Incidence and significance of treponemes in animals

The genus Treponema consists of various species. Currently most of them are not cultivable because respective cultivation conditions are unknown. Therefore the biodiversity of treponemes was only appreciated recently by applying comparative 16S rRNA sequence analysis. Treponemes are mainly representatives of the gastrointestinal autochthonal flora, especially in termites, but they have also been described in swine and cattle. On the other hand treponemes are involved in different infectious diseases, the most well known being syphilis in humans or venereal spirochetosis in rabbits. Furthermore, treponemes are associated with several infectious periodontal diseases, e.g. gingivitis or periodontitis, where they can be detected regularly. Culture has not been successful for most of the oral treponemes, so the major part can only be identified by their 16S rRNA sequence. Similar to these oral disorders treponemes are also associated with digital dermatitis (DD), a chronic inflammatory disease of the bovine skin, where different treponemal phylotypes were found in large numbers. Treponema brennaborense was first identified and isolated in DD biopsies. Unravelling the pathogenic potential and aetiological significance of treponemes in chronic infectious diseases like peridontitis or DD remains a costly task. Although treponemes can be frequently detected in such lesions, it is often unclear to what extent treponemes are involved in pathogenesis of these diseases. The possession of various virulence features like high motility, the ability to adhere and invade as well as to cause cytopathic effects in eukaryotic cells are highly indicative of the aetiological relevance of treponemes.     

Risk assessment of bluetongue disease incursion into Germany using geographic information system (GIS)

Using a geographic information system (GIS), by analysis of the relationship between the spatial distribution of cattle density and the risk factors temperature, altitude and rainfall, we defined geographical habitats enabling optimal development and competence of Culicoides spp. to transmit Bluetongue-Virus (BTV): Risk zones (low, high, highest risk) were identified mainly in Baden-Württemberg, Hessen and Rheinland-Pfalz if persistently infected ruminants are imported into these zones in summer (June to August mainly), based on the current climatic conditions, BTD outbreaks are considered a real possibility. Overwintering of the virus seems unlikely. However, global warming will lead to a steady increase of the size of the risk zones. In addition, the possibility of primary outbreaks increases. The reason for this is not only the expected northern shift of Culicoides imicola, but in addition an increasing vector competence of domestic Culicoides species. We therefore recommend the storage of vaccines as well as conducting ecological studies analysing the presence of Culicoides vectors. Using the data from these studies, it will be possible to produce updated quantitative risk assessment via GIS.     

High prevalence of treponemes in bovine digital dermatitis-a molecular epidemiology

To validate the epidemiology of Treponema spp. associated with digital dermatitis (DD) a large number of DD samples (n=56) were examined by DNA-DNA dot blot analyses using oligonucleotide probes specific for phylogenetic group I-VII of oral treponemes and DD-associated phylotypes DDKL-4, DDKL-12 and DDKL-20 as well as for T. brennaborense and T. socranskii. Positive hybridisation results were obtained for phylogenetic groups I, II and IV and phylotypes DDKL-4 and DDKL-12. While phylotype DDKL-4 was detected in 100% of the samples treponemes belonging to phylogenetic group TRE I, TRE II and TRE IV were prevalent in nearly 80% of the samples and phylotype DDKL-12 was detected in 66.1% of the samples. Analysis of Treponema groups present concurrently in the same sample revealed that a combination of TRE I-TRE II-TRE IV-DDKL-4 was most prevalent and could be detected in up to 71% of the samples. These data indicate that this combination of different Treponema spp. seems to be the most important one in the pathogenesis of DD. In contrast, T. brennaborense originally isolated from DD material this treponeme was not detected in any of the samples clearly indicating that this species is not absolutely associated with DD and therefore may represent only an incidental treponeme. Fluorescence in situ hybridisation (FISH) obviously highlights the invasive character of DD-associated treponemes. Mainly treponemes belonging to phylogenetic group TRE I and phylotype DDKL-4 were detected in high numbers compared to the total number of bacteria and also in deeper layers of the epithelium at the transition of unaffected and affected tissue. Our results confirm a high prevalence and diversity of Treponema spp. in DD lesions. In addition, our data indicate that certain combinations of Treponema spp. are detected much more frequently than others. Furthermore, Treponema spp. appears at the interface between healthy and diseased tissue underlining their importance for the pathogenesis of DD.     

Prevalence of enteropathogens in suckling and weaned piglets with diarrhoea in southern Germany

Faecal samples from suckling (n = 205) and weaned piglets (n = 82) with diarrhoea from 24 farms in Southern Germany were examined for shedding of important metazoic parasitic, viral and bacterial pathogens using culture, microscopic and electronmicroscopic methods. Escherichia coli isolates were tested further for the enterotoxin genes est-Ia and elt-I by colony blot hybridization. Isospora suis was diagnosed in 26.9% and Cryptosporidium parvum in 1.4% of the piglets investigated. The proportion of coronavirus-positive animals was 13.4% and 4% were positive for rotavirus. It was found that 17.6% of the animals were infected with enterotoxigenic E. coli (ETEC; 10.1% ETEC-ST-Ia and 8.6% ETEC-LT-I, respectively). The occurrence of the pathogens was significantly associated with the age of the animals examined (P < 0.001). Isospora suis was predominantly isolated from suckling piglets (in the second and third week of life), while in weaned piglets (fourth week of life) rotavirus and ETEC were most prevalent. On 22 of the 24 piglet production farms examined at least one of the investigated pathogens was detected. Coronavirus was diagnosed in 66.7%, I. suis in 62.5%, rotavirus in 20.8% and C. parvum in 8.3% of the farms. These results underline the fact that despite the hygienic, technical and immune preventive efforts during the last years, enteropathogens are still common in German piglet production units.     

