Comparison of the immune competence of Turopolje, German Landrace × Turopolje, and German Landrace × Pietrain pigs after PRRSV vaccination.

The competences of the immune systems of the ancient pig breed Turopolje (T×T), German Landrace × Turopolje (L×T) and 'modern' pig breed German Landrace × Pietrain (L×P) were compared in this study. All pigs were immunized with a modified live vaccine against 'Porcine Reproductive and Respiratory Syndrome' (PRRS) virus (Ingelvac PRRS MLV(®)) to simulate an infection. Antibody production against PRRS MLV was evaluated in serum. Elimination of the viral infectious fragments during the experimental period was monitored in serum, leukocytes and tonsils by RT-qPCR. Furthermore relevant immune marker genes were quantified either on gene expression level using RT-qPCR [toll like receptor (TLR) 7, TLR8, TRAF6, CD163, SIGLEC1, CD4, CD8, CD14, CD19, tumor necrosis factor alpha (TNFα), interleukin (IL) 1, IL2, IL6, IL12], and on protein level using ELISA [interleukin (IL)-1, IL-2, IL-6, and IL-12]. The three breeds showed individual inactivation efficiencies as a reaction to the PRRS MLV vaccination. T×T eliminated the virus in serum within 16 days, followed by L×T (28 days) and L×P (36 days). The antibody titers against PRRS MLV of L×T and L×P were significantly higher compared to T×T (p<0.05). The gene expression data and protein analysis of interleukins revealed that T×T reacted with a type 1 immune response. In contrast, the two other breeds (L×T and L×P) showed a type 2 immune response, which resulted in the higher synthesis of B-cells and an increased concentration of specific anti-PRRS MLV antibodies.

[Adipose-derived stromal cells (ASC) - basics and therapeutic approaches in otorhinolaryngology]. / Mesenchymale Stammzellen aus Fettgewebe (ASC) - Grundlagen und Anwendung in der HNO-Heilkunde.

Adipose-derived Stromal Cells (ASC) - Basics and Therapeutic Approaches in Otorhinolaryngology Mesenchymal stem cells from adipose tissue can be easily harvested with less discomfort, low donor-site morbidity and high amount compared to bone marrow-derived stem cells. Due to their multilineage differentiation potential in various cell types, immunmodulatory properties and their capability to enhance wound healing, ASC are a promising cell source for tissue engineering approaches and regenerative medicine. They are characterized by the expression of specific surface marker proteins and their differentiation potential into the mesenchymal lineages. Whereas only preclinical studies are published for otorhinolaryngology-related therapeutic options using ASC, various diseases, for instance graft-versus-host disease, have already been treated with ASC in single cases or clinical trials. Safety and genomic stability of ASC as well as the risk of spontaneous malignant transformation are still disputed. This review summarizes the current literature on characterization and anatomic localization of ASC. In addition, beside the presentation of preclinical studies concerning therapeutic approaches in otorhinolaryngology as well as of current clinical applications, the issue of safety of ASC in human stem cell therapy is discussed.

[Sudden death of outdoor housed pigs caused by Clostridium novyi. A case report]. / Plötzlicher Tod durch Clostridium novyi bei Schweinen in Freilandhaltung. Ein Fallbericht.

In an outdoor pig-breeding unit of the Tierpark Arche Warder e. V. (Germany), 16 pigs of different age and sex died in October 2011. Necropsy findings revealed tympany, liver emphysema, subcutaneous oedema, haemopericardium, haemothorax, and intense gas bubble infiltrations in muscles. The stomachs were filled. The initial anaerobic bacteriological investigations gave negative results. In further analyses of tissue samples, the flagellin gene of C. novyi types A and B was detected using PCR. Based on the anatomical-pathological and bacteriological findings as well as PCR testing, a C. novyi infection was assumed to be the cause of the pig mortality.

[Unexpected respiratory distress in the delivery room--a rare differential diagnosis for the obstetrician and neonatologist]. / Unerwartete Atemnot im Kreißsaal--eine seltene Differenzialdiagnose für Geburtshelfer und Neonatologen.

