[Deer farming in Switzerland - Current epidemiological situation with focus on husbandry, management and nutrition]. / Gehegewild in der Schweiz ­ Aktuelle epidemiologische Situation mit Schwerpunkt Haltung, Management und Fütterung.

INTRODUCTION: Between September 2016 and February 2017 a survey in Swiss deer farms were conducted to gain information about their husbandry. Questions about the business, feeding, management, health and deworming strategies were asked. 98 (19%) out of 527 registered farms (2016) participated in the survey. The farms were often run on a sideline business, had an average used agricultural area of 7.3 ha with an average of 38 deer. Pasture access was the preferred feeding strategy followed by offering first and second cut hay. Between 2013-2015 the most common causes of death were sudden death and injuries. Parasites were classified as no or rather small problem by 91 out of 102 deer owner. Fecal parasitological examinations of fecal samples were conducted in 36 (35%) of the responding farms. Gastrointestinal roundworms (Trichostrongylidae) were identified as the most common pathogens (in 42-59% of sampled farms), in addition large lungworms (Dictyocaulus sp.) and coccidia were detected. 45% of the participating farmers conducted at least one treatment against parasites between 2013 and 2015.

INTRODUCTION: Dans le but d'avoir une vue d'ensemble sur la détention du gibier d'élevage en Suisse, une enquête a été menée entre septembre 2016 et février 2017, comprenant des questions relatives à l'exploitation, à l'alimentation, à la situation sanitaire et aux stratégies en matière de vermifugation. 98 des 527 exploitations annoncées en 2016 (19%) ont participé à cette étude. Ces exploitations, qui constituent fréquemment un gain accessoire, avaient une surface agricole d'en moyenne 7,3 ha avec 38 cervidés. En matière d'alimentation, c'est le foin et le regain qui étaient le plus souvent utilisés en complément du pâturage. Les causes de pertes dans les troupeaux entre 2013 et 2015 étaient principalement les cas de mort subite ainsi que les blessures. 91 de 102 détenteurs de cervidés considéraient les parasites comme n'étant pas un problème ou n'étant qu'un faible problème. Des échantillons de selles, prélevés dans 36 (35%) des exploitations ayant répondu au questionnaire, montraient que les nématodes gastro-intestinaux (Trichostrongylidae) étaient les plus fréquents (présents dans 42-59% des exploitations testées); des vers pulmonaires (Dictyocaulus sp.) et des coccidies ont également été trouvés. Environ 45 % des détenteurs de cervidés ayant participé à l'enquête avaient effectué, dans la période comprise entre 2013 et 2015 au moins un traitement antiparasitaire.

[Displaced pain pump because of a pendulous abdomen - dermatolipectomy as solution of the problem]. / Rezidivierende Luxation einer Schmerzpumpe aufgrund einer Fettschürze - Problemlösung durch Dermatolipektomie.

A female patient with a chronic pain syndrome after multiple operation on her muskuloskeletal system presented with a displaced pain pump in her lower left abdominal region. After an excessive weight loss due to gastric banding the patient developed a massive pendulous abdomen causing the complication. An interdisciplinary dermatolipectomy together with a refixation of the pump was performed. Since this operation a recurrence of the displacement has not been observed and consecutively the filling of the pump could be accomplished without radiological investigations. This rare case demonstrates the significance of interdisciplinary treatment in which a primarily aesthetic intervention was necessary for a successful therapeutic outcome.

Single step fibula-pro-tibia transfer and soft tissue coverage with free myocutaneous latissimus dorsi flap after extensive osteomyelitis and soft tissue necrosis--a 3 year follow up.

A 10-year-old girl from Ghana, Africa, developed chronic osteomyelitis of her right tibia and a large soft tissue defect on the ventral lower leg after a minor injury. She was treated more than 6 months after trauma without any improvement. We report a follow up of 3 years after reconstruction with a single-step pedicled fibula-pro-tibia transfer and wound coverage with a free myocutaneous latissimus dorsi flap.

Decellularized dermis in combination with cultivated keratinocytes in a short- and long-term animal experimental investigation.

