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Marc C Lavoie

Publications and source records attributed to Marc C Lavoie.

9 recordsLinked to original sources

Mutacin H-29B is identical to mutacin II (J-T8).

BACKGROUND: Streptococcus mutans produces bacteriocins named mutacins. Studies of mutacins have always been hampered by the difficulties in obtaining active liquid preparations of these substances. Some of them were found to be lantibiotics, defined as bacterial ribosomally synthesised lanthionine-containing peptides with antimicrobial activity. The goal of this study was to produce and characterize a new mutacin from S. mutans strain 29B, as it shows a promising activity spectrum against current human pathogens. RESULTS: Mutacin H-29B, produced by S. mutans strain 29B, was purified by successive hydrophobic chromatography from a liquid preparation consisting of cheese whey permeate (6% w/v) supplemented with yeast extract (2%) and CaCO3 (1%). Edman degradation revealed 24 amino acids identical to those of mutacin II (also known as J-T8). The molecular mass of the purified peptide was evaluated at 3246.08 +/- 0.1 Da by MALDI-TOF MS. CONCLUSION: A simple procedure for production and purification of mutacins along with its characterization is presented. Our results show that the amino acid sequence of mutacin H-29B is identical to the already known mutacin II (J-T8) over the first 24 residues. S. mutans strains of widely different origins may thus produce very similar bacteriocins.

Amino Acid Sequence↗

Identification of Streptococcus iniae by commercial bacterial identification systems.

The fish pathogen Streptococcus iniae cannot be identified by most commercial bacterial identification systems. The results presented here indicate that over 70% of our S. iniae isolates have been identified using the Biolog(R) GP microplate panels and Microlog(R) database. The isolates were confirmed as S. iniae by specific PCR methods and have been found to conform to the result obtained with the type strain S. iniae ATCC 29178.

Animals↗

Human Campylobacter-associated enteritis on the Caribbean island of Barbados.

A longitudinal study of the incidence of Campylobacter enteritis in Barbados was undertaken from January 2000 to August 2003. Diarrheal stools received by the central public health laboratory were cultured for Campylobacter. The number of reported Campylobacter cases exceeded those of Shigella but were less than those of Salmonella, and increased steadily with each year. Isolates from stools were mainly C. jejuni (63.6%) and C. coli (31.8%). The highest isolation rate was found in children 1-4 years of age (40.8%), followed by infants less than 1 year of age (16.9%) and those 5-9 years of age (11.3%). The number of reported cases was higher in March, from June to August, and in November and December. There was no correlation between incidence and either rainfall, temperature, or humidity. Further epidemiologic investigation of this disease is needed to evaluate risk factors for Campylobacter infection and determine routes of transmission in Barbados.

Adolescent↗

In vivo activity of mutacin B-Ny266.

OBJECTIVES: The objective of this study was to assess the in vivo activity of mutacin B-Ny266 (a bacteriocin produced by Streptococcus mutans) in order to eventually use it as an antibiotic. METHODS: Intraperitoneal infection was induced with a methicillin-susceptible Staphylococcus aureus strain in mice. Some of the mice were simultaneously injected intraperitoneally with mutacin B-Ny266, some with the vehicle only and some with vancomycin. RESULTS: While there was 70 and 100% mortality in the control groups of mice, no mortality was observed in the mice injected with vancomycin or mutacin B-Ny266. CONCLUSIONS: The results presented here show, for the first time, the in vivo efficacy of a mutacin (B-Ny266) against an experimental intraperitoneal infection by S. aureus in a mouse model.

Animals↗

Pet dogs and chicken meat as reservoirs of Campylobacter spp. in Barbados.

