Retrospective analysis of the oral immunisation of wild boar populations against classical swine fever virus (CSFV) in region Eifel of Rhineland-Palatinate.

In the present study the effect of control measures implemented during the classical swine fever (CSF) epidemic in wild boar in the Eifel region of the German federal state of Rhineland-Palatinate from 1999 to 2005 was assessed. During the first 3 years after official confirmation of virus detection these measures comprised intensive hunting, especially of young animals and hygiene measures. Subsequently oral immunisation (o.i.) using a modified live virus vaccine was introduced as an additional control tool. All shot wild boar from the restricted area were tested virologically and serologically for CSF. The laboratory results from over 110,000 animals accompanied by information about age, gender and geographical origin of the animals were collected in a relational database. In total about 82% of all virologically positive wild boars were piglets, thus confirming the importance of this age group in the perpetuation of the epidemic. An analysis of the hunting bag showed that piglets were underrepresented compared to older animals throughout the eradication programme. This finding indicated that hunters did not comply with the control strategy of intense targeting of young animals. Before as well as after the implementation of o.i. a significantly higher virological prevalence and a significantly lower serological prevalence were observed in piglets compared to yearlings and adults. Shortly after the beginning of the vaccination campaign in February 2002 CSFV prevalence decreased significantly whereas the serological prevalence increased markedly in all age classes. In order to test the influence of age and vaccination on the serological prevalence a logistic regression model was used. Our results strongly suggest that under the field conditions in the Eifel region vaccination against CSFV had a crucial influence on the increase of seroprevalence rate and the elimination of CSFV. The last virus-positive pig was found 13 months after start of o.i.

[The suitability of ELISA for the detection of antibodies against Mycobacterium avium ssp. paratuberculosis in bulk milk samples from Rhineland-Palatinate]. / Untersuchungen zur Eignung von ELISAs zum Nachweis von Antikörpern gegen Mycobacterium avium ssp. paratuberculosis in Tankmilchproben aus Rheinland-Pfalz.

Paratuberculosis or Johne's Disease, caused by Mycobacterium avium subspecies paratuberculosis (MAP), is a notifiable disease in Germany which produces enormous economical losses in dairy farms. At present,there is no confirmed data about the actual number of infected livestock herds in Germany. A countrywide monitoring program to evaluate the prevalence in dairy herds would only be economically feasible on the basis of bulk milk testing. In this study, we evaluated two ELISA test kits (SVANOVIR Ptb-ELISA, IDEXX-M.pt. Milk test kit) for the detection of antibodies against MAP in bulk milk. First, the Paratuberculosis-status of the herd derived from the history of the farm was used as a gold standard. Paratuberculosis-negative farms were tested negative with each test, but paratuberculosis-positive or Paratuberculosis-serologically-positive farms were detected only in one case (Svanovir) or three cases (IDEXX), respectively. Even if inconclusive results are counted as positive, 82.9 % (Svanovir) or 80 % (IDEXX) of the paratuberculosis-positive or serologically paratuberculosis positive farms were not detected. Nevertheless, a re-validation of both ELISAs by means of ROC and TG-ROC analyses was attempted by searching for ideal cut-offs, optimised for bulk milk. If a high specificity was selected, no acceptable sensitivity could be reached. The best results were obtained using a sensitivity of 32.3 % at a specificity of 100 % (Svanovir). With a small change of the cut-off value, the sensitivity increased to still 57 %, but this reduced the specificity to 67 %. Similar results were obtained with the IDEXX-ELISA. We then evaluated the Svanovir-ELISA for the detection of bulk milk samples on the basis of the current paratuberculosis prevalence within 69 dairy herds from Rhineland-Palatinate using individual milk samples.When the bulk milk samples were tested in two different laboratories using the same ELISA, considerable differences in the results became evident. Nearly all samples were tested with a higher relative test result in one laboratory, which often led to differences in the classification of the prevalence levels. The estimated within-herd seroprevalences ranged between 0 % and 37 %. There was little agreement between the historical paratuberculosis herd status and the within-herd prevalence in milk serum, as reflected in a kappa-index of 0.146. To determine the sensitivity and specificity of the bulk milk ELISA by ROC and TG-ROC analysis, 116 bulk milk samples were used that had been obtained from the 69 dairy herds participating in the study. The optimal ratio of sensitivity (81 %) and specificity (77 %) relative to a "gold standard" was obtained when the cut-off was set at the 10 % level. These values for sensitivity and specificity were better than those obtained in an evaluation of the same ELISA in which the historical Paratuberculosis herd-status was used as a "gold standard." The results of this study question the suitability of the available ELISAs for bulk milk testing. Taking into account that the Svanovir-ELISA for individual milk samples has a sensitivity of 60 96% relative to the blood serum variant of the test, and that the latter has also a limited sensitivity due to the pathogenesis of paratuberculosis, the available test systems examined in this Study do not seem to be suitable for herd diagnosis by using bulk milk samples.

