High and novel genetic diversity of Francisella tularensis in Germany and indication of environmental persistence.

In Germany tularemia is a re-emerging zoonotic disease. Therefore, we investigated wild animals and environmental water samples for the presence and phylogenetic diversity of Francisella tularensis in the poorly studied Berlin/Brandenburg region. The phylogenomic analysis of three isolates from wild animals revealed three new subclades within the phylogenetic tree of F. tularensis [B.71 from a raccoon dog (Nyctereutes procyonoides); B.74 from a red fox (Vulpes vulpes), and B.75 from a Eurasian beaver (Castor fiber albicus)]. The results from histological, PCR, and genomic investigations on the dead beaver showed that the animal suffered from a systemic infection. Indications were found that the bacteria were released from the beaver carcass into the surrounding environment. We demonstrated unexpectedly high and novel phylogenetic diversity of F. tularensis in Germany and the fact that the bacteria persist in the environment for at least one climatic season. These findings support a broader host species diversity than previously known regarding Germany. Our data further support the assumption derived from previous serological studies of an underestimated frequency of occurrence of the pathogen in the environment and in wild animals. F. tularensis was isolated from animal species not previously reported as natural hosts in Germany.

Tularaemia seroprevalence of captured and wild animals in Germany: the fox (Vulpes vulpes) as a biological indicator.

A total of 2475 animals from Germany, both captive and wild, were tested for antibodies against Francisella tularensis to obtain more knowledge about the presence of this pathogen in Germany. An indirect and a competitive ELISA served as screening methods, positive and inconclusive samples were confirmed by Western blot. Of the zoo animals sampled between 1992 and 2007 (n = 1122), three (0·3%) were seropositive. The seroconversion of a hippopotamus in Berlin Zoo was documented. From 1353 serum samples of wild foxes (Vulpes vulpes), raccoon dogs (Nyctereutes procyonoides) and wild boars (Sus scrofa), collected between 2005 and 2009 in the federal state of Brandenburg (surrounding Berlin), a total of 101 (7·5%) tested positive for antibodies to F. tularensis lipopolysaccharide. Our results indicate a higher seroprevalence of F. tularensis in wildlife in eastern Germany than commonly assumed. Furthermore, we found foxes and raccoon dogs to be biological indicators for tularaemia.

[Tracheal collapse in a four month old Uckermaerker heifer]. / Trachealkollaps bei einem vier Monate alten Uckermärker Jungrind.

This report describes a case of tracheal collapse in a four month old Uckermaerker heifer. The animal was born spontaneous without signs of dystocia. It was kept outside on the pasture and died in the summer at high temperatures under signs of acute dyspnea and respiratory failure. Clinical symptoms were not observed by the owner until a few hours before death. At necropsy, the middle and caudal third of the trachea was found to be collapsed due to an inward bending of the dorsal ends of the cartilage rings. The most severe luminal reduction was located in the thoracic part of the trachea approximately 5 cm cranial the tracheal bronchus. Additional pathological lesions were absent except for signs of circulatory failure (lung edema, congestion of the shock organs). Based on clinical history and pathological findings, a traumatic cause of the tracheal collapse is unlikely. However, a hereditary influence seems possible since analysis of the breeding scheme revealed inbreeding. The mother of the affected animal was mated with her grandson (inbreeding coefficient 0,125).

[Do lactoferrin, lysozyme and the lactoperoxidase-thiocyanate-hydrogen peroxide-system cause negative microbiological results in mastitis secretions?]. / Laktoferrin, Lysozym und das Laktoperoxidase-Thiozyanat-Wasserstoffperoxid-System (LPS) als Ursache negativer mikrobiologischer Befunde aus Mastitissekreten?