Campylobacter spp. are the second most common pathogen isolated from stools of patients with gastroenteritis in Barbados. The aim of this study was to identify reservoirs of Campylobacter and the likely source(s) of human infection. Fecal specimens from 596 animals and 311 samples of animal food products were analyzed for the presence of Campylobacter spp. by standard culture techniques. Isolates were characterized by conventional phenotypic tests, confirmed by latex agglutination and PCR with genus-specific primers, and identified by the use of species-specific primers. High isolation rates were obtained for chickens (94.2%), pigs (90.5%), dogs (46.9%), cats (37.3%), and wild birds (39.3%). Campylobacter was also recovered from monkeys (17.1%) and sheep (4.2%) but not from cows. Chicken meat was frequently contaminated with Campylobacter (58.4%), but its recovery from other animal food products was rare. Campylobacter jejuni was the most commonly identified species in humans (63.6%), chickens (86.6%), dogs (51.5%), and chicken meat (79.8%). Porcine isolates were predominantly C. coli (98.4%), while cats harbored mainly C. upsaliensis and C. helveticus. Wild birds alone carried urease-positive thermophilic campylobacters. C. jejuni and C. coli isolates from different sources were compared with isolates from humans by randomly amplified polymorphic DNA typing with the primers OPA 11 and HLWL 85. Genotyping revealed similarities between isolates from chicken meat and those from humans and could not distinguish between two clinical isolates and four canine strains. Our results suggest that dogs are significant reservoirs of Campylobacter and contribute to human enteric infections and that chicken meat is a likely vehicle for the transmission of campylobacters to humans.

Animals↗

Improved methods for mutacin detection and production.

Studies of mutacins have always been hampered by the difficulties in obtaining active liquid preparations of these substances. In order to be commercially produced, good mutacin yields have to be obtained, preferably in inexpensive media. The results presented here indicate that mutacins can be produced in supplemented cheese whey permeate. The influence of carbon and nitrogen supplements on mutacin production varied according to the producer strain. The use of CaCO3 as a buffer in batch cultures resulted in improved yields of mutacin in the supernatants. Antimicrobial activity assays were improve by acidification of the diluent (pH 2) and were less variable in peptone water (0.5%). The culture medium consisting of cheese whey permeate (6% w/v), yeast extract (2% w/v) and CaCO3 (1% w/v) was found to be an inexpensive medium for the efficient production of mutacins.

Bacteriocins↗

Diversity of Streptococcus mutans bacteriocins as confirmed by DNA analysis using specific molecular probes.

Mutacin-producing strains have been classified into 24 groups (designated by letters A to X) by similarity in activity spectra and cross-immunity. Similarity in primary structure among these groups can be revealed using DNA hybridization. The amino acid sequences of four mutacins (B-Ny266, 1140/mutacin III and mutacin II) were used to design two DNA probes in order to detect similar genes among groups of Streptococcus mutans strains demonstrating inhibitory activity. In addition to the appropriate parent strain, each probe hybridized with the total DNA from only two out of the 24 mutacin group type strains. Thus, the remaining 18 groups of strains produce mutacins that differ from the mutacins sequenced to date. In order to explore the similarity between genes coding for mutacins B-Ny266 and JH1140, the group B specific probe was utilized to detect a DNA fragment of 1.9 kb in the genome of S. mutans strain Ny266. The sequence of the cloned fragment codes for three open reading frames (lanA, lanA' and lanB) similar to those of strains JH1140 and UA787. The gene lanA' is strongly similar to the structural gene lanA (67%), but only one RNA transcript of about 300 bases was detected by Northern hybridization using the lanA-lanA' probe. Transcription of lanA alone was verified by RT-PCR.

Amino Acid Sequence↗

Replacement of trifluoroacetic acid with HCl in the hydrophobic purification steps of pediocin PA-1: a structural effect.

Trifluoroacetic acid (TFA) is a purification contaminant associated with pediocin PA-1 that interferes with Fourier transform infrared spectroscopy structural analysis. As revealed by circular dichroism, its presence affects the structural folding of pediocin. Consequently, we propose a new pediocin PA-1 purification procedure using HCl instead of TFA in all of the hydrophobic steps. This procedural change does not affect the purification yield or the amount of pediocin PA-1 purified. Furthermore, removing HCl, as opposed to TFA, after purification is an easier procedure to carry out. In fact, the removal of TFA requires more experimentation and results in protein loss. Thus, HCl is a good alternative to TFA in pediocin PA-1 purification and can be extended to the purification of other proteins. We also show that TFA-induced structural modifications do not significantly affect the antimicrobial activity of pediocin PA-1.

Bacteriocins↗