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The Swedish Forum for Biopreparedness Diagnostics (FBD) is a network that fosters collaboration among the 4 agencies with responsibility for the laboratory diagnostics of high-consequence pathogens, covering animal health and feed safety, food safety, public health and biodefense, and security. The aim of the network is to strengthen capabilities and capacities for diagnostics at the national biosafety level-3 (BSL-3) laboratories to improve Sweden's biopreparedness, in line with recommendations from the EU and WHO. Since forming in 2007, the FBD network has contributed to the harmonization of diagnostic methods, equipment, quality assurance protocols, and biosafety practices among the national BSL-3 laboratories. Lessons learned from the network include: (1) conducting joint projects with activities such as method development and validation, ring trials, exercises, and audits has helped to build trust and improve communication among participating agencies; (2) rotating the presidency of the network steering committee has fostered trust and commitment from all agencies involved; and (3) planning for the implementation of project outcomes is important to maintain gained competencies in the agencies over time. Contacts have now been established with national agencies of the other Nordic countries, with an aim to expanding the collaboration, broadening the network, finding synergies in new areas, strengthening the ability to share resources, and consolidating long-term financing in the context of harmonized European biopreparedness., The Swedish Forum for Biopreparedness Diagnostics (FBD) is a network that fosters collaboration among the 4 agencies with responsibility for the laboratory diagnostics of high-consequence pathogens, covering animal health and feed safety, food safety, public health and biodefense, and security.

Botulism is a severe neuroparalytic disease that affects humans, all warm-blooded animals, and some fishes. The disease is caused by exposure to toxins produced by Clostridium botulinum and other botulinum toxin–producing clostridia. Botulism in animals represents a severe environmental and economic concern because of its high mortality rate. Moreover, meat or other products from affected animals entering the food chain may result in a public health problem. To this end, early diagnosis is crucial to define and apply appropriate veterinary public health measures. Clinical diagnosis is based on clinical findings eliminating other causes of neuromuscular disorders and on the absence of internal lesions observed during postmortem examination. Since clinical signs alone are often insufficient to make a definitive diagnosis, laboratory confirmation is required. Botulinum antitoxin administration and supportive therapies are used to treat sick animals. Once the diagnosis has been made, euthanasia is frequently advisable. Vaccine administration is subject to health authorities' permission, and it is restricted to a small number of animal species. Several measures can be adopted to prevent or minimize outbreaks. In this article we outline all phases of management of animal botulism outbreaks occurring in wet wild birds, poultry, cattle, horses, and fur farm animals.

C difficile has emerged as an important cause of diarrheic disease in horses. C difficile diarrhea is usually diagnosed in mature horses, mostly when they are treated with antimicrobials and hospitalized. It is important for clinicians at veterinary hospitals to have knowledge about the organism and the infection. To prevent C difficile diarrhea, judicious use of antimicrobials is important, as is minimizing different stress factors at the animal hospital or clinic. Infected horses must be isolated. Routine examination for C difficile and toxin A or B is recommended in horses with antibiotic-associated diarrhea. When treating foals for R equi pneumonia, it is important to avoid accidental ingestion of erythromycin by the dams. To reduce the number of environmental spores, thorough cleaning and surface disinfection of the animal hospital and clinic are important. Routine handwashing should be performed by all staff.

Background Strangles is a contagious equine-specific disease caused by Streptococcus equi subsp. equi. Unfortunately, detection of S. equi can fail in up to 40% of horses with strangles. Whereas recent molecular biologic methods and sampling techniques have improved recovery of S. equi optimal sampling methods and laboratory analyses remain ill-defined. Objectives To determine the yield of S. equi from horses with acute strangles in confirmed outbreaks by field-sampling methods subjected to culture and biochemical identification, and real-time PCR directly and after culture. Animals Fifty-seven horses of varying breeds and ages from 8 strangles outbreaks. Methods Prospective study. Culture with biochemical identification and real-time PCR directly, and from culture, were performed on nasal swabs, nasopharyngeal swabs, and nasopharyngeal lavages. Results Real-time PCR directly from samples identified the highest number of infected horses, with 45/57 nasal swabs, 41/57 nasopharyngeal swabs, and 48/57 nasopharyngeal lavages S. equi positive. Biochemical identification (highest positives 22/57) was inferior to real-time PCR for S. equi recovery regardless of sampling method. Real-time PCR of nasopharyngeal lavage directly and after culture yielded 52/57 positives whereas direct real-time PCR of nasopharyngeal lavage combined with either nasopharyngeal swabs or nasal swabs yielded 53/57 positives. Three horses were negative on all samples. Conclusions and Clinical Importance Nasopharyngeal lavage analyzed by a combination of real-time PCR directly and after culture or, alternatively, real-time PCR directly on a nasopharyngeal lavage and a nasal/nasopharyngeal swab can identify S. equi in over 90% of acute strangles cases.