Molecular epidemiology of avian pathogenic Escherichia coli (APEC) isolated from colisepticemia in poultry

The molecular biology and epidemiology of 150 avian pathogenic Escherichia coli strains (APEC) isolated from septicemic poultry in Germany was investigated by serotyping, pulsed field gel electrophoresis (PFGE), and polymerase chain reaction (PCR). Only 49.6% of the isolates could be grouped to serogroups O1, O2, and O78. Macrorestriction analyses data revealed two large clonal groups (clusters I and II) among the APEC strains with a similarity of 60.9% to each other. An association between restriction pattern and serogroup or origin of the strains was only present in a few subgroups of each clusters I and II, but was not evident. In contrast, our data revealed distinct combinations of virulence-associated genes in that 51.2% of the O2-strains harboured a combination of the genes fyuA, irp2, iucD, tsh, vat, fimC, and colV and 36.4% of the O78-strains possessed the same gene combination with exception of vat. With 34 different gene combinations the non-O1, -O2, -O78 isolates revealed a higher variability in their virulence gene pattern than O1-, O2-, and O78-strains with 6, 13, and 9 patterns, respectively. Our data indicate only a limited association between the virulence gene pattern and the serogroup of APEC strains and question the sensitivity of O-typing for APEC identification without the application of further diagnostic tools. Although a limited number of APEC clones exist, horizontal gene transfer seems to be common in these pathogens. These findings strengthen further research on the population structure of APEC and may be the reason for the lack of clear definition of this common E. coli pathotype.     

Systemic Immunosuppression by Methylprednisolone and Pregnancy Rates in Goats Undergoing the Transfer of Cloned Embryos

The presence of the zona pellucida has been perceived as a requirement for the oviductal transfer of cloned embryos at early stages of development while protecting the embryo from an immune system response. We hypothesized that steroid hormone therapy could reduce a potential cellular immune response after the transfer of zona‐free cloned embryos into the oviduct of recipient female goats. In Experiment 1, seven does were used to study the systemic immunosuppressant effect of the methylprednisolone administration (for 3 days) on blood cell counts. Whole blood was collected prior to treatment with methyprednisolone and then on Days 1, 2, 3, 4, 7, 14, 21 and 28 after the first dose of methylprednisolone for the analysis of haematological parameters. Methylprednisolone treatment significantly reduced circulating white blood cells and neutrophils in comparison with pre‐treatment levels, demonstrating a systemic immunosuppressant effect. In Experiment 2, a group of 58 does were used as recipient females to study the effect of administration of methylprednisolone for 3 days on the establishment of pregnancies after the transfer of zona‐free cloned embryos into the oviducts. No effects on pregnancy rates on Day 30 were observed regarding the distinct treatment groups (control vs. methylprednisolone), the source of oocytes (in vivo‐ vs in vitro‐matured) or the presence or absence of the zona pellucida in embryos. In summary, methylprednisolone was effective at inducing a systemic immunosuppressed state in goats, but the treatment prior to embryo transfer did not affect pregnancy rates. Moreover, pregnancy rates were similar between zona‐free and zona‐intact goat cloned embryos.     

Antimicrobial susceptibility of Klebsiella spp. and Proteus spp. from various organ systems of horses, dogs and cats as determined in the BfT-GermVet monitoring program 2004-2006

A total of 120 isolates of Klebsiella spp. and Proteus spp. collected from horses and small animals (dogs and cats) were screened for their susceptibility to 24 different antimicrobial agents. Klebsiella spp. were included from infections of the genital tract (GT) of horses (36 isolates) and the urinary/genital tract (UGT) from dogs and cats (17 isolates), while Proteus spp. were from small animal (dogs and cats) infections of the UGT (37 strains) and the skin (incl. ear/mouth) (30 isolates). In Klebsiella spp. resistance appeared most frequently to ampicillin (53-67%), sulfamethoxazole (19-29%) and potentiated sulfonamides (trimethoprim/sulfamethoxazole 1/19 combination) (19-24%). A further 29% of enrofloxacin resistant Klebsiella isolates were observed for the UGT of small animals. From the GT of horses for this antimicrobial agent there was no isolate detected with a comparably high minimum inhibitory concentration (MIC) value. In Proteus spp. highest percentages of resistance occurred against tetracycline (90-92%). Due to drug efflux proteins, high MIC values against this antimicrobial agent have been frequently reported in literature. In Proteus spp. relevant resistance percentages also occurred for potentiated sulfonamides (27-37%), sulfamethoxazole (24-37%) and chloramphenicol (24-37%).