Decellularized human dermis as a potentially ideal scaffold for dermal substitution in severe burns was examined in a two-staged animal experiment. In an initial step, an in vitro generated composite graft consisting of human keratinocytes and decellularized dermis (AlloDerm) was transplanted onto nude mice in a short-term trial (n = 20, 14 days). Subsequently, a combined one-step grafting of full thickness wounds with both decellularized dermis (in part preincubated with fibroblasts) and cultivated autologous keratinocytes as a cell suspension in fibrin glue was done in a long-term porcine animal model (n = 10, 6 months). In both series, macroscopic wound healing was evaluated by planimetry. Histological investigations included morphological as well as immunohistochemical parameters. The short-term study showed both successful integration of the composite grafts and reduction of wound contraction compared with the control group (epithelial grafts). The long-term porcine study displayed reduced myofibroblast formation and contraction in the wounds that had been treated with fibroblast-preincubated dermis. After 4 weeks, a decline of the structural integrity of the dermal matrix could be noticed. The utility of decellularized dermis as template for both dermal reconstitution and keratinocyte delivery vehicle was shown. The closure of full thickness wounds by a single-step combination of an autologous keratinocyte fibrin sealant suspension and acellular dermis in a pig animal model could be shown. Incorporation of fibroblasts led to reduced wound contraction but could not prevent the loss of dermal integrity. The engineered 'skin' remained viable and stable over a period of 6 months.

Artificial skin.

Replacement of skin has been one of the most challenging aims for surgeons ever since the introduction of skin grafts in 1871. It took more than one century until the breakthrough of Rheinwald and Green in 1975 that opened new possibilities of skin replacement. The combination of cell culture and polymer chemistry finally led to the field of tissue engineering. Many researchers all over the world have been fascinated by the chance of creating a skin-like substitute ex vivo without any further harm to the patients, especially those with massive burns. Many different approaches to create new substitutes and further improvements in genetical and stem cell research led to today's skin equivalents. But still, the "gold standard" for wound coverage is the autologous split-thickness skin graft. Future research will aim at originating biologically and physiologically equal skin substitutes for the treatment of severe burns and chronic ulcers.

Tissue engineering of skin substitutes.

Cultivated epithelial autografts as multilayered thin sheets represent common standard in clinically applied tissue engineering substitutes, outnumbering all experimental alternatives. However, the unsatisfying short and long term results concerning mechanical stability and scarring demand for alternatives. Our group investigated cultivation and transplantation of cultured autologous keratinocytes as a single cell suspension in a fibrin sealant matrix in athymic mice in combination with allogenic skin grafting. We observed reliable wound reepithialization after a cultivation period of only 2 weeks. Additionally, we could allocate successful combination of a keratinocyte fibrin sealant suspension and acellular dermis in an attempt to regenerate full thickness skin defects in a pig animal model. The potential clinical implication of subconfluently cultured keratinocytes is enhanced by the possibility of co-transplantation with decellularized dermis.

Skin tissue engineering.

The coverage of extensive wounds with viable autologous keratinocytes remains the only option of treatment if autologous donor skin is not obtainable. There is evidence that proliferating keratinocytes, as suspended cells or as a single layer, are adequate for wound closure. Understanding keratinocyte-matrix interactions not only allows us to influence keratinocyte outgrowth, adhesion, and migration, but may also guide us to modify matrix molecules for enhancing keratinocyte take. Further approaches may include the generation of genetically manipulated keratinocytes, which allow the use of an off-the-shelf epidermal replacement. As surgeons, our goal is to help burn patients with the best quality of skin in the shortest time possible. As tissue engineers, we have not achieved the goal of a universal skin product. By continually reviewing the options and using them, we can at least use the proper material in the adequate situation. Because of the limited resources, the need for comparisons of clinical effectiveness and cost are ever more important. As anatomy and physiology of engineered skin substitutes improve, they will become more similar to native skin autografts. Improvement of skin substitutes will result from inclusion of additional cell types (eg, melanocytes) and from modifications of culture media and scaffolds. Skin-substitute materials may be able to stimulate regeneration rather than repair, and tissue-engineered skin may match the quality of split-skin autografts, our present gold standard.

[Skin tissue engineering]. / Gewebeersatz (tissue engineering) von Dermis und Epidermis.