This chapter describes an unexpected anthrax outbreak in a group of Swedish beef cattle kept indoors during the winter season of 2008. Anthrax is a serious zoonotic disease, caused by the pathogenic agent Bacillus anthracis. The way in which the outbreak was managed, the costs involved and the lessons learned are discussed. Research needs, crisis management and preparedness gaps are also identified.

Background Treatment and protection of wounds in horses can be challenging; protecting bandages may be difficult to apply on the proximal extremities and the body. Unprotected wounds carry an increased risk of bacterial contamination and subsequent infection which can lead to delayed wound healing. Topical treatment with antimicrobials is one possibility to prevent bacterial colonization or infection, but the frequent use of antimicrobials ultimately leads to development of bacterial resistance which is an increasing concern in both human and veterinary medicine. Methods Standardized wounds were created in 10 Standardbred mares. Three wounds were made in each horse. Two wounds were randomly treated with LHP® or petrolatum and the third wound served as untreated control. All wounds were assessed daily until complete epithelization. Protocol data were recorded on day 2, 6, 11, 16, 21 and 28. Data included clinical scores for inflammation and healing, photoplanimetry for calculating wound areas and swab cytology to assess bacterial colonization and inflammation. Bacterial cultures were obtained on day 2, 6 and 16. Results Mean time to complete healing for LHP® treated wounds was 32 days (95%CI = 26.9-37.7). Mean time to complete healing for petrolatum and untreated control wounds were 41.6 days (95%CI = 36.2-47.0) and 44.0 days (95%CI = 38.6-49.4) respectively. Wound healing occurred significantly faster in LHP® wounds compared to both petrolatum (p = 0.0004) and untreated controls (p < 0.0001). There was no significant difference in time for healing between petrolatum and untreated controls. Total scores for bacteria and neutrophils were significantly (p < 0.0001) lower for LHP® treated wounds compared to petrolatum from day 16 and onwards. Staphylococcus aureus and Streptococcus zooepidemicus were only found in cultures from petrolatum treated wounds and untreated controls. Conclusions Treatment with LHP® reduced bacterial colonization and was associated with earlier complete wound healing. LHP® cream appears to be safe and effective for topical wound treatment or wound protection.

In this study we examined seven queens with normal oestrous cycles and a history of infertility after normal matings. We performed clinical examination, vaginal cytology, evaluation of oestradiol, progesterone and total T4 levels, vaginal bacterial culture, ultrasonography, and serum analyses for detection of antibodies against chlamydia and feline leukaemia virus (FeLV) antigenemia. Ovariohysterectomy (OHE) was recommended for 1/7 queens because of pathological uterine changes detected at ultrasonography and clinical examination. Four out of seven queens were treated with antibiotics and two of these had more litters. One of the queens that were treated was not mated again and one was mated without conceiving and was at a later OHE found to have degenerative uterine changes. No treatment was given and no diagnosis could be established in 2/7 queens. Both of them were later ovariohysterectomised and one showed degenerative uterine changes while the uterus of the other queen could not be obtained for follow-up. In summary, 4/7 cats were diagnosed with uterine pathology and no definitive diagnosis could be established for 3/7 queens.

