Reduction of antibiotics in the world Speksnijder, David C; Wagenaar, Jaap A
Tijdschrift voor diergeneeskunde,
2013-Sep-01, Letnik:
138, Številka:
9
Magazine Article
To compare sensitivity and specificity of various polymerase chain reaction (PCR) assays for detection of Leptospira borgpetersenii serovar hardjo in bovine urine and to compare results of the ...optimal PCR assay with results of immunofluorescence, nucleic acid hybridization, and bacteriologic culture.
6 heifers.
Heifers were exposed to serovar hardjo type hardjo-bovis by conjunctival instillation of 10(6) leptospires on 3 successive days. Urine samples were collected before and after infection. Sensitivity and specificity of 5 PCR assays were compared, to determine the optimal assay for use with bovine urine samples. The optimal PCR assay was then compared with results of bacteriologic culture, nucleic acid hybridization, and immunofluorescence.
A PCR assay with the best combination of specificity (100%) and sensitivity (91%) was selected for comparison with the other diagnostic tests. Sensitivity for nucleic acid hybridization was 55%, whereas sensitivity for bacteriologic culture and immunofluorescence was 89 to 93%.
Bacteriologic culture, PCR, and immunofluorescence were sensitive for detection of L borgpetersenii serovar hardjo type hardjo-bovis in urine specimens of cattle, but a single technique was not the most sensitive for each animal tested. Therefore, the use of 2 techniques in combination is warranted for maximal sensitivity for diagnosis.
A European Union‐wide baseline survey on Campylobacter in broiler batches and on Campylobacter and Salmonella on broiler carcasses was carried out in 2008. In the Salmonella sub‐survey a total of ...10,035 broiler batches were sampled from 561 slaughterhouses in 26 European Union Member States and two countries not belonging to the European Union. From each randomly selected batch one carcass was collected after chilling and the neck skin together with the breast skin was examined for the presence of Salmonella. Multivariable regression analysis showed that the risk for Salmonella‐contaminated carcasses increased with the slaughter capacity of the slaughterhouse and with processing of the carcass later during the day. The risk for contamination of carcasses with Salmonella varied significantly between countries and between slaughterhouses within a country, even when other associated factors were accounted for. The Salmonella serovar distribution varied among Member States, many of them having a specific distribution pattern of their own and no specific serovar was predominant in all countries in the survey. The most commonly reported serovars were S. Infantis, S. Enteritidis and S. Typhimurium. Many of the reported serovars seem to have become well‐established in broiler production.
Monoclonal antibodies (MAbs) recognizing external epitopes of the human MDR1 P-glycoprotein have been used both for the detection of multidrug-resistant cells and as specific inhibitors of ...P-glycoprotein-mediated multidrug resistance. Using a panel of recently developed transfected or transgenic cell lines containing variants of the human MDR1 and MDR3 P-glycoproteins, we have compared the specificity and binding properties of the previously isolated MAbs MRK16, HYB-241, UIC2 and 4E3, and of the newly isolated MAb 7G4. The removal of 1, 2 or all 3 of the N-glycosylation sites present in the first extracellular loop of MDR1 P-glycoprotein did not significantly affect the binding of these MAbs. In contrast, 20 amino acid deletion in the first extracellular loop of MDR1 P-glycoprotein completely abolished binding of UIC2, whereas the binding of all other MAbs was hardly affected. None of the MAbs tested bound detectably to cell lines containing a high level of the human MDR3 P-glycoprotein. The differences in the binding specificity between UIC2 and the other tested antibodies parallel the reported functional differences in the ability of these antibodies to inhibit P-glycoprotein-mediated drug efflux.
The human MDR3 (or MDR2) P-glycoprotein is probably involved in the transport of phospholipids from liver hepatocytes into bile (Smit et al. (1993) Cell 75, 451–462). In accordance with this ...function,
MDR3 is highly expressed in human liver, but lower mRNA levels were also found in adrenal, heart, muscle and cells of the B-cell compartment. We have cloned and analyzed the
MDR3 promoter region. It is GC-rich, and contains neither a TATA nor a CART box, but it does contain multiple putative SP1 binding sites, features also found in so-called housekeeping genes. RNase protection and primer extension analyses indicate that the
MDR3 gene has multiple transcription start sites in a GC-rich region with considerable homology to the putative mouse
mdr2 promoter. A 3 kb genomic fragment containing the
MDR3 start sites directs transcription of a chloramphenicol acetyltransferase (CAT) reporter gene upon transient transfection in the human hepatoma cell line HepG2. This transcription is orientation dependent, and stimulated by a SV40 enhancer, indicating that the 3 kb insert contains the core promoter elements of the
MDR3 gene. The promoter region contains several consensus sequences where known or putative liver-specific (C/EBP, HNF5) or lymphoid specific (Pu.1, ets-1) transcription factors may bind.
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