The suitability of ELISA for the detection of antibodies against Mycobacterium avium ssp. paratuberculosis in bulk milk samples from Rhineland-Palatinate     

Geue L  Köhler H  Klawonn W  Dräger K  Hess RG  Conraths FJ 《Berliner und Münchener tier?rztliche Wochenschrift》2007,120(1-2):67-78

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.  相似文献   

Dietary B vitamin needs of strains of pigs with high and moderate lean growth     

Stahly TS  Williams NH  Lutz TR  Ewan RC  Swenson SG 《Journal of animal science》2007,85(1):188-195

Ten sets of 5 littermate pigs from each of 2 genetic strains were utilized to determine the impact of the dietary concentration of 5 B vitamins (riboflavin, niacin, pantothenic acid, cobalamin, and folacin) on growth from 9 to 28 kg of BW in pigs with high or moderate capacity for lean growth. All pigs (penned individually) were reared via a segregated, early weaning scheme, so that the lean growth potential of each strain could be expressed. The basal diet provided the 5 test vitamins at concentrations of total and estimated bioavailability equivalent to a minimum of 100 and 70%, respectively, of their estimated requirements (NRC, 1998) for 5- to 10-kg pigs. At a BW of 9 +/- 0.9 kg, pigs within each litter were allotted to the basal diet supplemented with sources of the 5 test vitamins equivalent to an additional 0, 100, 200, 300, or 400% (bioavailable) of the NRC requirements. Pigs from the high lean strain consumed less feed (P < 0.05) and gained BW faster (P < 0.02) and more efficiently (P < 0.01) than pigs of the moderate lean strain. In both lean strains, the rate and efficiency of growth were improved (P < 0.01) as dietary B vitamin concentrations were increased. However, the dietary B vitamin concentrations needed to optimize G:F were greater (P < 0.03) in the high (>470% of NRC, 1998) vs. moderate (270%) lean strain. Based on these data, the dietary needs for 1 or more of the 5 B vitamins are greater than current NRC (1998) estimates, particularly in pigs expressing a high rate of lean tissue growth. The greater need for these vitamins is not associated with greater dietary energy intake or body energy accretion rate but is potentially due to shifts in the predominant metabolic pathways.  相似文献   

Field survival of the phytostimulator Azospirillum lipoferum CRT1 and functional impact on maize crop, biodegradation of crop residues, and soil faunal indicators in a context of decreasing nitrogen fertilisation     

Hamdy El Zemrany  Jérôme Cortet  Matthias Peter Lutz  Ezékiel Baudoin  Jacqueline Haurat  Denis Félix  René Bally 《Soil biology & biochemistry》2006,38(7):1712-1726

Heavy nitrogen fertilisation is often implemented in maize cropping systems, but it can have negative environmental effects. Nitrogen-fixing, phytohormone-producing Azospirillum plant growth-promoting rhizobacteria (PGPR) have been proposed as crop inoculants to maintain high yield when decreasing nitrogen fertilisation. In this context, agronomic and ecological effects of the inoculation of maize seeds with the PGPR Azospirillum lipoferum CRT1 were studied in two consecutive years. The inoculant was recovered from maize at 10⁵ CFU g⁻¹ root or higher. Inoculation enhanced root growth and development based on results of root biomass, rooting depth and/or parameters describing root system architecture, and a transient positive effect on shoot height was observed in the first year. Inoculation did not increase yield, but reducing mineral nitrogen fertilisation had only a minor effect on yield. This suggests that the lack of positive effect of the PGPR on yield was due to the fact that the whole field was heavily fertilised in years prior to the start of the experiment. Soil nitrogen levels decreased during the 2 years of the study, and the inoculant had no effect on residual soil nitrogen levels at harvest. Inoculation had no impact on Fusarium symptoms and concentration of the mycotoxin deoxynivalenol in maize kernels, but both were influenced by the interaction between inoculation and nitrogen fertilisation level. Inoculation did not influence meso/macrofaunal soil populations, but had a small but significant effect (smaller than the effect of added nitrogen) on decomposition, nitrogen mineralisation and mesofaunal colonisation of maize leaves (in litter bags). Overall, the ecological impact of seed inoculation with the PGPR A. lipoferum CRT1 was small, and its magnitude was smaller than that of chemical nitrogen fertilisation.  相似文献   