Cultivated epithelial autografts as multilayered, thin sheets represent a common standard in clinically applied tissue engineering substitutes, outnumbering all experimental alternatives. However, the unsatisfying short- and long-term results concerning mechanical stability and scarring require alternatives. The cultivation and transplantation of cultured autologous keratinocytes as a single cell suspension in a fibrin matrix, combined with allogenic skin grafting, has been investigated extensively in athymic nude mice. Wounds can be reliably reepithelialized after a cultivation period of only 14 days. Moreover, the successful combination of keratinocyte fibrin suspension and acellular dermis in an attempt to regenerate full thickness skin defects in a pig model has been demonstrated. The usefulness of subconfluently cultured keratinocytes-which can be harvested very early and are easy to handle-is enhanced by cotransplantation with decellularized dermis.

Feasibility of biolistic gene therapy in burns.

Skin is an especially attractive target for genetic manipulation because it is readily accessible and easily monitored for both the presence and the expression of inserted genes. This study was designed to assess the feasibility of particle mediated gene transfer to burned skin and to compare the transfection efficiency, anatomic distribution, and duration of transgene expression achievable in normal versus burned skin. Two days following scald injury of varying depths in 60 degrees C water (10 s: superficial partial; 20 s: deep partial; 40 s: full thickness) reporter gene (beta-galactosidase) constructs were delivered using a gene gun at various helium pressures (200-600 psi) to normal and burned skin. A time course study was performed to examine the kinetics of transgene expression. Animals received a superficial partial thickness burn and were sacrificed 12 h, 1, 3, 5, 7, 14, or 21 days after gene transfer. India Ink injection and immunohistochemistry were used to assess the depth of the scald injury. Transfection efficiency was measured in skin homogenates 24 h after gene transfer by morphometric and chemoluminescent assays. We found that the extent of tissue damage was directly related to the duration of heat source exposure. Reporter gene activity was significantly higher in superficial partial thickness burns compared to normal controls and gradually declined with increasing tissue injury. No activity was seen in the full thickness burn group. Beta-galactosidase activity reached a maximum level 12 h after gene transfer in both normal and superficial partial thickness burned skin with no levels seen after 5 days post-transfection. These findings indicate that particle-mediated gene transfer in thermally injured skin is feasible and may provide a means of introducing biologic agents into injured tissue capable of enhancing bacterial clearance and improving wound healing.

Induction of alpha-N-acetylgalactosamine-O-serine/threonine (Tn) antigen-mediated cellular immune response for active immunotherapy in mice.

A block in carbohydrate chain elongation of O-glycosylated mucins results in accumulation of alpha-GalNAc O linked to serine or threonine (Tn antigen) in a large percentage of human adenocarcinomas. Immunization of mice with desialylated ovine submaxillary mucin (A-OSM), which contains a large concentration of Tn antigen, provided protection against challenge of a highly invasive Tn expressing syngeneic mouse mammary tumor, TA3-Ha. A similar protective effect was not observed in mice immunized with the deglycosylated mucin or irradiated TA3-Ha cells. Immunization with A-OSM but not with deglycosylated mucin resulted in high anti-Tn antibody response in mice. A-OSM induced in vitro proliferation of T-lymphocytes obtained from mice preimmunized with A-OSM or irradiated TA3-Ha cells. This antigen-specific T-cell response was significantly lower if lymphocytes were stimulated with either the deglycosylated or sialylated form of mucin. A-OSM stimulation induced primarily a CD4+ T-cell population, and these cells secreted interleukin 2 in a dose-dependent fashion. A-OSM was also able to induce delayed-type hypersensitivity in mice in response to footpad injections with irradiated TA3-Ha cells. These studies indicate that Tn antigen presented on a protein backbone is capable of providing cellular immunity and protection against tumor in mice.

Demonstration of heterogeneity among the antigenic proteins of Mobiluncus species.

The protein and antigenic profiles of the American Type Culture Collection type strains of Mobiluncus species and those of 114 clinical isolates were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and immunoblotting with homologous polyvalent antisera. The majority of isolates (82%) possessed characteristic protein profiles and could be identified to the species level by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. The major protein bands were also antigenic, and some antigenic cross-reactivity was noted between the two Mobiluncus species. All of the isolates were examined for reactivity with a panel of 12 monoclonal antibodies previously prepared against the type strains. While 56 of 60 clinical isolates of Mobiluncus curtisii (93%) reacted with one or more of the monoclonal antibodies, only 23 of 54 clinical isolates which were identified as Mobiluncus mulieris by biochemical methods (48%) reacted with one or more of the monoclonal antibodies. One of the 4 M. curtisii isolates (25%) and 11 of the 31 M. mulieris isolates (35%) which did not react with the monoclonal antibodies also had atypical protein profiles. These results demonstrate a high degree of heterogeneity in the protein and antigenic profiles of Mobiluncus isolates and suggest that further taxonomic division may be appropriate.