Nombre de pages de l'ouvrage: 240 p.; International audience

Reasons for performing study When managing strangles in horses, it is crucial to detect chronic infection with Streptococcus equi (SE), i.e. silent carriers. Objectives Evaluate diagnostics for SE carriers over time in a farm. Study design Longitudinal observational study. Methods Sixty-three Icelandic horses isolated on an island were studied 4 to 26 months after remission of acute strangles, including repeated clinical examination and collection of blood, nasopharyngeal lavage (NPL), and guttural pouch lavage (GPL) samples. Twenty-two horses were treated with penicillin locally and systemically. Serology for S. equi was examined by iELISA [1]. Nasopharyngeal lavage and GPL samples were investigated for S. equi and S. zooepidemicus by real-time PCR [2]. Results Thirty-three per cent were SE carriers after 15 months, despite repeated penicillin treatment. In 16/18 carriers, GPL samples were PCR-positive, but not NPL samples, whereas the opposite was true in 2 horses. Several carriers with persistent aerocystitis were not detected by 3 consecutive NPL samples. Five of 18 carriers were seronegative (27.8%) at 15 months. Following conservative treatment, 7 of these 18 carriers were still carriers at 20 months, 8 were negative, and 3 were lost for sampling. Only GPL samples were positive at this point, and only 1/7 carriers were seropositive. Conclusions To detect chronic carriers of strangles, RT-PCR analysis from both GPL and NPL samples may be necessary. Serological screening at individual level often misses individual carriers in long-term cases. Ethical animal research: The testing was approved by the Swedish Ethical Committee on Animal Experiments and horse owners gave their informed consent for inclusion of animals in the study. Source of funding: The Swedish-Norwegian Foundation for Equine Research. Competing interests: None declared.

Summary In Sweden there are several reports of mares developing acute colitis while their foals were being treated orally for Rhodococcus equi pneumonia with the combination of erythromycin and rifampicin. In this study 6 adult horses were given low oral dosages of these antibiotics, singly or in combination. Within 3 days post administration of erythromycin, in one case in combination with rifampicin, 2 horses developed severe colitis (one fatal). Clostridium difficile was isolated from one of the horses, whereas no specific pathogens were isolated from the other. Both horses had typical changes in blood parameters seen in acute colitis. Clostridium difficile was also isolated from the faeces of a third horse given an even lower dosage of erythromycin in combination with rifampicin. This horse developed very mild clinical symptoms and recovered spontaneously. In the fourth horse given erythromycin only, very high numbers of Clostridium perfringens were isolated. The horses given rifampicin only did not develop any clinical symptoms and there were no major changes in their faecal flora. In conclusion, it has been demonstrated that low dosages of erythromycin ethylsuccinate can induce severe colitis in horses associated with major changes of the intestinal microflora. Clostridium difficile has been demonstrated as a potential aetiological agent in antibiotic-induced acute colitis.

Summary Clostridium (C.) difficile, or its cytoxin, was demonstrated in faecal samples from 10 of 25 (40%) mature horses investigated with acute colitis treated primarily with antibiotics for disorders other than diarrhoea. C. difficile was not found in faecal samples from 140 horses without signs of enteric disorders, from 21 nondiarrhoeic horses treated with antibiotics, nor from 22 horses with colitis untreated with antibiotics. Except for C. difficile neither Salmonella nor any other investigated intestinal pathogen was isolated in any of the diarrhoeic horses. The findings strongly support some earlier reports that C. difficile is associated with acute colitis in mature horses treated with antibiotics. Of the 10 horses, 4 proved positive for C. difficile both in culture and in the cytotoxin test, 4 in culture only and 2 only in the cytotoxin test. Eight of 10 horses positive for C. difficile were or had recently been hospitalised, indicating that C. difficile may be a nosocomial infection in horses. All horses positive for C. difficile were treated with β-lactam antibiotics. The authors are grateful to A. Hellander-Edman, B. Green and J. Skidell for fruitful cooperation and for providing specimens and to G. Holmstrom, G. Sigstam and H. Ljung for excellent technical assistance. This research was supported by ATG (the Swedish Horserace Totalizator Board) and the AGRIA Foundation for Research.