Freilandversuche zur eignung chemisch veränderter weichbraunkohle als bodenverbesserungsmittel bei der landwirtschaftlichen rekultivierung humusfreier kippenböden 1 : Field experiments with n-modified brown coal as humus fertilizer in agricultural revitalisation of humus free dump soils     

Joachim Katzur  Klaus Fischer  Lutz Böcker  Falk Liebner  Rainer Schiene 《Archives of Agronomy and Soil Science》2013,59(6):639-653

Die bodenmeliorativen und ertragssteigernden Effekte des Einsatzes von N-modifizierter Weichbraunkohle als Humusdüngestoff bei der landwirtschaftlichen Rekultivierung humusfreier Kippenböden werden in mehreren Freilandversuchen überprüft. In diesem Beitrag wird über die zu Rekultivierungsbeginn mit W.-Roggen, W.-Weizen, Kartoffeln und Luzerne erzielten Ergebnisse berichtet.  相似文献   

Fatal bovine anaplasmosis in a herd with new genotypes of Anaplasma marginale, Anaplasma ovis and concurrent haemoplasmosis   总被引：1，自引：0，他引：1  

Hornok S  Micsutka A  Fernández de Mera IG  Meli ML  Gönczi E  Tánczos B  Mangold AJ  Farkas R  Lutz H  Hofmann-Lehmann R  de la Fuente J 《Research in veterinary science》2012,92(1):30-35

Haematological and molecular analysis of blood samples was carried out during an outbreak of bovine anaplasmosis in Hungary. Acute disease was observed in five animals, two of which died. Anaplasma-carrier state was diagnosed in 69 (92%) of cattle. Further evaluation of 24 blood samples revealed concurrent infections with Mycoplasma wenyonii and 'CandidatusM. haemobos' in 22 and 21 animals, respectively. In addition, two cows were identified with rickettsaemia. Regarding molecular investigation of potential hard tick vectors, Haemaphysalis inermis and Dermacentor marginatus males collected from the animals were PCR-negative. However, in one pool (out of 18) of Ixodesricinus males, and in six pools (out of 18) of D. reticulatus males the msp4 gene of Anaplasma marginale was detected. In the same I. ricinus pool Anaplasma ovis was also identified. All ticks were negative for haemoplasmas. Anaplasma sequences yielded 97-99% homology to sequences deposited in the Genbank. This is the first report of fatal bovine anaplasmosis associated with divergent A. marginale genotypes and concurrent 'CandidatusM. haemobos' infection, as well as of an A. ovis strain in ticks collected from cattle.  相似文献   

Natural feline coronavirus infection: differences in cytokine patterns in association with the outcome of infection     

Kipar A  Meli ML  Failing K  Euler T  Gomes-Keller MA  Schwartz D  Lutz H  Reinacher M 《Veterinary immunology and immunopathology》2006,112(3-4):141-155

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Shedding of feline leukemia virus RNA in saliva is a consistent feature in viremic cats     

Gomes-Keller MA  Tandon R  Gönczi E  Meli ML  Hofmann-Lehmann R  Lutz H 《Veterinary microbiology》2006,112(1):11-21