We present the case of a female infant born prematurely at 34 weeks of gestation. Prenatally a midsized ventricular septal defect was diagnosed. Due to marked respiratory distress intubation was attempted but failed, since the tube could not be placed beyond the glottis. Oxygenation could be improved by nasopharyngeal bag ventilation. The clinical course as well as radiographic imaging was suggestive for a complete tracheal agenesis with broncho-oesophageal fistula which was confirmed at autopsy. Tracheal agenesis (TA) is a rare differential diagnosis of postnatal respiratory distress and the obstetrician or neonatologist will regularly be surprised by this malformation. Partial or complete absence of the trachea without associated malformations will be rarely diagnosed antenatally. In the case of the absence of an oesophageal fistula to the remaining airway a congenital high airway obstruction syndrome (CHAOS) ensues, leading to enlarged hyperechogenic lungs, dilated and fluid-filled trachea and bronchi and an absent tracheal flow during foetal breathing. Aetiology of TA is unknown, therapeutic options are limited thus making TA a usually fatal disorder.

Rapid detection of CWD PrP: comparison of tests designed for the detection of BSE or scrapie.

Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy (TSE) mainly affecting cervids in North America. The accumulation of an abnormal form of host-encoded prion protein (PrP(CWD) ) in the CNS and lymphoid tissues is characteristic of the disease and known to be caused by pathogenic prion proteins (PrP(res) ), which are thought to be transmitted mainly by contact with body fluids, such like saliva. Species known to be naturally infected by CWD include Rocky Mountain elk (Cervus elaphus nelsoni), white-tailed deer (Odocoileus virginianus) and mule deer (Odocoileus hemionus). Recently, large-scale disease eradication or control programs have been attempted to curtail the spread of disease. But reports of diseased free-ranging and farmed cervids in many locations in the USA and Canada are still continuing. The goal of this study was to find sensitive rapid test systems that are reliably able to detect CWD-associated PrP(CWD) in cervids, thereby reviewing an important control tool in case the disease spreads further and reaches Europe. Seven tests, originally developed for the detection of other TSE diseases such as Scrapie and bovine spongiform encephalopathy, including two Western blots, four enzyme-linked immunosorbent assays (ELISAs), and one lateral flow device, were included in this study. All seven tests evaluated were able to detect pathogenic prion proteins (PrP(CWD) ) in Northern American infected animals and distinguish physiologic prion protein (PrP(c) ) in brainstem (obex region) and lymph node samples from North American and European cervids, respectively. However, the specificity and sensitivity of the tests differed significantly. Highly sensitive tests for the detection of prion proteins are an important tool both for the design of effective disease surveillance and control strategies and the safety of the food chain. Thus, this study contributes to the emergency preparedness against CWD.

Short incubation with methyl aminolevulinate for photodynamic therapy of actinic keratoses.

BACKGROUND: Photodynamic therapy (PDT) using methyl aminolevulinate (MAL) is an effective first-line treatment for actinic keratoses. A reduced incubation period may have practical advantages. OBJECTIVE: This study aims to evaluate the effect of incubation time (1 vs. 3 h), MAL concentration (160 mg/g vs. 80 mg/g) and lesion preparation in the setting of MAL-PDT for treatment of actinic keratosis (AK). DESIGN: Open, randomized, parallel-group multicentre study. SETTING: Outpatient dermatology clinics. SUBJECTS: One hundred and twelve patients with 384 previously untreated AK. Most lesions (87%) were located on the face and scalp and were thin (55%) or moderately thick (34%). METHODS: Lesions were debrided, and MAL cream (160 mg/g or 80 mg/g) was applied before illumination with red light (570-670 nm; light dose, 75 J/cm2). Patients were followed up at 2 and 3 months. Sixty patients (54%) were re-treated and assessed at 6 months. MAIN OUTCOME: Complete lesion response rates 3 and 12 months after last treatment. RESULTS: For lesions on the face/scalp, lesion complete response rates were 78% for thin AK and 74% for moderately thick AK lesions after 1 h vs. 96% and 87% after 3 h incubation with MAL 160 mg/g. Lesion recurrence rates at 12 months after two treatments were similar [19% (3 of 16) with 1 h vs. 17% (3 of 18) with 3 h 160 mg/kg MAL-PDT] and lower than for 80 mg/g MAL-PDT (44-45%). CONCLUSION: MAL-PDT using a 1-h incubation may be sufficient for successful treatment of selected AK lesions.

Renin, endothelial NO synthase and endothelin gene expression in the 2kidney-1clip Goldblatt model of long-term renovascular hypertension.

