CODD) was first described in the UK in 1997. The disease is characterised by severe lameness associated with initial inflammation at the coronary band, followed by progressive separation of the hoof capsule from the underlying tissue. On microbiological examination, treponeme bacteria have been frequently isolated from cases of CODD, including treponemes phylogenetically identical to those associated with bovine digital dermatitis (BDD). Dichelobacter nodosus and Fusobacterium necrophorum have also been isolated from CODD lesions although their role in the pathogenesis remains uncertain. While epidemiological data indicate that the prevalence of CODD is increasing in the UK, the routes of transmission and associated risk factors have not been clearly elucidated. Evidenced-based treatment trials indicate that parenteral administration of long-acting amoxicillin is an efficacious treatment for CODD, while anecdotal evidence suggests other antibiotics, given locally and/or parenterally, may also be beneficial. Further microbiological and epidemiological research is urgently required to develop sustainable control strategies, including the development of vaccines and appropriate biosecurity and farm management protocols. In this review current knowledge of the clinical, aetiological, and epidemiological aspects of CODD is assessed as well as approaches to its control.


Asunto(s)
Bacterias/aislamiento & purificación , Dermatitis Digital , Panadizo Interdigital , Enfermedades de las Ovejas , Animales , Dermatitis Digital/diagnóstico , Dermatitis Digital/epidemiología , Dermatitis Digital/etiología , Dermatitis Digital/prevención & control , Panadizo Interdigital/diagnóstico , Panadizo Interdigital/epidemiología , Panadizo Interdigital/etiología , Panadizo Interdigital/prevención & control , Prevalencia , Ovinos , Enfermedades de las Ovejas/diagnóstico , Enfermedades de las Ovejas/epidemiología , Enfermedades de las Ovejas/etiología , Enfermedades de las Ovejas/prevención & control
16.
J Clin Microbiol ; 52(4): 1228-31, 2014 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-24452162

RESUMEN

Ovine foot rot caused by Dichelobacter nodosus is affecting sheep worldwide. The current diagnostic methods are difficult and cumbersome. Here, we present a competitive real-time PCR based on allelic discrimination of the protease genes aprV2 and aprB2. This method allows direct detection and differentiation of virulent and benign D. nodosus from interdigital skin swabs in a single test. Clinically affected sheep harbored high loads of only virulent strains, whereas healthy sheep had lower loads of predominantly benign strains.


Asunto(s)
Proteínas Bacterianas/análisis , Técnicas de Tipificación Bacteriana/métodos , Dichelobacter nodosus/aislamiento & purificación , Panadizo Interdigital/diagnóstico , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Serina Endopeptidasas/análisis , Enfermedades de las Ovejas/diagnóstico , Animales , Proteínas Bacterianas/genética , Panadizo Interdigital/microbiología , Serina Endopeptidasas/genética , Ovinos , Enfermedades de las Ovejas/microbiología
17.
Vet Microbiol ; 162(2-4): 756-760, 2013 Mar 23.
Artículo en Inglés | MEDLINE | ID: mdl-23084506

RESUMEN

Footrot in sheep and goats is expressed as a spectrum of clinical entities ranging from benign, which is a self limiting interdigital dermatitis to highly virulent, in which severe under running of the horn of the hoof occurs. Interactions between the host, the virulence of the causative strain of Dichelobacter nodosus and environmental conditions determine the severity of the disease. Clinical diagnosis of virulent footrot, which a notifiable disease in some states of Australia, is not always straightforward. Therefore, the gelatin gel and elastase tests for protease activity, and the intA PCR test for an inserted genetic element in D. nodosus are commonly used to support or to confirm a clinical diagnosis. A comparative study of these laboratory tests with a large number of samples collected from 12 flocks of sheep with clinically virulent footrot was conducted. Based on the elastase test, 64% of the isolates tested were classified as virulent compared to 91% on the gelatin gel test and 41% according to the intA test. The agreement between the elastase and the gelatin gel test was low (kappa=0.12) as were the agreements between other tests. Only about 21% of the isolates were virulent in all 3 tests. Therefore these tests on their own may not provide standard and reliable results and are likely to remain as supplementary tests for clinical diagnosis of the disease.


Asunto(s)
Dichelobacter nodosus/aislamiento & purificación , Panadizo Interdigital/microbiología , Enfermedades de las Ovejas/microbiología , Animales , Australia , Dichelobacter nodosus/enzimología , Dichelobacter nodosus/genética , Dichelobacter nodosus/patogenicidad , Panadizo Interdigital/diagnóstico , Panadizo Interdigital/enzimología , Pezuñas y Garras/microbiología , Elastasa Pancreática/análisis , Péptido Hidrolasas/análisis , Reacción en Cadena de la Polimerasa , Ovinos , Enfermedades de las Ovejas/diagnóstico , Enfermedades de las Ovejas/enzimología , Oveja Doméstica , Virulencia
18.
Pesqui. vet. bras ; 32(12): 1289-1296, Dec. 2012. ilus, tab
Artículo en Portugués | LILACS | ID: lil-662562

