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1.
BMC Res Notes ; 12(1): 534, 2019 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-31439030

RESUMO

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.


Assuntos
Técnicos em Manejo de Animais/estatística & dados numéricos , Proteínas de Bactérias/genética , Dichelobacter nodosus/genética , Pododermatite Necrótica dos Ovinos/diagnóstico , Infecções por Bactérias Gram-Negativas/veterinária , Técnicas de Amplificação de Ácido Nucleico/métodos , Serina Endopeptidases/genética , Doenças dos Ovinos/diagnóstico , Técnicos em Manejo de Animais/normas , Animais , Dichelobacter nodosus/fisiologia , Pododermatite Necrótica dos Ovinos/microbiologia , Infecções por Bactérias Gram-Negativas/diagnóstico , Infecções por Bactérias Gram-Negativas/microbiologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Ovinos , Doenças dos Ovinos/microbiologia , Austrália do Sul
2.
Schweiz Arch Tierheilkd ; 161(7): 465-472, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31298215

RESUMO

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.


Assuntos
Infecções por Bactérias Gram-Negativas/veterinária , Animais , Carga Bacteriana , Dichelobacter nodosus/fisiologia , Feminino , Pododermatite Necrótica dos Ovinos/diagnóstico , Pododermatite Necrótica dos Ovinos/epidemiologia , Pododermatite Necrótica dos Ovinos/microbiologia , Pododermatite Necrótica dos Ovinos/transmissão , Infecções por Bactérias Gram-Negativas/diagnóstico , Infecções por Bactérias Gram-Negativas/epidemiologia , Infecções por Bactérias Gram-Negativas/transmissão , Reação em Cadeia da Polimerase , Ovinos , Doenças dos Ovinos/diagnóstico , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/microbiologia , Doenças dos Ovinos/transmissão , Suíça/epidemiologia
3.
Sci Rep ; 9(1): 10055, 2019 07 11.
Artigo em Inglês | MEDLINE | ID: mdl-31296905

RESUMO

The Gram-negative anaerobic bacterium Dichelobacter nodosus (Dn) causes footrot in ruminants, a debilitating and highly contagious disease that results in necrotic hooves and significant economic losses in agriculture. Vaccination with crude whole-cell vaccine mixed with multiple recombinant fimbrial proteins can provide protection during species-specific outbreaks, but subunit vaccines containing broadly cross-protective antigens are desirable. We have investigated two D. nodosus candidate vaccine antigens. Macrophage Infectivity Potentiator Dn-MIP (DNO_0012, DNO_RS00050) and Adhesin Complex Protein Dn-ACP (DNO_0725, DNO_RS06795) are highly conserved amongst ~170 D. nodosus isolates in the https://pubmlst.org/dnodosus/ database. We describe the presence of two homologous ACP domains in Dn-ACP with potent C-type lysozyme inhibitor function, and homology of Dn-MIP to other putative cell-surface and membrane-anchored MIP virulence factors. Immunization of mice with recombinant proteins with a variety of adjuvants induced antibodies that recognised both proteins in D. nodosus. Notably, immunization with fimbrial-whole-cell Footvax vaccine induced anti-Dn-ACP and anti-Dn-MIP antibodies. Although all adjuvants induced high titre antibody responses, only antisera to rDn-ACP-QuilA and rDn-ACP-Al(OH)3 significantly prevented rDn-ACP protein from inhibiting lysozyme activity in vitro. Therefore, a vaccine incorporating rDn-ACP in particular could contribute to protection by enabling normal innate immune lysozyme function to aid bacterial clearance.


Assuntos
Adesinas Bacterianas/imunologia , Proteínas de Bactérias/imunologia , Vacinas Bacterianas/imunologia , Dichelobacter nodosus/fisiologia , Pododermatite Necrótica dos Ovinos/imunologia , Peptidilprolil Isomerase/imunologia , Proteínas Recombinantes/imunologia , Animais , Anticorpos Antibacterianos/sangue , Formação de Anticorpos , Fímbrias Bacterianas/imunologia , Camundongos , Muramidase/antagonistas & inibidores , Filogenia , Conformação Proteica , Ruminantes , Vacinação
4.
Trop Anim Health Prod ; 51(5): 1089-1095, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30627862