Potential risk factors for bovine Neospora caninum infection in Germany are not under the control of the farmers.

In the German state of Rhineland-Palatinate, herds were identified that were likely to have a Neospora caninum sero-prevalence > or = 10% by using a bulk milk ELISA. Individual herd data were obtained by a questionnaire. Univariate logistic regression showed that bulk milk positive farms had a significantly higher chance to report an increased abortion rate than negative farms (P(Wald)<0.1). The chance to have a bulk milk positive herd increased with the minimum number of years a farm had reported an increased abortion rate (P(Wald)<0.1). Questionnaire data, population and dog density as well as climatic data specific for the farm localization were used to identify potential risk factors for a herd to have acquired N. caninum infections. Within an optimized multiple logistic regression model 'Number of farm dogs', 'Herd size', and factors related to the municipality the farm was localized, i.e. 'Mean temperature in July', and 'Dog density' were significant risk factors (P(Wald)<0.1). The present study underlines the role farm dogs have in the epidemiology of neosporosis. In addition, it suggests that the risk a herd has to acquire N. caninum infections is also associated with factors related to the farm location, i.e. factors that are largely out of the control of farmers.

Regional distribution of bovine Neospora caninum infection in the German state of Rhineland-Palatinate modelled by Logistic regression.

To obtain a rapid overview over the distribution of bovine Neospora caninum-infections in the German state of Rhineland-Palatinate, an ELISA to determine specific bovine antibodies against a p38 surface antigen of N. caninum tachyzoites was modified to examine bulk milk samples from cattle herds. Experimental bulk milk samples were used to demonstrate that the seroprevalence in a group of animals can be estimated with this ELISA. A cut-off was selected for the specific detection of herds having a seroprevalence > or =10%. About 90% of the dairy herds located in Rhineland-Palatinate were examined. An overall prevalence of bulk milk-positive herds of 7.9% (95% confidence interval 7.0-8.9%), respectively, was determined. Major regional differences in the distribution of bulk milk-positive herds were observed. Prevalences were higher in regions with an increased degree of urbanisation. Logistic regression was applied to model the prevalence of bulk milk-positive herds on a district and city level. Variables describing the dog density, mean temperature in July, mean temperature in January and the total yearly precipitation in districts and cities were able to explain most of the observed variability in the regional prevalences. Our results provide evidence that in addition to risk factors related to individual farms also risk factors related to the farm location such as dog density in the surrounding and climate factors are important in the epidemiology of bovine neosporosis.

[The importance of allergic skin test with Johnin, antibody ELISA, cultural fecal test as well as vaccination for the sanitation of three chronically paratuberculosis-infected dairy herds in Rhineland-Palatinate]. / Zur Bedeutung von allergischem Hauttest mit Johnin, Antikörper-ELISA, kultureller Kotuntersuchung sowie der Impfung für die Sanierung dreier chronisch Paratuberkulose-infizierter Milchviehherden in Rheinland-Pfalz.