RESUMEN

Neste artigo realizou-se a avaliação de ovinos mestiços Santa Inês com pododermatite infecciosa, verificando as alterações no leucograma e proteínas de fase aguda. No primeiro experimento, 70 fêmeas foram separadas em três grupos de acordo com o seu escore podal: Grupo controle (G1) com escore 0; Dermatite interdigital (G2) com escore 1 ou 2 e Pododermatite necrosante (G3) com escore 3, 4 ou 5. Durante dois meses observou-se a evolução clínica e efeitos no leucograma em cinco momentos: M1 (dia 0), M2 (dia 15), M3 (dia 30), M4 (dia 45) e M5 (dia 60). A intensidade do resultado do leucograma foi de baixa magnitude, observando-se alterações significativas (p<0,05) como uma leve leucocitose (G3, M4), atribuída a neutrofilia e um discreto aumento no número total de monócitos (G3, M2 e M3) apesar do extenso dano e necrose tecidual existentes na última fase da doença. No segundo experimento, utilizaram-se 105 animais de sete propriedades com objetivo de isolar o agente etiológico e avaliar os efeitos da doença sobre proteína plasmática total e proteínas de fase aguda. Os animais também foram separados em três grupos: controle, dermatite interdigital e pododermatite necrosante. Em todas as propriedades foi realizado o isolamento de Dichelobacter nodosus. Não houve correlação significativa (p<0,05) das diferentes fases da doença sobre as proteínas estudadas, porém a haptoglobina dos grupos com animais doentes apresentou médias superiores ao grupo controle. De acordo com a metodologia utilizada e resultados obtidos, conclui-se que as proteínas de fase aguda estudadas não foram eficientes na caracterização das fases da pododermatite infecciosa ovina e que a resposta leucocitária foi branda, tornando difícil sua utilização para este fim.


In this paper, Santa Inês crossbred sheep with footrot were evaluated, checking the changes on leukogram and the acute phase proteins. In the first experiment, 70 females were divided into three groups according to their podal scores: a control group (G1) with score 0, scald group (G2) with score 1 or 2 and a footrot group (G3) with score 3, 4 or 5. During two months, the clinical course and its effect on leukogram were observed at five moments: M1 (day 0), M2 (day 15), M3 (day 30), M4 (day 45) and M5 (day 60). The intensity of the white blood cell count was low magnitude, observing significant changes (p <0.05) as a mild leukocytosis (G3, M4), attributed to neutrophilia and a slight increase in the total number of monocytes (G3, M2 and M3) despite the extensive damage and tissue necrosis existed during the final stage of the disease. In the second experiment, 105 animals from seven farms were used with the objective of isolating the causative agent and to evaluate the effect of disease on total plasma protein and acute phase proteins. The animals were also divided into three groups: control, scald and footrot. In all farms, the isolation of Dichelobacter nodosus was regarded. For the different stages of the disease on the proteins studied there was no significant correlation (p<0.05) but the haptoglobin in scald and footrot groups showed higher average than the control group. Based on these results, we conclude that the acute phase proteins studied were not efficient in the characterization of the phases of the disease and the leukocyte response was mild, making it difficult to use for this purpose.


Asunto(s)
Animales , Recuento de Leucocitos , Ovinos/parasitología , Panadizo Interdigital/diagnóstico , Proteínas/análisis , Dichelobacter nodosus , Fusobacterium necrophorum
19.
Prev Vet Med ; 95(1-2): 127-36, 2010 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-20303191

RESUMEN

Dichelobacter nodosus is a slow-growing anaerobic bacterium that is the causative agent of virulent ovine footrot. Vaccination targeted at up to two specific serogroups can eliminate those serogroups from infected flocks, but requires identification of serogroups present in infected flocks. Serogroups can be identified using slide agglutination or polymerase chain reaction (PCR) methods. The objectives of this project were to use stochastic simulation modeling to estimate the efficacy of sampling strategies encompassing 5-40 sheep per flock and 2-4 colonies per sheep, and to compare efficacies based on slide agglutination or multiplex PCR test results. Foot swabs collected from sheep in 12 flocks were used as the basis for a sampling strategy simulation model. None of the evaluated sampling strategies identified the two most common serogroups in the flock, or all serogroups present in the flock, in 95% of iterations. However, a simulated sample of 22 sheep/flock and 2 colonies/sheep resulted in a simulated vaccine that protected 95% of the sheep that could be protected by a single bivalent vaccine, while a sample of 24 sheep/flock and 2 colonies/sheep resulted in a series of simulated bivalent vaccines that protected 95% of diseased infected sheep. The difference in outcome was due to the distribution and frequency of serogroups within certain flocks where some serogroups were uncommon and others dominant. A sampling strategy (>40 sheep/flock, 4 colonies/sheep) that will identify the two most common serogroups in a flock 95% of the time may not be cost effective. Evaluating efficacy based on the expected effect on the flock may be more useful than one which seeks to determine the most common serogroups. These findings are broadly applicable to diseases where more than one strain or type of pathogen may be present and must be represented in a vaccine.


Asunto(s)
Vacunas Bacterianas/uso terapéutico , Dichelobacter nodosus , Panadizo Interdigital/diagnóstico , Infecciones por Bacterias Gramnegativas/veterinaria , Enfermedades de las Ovejas/diagnóstico , Animales , Dichelobacter nodosus/clasificación , Dichelobacter nodosus/inmunología , Dichelobacter nodosus/patogenicidad , Femenino , Panadizo Interdigital/prevención & control , Infecciones por Bacterias Gramnegativas/diagnóstico , Infecciones por Bacterias Gramnegativas/prevención & control , Masculino , Método de Montecarlo , Serotipificación/veterinaria , Ovinos , Enfermedades de las Ovejas/prevención & control , Procesos Estocásticos , Virulencia
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