Spider silk nerve graft promotes axonal regeneration on long distance nerve defect in a sheep model.

Peripheral nerve injuries with substantial tissue loss require autologous nerve transplantation or alternatively reconstruction with nerve conduits. Axonal elongation after nerve transection is about 1 mm/day. The precise time course of axonal regeneration on an ultrastructural level in nerve gap repair using either autologous or artificial implants has not been described. As peripheral nerve regeneration is a highly time critical process due to deterioration of the neuromuscular junction, this in vivo examination in a large animal model was performed in order to investigate axonal elongation rates and spider silk material degradation in a narrowly delimited time series (20, 30, 40, 50, 90, 120, 150 and 180 days) by using a novel spider silk based artificial nerve graft as a critical prerequisite for clinical translation. Autologous nerves or artificial nerve conduits based on spider silk of the spider species Trichonephila edulis were transplanted in a 6.0 cm nerve defect model in the black headed mutton. At each of the post-implant time point, electrophysiology recordings were performed to assess functional reinnervation of axonal fibers into the implants. Samples were analyzed by histology and immunofluorescence in order to verify the timeline of axonal regeneration including axonal regeneration rates of the spider silk implant and the autologous transplant groups. Spider silk was degraded within 3 month by a light immune response mainly mediated by Langhans Giant cells. In conjunction with behavioral analysis and electrophysiological measurements, the results indicate that the spider silk nerve implant supported an axonal regeneration comparable to an autologous nerve graft which is the current gold standard in nerve repair surgery. These findings indicate that a biomaterial based spider silk nerve conduit is as effective as autologous nerve implants and may be an important approach for long nerve defects.

Analysis of porcine body size variation using re-sequencing data of miniature and large pigs.

BACKGROUND: Domestication has led to substantial phenotypic and genetic variation in domestic animals. In pigs, the size of so called minipigs differs by one order of magnitude compared to breeds of large body size. We used biallelic SNPs identified from re-sequencing data to compare various publicly available wild and domestic populations against two minipig breeds to gain better understanding of the genetic background of the extensive body size variation. We combined two complementary measures, expected heterozygosity and the composite likelihood ratio test implemented in "SweepFinder", to identify signatures of selection in Minipigs. We intersected these sweep regions with a measure of differentiation, namely FST, to remove regions of low variation across pigs. An extraordinary large sweep between 52 and 61 Mb on chromosome X was separately analyzed based on SNP-array data of F2 individuals from a cross of Goettingen Minipigs and large pigs. RESULTS: Selective sweep analysis identified putative sweep regions for growth and subsequent gene annotation provided a comprehensive set of putative candidate genes. A long swept haplotype on chromosome X, descending from the Goettingen Minipig founders was associated with a reduction of adult body length by 3% in F2 cross-breds. CONCLUSION: The resulting set of genes in putative sweep regions implies that the genetic background of body size variation in pigs is polygenic rather than mono- or oligogenic. Identified genes suggest alterations in metabolic functions and a possible insulin resistance to contribute to miniaturization. A size QTL located within the sweep on chromosome X, with an estimated effect of 3% on body length, is comparable to the largest known in pigs or other species. The androgen receptor AR, previously known to influence pig performance and carcass traits, is the most obvious potential candidate gene within this region.

Detection of PR-39, a porcine host defence peptide, in different cell sub-linages in pigs infected with Actinobacillus pleuropneumoniae.