OBJECTIVE: Numerous reports have shown the influence of renin, nitric oxide (NO) and the endothelin (ET) systems for regulation of blood pressure and renal function. Furthermore, interactions between these peptides have been reported. Aim of our study was to investigate the relative contribution of these compounds in long-term renovascular hypertension / renal ischemia. METHODS: Hypertension / left-sided renal ischemia was induced using the 2K1C-Goldblatt rat model. Renal renin, ET-1, ET-3 and endothelial NO synthase (eNOS) gene expression was measured by means of RNAse protection assay at different timepoints up to 10 weeks after induction of renal artery stenosis. RESULTS: Plasma renin activity and renal renin gene expression in the left kidney were increased in the clipped animals while eNOS expression was unchanged. Furthermore, an increase in ET-1 expression and a decrease of ET-3 expression was detected in early stenosis. CONCLUSIONS: While renin is obviously involved in regulation of blood pressure and renal function in unilateral renal artery stenosis, ET-1, ET-3 and endothelium derived NO do not appear to play an important role in renal adaptation processes in long-term renal artery stenosis, although ET-1 and ET-3 might be involved in short-term adaptation processes.

Single and dual cation sites in zeolites: theoretical calculations and FTIR spectroscopic studies on CO adsorption on K-FER.

Interaction of CO with K-FER zeolite was investigated by a combination of variable-temperature IR spectroscopy and computational study. Calculations were performed using omega(CO)/r(CO) correlation method in combination with a periodic density functional theory model. On the basis of agreement between experimental and calculated results, the following carbonyl complexes were identified: (i) mono- and dicarbonyl C-down complexes on single K(+) sites characterized by IR absorption bands at 2163 and 2161 cm(-1), respectively; (ii) complexes formed by CO bridging two K(+) ions separated by about 7-8 A (dual sites) characterized by a band at 2148 cm(-1); and (iii) isocarbonyl (O-down) complexes characterized by a band at 2116 cm(-1). The bridged carbonyl complexes on dual K(+) sites are about 5 kJ/mol more stable than monodentate (monocarbonyl) CO complexes. The C-O stretching frequency of monocarbonyl species in K-FER depends on K(+) location in the zeolite, and not on K(+) coordination to the framework. A combination of theoretical calculations using a periodic density functional model and experimental results showed formation of two types of monocarbonyls. The most abundant type appears at 2163 cm(-1), and the less abundant one at 2172 cm(-1). These experimentally determined wavenumber values coincide, within +/-2 cm(-1), with those derived from theoretical calculations.

Navigation-assisted construction of an external ear canal using an autogenous foreskin graft.

Classic microtia is combined with external ear canal atresia and middle ear malformation. In order to evaluate whether an operation to improve hearing ability and the use of computer-assisted surgery are indicated, preoperative high-resolution navigation CT is mandatory. We combined atresia surgery and tympanoplasty with auricular reconstruction in the case of an 8-year-old boy with bilateral microtia, aural atresia and malformation of the middle ear. After creating an auricle framework with rib cartilage and transplanting it under the skin of the mastoid plane, we shifted it forward in the second step, and the new auditory canal was drilled under computer-assisted navigation and facial nerve monitoring. In the same operation, tympanoplasty was accomplished, and a silastic cylinder, wrapped into pieces of rib cartilage, was inserted into the constructed canal and removed 2 months later. For lining the new auditory canal, we used the patient's prepuce, harvested by elective circumcision.

A review of mutual transmission of important infectious diseases between livestock and wildlife in Europe.

Oral vaccination of red foxes against rabies has been practiced in Europe since 1978 and has succeeded in greatly reducing the occurrence of this disease in foxes: this is an example of coordinated activity against a disease that affects both wild and domestic animals as well as humans. Some examples of diseases that affect both domestic and wild animals in Europe are: classical swine fever (hog cholera) in wild boars and domestic swine; myxomatosis and rabbit hemorrhagic disease in domestic and wild rabbits; bovine viral diarrhea (BVD) in cattle and roe deer; contagious ecthyma in domestic sheep and goats and also in, e.g., chamois, muskox, and reindeer; Mycobacterium bovis in cattle, wild boars, badgers, and deer; and brucellosis in a broad range of livestock and wildlife in all European countries. In addition, serological surveys performed in different free-ranging ungulate species revealed the presence of alphaherpesviruses related to bovine herpesvirus-1 in 7 European countries; and a study of malignant catarrhal fever in deer in Germany might indicate that in this case sheep are the main reservoir species. Although many data on infectious diseases are available in various European countries, there is more need for systematic surveillance and coordinated research.

Identification and analysis of a new hepadnavirus in white storks.