Efficacy of azithromycin for therapy of active syphilis in the rabbit model.

Azithromycin was shown to be as effective as standard benzathine penicillin and erythromycin in the therapy of active syphilis in the rabbit model. Following production of primary chancres by intradermal inoculation of 10(6) Treponema pallidum, groups of six rabbits were treated with benzathine penicillin (200,000 units im weekly for two weeks), erythromycin base (30 mg/kg/day orally four times daily for 15 days) or azithromycin (30 mg/kg/day given orally once or twice daily for 15 days); one group was untreated. Daily darkfield (DF) microscopic examinations of chancre aspirates were conducted to identify motile organisms. Although all treated animals became DF negative prior to completion of therapy, the median time to DF negativity was longer in animals given azithromycin once daily, compared with animals receiving benzathine penicillin (P less than 0.01); no difference was seen in comparison with animals receiving erythromycin. Untreated animals remained DF positive for greater than 15 days. The mean maximum lesion diameters for all treated animals were similar and were significantly smaller than in untreated rabbits; fewer lesions ulcerated in treated than in untreated animals. Subsequent dose-ranging studies indicated that administration of lower doses of azithromycin (15 mg/kg/day given orally either once or twice daily, or 7.5 mg/kg/day given once daily) was as effective as benzathine penicillin for therapy of active syphilis in this model, though the median time to darkfield negativity was significantly longer in the azithromycin-treated animals (P less than 0.01). Persistent infection was demonstrable in lymph nodes of untreated animals, but no evidence of virulent T. pallidum was found three months following transfer of tissue from any animal treated with penicillin, erythromycin, or azithromycin.

Production and characterization of monoclonal antibodies to Mobiluncus species.

Members of the genus Mobiluncus are anaerobic motile curved rods which are associated with bacterial vaginosis (BV). Murine monoclonal antibodies (MAbs) to the ATCC type strains of M. curtisii subsp. curtisii, M. curtisii subsp. holmesii, and M. mulieris were produced and characterized by enzyme-linked immunosorbent assay and indirect immunofluorescence assay. Four MAbs were subspecies specific and reacted with M. curtisii subsp. curtisii but not with M. curtisii subsp. holmesii; four were specific for M. mulieris. The remaining antibodies demonstrated some cross-reactivity: three were species specific and reacted with both subspecies of M. curtisii, and one defined an epitope shared by M. curtisii subsp. holmesii and M. mulieris but not by M. curtisii subsp. curtisii. None of the MAbs reacted with a panel of other bacteria commonly present in the vaginas of normal women or women with BV. Examination of the molecular specificities of the antibodies by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting revealed four antibodies which were specific for an 82,000-dalton molecule of M. curtisii subsp. curtisii and five antibodies which bound a major band of M. mulieris at 93,000 daltons. Selected MAbs reacted in the indirect immunofluorescence assay with 24 of 25 Mobiluncus spp. clinical isolates from local women with BV and could be used for direct detection of Mobiluncus spp. in vaginal fluid from a patient with BV.

Development of cellular immunity to individual soluble antigens of Treponema pallidum during experimental syphilis.

The contribution of individual specific molecules of Treponema pallidum subspecies pallidum to cellular immunity in experimental syphilis was evaluated by combining the techniques of Ag identification and purification with the lymphocyte proliferation assay. Proliferative responses of splenic lymphocytes from syphilitic rabbits to complex treponemal Ag and Con A were vigorous throughout the course of intratesticular infection (6, 10, 17, 30, and 210 days). Normal rabbits did not respond to any treponemal preparations and all rabbits failed to respond to normal rabbit testicular Ag (NRT). Seven defined treponemal Ag (47 kDa, 37 kDa, 35, 33-kDa, 30-kDa, 14 kDa, and 12 kDa) stimulated lymphocytes from infected rabbits. Cellular responses to the 37-kDa and 30-kDa fractions were evident by day 6 of infection and responses to the 35, 33-kDa and 14-kDa Ag were first detected on day 10; responsiveness to these Ag continued throughout the observation period. Cellular responses to the 47-kDa molecule were detectable but lower when compared with other individual Ag. Responsiveness to the 12-kDa Ag was not evident until 7 mo postinfection. Specific immunoblot reactivity of serum from rabbits used in this study generally correlated with the development of cellular reactivity to individual Ag of T. pallidum.