Innate immunity is critically important for the outcome of infection in many diseases. It was previously shown that cathelicidin PR-39, an important porcine multifunctional host defence peptide, is elevated in bronchoalveolar lavage fluid and respiratory tract tissue after experimental infection with Actinobacillus pleuropneumoniae (A.pp.). To date, neutrophil polymorphonuclear leukocytes (PMNs) are thought to be the only source of PR-39. The aim of this study was to further characterize PR-39⁺ cells and selected immune cell populations in lung tissue during the peracute (7-10 hours), acute (2 days), reconvalescent (7 days) and chronic (21 days) stages of experimental infection with A.pp. serotype 2. In total, six mock-infected control pigs and 12 infected pigs were examined. Using immunofluorescence double-labeling, antibodies against PR-39 were combined with antibodies against CD3 (T-cells), CD79 (B-cells), Iba1 (activated macrophages), TTF-1 (lung epithelial cells expressing surfactant proteins), macrophage/L1 protein and myeloperoxidase (MPO, cells of the myeloid linage). In the peracute and acute phases of infection, total PR-39⁺ cells and myeloid linage cells increased, whereas CD3⁺ cells and TTF-1⁺ cells decreased. Double labeling revealed that most Macrophage/L1 protein+ cells and to a lesser extent MPO⁺ cells co-expressed PR-39. In addition, few bronchial epithelial cells and type 2 alveolar epithelial cells (both identified with TTF-1) produced PR-39. Occasionally, CD3⁺ T cells expressing PR-39 were seen in infected animals. Taken together, this study identifies cell types, other than PMNs, in lungs of A.pp.-infected pigs that are capable of producing PR-39. In addition, these findings provide further insights into the dynamics of different immune cell populations during A.pp.-infection.

Concept and evaluation of the German War Surgery Course - Einsatzchirurgie-Kurs der Bundeswehr.

INTRODUCTION: Military surgeons must be prepared to care for severe and complex life-threatening injuries rarely seen in the civilian setting. Typical civilian training and practice do not provide adequate exposure to the broad set of surgical skills required. The German Bundeswehr Medical Service has developed and refined the War Surgery Course (WSC) to meet this training gap. This article describes the recent experience with this readiness curriculum. METHODS: Run annually since 1998, WSC consists nowadays of 5 days with 20 theoretical modules. Four sessions with standardised practical skills training use a live tissue porcine model, and the recently added cadaver-based Advanced Surgical Skills for Exposure in Trauma course. Sixteen military surgeons who participated in the WSC in January 2016 completed a survey of their self-rated readiness for 114 predefined emergency skills before and after completion, and provided an overall evaluation of the course. RESULTS: Self-assessed readiness improved significantly over baseline for all areas covered in both the practical skills and theoretical knowledge portions of the WSC curriculum. Additionally, all participants rated the course as important and universally recommended it to other military surgeons preparing for missions. CONCLUSIONS: The WSC course format was well received and perceived by learners as a valuable readiness platform. Ongoing evaluation of this course will enable data-driven evolution to ensure a maximum learning benefit for participants. With the increasing multinational nature of modern military missions, surgeons' training should follow international standards. Continuing evolution of military surgical training courses should further encourage the sharing and adoption of best educational practices.

[Uterine neoplasia in two geriatric miniature pet pigs - diagnostics and therapy. Two case reports]. / Uterustumore bei älteren Miniaturschweinen - Diagnostik und Therapie. Zwei Fallberichte.

Miniature pet pigs are becoming ever more common among the patients of small animal practices. Due to an advanced age attained in these animals in comparison with conventional pigs in pork production, miniature pet pigs develop age-related diseases that are infrequently observed in farm pigs, including neoplastic alterations of the uterus. We describe two cases of a uterine adenocarcinoma and a leiomyoma, respectively, in two miniature pet pig sows. Options and limits of diagnostic measures (unspecific clinical symptoms, x-ray, sonography and computed tomography) and therapy within the legal limits as well as the risk of an incorrect diagnosis based upon the more common differential diagnoses in swine medicine, including obstipation, cystitis and endometritis, are discussed.

[Septicaemia in piglets associated with a positive finding of a methicillin-resistant S. aureus strain]. / Septikämisches Krankheitsbild bei Saugferkeln mit MRSA-Nachweis.

Staphylococcus (S.) aureus has been associated with septicaemia, mastitis, vaginitis, metritis, osteomyelitis, and endocarditis. This case report demonstrates S. aureus-induced septicaemia in suckling pig- lets. Three days after birth, littermates displayed severe ecchymosis and cyanosis, with a 50% mortality rate. The surviving littermates were cross-fostered, but died 1 day later. Other piglets, which were allowed to suck at the sow, developed similar clinical signs. Haematological findings were anaemia, thrombocytopenia, and leukopenia; therefore, neither isohaemolytic anaemia nor septicaemia could be excluded as differential diagnoses. At necropsy, petechial bleeding on inner organ surfaces and free blood in body cavities were found. Bacteriological examination of the sow's milk and of the spleen of one piglet detected a methicillin-resistent S. aureus strain (MRSA CC398), which was in all likelihood the cause of the disease. Potential differential diagnoses are discussed.