RESUMO

The aim of this study was to determine the prevalence, serological diversity, and virulence of Dichelobacter nodosus in footrot lesions of sheep and identification of its predominant serotype as a potential vaccine candidate. The overall prevalence of footrot in sheep was 16.19%, and ranged from 13.69 to 19.71%, respectively. A total of 759 flocks with 22,698 sheep were investigated for footrot and 2374 clinical samples were collected from naturally infected sheep exhibiting footrot lesions. Of the 2374 samples collected, 1446 (60.90%) were positive for D. nodosus by polymerase chain reaction (PCR). These positive samples when subjected to serogroup-specific multiplex PCR, 1337 (92.46%) samples carried serogroup B, 247 (17.08%) possessed serogroup E, 86 (5.94%) serogroup I, and one (0.069%) serogroup G of D. nodosus. While mixed infection of serogroups B and E was detected in 127 (8.78%), B and I in 46 (3.18%) and B, E, and I in 26 (1.79%) samples, respectively. The serogroup B of D. nodosus was the predominant (92.47%) serogroup affecting sheep population with footrot followed by serogroup E (19.91%) and serogroup I (4.57%), respectively. Virulent status of D. nodosus strains were confirmed by presence of virulence-specific integrase A (intA) gene and the production of thermostable proteases. The intA gene was detected in 709 (72.79%) samples while gelatin gel test carried out on 246 representative isolates all positive for intA gene produced thermostable proteases, confirming their virulence nature. The PCR-restriction fragment length polymorphism (PCR-RFLP) of whole fimA gene of serogroup B revealed the predominance of serotype B5 (82.97%) of serogroup B. This information suggests that serotype B5 is the predominant serotype of D. nodosus associated with severe footrot lesions in sheep in Jammu & Kashmir (J&K), India. Hence, this serotype can be a potential vaccine candidate for the effective control and treatment of ovine footrot.


Assuntos
Vacinas Bacterianas/imunologia , Dichelobacter nodosus/fisiologia , Dichelobacter nodosus/patogenicidade , Pododermatite Necrótica dos Ovinos/prevenção & controle , Infecções por Bactérias Gram-Negativas/veterinária , Doenças dos Ovinos/prevenção & controle , Vacinação/veterinária , Animais , Dichelobacter nodosus/genética , Dichelobacter nodosus/imunologia , Pododermatite Necrótica dos Ovinos/microbiologia , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/prevenção & controle , Índia/epidemiologia , Prevalência , Estudos Soroepidemiológicos , Sorogrupo , Ovinos , Doenças dos Ovinos/microbiologia , Virulência
5.
Vet J ; 220: 1-6, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28190485

RESUMO

The aim of this study was to investigate the cost-benefit of different strategies to treat and control ovine footrot. In November 2006, 162 sheep farmers in England responded to a survey on prevalence and management of lameness. The costs of lameness per ewe per year (PEPY) were calculated for 116 flocks. Linear regression was used to model the overall cost of lameness PEPY by management method. Associations between farmer satisfaction and time and money spent managing lameness were investigated. The median prevalence of lameness was 5% (inter-quartile range, IQR, 4-10%). The overall cost of lameness PEPY in flocks with ≥10% lameness was UK £6.35 versus £3.90 for flocks with <5% lameness. Parenteral antibiotic treatment was associated with a significantly lower overall cost of lameness by £0.79 PEPY. Routine foot trimming and foot bathing were associated with significantly higher overall costs of lameness PEPY of £2.96 and £0.90, respectively. Farmers satisfied with time managing lameness spent significantly less time (1.46 h PEPY) than unsatisfied farmers (1.90 h PEPY). Farmers satisfied with money spent managing lameness had significantly lower treatment (£2.94 PEPY) and overall (£5.00 PEPY) costs than dissatisfied farmers (£5.50 and £7.60 PEPY, respectively). If the farmers in this study adopted best practice of parenteral antibiotic treatment with no routine foot trimming, and minimised foot bathing to treatment/prevention of interdigital dermatitis, the financial benefits would be approximately £4.65 PEPY. If these costs are similar on other farms the management changes would lead to significant economic benefits for the sheep industry.


Assuntos
Criação de Animais Domésticos/métodos , Análise Custo-Benefício , Pododermatite Necrótica dos Ovinos/economia , Doenças dos Ovinos/economia , Animais , Dichelobacter nodosus/fisiologia , Inglaterra/epidemiologia , Feminino , Pododermatite Necrótica dos Ovinos/epidemiologia , Pododermatite Necrótica dos Ovinos/microbiologia , Modelos Teóricos , Ovinos , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/microbiologia
6.
Vet J ; 216: 25-32, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27687922