Three chronically paratuberculosis infected herds were tested for six years twice a year (intradermal Johnin test, antibody ELISA (IDEXX Corp.), microbial culture) according to a sanitary program. Culling of shedding animals and vaccination of calves with NEOPARASEC (Merial Corp.) were part of the program. In course of experiment, 1015 samples of 228 non vaccinated cows and 1502 samples of 293 vaccinated cattle have been tested. 3.8% of the vaccinated animals proved positive in microbial culture. Nearly all vaccinated calves developed granulomas sized from hazelnut to loaf at the injection site. Positive reactions in intradermal test as well as in antibody ELISA were found in very young calves. 24.3%, 33.7%, 25.9%, respectively of the non vaccinated animals were identified as shedders of M. avium subsp. paratuberculosis (MAP) by microbial culture. In the first and in the second herd most shedders of MAP were found in the first herd examination (66.7%, 42.9%, respectively), whereas in the third herd they were detected in the fifth examination (31.0%). At the beginning, 17.9% of non vaccinated animals proved positive in intradermal test, 14.4% in antibody ELISA. Afterwards, the number of positive test results decreased but increased again towards the end of the experiment. 48.5% of the 66 shedders showed positive reactions in intradermal test, 57.6% in antibody ELISA, 77.3% in at least one of these both tests. Antibodies in ELISA were found in rising frequency from two years before the time of shedding. 50.0% of the shedders reacted positive in ELISA at the time of shedding. In selected shedders first positive results were found at the age of about two years. Unfortunately, only incomplete hygienic measures were realized by the farmers. Under field conditions the realisation of attending sanitary programs is difficult. MAP is spread mainly by buying of animals, therefore a certification program for paratuberculosis free herds is urgently necessary as well as an improvement of diagnostic methods.

Immune reactions in cattle after immunization with a Mycobacterium paratuberculosis vaccine and implications for the diagnosis of M. paratuberculosis and M. bovis infections.

After immunization of four calves with a live modified Mycobacterium paratuberculosis vaccine the course of the humoral and cell mediated immune reactions was studied during a 2-year clinical investigation. Furthermore, the possibility of shedding of the vaccine strain and the influence of the vaccination on the tuberculin skin test was determined. In addition to standard procedures recently developed diagnostic methods (antibody enzyme-linked immunosorbent assay, interferon-gamma test, polymerase chain reaction) were used. A cell-mediated immune reaction, reflected in an increased, specifically induced, interferon-gamma production developed much earlier (1-2 weeks post-immunization) than humoral immunity (8-16 weeks post-gamma immunization). While the increase in antibody titres was transient, declining to extremely low levels 48-60 weeks post-immunization, cell-mediated immunity remained detectable until the end of the investigation. Spread of the vaccine strain into the body and shedding were never detected during the whole course of the study except for one colon site in one calf. As late as 2 years after vaccine application positive or doubtful skin reactions against M. bovis purified protein derivative were measured, reflecting possible interference of the immunization with the diagnosis of bovine tuberculosis. At the end of the investigation, a positive cell-mediated immune reaction was detected the control animal although clinical, pathological and bacteriological examinations gave no indication for a mycobacterial infection.

[Detection of infection with classical swine fever virus in wild boar: a comparison of different laboratory diagnostic methods]. / Nachweis der Infektion mit dem Virus der Klassischen Schweinepest bei Schwarzwild: ein Vergleich verschiedener labordiagnostischer Methoden.

The aim of this study was to evaluate two commercially available ELISAs for routine diagnosis of classical swine fever virus (CSFV) in wild boar. For this, 222 tissue samples from wild boar were tested in the ELISAs and the results were compared to those obtained using standard methods. First, frozen spleen sections were examined by direct immunofluorescence, and organ suspensions were prepared and tested for CSFV antigen samples were simultaneously examined with the Chekit-ELISA (Dr. Bommeli AG) and the Herd-Chek-ELISA (IDEXX). From the 222 organ suspensions examined in cell culture 102 were positive for CSFV, while no virus could be isolated from the remaining 120 samples. Taking virus isolation as a standard, the Chekit-ELISAs showed a sensitivity of 97%, and the Herd-Chek-ELISA of 72.5%. Both ELISAs revealed high specificities ranging between 99 and 100%. No correlation was found between false negative results obtained in one or in both of the ELISAs with the positive findings in the immunofluorescence test and in the PLA, nor with the clinical reports. Due to the fact that a big number of samples can be processed in a short time with accurate results, the Chekit-ELISA may be considered useful for routine testing of wild boar samples for CSFV.

