Macrolide-resistant Campylobacter: the meat of the matter.

The use of macrolide antibiotics in food animals has the potential to select for macrolide-resistant strains of resident bacterial flora. This may include the animal pathogens that are the intended targets of macrolide antibiotic intervention and Campylobacter, common inhabitants of the intestinal tract of food animals that are zoonotic pathogens in man. Such Campylobacter strains are not only resistant to the macrolide antibiotics used in food animals, e.g. tylosin, tilmicosin and tulathromycin, but to the macrolide antibiotics used in human medicine, e.g. erythromycin, azithromycin and clarithromycin, as well. Retail meat is a possible source of Campylobacter and persons consuming the meat derived from macrolide-treated food animals could acquire infections due to macrolide-resistant strains of this organism. Erythromycin is sometimes used to treat human cases of campylobacteriosis and those infected with animal-derived macrolide-resistant Campylobacter may not respond to treatment. The actual risk to human health from the use of macrolide antibiotics in food animals has been difficult to determine because of a lack of information about the macrolide-resistant Campylobacter found on the farm and in the clinic. Recently, however, a plethora of new information has become available on this topic. This review discusses what is currently known about the selection of macrolide-resistant Campylobacter in food animals, the prevalence of macrolide-resistant Campylobacter on retail meat, the prevalence of animal-derived macrolide-resistant Campylobacter in the clinic and the human health consequences associated with macrolide-resistant Campylobacter infection. This work will emphasize the comprehensive body of data generated in Denmark and the US as part of government-sponsored research studies over the last 10 years. These scientific findings may allow informed decisions to be made in the future about how macrolide antibiotics should be used in food animals while still safeguarding human health.

Pyruvate oxidase is a determinant of Avery's rough morphology.

In pioneering studies, Avery et al. identified DNA as the hereditary material (A. T. Avery, C. M. MacLeod, and M. McCarty, J. Exp. Med. 79:137-158, 1944). They demonstrated, by means of variation in colony morphology, that this substance could transform their rough type 2 Streptococcus pneumoniae strain R36A into a smooth type 3 strain. It has become accepted as fact, from modern textbook accounts of these experiments, that smooth pneumococci make capsule, while rough strains do not. We found that rough-to-smooth morphology conversion did not occur in rough strains R36A and R6 when the ability to synthesize native type 2 capsule was restored. The continued rough morphology of these encapsulated strains was attributed to a second, since-forgotten, morphology-affecting mutation that was sustained by R36A during strain development. We used a new genome-PCR-based approach to identify spxB, the gene encoding pyruvate oxidase, as the mutated locus in R36A and R6 that, with unencapsulation, gives rise to rough colony morphology, as we know it. The variant spxB allele of R36A and R6 is associated with increased cellular pyruvate oxidase activity relative to the ancestral strain D39. Increased pyruvate oxidase activity alters colony shape by mediating cell death. R36A requires a wild-type spxB allele for the expression of smooth type 2 morphology but not for the expression of smooth type 3 morphology, the phenotype monitored by Avery et al. Thus, the mutated spxB allele did not impact their use of smooth morphology to identify the transforming principle.

PCR-based ordered genomic libraries: a new approach to drug target identification for Streptococcus pneumoniae.

Described here are the development and validation of a novel approach to identify genes encoding drug targets in Streptococcus pneumoniae. The method relies on the use of an ordered genomic library composed of PCR amplicons that were generated under error-prone conditions so as to introduce random mutations into the DNA. Since some of the mutations occur in drug target-encoding genes and subsequently affect the binding of the drug to its respective cellular target, amplicons containing drug targets can be identified as those producing drug-resistant colonies when transformed into S. pneumoniae. Examination of the genetic content of the amplicon giving resistance coupled with bioinformatics and additional genetic approaches could be used to rapidly identify candidate drug target genes. The utility of this approach was verified by using a number of known antibiotics. For drugs with single protein targets, amplicons were identified that rendered S. pneumoniae drug resistant. Assessment of amplicon composition revealed that each of the relevant amplicons contained the gene encoding the known target for the particular drug tested. Fusidic acid-resistant mutants that resulted from the transformation of S. pneumoniae with amplicons containing fusA were further characterized by sequence analysis. A single mutation was found to occur in a region of the S. pneumoniae elongation factor G protein that is analogous to that already implicated in other bacteria as being associated with fusidic acid resistance. Thus, in addition to facilitating the identification of genes encoding drug targets, this method could provide strains that aid future mechanistic studies.

Streptococcus pneumoniae as a genomics platform for broad-spectrum antibiotic discovery.

Streptococcus pneumoniae is a useful tool for the discovery of broad-spectrum antibiotics because of its genetic malleability and importance as a pathogen. Recent publications of complete chromosomal DNA sequences for S. pneumoniae facilitate rapid and effective use of genomics-based technology to identify essential genes encoding new targets for antibacterial drugs. These methods include computational comparative genomics, gene disruption studies to determine essentiality or identify essential genes, and gene expression analysis using microarrays and gel-based proteomics. We review how genomics has transformed the use of the pneumococcus for the pursuit of new antibiotics, and made it the best species for the identification and validation of new antibiotic targets.