Tagging and elimination of plasmids in Salmonella of avian origin

This study compared the effectiveness of a number of procedures designed to label and eliminate plasmids that may play a role in virulence in Salmonella. Twenty strains of Salmonella of 9 serovars were subjected to 3 methods for labelling plasmids with transposons. Strains containing labelled and unlabelled plasmids were exposed to physical and chemical curing agents. Plasmids in 9 of 20 strains of Salmonella were tagged by conjugation with a donor Escherichia coli containing a temperature-sensitive RP4 plasmid that carried the Tn1 transposon. Plasmids in 2 of 5 strains of Salmonella were labelled by conjugation with a donor E. coli that contained a F' tslac::Tn5 plasmid. Transduction of Salmonella with a P22 bacteriophage that carried a temperature-sensitive Tn10 transposon resulted in chromosomal insertion of Tn10 in 2 of 10 strains. Use of chemical curing agents resulted in curing of plasmids in only 6 of 17 strains. Two strains were cured by ethidium bromide, two by a combination of ethidium bromide and novobiocin, two by a combination of imipramine and methylene blue, and none by acridine orange, novobiocin, sodium dodecyl sulfate or rifampicin. In contrast, plasmids in 14 of 17 Salmonella strains were eliminated by incubation at 45.5 degrees C.     

An evaluation of 1-hexadecylpyridinium chloride as a decontaminant in the primary isolation of Mycobacterium bovis from bovine lesions

For the primary isolation of Mycobacterium bovis from bovine lesions, 1-hexadecylpyridinium chloride (HPC) at a concentration of 0.75% was as effective as 2% NaOH in controlling the growth of contamination. The advantages of using HPC over NaOH are that it is a rapid one-step procedure not requiring neutralisation with acid, it is less toxic to M. bovis thus increasing isolation rates, and it promotes the earlier appearance of colonies.     

Monoclonal antibodies to canine adenoviruses 1 and 2 that are type-specific by virus neutralization and immunofluorescence

Monoclonal antibodies were produced against the Mirandola strain of canine adenovirus Type 1 (CAV-1) and the Manhattan strain of canine adenovirus Type 2 (CAV-2). The monoclonal antibodies were used in vitro in virus neutralization (VN) assays and in indirect fluorescent antibody (IFA) tests to examine several strains of each virus type. Out of 36 monoclonal antibodies produced against the Mirandola strain, 18 were type-specific for CAV-1 by IFA and 13 of those neutralized the virus in vitro. The other 18 antibodies bound both CAV-1 and CAV-2 by IFA; however, 7 of those specifically neutralized only CAV-1. The 160 monoclonal antibodies made against the Manhattan strain of CAV-2 yielded 77 type-specific antibodies by IFA, of which 39 neutralized only CAV-2 in vitro. The remaining 83 antibodies recognized both CAV-1 and CAV-2 by IFA, with 3 of those neutralizing both viral types. The hemagglutination inhibition (HI) test was performed on a selected monoclonal antibody from each specificity group. Although Type 1 CAV could be readily differentiated from Type 2 CAV by using type-specific monoclonal antibodies in the IFA or VN tests, strains within each type could not be differentiated. This is the first report of neutralizing monoclonal antibodies for a mammalian adenovirus.     

Feline immunodeficiency virus, feline leukaemia virus, Toxoplasma gondii, and intestinal parasitic infections in Taiwanese cats

A population consisting of 70 breeder cats, 43 clinical cases, and 16 feral cats was examined for the presence of Toxoplasma gondii, feline immunodeficiency virus (FIV), and feline leukaemia virus (FeLV). No oocysts of T. gondii were observed in 96 faecal samples; faecal samples were not available from the feral cats. Other intestinal parasites identified included Isospora felis (three cats), Isospora rivolta (five), Dipylidium canium (two), Toxocara cati (four), Toxascaris leonina (one), and Ancylostoma sp. (two). Using a kinetics-based enzyme-linked immunosorbent assay on 117 sera including all the feral cats, nine had antibody to T. gondii antigen, three for antigens to FIV, and seven to the p27 antigen of FeLV. Of the nine cats with antibody to T. gondii, only one was also infected with FIV.     

Recent advances in laboratory models for evaluation of helminth chemotherapy

Extensive research on nematodes of veterinary importance has resulted in new tests based on incubation of parasitic nematodes in culture media, with a variety of detection systems including visual observation, inhibition of aggregation, reduced secretion of acetylcholinesterase and electronic motility measurements. There has been a proliferation of tests to detect benzimidazole resistant nematodes including egg embryonation, egg hatch, larval motility, differences in esterases, and tubulin binding by labelled benzimidazoles. A simple larval development test, which detects resistance to all types of anthelmintics, is likely to become the preferred test for evaluation of resistance in nematodes of grazing animals. By contrast the relative unimportance of trematodes and cestodes from a commercial point of view has resulted in few recent developments in models for evaluating chemotherapy. But the emergence of drug resistant strains of both schistosomes and liver fluke indicates the need for simple tests to detect resistance.     

A biotyping scheme for Salmonella livingstone

Salmonella livingstone is one of the more common salmonella serotypes isolated in the United Kingdom. The characterization of 70 different isolates of S. livingstone using biochemical tests and plasmid profile analysis is described. The isolates could be divided into four groups by their ability to grow in d-tartrate, l-tartrate and Stern's glycerol. Further subdivision was achieved by the use of plasmid profile analysis; 24 of the isolates possessed light plasmids (less than 9.5 MD) and four possessed heavy plasmids (greater than 30 MD). The combination of biotyping and plasmid profile analysis can be used as the basis of a scheme to 'fingerprint' S. livingstone for epidemiological studies.     

Observations on the treatment of necrobacillosis in wallabies

Clinical experience of 73 cases of necrobacillosis in red-necked wallabies (Macropus rufogriseus) over a 6-year period is reviewed. Diagnostic methods and the medical and surgical treatment options are detailed. It is suggested that survival curves are necessary to assess the outcome of treatment.     

Comparative proteomic analysis of mushroom cell wall proteins among the different developmental stages of Pleurotus tuber-regium

Cell wall proteins (CWPs) play a vital role in the development of the different morphological stages including mycelium, fruiting body, and sclerotium in mushrooms which are important human food sources. Using fractionation by detergents and reducing agents, mushroom cell wall fractions from the different developmental stages of Pleurotus tuber-regium (PTR) were prepared. Using one-dimensional gel electrophoresis coupled with LC-MS, there were 103, 91, and 48 noncovalently linked CWPs identified in the cell wall fractions of the PTR mycelium, fruiting body, and sclerotium, respectively. Comparing the CWPs in these cell wall fractions, 19 of them were in common, among which 17 belonged to the functional categories of translation, ribosomal structure, and biogenesis. This is the first study to provide important biochemical insights into the different developmental stages of PTR mediated by CWPs, and the identified CWPs helped to explain the morphological changes of PTR mushrooms during cultivation.     

Creatine phosphokinase in thyroid: isoenzyme composition compared with other tissues

Considerable creatine phosphokinase activity was demonstrated in thyroid glands of several species. After starch gel electrophoresis, thyroid exhibited a creatine phosphokinase isoenzyme pattern different from that of muscle, heart, and brain. The isoenzyme repertory of various tissues and a hybrid enzyme produced from muscle and brain enzyme are consistent with the postulated dimeric structure of creatine phosphokinase.