Horses that lack obvious clinical signs of respiratory disease can be silent carries of Streptococcus equi subsp. equi (S. equi) in their guttural pouches. Guttural pouch lavage and repeated nasopharyngeal lavages(1) analyzed by real-time PCR(2) are sensitive methods for analysis for S.equi and are considered to be a standard to identify carrier stage. The carrier screening, identifying and treatment of carriers, in the end of a disease outbreak by sampling guttural pouches is expensive and time consuming. According guidelines carriers that have endoscopic findings of guttural pouch empyema, chondroids or discharging lymph nodules are sampled and treated. The sensitivity of using macroscopic appearance of guttural pouches as an indication of persistent S. equi colonization is poorly studied. The aim of this study was to investigate if there is a correlation between endoscopic findings in guttural pouches and positive qPCR analysis for S.equi. In total 179 endoscopic findings from upper airways from 91 individual horses after a strangles outbreak were compared with positive qPCR results from nasopharyngeal lavage and guttural pouch lavage samples. Nasopharyngeal lavage was performed with 120 ml saline and guttural pouch lavage with 40 ml saline for each pouch and analyzed separately. The nasopharyngeal lavage was performed with 120 ml saline prior endoscopy and endoscopic guttural pouch lavages using 40 ml saline for each pouch and analyzed separately. The endoscopic findings of guttural pouches, including empyema, chondroids or discharging retropharyngeal lymph nodules were recorded and compared to the qPCR analysis for S. equi. Therewas no statistical correlation between endoscopy findings and qPCR results. However, if the first guttural pouch lavage was positive by qPCR for S. equi, it was statistically more likely that the contralateral guttural pouch also was positive, indicating a risk for contamination of the second sample. Horses that lack obvious clinical abnormalities on upper airway endoscopy including the guttural pouches can be silent carriers of S. equi. In conclusion, negative endoscopic findings of the guttural pouches of recovered strangles cases cannot rule out silent carrier status. This study was approved by the Swedish Ethical Committee on Animal Experiments and each horse owner had given their informed consent to participate in the study. Financial support was achieved from The Swedish-Norwegian Foundation for Equine Research and Swedish Research Council Formas.

Background A cross-sectional study was carried out to determine the seroprevalence of different serovars of Leptospira spp. and their association with clinical disease and host factors in Swedish horses. Methods Sera from 2017 horses brought to equine clinics during 1997–98 were investigated. The sera were examined by microscopic agglutination test for the presence of antibodies against the following L. interrogans serovars: Bratislava strain Jez, Icterohaemorrhagiae strain Kantorowicz and Pomona strain Pomona and also L. kirschneri sv Grippotyphosa strain Duyster and L. borgpetersenii sv Sejroe strain M 84. Host factors, disease factors, season, pasture access and outdoor confinement variables were analysed with respect to seropositivity to sv Bratislava and Icterohaemorrhagiae. Multivariable logistic regression was used to model seropositivity to sv Bratislava and Icterohaemorrhagiae (seroprevalence > 8%). Results The seroprevalence, at a cut-off 1:100, were for sv Bratislava (16.6%), Icterohaemorrhagiae (8.3%), Sejroe (1.2%), Pomona (0.5%) and Grippotyphosa (0.4%). In the multivariable analysis, it was demonstrated that seroprevalence increased with age for sv Bratislava and Icterohaemorrhagiae. For sv Bratislava the seasons April – June and October – December and for sv Icterohaemorrhagiae October – December had higher seroprevalences than other seasons. Horses not used for racing had higher levels of seropositivity to sv Bratislava. Furthermore, horses with respiratory problems as well as horses with fatigue had higher levels of seropositivity to sv Bratislava. Ponies and coldbloods, and horses with access to pasture, had lower seroprevalence for sv Icterohaemorrhagiae. Healthy horses had lower seroprevalence for sv Icterohaemorrhagiae, than non-healthy horses. Conclusion There was no significant association between clinical signs and disease and positive titres to sv Bratislava (except for the association between respiratory problems and fatigue and seropositivity to sv Bratislava). The results suggest that horses with increasing age and exposed to factors associated with outdoor life had an increased seroprevalence for sv Bratislava, indicating that horses get infected from outdoor and/or are exposed to shedding from other horses (management dependent). For sv Icterohaemorrhagiae, management possibly plays a role as ponies and coldbloods as well as healthy horses had lower seroprevalence. Overall, the age of the horse should be taken into consideration when evaluating the titre as the average healthy horse has a higher titre than a young horse.