We identified, cloned, and functionally characterized a new avian hepadnavirus infecting storks (STHBV). STHBV has the largest DNA genome of all avian hepadnaviruses and, based on sequence and phylogenetic analysis, is most closely related to, but distinct from, heron hepatitis B virus (HHBV). Unique for STHBV among the other avian hepadnaviruses is a potential HNF1 binding site in the preS promoter. In common only with HHBV, STHBV has a myristylation signal on the S and not the preS protein, two C terminally located glycosylation sites on the precore/core proteins and lacks the phosphorylation site essential for the transcriptional transactivation activity of duck-HBV preS protein. The cloned STHBV genomes were competent in gene expression, replication, and viral particle secretion. STHBV infected primary duck hepatocytes very inefficiently suggesting a restricted host range, similar to other hepadnaviruses. This discovery of stork infections unravels novel evolutionary aspects of hepadnaviruses and provides new opportunities for hepadnavirus research.

Seroepizootiology of selected infectious disease agents in free-living birds of prey in Germany.

Four hundred forty-eight blood plasma samples from free-living birds of prey from Berlin and the Brandenburg area in eastern Germany were tested for antibodies against Newcastle disease virus (NDV), falcon herpesvirus (FHV), owl herpesvirus (OHV), and Chlamydia psittaci. Antibodies to NDV were detected in 6 (2%) of 346 tested diurnal birds of prey, whereas none of the owls (n = 55) was positive. The positive samples originated from two common buzzards (Buteo buteo), three ospreys (Pandion haliactus) and one marsh harrier (Circus aeruginosus). Titers varied between 1:8 and 1:32. Of 253 birds of prey one osprey (<1%) tested positive for antibodies to FHV with low titer of 1:6. This is the first detection of antibodies against FHV in an osprey. Furthermore, antibodies against OHV could be found in one tawny owl (Strix aluco) and one common buzzard (2 of 253, 1%) with low titers of 1:6. Of 422 birds of prey 267 (63%) tested positive for antibodies to Chlamydia psittaci with titers varying between 1:5 and 1:256 which reflects the ubiquitous occurrence of Chlamydia psittaci in these birds of prey.

European brown hare syndrome in free-ranging European brown and mountain hares from Switzerland.

From 1997 to 2000, complete necropsy and histopathologic investigations were performed on 157 free-ranging European brown hares (Lepus europaeus) found dead throughout Switzerland. Organ samples of all these individuals (157 livers and 107 spleens available) were tested for European brown hare syndrome virus (EBHSV)-antigen by enzyme-linked immunosorbent assay (ELISA) test kit. Furthermore, 60 additional blood samples were tested for antibodies against EBHSV by ELISA. In addition, liver samples of 87 free-ranging mountain hares (Lepus timidus) hunted in 1996 were tested for EBHSV-antigen. In two European brown hares from southern Switzerland lesions suggestive of changes induced by EBHSV were present, and high titers of EBHSV-antigen were detected in both liver and spleen samples of these animals. Based on negative staining electron microscopy investigations of liver and spleen homogenates, we observed calicivirus in one antigen-positive hare. Low EBHSV-antigen titers were found in three additional European brown hares from central and western Switzerland, but EBHS-lesions were absent. Antibodies against EBHSV were not detected in any of the sera of European brown hares, and EBHSV-antigen was not found in the samples of mountain hares. This is the first report of EBHS in European brown hares from Switzerland.

Absence of antibodies against canine distemper virus in free-ranging populations of the Eurasian badger in Great Britain.

Canine distemper virus (CDV) is a serious disease of wild carnivores throughout the world. In Europe, infection has been detected in several carnivores including the Eurasian badger (Meles meles). In the present study 182 badger blood samples were collected from an intensively studied population of wild badgers in southwestern England (January-July, 1997), and a further 286 from throughout southern Britain (June 1996-November 1998). A neutralizing peroxidase-linked antibody test was used for the detection of antibodies against CDV. All the samples were negative for CDV antibodies, suggesting that in contrast to mainland Europe, the disease may be either absent or maintained at low levels in British badgers.

Epizootiological investigations of canine distemper virus in free-ranging carnivores from Germany.

Canine distemper virus (CDV) infects a broad range of carnivores. To assess whether wild carnivores may play a role in the epidemiology of CDV in domestic dogs in Germany, the seroprevalence of CDV was determined. In sera from red foxes (30 of 591 (5%)) and stone martens (2 of 10 (20%)) antiviral antibodies were detected using a neutralization assay, whereas sera of raccoons, two mink, one pine marten and one raccoon dog were negative. In foxes, there was a significantly higher prevalence in urban and suburban compared to rural regions. When testing lung and spleen tissue samples (fox, badger, stone marten, polecat, raccoon dog) 13 of 253 (5.1%) foxes, 2 of 13 (15.4%) stone martens and 2 of 6 (33%) badgers were virus positive using RT-PCR. Phylogenetic analysis based on partial sequences of the F gene revealed a distinct relatedness to canine CDV isolates. Together, the data support the concept of transmission of CDV between domestic dogs and wild carnivores.