Contribution to the diagnosis of Johne's disease in cattle. Comparative studies on the validity of Ziehl-Neelsen staining, faecal culture and a commercially available DNA-Probe test in detecting Mycobacterium paratuberculosis in faeces from cattle.

In the present study, 132 selected faecal samples from clinically affected and subclinically infected cattle from dairy herds known to be affected by Johne's disease were investigated for the presence of Mycobacterium paratuberculosis using Ziehl-Neelsen staining, faecal culture and a commercially available DNA-Probe test. The sensitivity was 36.4% for Ziehl-Neelsen staining, 85.6% for faecal culture and 47.7% for the DNA-Probe test. Proving the presence of acid-fast bacteria in 49.3% of the samples from clinically affected cattle and 19.3% of those from subclinically infected cattle, Ziehl-Neelsen staining had the lowest detection rate of the three tests under investigation. Faecal culture showed the highest detection rate of M. paratuberculosis in samples from both clinically affected (84.0%) and subclinically infected (87.7%) animals. The DNA-Probe test showed a positive result in 68.0% of the samples from clinically affected cattle and 21.1% of those from subclinically infected cattle. Ziehl-Neelsen staining proved unreliable in diagnosing Johne's disease. Faecal culture was the most sensitive method for detecting M. paratuberculosis both in clinically affected and subclinically infected cattle. The sensitivity of a commercially available DNA-Probe test has to be enhanced to enable a quick and reliable diagnosis of Johne's disease.

Measuring nursing governance.

BACKGROUND: Despite outcomes studies measuring such variables as job satisfaction, autonomy, professionalism, turnover, leadership styles, and more recently, cost-effectiveness, no consistent relationships have been established between shared governance models and outcomes. OBJECTIVES: To define and develop an instrument to measure the governance of hospital-based nurses. METHODS: The 88-item Index of Professional Nursing Governance (IPNG) was developed to measure professional nursing governance of hospital-based nurses. Psychometric properties were tested with 1,162 registered nurses from 10 hospitals. RESULTS: Content validity after item generation was .95, using Popham's average congruency procedure. Six factors explained 42% of the variance with subscale intercorrelations between .43 and .67. All subscales had a high degree of internal consistency (alphas .87 to .91); test-retest reliability was .77. Construct validity testing showed that scores between shared governance and traditionally governed hospitals were significantly different. A correlation was found between scores on the IPNG and the Hague and Aiken Index of Centralization. CONCLUSIONS: The results of this study support the validity of the 88-item IPNG as a reliable instrument for measuring the distribution of professional nursing governance of hospitals.

[Veterinary medical offer for udder health and milking hygiene: results in Rheinland-Pfalz]. / Amtlicher Tierarzt für Eutergesundheit und Milchhygiene: Arbeitsergebnisse in Rheinland-Pfalz.

A sanitation programme was installed by veterinary medical officers on 261 dairy farms. All farms had been selected because of problems with somatic cell counts in milk for a long period of time. Quarter milk samples were taken from all lactating cows, and management factors (including milking equipment, -hygiene, housing and feeding) were assessed. Contagious mastitis pathogens like Sc. agalactiae and Staphylococci represented the main bacteriological problem in 79% of the herds. Only minor problems were caused by environmental pathogens like esculin-positive Streptococci and Coliforms. The sanitation programmes were mainly based on improvements with milking hygiene and techniques. Recommendations for antibiotic therapy were also given. Farmers and veterinary surgeons were entirely responsible for the implementation of these programmes on the farm. Success was controlled by monitoring somatic cell counts in bulk milk four months before to 18 months after the veterinary medical officer visited the farm. As early as one month after the visit cell counts decreased significantly (p < 0.01) and continued decreasing during the second (p < 0.05) and following months. Somatic cell counts of bulk milk stabilised on a significantly lower level for all over the period monitored.