Antibiotic susceptibility of Salmonella, Campylobacter coli, and Campylobacter jejuni isolated from Northern German fattening pigs.

This study was conducted to assess the antimicrobial susceptibility rate of Salmonella and Campylobacter spp. isolated from Northern German fattening pigs. From 540 lymph node samples, 16 Salmonella Typhimurium, 1 Salmonella Brandenburg, 37 Campylobacter coli, and 11 Campylobacter jejuni strains were isolated. Antibiotic susceptibility testing was carried out by the broth dilution method. The 14 tested antibiotics for Salmonella were ampicillin, cefotaxime, ceftazidime, chloramphenicol, ciprofloxacin, colistin, florfenicol, gentamicin, kanamycin, nalidixic acid, streptomycin, sulfamethoxazole, tetracycline, and trimethoprim. The eight tested antibiotics for Campylobacter spp. were ampicillin, ampicillin-sulbactam (2:1), ciprofloxacin, erythromycin, gentamicin, nalidixic acid, sulfamethoxazole-trimethoprim (1:19), and tetracycline. In total, 93.7% (n = 16) of Salmonella Typhimurium, 75.7% (n = 37) of C. coli, and 54.5% (n = 11) of C. jejuni isolates were resistant to at least one of the tested antibiotics. Multiresistance to three antibiotics was observed in 75% of Salmonella Typhimurium, 16.2% of C. coli, and 0% of C. jejuni isolates. Pansusceptibility was detected in 6.3% of Salmonella Typhimurium, 24.3% of C. coli, and 45.5% of C. jejuni isolates. Multiresistance is defined as resistance to three or more antibiotics, and pansusceptibility is defined as not having resistance to any antibiotic. Regarding drugs of last resort--cefotaxime, ciprofloxacin, and nalidixic acid--resistance was not common among Salmonella (6.3%). The resistance rate of Campylobacter spp. to last-resort drugs--erythromycin, ciprofloxacin, and nalidixic acid--varied between species. The observed trend was not statistically significant. No C. coli isolates and few C. jejuni isolates (9.1%) were resistant to erythromycin. In contrast to C. jejuni, the C. coli isolates were more likely to be resistant to ciprofloxacin (9.1 and 18.9%, respectively) and nalidixic acid (0 and 13.5%, respectively). The same phenomenon was detected for tetracycline (27.3 and 62.2%, respectively), sulfamethoxazole (9.1 and 43.2%, respectively), and ampicillin (9.1 and 21.6%, respectively).

[Computed tomography of the pig lung. An innovative approach to the definition of the pulmonary health status]. / Computertomographie der Lunge des Schweines. Ein innovativer Ansatz zur Definition des Lungengesundheitsstatus.

OBJECTIVE: In veterinary medicine computed tomography (CT) imaging has gained importance in recent years, especially for diagnostics in pets, but also during the course of experimental studies in animal models for human medicine. In this study the applicability of CT as an imaging method for the depiction of the porcine thorax and in particular of the pig lung was evaluated. MATERIAL AND METHODS: CT examinations were performed with 11 healthy pigs of two age groups. For evaluation, CT findings were related to clinical, radiological, macroscopical, microscopical, and microbiological findings. RESULTS: Clinically relevant anatomical structures were determined and recorded using transverse slices. In ventral recumbency, lung parenchyma density measurements at the levels of the second, fourth and seventh thoracic vertebrae resulted in significantly higher densities of the ventral in comparison to those of the dorsal lung quadrants. CONCLUSION AND CLINICAL RELEVANCE: Computed tomography is a valuable tool for the high-contrast depiction of the porcine lung without superposition. In future studies this CT reference base for unaltered pig lungs may facilitate the identification of anatomical structures within the porcine lung as well as the assessment of pathological lung alterations.

Prevalence of pathogenic Yersinia enterocolitica strains on liver surfaces of pigs and their antimicrobial susceptibility.