The purpose of this investigation was to characterize the shedding pattern of feline leukemia virus (FeLV) RNA in saliva, and to correlate it with the proviral load in whole blood, viral load in plasma, levels of p27 in saliva and plasma, the isolation of infectious FeLV from saliva, and the titers of FeLV-specific antibodies of the IgG and IgA isotypes. We evaluated 24 experimentally FeLV-infected cats for these parameters using real-time RT-PCR and PCR, cell culture assay and sandwich ELISA. We observed that shedding of viral RNA in saliva was a consistent feature in viremic cats. Latently FeLV-infected cats, displaying a very low proviral load, did not shed infectious virus in saliva, but occasionally shed viral RNA. Consequently, salivary shedding of FeLV RNA may not necessarily indicate a transmission potential for susceptible cats. This study also confirmed previous results from our laboratory, showing that a negative result for p27 in plasma, or for viral RNA in plasma or saliva does not exclude FeLV infection, considering that blood cells from those cats contained provirus. We also showed that FeLV RNA and DNA were stable for more than 64 days in saliva samples stored at room temperature. We conclude that the detection of FeLV RNA in saliva may be a useful indicator of viremia, and that the detection of salivary viral RNA by RT-PCR could become a reliable tool for the diagnosis of FeLV infection, which is facilitated by the low invasive method of collection of the samples.  相似文献   

Equine herpesvirus type 1-associated myeloencephalopathy in The Netherlands: a four-year retrospective study (1999-2003)     

Goehring LS  van Winden SC  van Maanen C  Sloet van Oldruitenborgh-Oosterbaan MM 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2006,20(3):601-607

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Degradation of transgenic Cry1Ab DNA and protein in Bt-176 maize during the ensiling process     

Lutz B  Wiedemann S  Albrecht C 《Journal of animal physiology and animal nutrition》2006,90(3-4):116-123

Maize silage is commonly used as feed for farm animals. The aim of this study was to monitor the time-dependent degradation of non-recombinant chloroplast DNA (exemplified by the rubisco gene) in comparison with the recombinant cry1Ab gene in the course of the ensiling process. In parallel, the Cry1Ab protein content and fragment sizes were determined. Fragments of the rubisco (173, 896, 1197, 1753 and 2521 bp) and of the cry1Ab gene (211, 420, 727 and 1,423 bp) were selected to investigate the DNA degradation process. The detection of the Cry1Ab protein was performed using an enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Rubisco gene fragments of 173 bp were still detectable after 61 days, while fragments of 1,197 and 2,521 bp were detectable up to 30 days and on the first day only respectively. Polymerase chain reaction (PCR) analyses revealed that fragments of the cry1Ab gene with sizes of 211 and 420 bp were detectable up to 61 days, fragments with sizes of 727 and 1,423 bp, 30 and 6 days respectively. The ELISA showed a decrease of the Cry1Ab protein in maize silage during the ensiling process. No marked degradation was observed during the first 43 h. Thereafter, a sharp decrease was measured. After 61 days, 23.6 +/- 0.9% of the initial Cry1Ab protein was still detectable. Immunoblotting confirmed the results of the ELISA showing a positive signal of approximately 60 kDa size for 8 days of ensiling; no further immunoactive fragments were detectable by immunoblotting. In conclusion, the ensiling process markedly decreases the presence of long functional cry1Ab gene fragments and full size Cry1Ab protein.  相似文献   

MODELING CARRYING CAPACITIES FOR BIVALVE MOLLUSCS IN OPEN,SUSPENDED-CULTURE SYSTEMS     

Lewis S. Incze  Richard A. Lutz  Ernest True 《Journal of the World Aquaculture Society》1981,12(1):141-155

Standard accounting procedures in ecological energetics would approach the carrying capacity problem by attempting to quantify gross energy needs of a cultivated population and comparing these with “available” energy (seston) flow through the culture area. Inability to distinguish between “available” and “utilizable” particles is only one of several difficulties encountered in formulating energy budgets for large-scale cultivation in natural waters, however. A model based on maintaining critical levels of particle flow through culture areas is offered as an alternative. This approach reduces the problem to terms which are easier to obtain than those involved in equations of energy flow. Application of the model depends on a priori knowledge that seasonal patterns in seston composition and other environmental conditions are conducive to growth of the cultivated species.  相似文献