RESUMO

Virulent ovine footrot caused by Dichelobacter nodosus is an endemic disease worldwide. It is associated with severe pain, impaired animal welfare and economic losses. The competitive real-time PCR for the differentiation of virulent aprV2 and benign aprB2 strains of Dichelobacter nodosus provides an objective, rapid and sensitive diagnostic tool for footrot surveillance, especially as it enables early detection of subclinical carriers of virulent strains. The aim of this study was to evaluate the feasibility of complete elimination of Dichelobacter nodosus strains carrying the aprV2 gene from sheep flocks. The treatment protocol was based on careful removal of loose and severely overgrown claw horn, weekly stand-in foot baths of the entire flock for 10 min per sheep, using a 10% zinc sulphate disinfectant solution, clinical and PCR follow-up and isolation or culling of non-responders. Dichelobacter nodosus strains carrying the aprV2 gene were successfully eliminated from the feet of the sheep of all 28 flocks (100%) participating in the study within 6-19 weeks of weekly foot bathing. A strong correlation between the length of time for weekly foot bathing to eliminate the virulent strains and the within-flock prevalence of clinical footrot at the beginning was observed (rho, 0.68; P <0.001). A statistically significant correlation was not detected between flock size and the length of time for weekly foot bathing (rho, 0.28; P = 0.14), or the prevalence of clinical footrot at study commencement (rho, -0.04; P = 0.82), respectively. In conclusion, a complete elimination of Dichelobacter nodosus strains carrying the aprV2 gene in sheep flocks was possible with a protocol based on careful trimming, weekly stand-in foot baths, and identification of infection using a specific PCR-test and isolation or culling of non-responders.


Assuntos
Dichelobacter nodosus/fisiologia , Desinfetantes/uso terapêutico , Pododermatite Necrótica dos Ovinos/terapia , Infecções por Bactérias Gram-Negativas/veterinária , Doenças dos Ovinos/tratamento farmacológico , Sulfato de Zinco/uso terapêutico , Criação de Animais Domésticos , Animais , Proteínas de Bactérias/análise , Dichelobacter nodosus/efeitos dos fármacos , Dichelobacter nodosus/patogenicidade , Estudos de Viabilidade , Feminino , Pododermatite Necrótica dos Ovinos/tratamento farmacológico , Pododermatite Necrótica dos Ovinos/microbiologia , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/terapia , Masculino , Estudos Prospectivos , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Serina Endopeptidases/análise , Ovinos , Doenças dos Ovinos/microbiologia , Doenças dos Ovinos/terapia , Suíça , Virulência
7.
Anaerobe ; 38: 81-87, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26746387

RESUMO

Dichelobacter nodosus (D. nodosus) is the causative agent of footrot in sheep; one of the most important health and welfare issues of sheep worldwide. For control programmes to be effective, it is essential that the transmission cycle of D. nodosus is understood and bacterial reservoirs in the environment are better defined. This study evaluated the survival of D. nodosus in different soils using soil microcosms. Cultivation independent and dependent methods were used to detect D. nodosus over 40 days from seeding in soil. A D. nodosus specific probe was used for quantification by qPCR and viability was assessed by cell permeability to an intercalating dye, PMA, and by culture. Survival varied dramatically depending on soil type, matric potential (MP) and temperature. Our findings indicate that D. nodosus survival was higher at 5 °C compared with 25 °C in all soils and significantly longer at both temperatures in clay soil (>44% clay) compared with other soil types. Survival under all conditions was longer than 30 days for both culture independent and dependent methods, this is substantially longer than previous studies and, if this is an infectious dose, longer than the current recommendation of resting a field for 14 days to prevent onward infection.


Assuntos
Dichelobacter nodosus/fisiologia , Pododermatite Necrótica dos Ovinos/microbiologia , Viabilidade Microbiana , Doenças dos Ovinos/microbiologia , Microbiologia do Solo , Animais , Anti-Infecciosos/farmacologia , Azidas/farmacologia , DNA Bacteriano , Dichelobacter nodosus/classificação , Dichelobacter nodosus/isolamento & purificação , Propídio/análogos & derivados , Propídio/farmacologia , Ovinos
8.
Acta Vet Scand ; 57: 55, 2015 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-26407552

RESUMO

BACKGROUND: Dichelobacter nodosus is the main causative agent of ovine footrot, and there are strong indications that the bacterium can be transferred to cattle grazing on the same pasture as sheep. The aim of this study was to investigate if benign and virulent D. nodosus strains isolated from sheep can be transferred to the interdigital skin of cattle under experimental conditions. Further, we wanted to observe the impact of such infection on bovine foot health, and test the effect of topical chlortetracycline (Cyclo spray(®): Eurovet) on the infection. FINDINGS: Six heifers were included in the study. After an initial 18-day maceration period, three heifers were inoculated on one single foot with a benign strain and three with a virulent strain by adding bacterial suspension in a bandage. The bandages were left on for 17 days, and when removed, D. nodosus was isolated from all six heifers. All six heifers developed interdigital dermatitis. In five of the heifers D. nodosus organisms were demonstrated within the epidermis. Twenty-four days after treatment with chlortetracycline all heifers were negative by cultivation, but tested positive for D. nodosus by polymerase chain reaction (PCR). Two of the six heifers still tested positive for D. nodosus by PCR 49 days after treatment. After 70 days, all heifers tested negative for D. nodosus. CONCLUSIONS: This study shows that both virulent and benign D. nodosus strains originating from sheep can be transferred to naïve heifers under experimental conditions. Further, the study supports the hypothesis that infections with virulent D. nodosus in cattle are associated with interdigital dermatitis. No conclusion regarding the treatment of D. nodosus infection with chlortetracycline was possible.