A study to determine the occurrence of pathogenic Yersinia enterocolitica on surfaces of slaughtered pig livers and the antimicrobial resistant pattern of the isolates was carried out in a slaughterhouse in Lower Saxony, Germany. During the slaughtering process, 1,500 surfaces of pig livers from 50 fattening herds were swabbed in order to isolate and characterize Y. enterocolitica isolates by serotyping, detecting the virulence plasmid coding the yopT gene, and resistance testing. Of the livers tested, 4.7% were positive for Y. enterocolitica O:3, which was the only identified serotype. The virulence gene yopT was found in 90.0% of these isolates. Antimicrobial susceptibility was tested by the broth dilution method, and the MICs were determined for 13 antimicrobials. All isolates were resistant to ampicillin and sulfamethoxazole but were susceptible to ciprofloxacin, nalidixic acid, gentamicin, ceftiofur, tetracycline, kanamycin, cefotaxime, and chlorphenicol. Up to now, resistance to florfenicol has always been described in combination with resistance to chloramphenicol. In the present study, 15.3% of the isolates were resistant to florfenicol, while no chloramphenicol-resistant strains could be identified. Multiresistance to three or more antimicrobials was detected in 22 strains (27.3%). Nevertheless, third-generation cephalosporines or fluoroquinolones, which were recommended for extraintestinal Y. enterocolitica infection in humans, were not affected.

Characterization of the porcine transferrin gene (TF) and its association with disease severity following an experimental Actinobacillus pleuropneumoniae infection.

Transferrin (TF)-mediated provision of iron is essential for a productive infection by many bacterial pathogens, and iron-depletion of TF is a first line defence against bacterial infections. Therefore, the transferrin (TF) gene can be considered a candidate gene for disease resistance. We obtained the complete DNA sequence of the porcine TF gene, which spans 40 kb and contains 17 exons. We identified polymorphisms on a panel of 10 different pig breeds. Comparative intra- and interbreed sequence analysis revealed 62 polymorphisms in the TF gene including one microsatellite. Ten polymorphisms were located in the coding sequence of the TF gene. Four SNPs (c.902A>T, c.980G>A, c.1417A>G, c.1810A>C) were predicted to cause amino acid exchanges (p.Lys301Ile, p.Arg327Lys, p.Lys473Glu, p.Asn604His). We performed association analyses using six selected TF markers and 116 pigs experimentally infected with Actinobacillus pleuropneumoniae serotype 7. The analysis showed breed-specific TF allele frequencies. In German Landrace, we found evidence for a possible association of the severity of A. pleuropneumoniae infection with TF genotypes.

[Comparison of the diagnostic significance of clinical, radiographic and ultrasonographic results after an experimental aerosol infection of pigs with Actinobacillus pleuropneumoniae]. / Vergleich der diagnostischen Aussagekraft klinischer, röntgenologischer und sonographischer Befunde bei der experimentellen Infektion des Schweines mit Actinobacillus pleuropneumoniae.

Scoring schemes for clinical, ultrasonographic and radiographic findings in pigs were developed based upon a standardized animal model for Actinobacillus pleuropneumoniae infection.The results of these methods were compared to each other as well as with the corresponding pathomorphological findings during necropsy. Altogether 69 pigs of different breeding lines (Hampshire, Pietrain and German Landrace were examined. Positive correlations were found between the results of all three methods as well as with the necropsy scores (p <0.0001). Different pathomorphological findings were detected either by radiographic or by ultrasonographic examination dependent upon the type of lung tissue alterations: Alterations of the pleura as well as sequestration of lung tissue on the lung surface could be clearly identified during the ultrasonographic examination while deep tissue alterations with no contact to the lung surface could be detected reliably by radiographic examination. Both methods complement each other, and the application of a combined ultrasonographic and radiographic examination of the thorax allows a comprehensive inspection of the lung condition. Particularly during the acute phase of the disease the extent of lung tissue damage can be estimated more precisely than by clinical examination alone.

Comparison of three common methods of lung lavage in healthy pigs.