Necrobacillosis in free-living male European bison in Poland.

In the Bialowieza Primeval Forest (Poland) a chronic disease of the external genital organs has been observed in free-living male European bison (Bison bonasus) since 1980. Investigations on this disease started in the late 1980s. The most striking findings are necrotic and ulcerative lesions of the prepuce and penis of bison aged from 6 mo to >10 yr. Histologic examination of tissue samples from the prepuce of six bison (9-mo- to 8-yr-old), and from the penis of two bison (3- and 8-yr-old), were characteristic of necrobacillosis. Masses of slender, Gram-negative, rod-like or filamentous bacteria occurred in necrotic tissue. At the periphery of necrotic tissue filamentous bacteria were often arranged in large clusters and strands that advanced towards healthy tissue. Immunolabeling and electron microscopy also suggest that these organisms are Fusobacterium sp.

Serosurvey for selected infectious disease agents in free-ranging black and white rhinoceros in Africa.

Two hundred and eighty one serum samples collected from free-ranging black (Diceros bicornis) and white (Ceratotherium simum) rhinoceros, in the Republic of South Africa (RSA), Namibia, and Kenya from 1987-97, were examined for antibody to 16 different infectious agents. Positive antibody titers were detected against Akabane (59.8%), bluetongue (55%), African horse sickness (27.9%), epizootic haemorrhagic disease of deer (19.4%), parainfluenza type 3 (25.3%), bovine herpes virus 1 (3.1%), equine herpes virus 1 (8.8%) and bovine viral diarrhea (1.2%) viruses, and four serovars of Leptospira interrogans, (ranging 1.2 to 8.8%). No antibody was detected against Rift Valley fever virus, encephalomyocarditis virus, Brucella abortus, and Trypanosoma equiperdum. Interspecies differences were detected for African horse sickness, epizootic haemorrhagic disease of deer and parainfluenza type 3 viruses. There appeared to be some geographic variation in the prevalence of antibody for African horse sickness, bluetongue, epizootic haemorrhagic disease of deer, parainfluenza type 3, equine herpes virus 1 and Leptospira interrogans serovar bratislava.

A new avian hepadnavirus infecting snow geese (Anser caerulescens) produces a significant fraction of virions containing single-stranded DNA.

We describe the identification and functional analysis of an evolutionary distinct new avian hepadnavirus. Infection of snow geese (Anser caerulescens) with a duck hepatitis B virus (DHBV)-related virus, designated SGHBV, was demonstrated by detection of envelope proteins in sera with anti-DHBV preS and S antibodies. Comparative sequence analysis of the PCR-amplified SGHBV genomes revealed unique SGHBV sequence features compared with other avian hepadnaviruses. Unlike DHBV, SGHBV shows an open reading frame in an analogous position to orthohepadnavirus X genes. Four of five cloned genomes were competent in replication, gene expression, and virus particle secretion in chicken hepatoma cells. Primary duck hepatocytes were permissive for infection with SGHBV, suggesting a similar or identical host range. SGHBV was found to secrete a significant fraction of virion-like particles containing single-stranded viral DNA. This was observed both in cell culture medium of SGHBV DNA-transfected LMH cells and in viremic sera of several birds, suggesting that it is a stable trait of SGHBV. Taken together, SGHBV has several unique features that expand the knowledge of the functional and evolutionary diversity of hepadnaviruses and offers new experimental opportunities for studies on the life cycle of hepadnaviruses.

Influence of environmental cadmium on testicular proliferation in roe deer.

The influence of cadmium on spermatogenesis in roe deer was studied, comparing animals from areas with high (n = 37) and low (n = 23) cadmium exposure. Testes were evaluated during pre-rut (May) and rut (July/August). During these periods the cadmium accumulation (mg/kg kidney) in polluted regions averaged 3.41 and 3.52 in comparison with 1.61 and 1.81 in controls. Cadmium accumulation was positively correlated with testis weight during both seasons. In May the proliferation (units of tissue polypeptid specific antigen/g parenchyma) was lower in animals with higher cadmium (24.02 compared with 78.20; P < 0.01). During the rut, testicular proliferation and spermatozoa/g testis as well as apoptosis showed no significant changes with increased cadmium contamination. The results suggest delayed proliferation during the pre-rutting period in animals with high cadmium exposure, but other indications of effects on the testis were not significant.