Specificity of antibodies from patients with pinta for antigens of Treponema pallidum subspecies pallidum.

Inhabitants of a remote Panamanian village were examined for clinical and serological evidence of pinta infection. Of 104 persons examined, 21 (20%) had clinical evidence of active or inactive pinta, and 54 (52%) were seropositive. Sera were evaluated for antibody to individual Treponema pallidum antigens. Sera from all four patients with active pinta contained antibody to the 47-48 kilodalton major antigen; the intensity of reactivity and the number of antigens recognized increased with age and, presumably, duration of infection. Sera from six children with inactive pinta reacted strongly with multiple T. pallidum antigens, whereas adults with inactive pinta had less intense reactivity against fewer molecules. Seronegative controls demonstrated only weak reactivity to fewer than five molecules. The development of antibody reactivity to the full spectrum of T. pallidum antigens during the course of infection demonstrates the high degree of antigenic relatedness of T. pallidum and Treponema carateum and is similar to the development of humoral responsiveness during syphilis infection.

Human immunoglobulin G antibody response to the major gonococcal iron-regulated protein.

In humans, gonococcal infection occurs in environments limited with respect to free iron. Neisseria gonorrhoeae produces increased quantities of iron-regulated membrane proteins when grown under in vitro conditions which restrict the availability of free iron. Using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot (immunoblot) techniques, we studied the reactivity of specific antibodies to the 37-kilodalton (kDa) major iron-regulated protein (MIRP) of gonococci grown under iron-limiting conditions. Antibodies reactive with the 37-kDa MIRP were distinguished from those reactive with protein I by using purified 37-kDa MIRP or gonococcal protein preparations. Acute-phase sera from patients with disseminated gonococcal infection (DGI) reacted strongly to both the 37-kDa MIRP and protein I. Acute sera from nine patients with uncomplicated gonorrhea did not exhibit strong reactivity with the 37-kDa MIRP and were indistinguishable from five control sera. When compared with acute-phase sera, convalescent-phase sera from patients with DGI failed to demonstrate increased reactivity, whereas convalescent-phase sera from one of nine patients with uncomplicated gonorrhea developed reactivity to the 37-kDa MIRP. These data indicate that (i) the 37-kDa MIRP is expressed and antigenic in vivo and (ii) humans with DGI consistently develop a systemic antibody response to the 37-kDa MIRP.

Monoclonal antibodies to a 28,000 mol. wt protein antigen of Mycobacterium leprae.

Monoclonal antibodies (MoAb) have been used to analyse a protein antigen from Mycobacterium leprae with a subunit mol. wt of 28,000 daltons. Three different patterns of species specificity were observed with two antibodies being specific for M. leprae, two partially specific, and one broadly cross-reactive amongst all mycobacteria. Competitive binding and sandwich assays demonstrated that the specific and partially specific antibodies recognized closely related regions of the molecule while the cross-reactive antibody recognized a spatially separate epitope on the same polypeptide chain. Identification of specific and cross-reactive epitopes on a single antigenic molecule may be of considerable importance for understanding the functioning of the cell-mediated immune system during leprosy infection and the use of MoAb for such analyses is discussed.

Use of a polysulfone membrane support for immunochemical analysis of a glycolipid from Mycobacterium leprae.

Polysulfone membranes have been used as a solid support for chromatography and immunoblotting of phenolic glycolipid I from Mycobacterium leprae. These membranes have an advantage over other supports such as nitrocellulose and silica gel in that very little non-specific background binding of antibodies occurs and assays can readily be carried out with IgM antibodies from human sera. An example of use of the polysulfone chromatography system for detection of phenolic glycolipid I in sera from leprosy patients is described.