Bronchoscopic, endotracheal and transtracheal lung lavage were evaluated in 38 healthy pigs taken from a nucleus herd in a good state of health with respect to their applicability in practice and the traceability of bacteria, cellular parameters and the antimicrobial peptide PR-39 in the respective lavage fluid samples. The total cell count, qualitative morphological cellular characteristics as well as PR-39 could be determined in all lavage fluid samples, while quantitative cell differentiation was not possible in endotracheal lavage samples. The comparison of the three methods resulted in a higher proportion of polymorphonuclear neutrophil granulocytes (PMNs) and higher concentrations of PR-39 in transtracheal samples. For this reason different valuation standards with respect to PMNs and PR-39 concentrations are presupposed for transtracheal lavage samples. The occurrence of pavement epithelial cells as well as the number of contaminating bacterial species per sample was the lowest in transtracheal lavage. Mycoplasma hyopneumoniae polymerase chain reaction appeared to have the highest diagnostic sensitivity in combination with bronchoscopic lavage. In conclusion, bronchoscopic and transtracheal lavage were considered to be more appropriate for bacteriological and cytological diagnostics than endotracheal lavage.

Respiratory disease markers in porcine bronchoalveolar lavage fluid.

In bronchoalveolar lavage fluid (BALF) of pigs originating from different herds bacteria, cells and the antibacterial peptide PR-39 were examined to gain information about the lung health status. In a high health nucleus herd 56% and in low health herds 20-100% of the examined pigs were found positive for potentially pathogenic bacteria. Based on these findings, a novel definition for bacterial respiratory tract disease was established using an 8% cut-off for the relative number of neutrophils in bronchoscopic and a 40% cut-off in transtracheal BALF in combination with the occurrence of potentially pathogenic microorganisms. The antibacterial peptide PR-39 was highly correlated to this definition of respiratory disease. An assessment of the bacteriological respiratory health status appears to be possibly based on the determination of PR-39 concentrations in BALF using different cut-off values according to the lavage method (2.5 nM for bronchoscopic and 5 nM for transtracheal BALF).

Immunization of sows in an integrated pig-breeding herd using a homologous inactivated Salmonella vaccine decreases the prevalence of Salmonella typhimurium infection in the offspring.

The efficacy of a homologous vaccination in preventing infection of suckling piglets with Salmonella (S.) Typhimurium was evaluated after an immunization of pregnant sows using an inactivated herd-specific S. Typhimurium vaccine. Twenty-five pregnant sows were vaccinated three times antepartum. The efficiency of this vaccine regime was assessed by comparison with a control group of 37 sows and their suckling piglets, which were daily treated with enrofloxacin from day 14 antepartum until the day of weaning. From the first day of life until day 142 post-partum, faecal samples of the piglets were collected and analysed for Salmonella shedding. In parallel, systemic antibody responses were monitored using a whole cell-based isotype-specific enzyme-linked immunosorbent assay (ELISA). The bacteriological investigation showed marked effects of vaccination. Salmonella Typhimurium could not be detected in any of the faecal samples of the piglets from the vaccinated sows. In contrast, the piglets of the group with long-time antibiotic treatment shed salmonellae rating to 47.4% of the animals. Furthermore, the offspring from vaccinated sows showed significantly decreased antibody activities of immunoglobulin (Ig)A and IgG. These bacteriological and serological results indicate a significantly lower Salmonella prevalence in piglets of the vaccinated group. As this study shows, the presented strategy of vaccination of pregnant sows with an inactivated Salmonella vaccine seems to be a suitable measure in decreasing Salmonella prevalence in offspring of infected sows.

Effects of fluorequinolone treatment acidified feed, and improved hygiene measures on the occurrence of Salmonella Typhimurium DT104 in an integrated pig breeding herd.