Assuntos
Antibacterianos/uso terapêutico , Doenças dos Bovinos/microbiologia , Clortetraciclina/uso terapêutico , Dichelobacter nodosus/fisiologia , Dermatite Digital/microbiologia , Infecções por Bactérias Gram-Negativas/veterinária , Animais , Bovinos , Doenças dos Bovinos/tratamento farmacológico , Doenças dos Bovinos/transmissão , Dermatite Digital/tratamento farmacológico , Dermatite Digital/transmissão , Feminino , Pododermatite Necrótica dos Ovinos/microbiologia , Pododermatite Necrótica dos Ovinos/transmissão , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/transmissão , RNA Ribossômico 16S/genética , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Ovinos , Doenças dos Ovinos/microbiologia , Doenças dos Ovinos/transmissão , Carneiro Doméstico
9.
Vet Microbiol ; 179(1-2): 34-41, 2015 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-25891425

RESUMO

Dichelobacter nodosus is the essential causative agent of footrot in sheep and type IV fimbriae-mediated twitching motility has been shown to be essential for virulence. We have identified a two-component signal transduction system (TwmSR) that shows similarity to chemosensory systems from other bacteria. Insertional inactivation of the gene encoding the response regulator, TwmR, led to a twitching motility defect, with the mutant having a reduced rate of twitching motility when compared to the wild-type and a mutant complemented with the wild-type twmR gene. The reduced rate of twitching motility was not a consequence of a reduced growth rate or decreased production of surface located fimbriae, but video microscopy indicated that it appeared to result from an overall loss of twitching directionality. These results suggest that a chemotactic response to environmental factors may play an important role in the D. nodosus-mediated disease process.


Assuntos
Dichelobacter nodosus/genética , Fímbrias Bacterianas/genética , Pododermatite Necrótica dos Ovinos/microbiologia , Infecções por Bactérias Gram-Negativas/veterinária , Doenças dos Ovinos/microbiologia , Transdução de Sinais , Animais , Quimiotaxia , Dichelobacter nodosus/fisiologia , Fímbrias Bacterianas/metabolismo , Infecções por Bactérias Gram-Negativas/microbiologia , Mutagênese Insercional , Ovinos , Virulência
10.
Vet Res ; 46: 10, 2015 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-25828795

RESUMO

When severe footrot was detected in Norway in 2008, a surveillance programme was initiated and followed by an elimination programme. By 2013 the disease had spread to two of 19 counties and a total of 119 (1%) sheep flocks had been diagnosed with severe footrot. A simulation model was developed to estimate the potential spread of severe footrot in Norway and to estimate the relative importance of the different spreading routes. The model parameters were based on the rate of spread of the first 38 diagnosed cases and the management and climatic factors particular for Norway. The model showed that by 2013, severe footrot would have spread to six counties and infected 16% of the sheep flocks if no elimination programme had been initiated. If this is compared with the 1% of flocks that were diagnosed in Norway by 2013, there seems to be a large effect of the implemented footrot elimination programme. By 2035, it was estimated that severe footrot would have spread to 16 counties and 64% of the sheep flocks. Such an extensive spread would probably impose a large negative impact on the sheep industry and welfare of the sheep. The most effective way to curb the spread of severe footrot was by decreasing the within county infection rate. This could be achieved by decreasing the contact between flocks or by decreasing the environmental load of D. nodosus, for example by footbathing sheep, culling diseased sheep or eliminating severe footrot in the flock.