Lactoferrin, lysozyme and the lactoperoxidase-thiocyanate-peroxide-system are naturally occurring antimicrobial components of milk. The objective of this study was to examine, whether these components were responsible for negative results, when mastitis milk is cultured microbiologically. Quarter milk samples from 75 cows with clinical mastitis on a dairy farm in Brandenburg were submitted for microbiological culture and analysed for the content and the activities of the three components. Animals from all stages of lactation with clinical mastitis were included in the study. Animals were examined clinically and milk samples were collected prior to first treatment. Secretions from quarters with clinical mastitis were compared to those of neighbouring quarters without clinical mastitis. Secretions with positive cultural results were compared to those with negative results. The concentrations or activities of the three factors were significantly higher in the diseased quarters than in the quarters without clinical signs of mastitis. The concentration of lysozyme increased with severity of the clinical signs (local swelling and changes in secretion). The concentration of lactoferrin was significantly higher in quarters with slight alterations of glandular tissue than in quarters with medium or severe alterations (P < 0.05). LPS-activities did not correlate with the severity of clinical signs. No differences in the concentration of lactoferrin or LPS-activities were seen between mastitis with positive and negative culture results. The concentration of lysozyme was even higher in culturally positive samples than in negative samples (P < 0.05). Results from this study indicate that the three factors examined did not impair the results of microbiological culture of milk samples from quarters with clinical mastitis.

Evaluation of a systemic antibiotic treatment of toxic puerperal metritis in dairy cows.

The objective of this study was to evaluate the efficacy and economic efficiency of a systemic treatment of toxic puerperal metritis in dairy cows with ceftiofur. Cows with abnormal vaginal discharge at a postpartum examination (d 4 to 6 after calving) and a rectal temperature > or = 39.5 degrees C were assigned to three treatment groups. Cows in group 1 (n = 70) received 600 mg of ceftiofur intramuscularly (i.m.) on 3 consecutive days. Cows in group 2 (n = 79) received an intrauterine treatment with antibiotic pills consisting of 2500 mg of ampicillin and 2500 mg of cloxacillin and an additional 6000 mg (i.m.) of ampicillin. This treatment was performed on 3 consecutive days. Cows in group 3 (n = 78) received the same intrauterine treatment as in group 2. In addition, 600 mg of ceftiofur was administered i.m. on 3 consecutive days. Body temperature was recorded daily for 6 d after first treatment. There were no significant differences among the groups regarding clinical efficacy at d 6 after first treatment. The cure rates based on rectal temperatures declining to below 39.5 degrees C on d 6 after treatment were 82.9, 84.8, and 84.6% for groups 1, 2, and 3, respectively. Reproductive performance did not differ significantly between group 1 and groups 2 and 3 for any of the measures tested. A financial analysis with 87 different cost scenarios demonstrated that a systemic treatment of toxic puerperal metritis in cattle with ceftiofur is an effective alternative to the combination of local and systemic treatments.

Serum antibodies against particular antigens of Borrelia burgdorferi sensu stricto and their potential in the diagnosis of canine Lyme borreliosis.

Dog sera (n = 118) were tested for antibodies recognizing Borrelia (B.) burgdorferi sensu stricto strain B31 (ATCC 35210) antigens. In total, 18 of the dog sera gave positive results in a whole cell sonicate ELISA (WCS ELISA). These positive sera were further evaluated by immunoblot assay, utilizing a whole bacterial lysate as antigens. 94.4% (17 of 18) of the dog sera reacted with immunodominant antigens at 20-22 kDa (protein C, pC), 31 kDa (outer surface protein A, OspA), 34 kDa (outer surface protein B, OspB), 41 kDa (flagellin), 60 kDa ("common antigen"), and/or 100 kDa (presumably p100). Sera recognizing pC (20-22 kDa) and antigens > 94 kDa always detected the highest number of antigen bands, indicating the specificity of those antigens in serological diagnosis. The results clearly demonstrate that the WCS ELISA is a useful tool for testing sera of dogs for antibodies against B. burgdorferi. However, positive results should be confirmed by immunoblot, using WCS as antigen. According to the presented data, we recommend criteria for B. burgdorferi immunoblots using dog sera as follows: sera have to be considered as positive if they detect the 41 kDa flagellin, and two of the 5 immunodominant antigens, namely > 94 kDa (presumably p100), 60 kDa ("common antigen"), 34 kDa and 29-31 kDa (OspB and OspA, respectively) and 20-22 kDa (pC). If sera only recognize the 41 kDa flagellin, this result is equivocal, requiring testing a second serum sample 4 to 8 weeks later.

[Vaccination of pigeons against Salmonella infections]. / Zur Schutzimpfung gegen die Taubensalmonellose.

The efficiency of the live vaccine Zoosal T with a double marker mutant of Salmonella Typhimurium was tested on conditioned pigeons. For challenge infection we used a pigeon specific variation copenhagen strain in a defined state of virulence. The reduction of mortality and the persistence of Salmonella in organs were evaluated. An oral booster enhances the protection due to vaccination.