Worldwide, the use of antimicrobials in food production has been associated with drug resistance in foodborne pathogens such as Salmonella. However, little is known about the efficaciousness of fluorequinolone treatment on Salmonella Typhimurium T104 infections in pig breeding herds. A combined eradication procedure with enrofloxacin application on sows and piglets, feeding of encapsulated organic acids to sows, disinfection with peracetic acid, separation of the growers from the sows and serological discrimination using a new whole-cell-based enzyme-linked immnosorbent assay (ELISA) was evaluated for the suitability to eradicate and to control endemic S. Typhimurium DT104 infections in a closed herd. Thirty-seven sows and their piglets were treated everyday from day 14 ante partum until the day of weaning. Eighteen sows and their piglets served as controls. From the first day of life until day 168 after birth, faecal samples (n = 1671) of all piglets were analysed for Salmonella shedding. In parallel, systemic antibody responses were monitored by whole-cell-based isotype-specific ELISA systems. From birth to weaning the prevalence in both groups was between 2% and 9%. After weaning, intermittent shedding could be observed in both groups, and salmonellae could be found in up to 7.7% of the faecal samples. As a result, a dramatic increase in Salmonella-infected growers was observed, as of day 115 after birth, 47.4% of the animals of the treated group were tested positive for S. Typhimurium. Our results indicate that despite long-term antibiosis treatment and optimized hygiene measures, shedding of S. Typhimurium by the sows and the subsequent infection of their offspring could not be effectively prevented. Although it could be not shown that elimination of S. Typhimurium DT104 infection was achieved, the disinfection procedures described and the diagnostic test used are effective instruments to decrease the Salmonella load and to identify individual infected animals. Both of these are important factors for an improved consumer protection.

[Outlook for control of current zoonoses in swine--an issue on consumer health protection]. / Ausblick zur Bekämpfung aktueller Zoonosen des Schweines--ein Beitrag zum Verbraucherschutz.

With the development of a uniform European strategy for the control of zoonoses the European Commission has placed the main emphasis on the protection of consumer health. This direction is clearly marked by the White Paper on Food Safety and the attached proposals for directives and regulations. The present article considers the peculiarities of the epidemiological situation of the agents Campylobacter spp. and Yersinia enterocolitica in pork production. The situation of zoonoses in Europe is used as an opportunity to present the distinctive characteristics of these agents, results of epidemiological studies available, and the risk to consumer health in light of the literature. For the approaching transfer of the responsibility for food safety to the primary producers, i.e. the farmers, the most interesting data concern the prevalence, distribution and risk factors. But as there is a strong need for clarification of further questions about the main ways of entry of these agents on farms and into the production chain before a successful preventive strategy can be developed on pig farms, these issues are considered in more detail here.

[Housing- and management-specific effects on the health of swine]. / Haltungs- und managementspezifische Einflüsse auf die Gesundheit von Schweinen.

Important diseases with regard to inadequate housing and management conditions in pig production are given. Typical examples like claw lesions, cannibalism and infectious factorial diseases are described. Beside negative factors of pig housing and management which are of particular pathogenic importance some aspects of disease control and herd health are listed finally.

[Keeping sows outdoors during the winter season]. / Freilandhaltung von Sauen während des Winterhalbjahres.

During the last decade the number of outdoor breeding pig units increased in Northern Europe. With comparable results in reproduction and up to 70 per cent lower capital outlays in relation to the conventional indoor systems the interest of farmers in outdoor breeding grew also in Germany. There are a number of issues that are specific to outdoor systems that need to be considered during winter season.

Studies on the effect of specific egg antibodies against Escherichia coli infections in piglets.

The effect of chicken egg powder enriched with immunoglobulins specific for rotavirus antigen and fimbrial adhesions F4, F5, F6 of enterotoxigenic Escherichia coli (ETEC) (Globigen 66 S, Lohmann Animal Health, Cuxhaven, Germany) was studied in 465 sucking piglets on a commercial farm. Half of those piglets were given Globigen 66 S as an additive to milk replacer from day 2 until day 12 of life in addition to sows' milk. These piglets showed a higher intake of milk replacer and a lower prevalence of diarrhoea on days 2 and 3 of life. Statistical evaluation showed, that the effect of sows' milk on the duration of diarrhoea and on piglet weight gains was more pronounced than the effect of Globigen 66 S. Anti-ETEC F4-antibody-activities were measured using an indirect ELISA (enzyme-linked immunosorbent assay). There was an inverse relationship between the intensity of diarrhoea and colostral antibody-activity (r = -0.2). Comparison of binding affinities of avian and porcine antibodies for F4 showed only a limited common spectrum of epitopes, so, in all probability, they might complement each other in the intestine.