Assuntos
Dichelobacter nodosus/fisiologia , Pododermatite Necrótica dos Ovinos/transmissão , Infecções por Bactérias Gram-Negativas/veterinária , Doenças dos Ovinos/transmissão , Animais , Clima , Simulação por Computador , Pododermatite Necrótica dos Ovinos/microbiologia , Pododermatite Necrótica dos Ovinos/prevenção & controle , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/prevenção & controle , Infecções por Bactérias Gram-Negativas/transmissão , Modelos Teóricos , Noruega , Ovinos , Doenças dos Ovinos/microbiologia , Doenças dos Ovinos/prevenção & controle
11.
Vet Microbiol ; 176(3-4): 321-7, 2015 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-25742734

RESUMO

Analysis of bacterial populations in situ provides insights into pathogen population dynamics and potential reservoirs for disease. Here we report a culture-independent study of ovine footrot (FR); a debilitating bacterial disease that has significant economic impact on sheep farming worldwide. Disease begins as an interdigital dermatitis (ID), which may then progress to separation of the hoof horn from the underlying epidermis causing severe footrot (SFR). Dichelobacter nodosus is the causative agent of ovine FR, however, the role of Fusobacterium necrophorum and other bacteria present in the environment and on the feet of sheep is less clear. The objective of this study was to use fluorescence in situ hybridisation (FISH) to detect, localise and quantify D. nodosus, F. necrophorum and the domain Bacteria from interdigital skin biopsies of healthy, ID- and SFR-affected feet. D. nodosus and F. necrophorum populations were restricted primarily to the epidermis, but both were detected more frequently in feet with ID or SFR than in healthy feet. D. nodosus cell counts were significantly higher in feet with ID and SFR (p<0.05) than healthy feet, whereas F. necrophorum cell counts were significantly higher only in feet with SFR (p<0.05) than healthy feet. These results, together with other published data, indicate that D. nodosus likely drives pathogenesis of footrot from initiation of ID to SFR; with D. nodosus cell counts increasing prior to onset of ID and SFR. In contrast, F. necrophorum cell counts increase after SFR onset, which may suggest an accessory role in disease pathogenesis, possibly contributing to the severity and duration of SFR.


Assuntos
Dichelobacter nodosus/patogenicidade , Pododermatite Necrótica dos Ovinos/microbiologia , Fusobacterium necrophorum/patogenicidade , Doenças dos Ovinos/microbiologia , Carneiro Doméstico , Animais , Dichelobacter nodosus/fisiologia , Fusobacterium necrophorum/fisiologia , Casco e Garras/patologia , Hibridização in Situ Fluorescente/veterinária , Dinâmica Populacional , Ovinos , Pele/microbiologia
12.
Vet J ; 201(3): 295-301, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24973007

RESUMO

Footrot, including interdigital dermatitis, is caused by Dichelobacter nodosus cause the majority of lameness in sheep in the UK. Lame sheep often have overgrown hoof horn but recent evidence has indicated that trimming overgrown hoof horn increases recovery time, and that routine foot trimming of the flock does not reduce the prevalence or incidence of lameness. The objectives of this study were to investigate the temporal associations between hoof horn length, footrot and climate. Fifty multiparous ewes were monitored for 10 months. On eight occasions hoof horn length, foot lesions and body condition were recorded. At the first examination, ewes were assigned to one of two treatment groups. All ewes that became lame with footrot were treated at one time point per week, either by trimming hoof horn and applying a topical antibiotic spray or with parenteral antibiotic and topical antibiotic spray. Hoof horn length in ewes at pasture varied over the year and was associated with temperature and rainfall. New cases of footrot occurred all year round and were associated with prior prevalence of footrot in the flock and prior temperature and rainfall. Overgrown hoof horn did not precede lameness but occurred once the sheep were lame. One year of prompt treatment of footrot reduced the range in hoof horn length in the sheep in both treatment groups. At the end of the study the hoof lengths of ewes in both groups were not significantly different. On this farm, hoof horn length was self-regulating in both non-lame and treated lame sheep whether trimming was part of the treatment or not and there would have been no benefit from routine foot trimming of this flock.


Assuntos
Pododermatite Necrótica dos Ovinos/cirurgia , Infecções por Bactérias Gram-Negativas/veterinária , Casco e Garras/cirurgia , Doenças dos Ovinos/cirurgia , Animais , Dichelobacter nodosus/fisiologia , Inglaterra/epidemiologia , Feminino , Pododermatite Necrótica dos Ovinos/epidemiologia , Pododermatite Necrótica dos Ovinos/microbiologia , Infecções por Bactérias Gram-Negativas/epidemiologia , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/cirurgia , Casco e Garras/anatomia & histologia , Prevalência , Chuva , Distribuição Aleatória , Estações do Ano , Ovinos , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/microbiologia , Temperatura
13.
Vet Microbiol ; 170(3-4): 375-82, 2014 Jun 04.
Artigo em Inglês | MEDLINE | ID: mdl-24698131

RESUMO

Dichelobacter nodosus is the main aetiological agent of ovine footrot and the bacterium has also been associated with interdigital dermatitis is cattle. The aim of this study was to investigate possible cross-infection of virulent D. nodosus between sheep and co-grazing cattle. Five farms, where sheep previously diagnosed with virulent D. nodosus were co-grazing with cattle for different periods of time, were included. The study sample consisted of 200 cows and 50 sheep. All cows were examined for the presence of interdigital dermatitis, and ten ewes, preferably with symptoms of footrot, had the footrot scores recorded. On each farm, the same ten ewes and ten cows were chosen for bacterial analyses. Swabs were analysed for D. nodosus by PCR and culturing. D. nodosus isolates were virulence-tested and assigned to serogroups by fimA variant determination. Biopsies were evaluated histopathologically and analysed by fluorescent in situ hybridization for D. nodosus, Treponema spp. and Fusobacterium necrophorum. D. nodosus defined as virulent by the gelatin gel test were isolated from 16 sheep from four farms and from five cows from two of the same farms. All five cows had interdigital dermatitis. Two of the cows stayed infected for at least eight months. By pulsed-field gel electrophoresis (PFGE), the isolates from the five cows were found to be genetically indistinguishable or closely related to isolates from sheep from the same farm. This indicates that cross-infection between sheep and cows have occurred.


Assuntos
Doenças dos Bovinos/transmissão , Dichelobacter nodosus/fisiologia , Pododermatite Necrótica dos Ovinos/transmissão , Infecções por Bactérias Gram-Negativas/veterinária , Doenças dos Ovinos/transmissão , Animais , Bovinos , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/patologia , Dichelobacter nodosus/genética , Dichelobacter nodosus/isolamento & purificação , Eletroforese em Gel de Campo Pulsado , Pododermatite Necrótica dos Ovinos/microbiologia , Pododermatite Necrótica dos Ovinos/patologia , Infecções por Fusobacterium/veterinária , Fusobacterium necrophorum , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/transmissão , Reação em Cadeia da Polimerase , Ovinos , Doenças dos Ovinos/microbiologia , Doenças dos Ovinos/patologia , Treponema , Infecções por Treponema/veterinária
14.
Acta Vet Scand ; 55: 4, 2013 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-23343097

RESUMO

BACKGROUND: Dichelobacter nodosus is the causative agent of footrot in sheep. The survival of the bacterium in soil is of importance for the epidemiology of the disease. The investigation evaluates the survival of D. nodosus in soil with and without added hoof powder stored under different temperatures. RESULTS: An experimental setup was used with bacteriological culture and real-time polymerase chain reaction (PCR), and the results indicate that the bacteria can survive in soil for longer time than previously expected. The survival time was found to be dependent on temperature and the addition of hoof powder to the soil, with the longest survival time estimated to be 24 days in soil samples with hoof powder stored at 5°C. CONCLUSION: Our findings indicate that the survival time of D. nodosus and its ability to infect susceptible sheep on pasture under different climatic conditions should be studied further.


Assuntos
Dichelobacter nodosus/fisiologia , Microbiologia do Solo , Solo/química , Animais , Casco e Garras , Reação em Cadeia da Polimerase em Tempo Real , Ovinos , Doenças dos Ovinos/microbiologia , Temperatura , Fatores de Tempo
15.
Vet Res ; 42: 99, 2011 Sep 06.
Artigo em Inglês | MEDLINE | ID: mdl-21896161

RESUMO

We reported previously that sheep affected with footrot (FR) have lower whole-blood selenium (WB-Se) concentrations and that parenteral Se-supplementation in conjunction with routine control practices accelerates recovery from FR. The purpose of this follow-up study was to investigate the mechanisms by which Se facilitates recovery from FR. Sheep affected with FR (n = 38) were injected monthly for 15 months with either 5 mg Se (FR-Se) or saline (FR-Sal), whereas 19 healthy sheep received no treatment. Adaptive immune function was evaluated after 3 months of Se supplementation by immunizing all sheep with a novel protein, keyhole limpet hemocyanin (KLH). The antibody titer and delayed-type hypersensitivity (DTH) skin test to KLH were used to assess humoral immunity and cell-mediated immunity, respectively. Innate immunity was evaluated after 3 months of Se supplementation by measuring intradermal responses to histamine 30 min after injection compared to KLH and saline, and after 15 months of Se supplementation by isolating neutrophils and measuring their bacterial killing ability and relative abundance of mRNA for genes associated with neutrophil migration. Compared to healthy sheep, immune responses to a novel protein were suppressed in FR-affected sheep with smaller decreases in FR-affected sheep that received Se or had WB-Se concentrations above 250 ng/mL at the time of the immune assays. Neutrophil function was suppressed in FR-affected sheep, but was not changed by Se supplementation or WB-Se status. Sheep FR is associated with depressed immune responses to a novel protein, which may be partly restored by improving WB-Se status (> 250 ng/mL).


Assuntos
Pododermatite Necrótica dos Ovinos/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Selênio/uso terapêutico , Doenças dos Ovinos/imunologia , Ração Animal/análise , Animais , Anticorpos Antibacterianos/sangue , Dichelobacter nodosus/fisiologia , Dieta/veterinária , Suplementos Nutricionais/análise , Feminino , Pododermatite Necrótica dos Ovinos/tratamento farmacológico , Pododermatite Necrótica dos Ovinos/microbiologia , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/microbiologia , Hemocianinas/farmacologia , Histamina/administração & dosagem , Histamina/farmacologia , Hipersensibilidade Tardia/veterinária , Doenças do Sistema Imunitário/veterinária , Imunidade Celular/efeitos dos fármacos , Imunidade Humoral/efeitos dos fármacos , Imunidade Inata/efeitos dos fármacos , Testes Intradérmicos/veterinária , Transtornos Leucocíticos/veterinária , Neutrófilos/metabolismo , Reação em Cadeia da Polimerase/veterinária , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Selênio/administração & dosagem , Ovinos , Doenças dos Ovinos/tratamento farmacológico , Doenças dos Ovinos/microbiologia
16.
Vet Microbiol ; 153(1-2): 59-66, 2011 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-21596496

RESUMO

Ovine footrot is a contagious and debilitating disease that is of major economic significance to the sheep meat and wool industries. The causative bacterium is the gram negative anaerobe Dichelobacter nodosus. Research that has used a classical molecular genetics approach has led to major advances in our understanding of the role of the key virulence factors of D. nodosus in the disease process. D. nodosus strains produce polar type IV fimbriae and extracellular serine proteases. Mutagenesis of the fimbrial subunit gene fimA and the pilT gene, which is required for fimbrial retraction, and subsequent testing of these mutants in sheep virulence trials has shown that type IV fimbriae-mediated twitching motility is essential for virulence. The extracellular protease genes aprV2, aprV5 and bprV have also been mutated. Analysis of these mutants has shown that ArpV5 is the major extracellular protease and that AprV2 is the thermostable protease that is responsible for the extracellular elastase activity. Structural analysis of AprV2 has revealed that it contains several novel loops, one of which appears to act as an exosite that may modulate substrate accessibility. Finally, virulence experiments in sheep have shown that the AprV2 protease is required for virulence.


Assuntos
Dichelobacter nodosus/patogenicidade , Pododermatite Necrótica dos Ovinos/microbiologia , Doenças dos Ovinos/microbiologia , Animais , Dichelobacter nodosus/genética , Dichelobacter nodosus/fisiologia , Fímbrias Bacterianas/metabolismo , Humanos , Serina Endopeptidases/genética , Serina Endopeptidases/metabolismo , Ovinos , Virulência , Fatores de Virulência/genética , Fatores de Virulência/metabolismo
17.
J Bacteriol ; 188(13): 4801-11, 2006 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-16788189

RESUMO

Type IV fimbriae are expressed by several bacterial pathogens and are essential for virulence in Dichelobacter nodosus, which causes ovine footrot. We have identified a two-component signal transduction system (PilR/S) and an alternative sigma factor (sigma 54) that were shown by insertional inactivation to be required for the regulation of fimbrial biogenesis in D. nodosus. Western blots showed that in both pilR and rpoN mutants, fimbrial subunit production was significantly reduced by a process that was shown to occur at a PilR- and sigma 54-dependent promoter. The mutants lacked surface fimbriae, which were shown to be required for the adherence of D. nodosus cells to tissue culture monolayers. The reduction in fimbrial subunit production in these mutants also resulted in a concomitant loss of the ability to secrete extracellular proteases. A maltose binding protein-PilR fusion protein was purified and was shown to bind specifically to a region located 234 to 594 bp upstream of the fimA transcriptional start point. To determine additional targets of PilR and sigma 54, genome-wide transcriptional profiling was performed using a whole-genome oligonucleotide microarray. The results indicated that PilR and sigma 54 regulated genes other than fimA; these genes appear to encode surface-exposed proteins whose role in virulence is unknown. In conclusion, this study represents a significant advancement in our understanding of how the ability of D. nodosus to cause ovine footrot is regulated, as we have shown that the biogenesis of type IV fimbriae in D. nodosus is regulated by a sigma 54-dependent PilR/S system that also indirectly controls protease secretion.


Assuntos
Dichelobacter nodosus/genética , Fímbrias Bacterianas/genética , Regulação Bacteriana da Expressão Gênica , Genes Bacterianos , Transdução de Sinais/genética , Aderência Bacteriana , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/fisiologia , Dichelobacter nodosus/metabolismo , Dichelobacter nodosus/fisiologia , Proteínas de Fímbrias/genética , Fímbrias Bacterianas/metabolismo , Infecções por Bactérias Gram-Negativas/microbiologia , Dados de Sequência Molecular , Análise de Sequência com Séries de Oligonucleotídeos , Peptídeo Hidrolases/genética , Peptídeo Hidrolases/metabolismo , RNA Polimerase Sigma 54/genética , RNA Polimerase Sigma 54/fisiologia , Fatores de Transcrição/genética , Fatores de Transcrição/fisiologia
18.
Vet J ; 169(1): 28-41, 2005 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-15683762

RESUMO

Ovine footrot remains the most important cause of lameness in sheep flocks in the UK, despite the existence of proven methods for the control of the disease. Recent research suggests that sheep farmers may be unaware of these methods and may allocate greater resources to treatment of footrot rather than to its prevention. Foot paring, topical treatments, vaccination and parenteral antibiotic therapy all have a role in treating sheep with advanced footrot infections, but prevention of severe infections is best achieved by the timely implementation of control programmes. These are usually based on footbathing and vaccination. For control programmes to be effective it is essential that the pathogenesis and epidemiology of footrot is understood and that control methods are implemented at appropriate times in the season, depending on climatic and pasture conditions. This article reviews these strategies and makes recommendations for steps to reduce the spread of footrot between flocks and to reduce the incidence of footrot within UK flocks.


Assuntos
Pododermatite Necrótica dos Ovinos/prevenção & controle , Doenças dos Ovinos/prevenção & controle , Administração Tópica , Animais , Terapia Combinada , Dichelobacter nodosus/fisiologia , Pododermatite Necrótica dos Ovinos/diagnóstico , Pododermatite Necrótica dos Ovinos/tratamento farmacológico , Fusobacterium necrophorum/fisiologia , Casco e Garras/microbiologia , Casco e Garras/patologia , Ovinos , Doenças dos Ovinos/diagnóstico , Doenças dos Ovinos/tratamento farmacológico , Vacinação/veterinária , Sulfato de Zinco/uso terapêutico
19.
Aust Vet J ; 77(5): 318-21, 1999 May.
Artigo em Inglês | MEDLINE | ID: mdl-10376104

RESUMO

OBJECTIVE: To determine sheep farmers' attitudes to and beliefs about ovine footrot, and to improve their knowledge about the diagnosis, control and eradication of this disease. METHOD: Eighteen workshops, involving 291 farmers, were conducted across Victoria in the spring of 1996. The workshops were designed as small-group discussions with a maximum attendance of 20 farmers to encourage active participation. All participants completed questionnaires before each workshop and 12 to 40 weeks after the last workshop. RESULTS: Before the workshops the farmers had a poor understanding of the principles of diagnosis, control and eradication of footrot. For example, only 50% knew the footrot organism survived in soil for less than 7 days, over two-thirds did not know the reason for paring sheep's feet during an eradication program, and only 31% realised cattle were a potential source of footrot infection for sheep. After the workshops, understanding about footrot was significantly improved; 87% said Dichelobacter nodosus survived in soil less than 7 days, 71% knew the reason for paring sheep's feet and 64% realised that cattle were a potential source of footrot infection. As well as improved knowledge, change of attitude among farmers is fundamentally important if virulent footrot is to be successfully controlled and eradicated. The workshops successfully initiated this process; 40% of farmers thought the workshops changed their attitudes to footrot, while 37% said they gained an increased understanding of other people's opinions about the disease. CONCLUSION: Farmers' poor understanding of ovine footrot is a constraint to the programs aimed at controlling this disease. Small group workshops may be an effective way to influence farmers' attitudes and beliefs, and could facilitate the effectiveness of regulatory disease control programs.


Assuntos
Pododermatite Necrótica dos Ovinos/diagnóstico , Pododermatite Necrótica dos Ovinos/prevenção & controle , Doenças dos Ovinos/diagnóstico , Doenças dos Ovinos/prevenção & controle , Ensino , Animais , Bovinos , Dichelobacter nodosus/fisiologia , Pododermatite Necrótica dos Ovinos/microbiologia , Conhecimentos, Atitudes e Prática em Saúde , Humanos , Quarentena/veterinária , Ovinos , Doenças dos Ovinos/microbiologia , Microbiologia do Solo , Inquéritos e Questionários , Vitória
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