Marc C. Lavoie

Pet Dogs and Chicken Meat as Reservoirs of Campylobacter spp. in Barbados

ABSTRACT 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.

Isolation and Genetic Characterization of Avian Influenza Viruses and a Newcastle Disease Virus from Wild Birds in Barbados: 2003–2004

Abstract Zoonotic transmission of an H5N1 avian influenza A virus to humans in 2003–present has generated increased public health and scientific interest in the prevalence and variability of influenza A viruses in wild birds and their potential threat to human health. Migratory waterfowl and shorebirds are regarded as the primordial reservoir of all influenza A viral subtypes and have been repeatedly implicated in avian influenza outbreaks in domestic poultry and swine. All of the 16 hemagglutinin and nine neuraminidase influenza subtypes have been isolated from wild birds, but waterfowl of the order Anseriformes are the most commonly infected. Using 9-to-11-day-old embryonating chicken egg culture, virus isolation attempts were conducted on 168 cloacal swabs from various resident, imported, and migratory bird species in Barbados during the months of July to October of 2003 and 2004. Hemagglutination assay and reverse transcription–polymerase chain reaction were used to screen all allantoic fluids for the presence of hemagglutinating agents and influenza A virus. Hemagglutination positive–influenza negative samples were also tested for Newcastle disease virus (NDV), which is also found in waterfowl. Two influenza A viruses and one NDV were isolated from Anseriformes (40/168), with isolation rates of 5.0% (2/40) and 2.5% (1/40), respectively, for influenza A and NDV. Sequence analysis of the influenza A virus isolates showed them to be H4N3 viruses that clustered with other North American avian influenza viruses. This is the first report of the presence of influenza A virus and NDV in wild birds in the English-speaking Caribbean.

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

BackgroundStreptococcus 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.ResultsMutacin 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.ConclusionA 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.

Production, purification, sequencing and activity spectra of mutacins D-123.1 and F-59.1

BackgroundThe increase in bacterial resistance to antibiotics impels the development of new anti-bacterial substances. Mutacins (bacteriocins) are small antibacterial peptides produced by Streptococcus mutans showing activity against bacterial pathogens. The objective of the study was to produce and characterise additional mutacins in order to find new useful antibacterial substances.ResultsMutacin F-59.1 was produced in liquid media by S. mutans 59.1 while production of mutacin D-123.1 by S. mutans 123.1 was obtained in semi-solid media. Mutacins were purified by hydrophobic chromatography. The amino acid sequences of the mutacins were obtained by Edman degradation and their molecular mass was determined by mass spectrometry. Mutacin F-59.1 consists of 25 amino acids, containing the YGNGV consensus sequence of pediocin-like bacteriocins with a molecular mass calculated at 2719 Da. Mutacin D-123.1 has an identical molecular mass (2364 Da) with the same first 9 amino acids as mutacin I. Mutacins D-123.1 and F-59.1 have wide activity spectra inhibiting human and food-borne pathogens. The lantibiotic mutacin D-123.1 possesses a broader activity spectrum than mutacin F-59.1 against the bacterial strains tested.ConclusionMutacin F-59.1 is the first pediocin-like bacteriocin identified and characterised that is produced by Streptococcus mutans. Mutacin D-123.1 appears to be identical to mutacin I previously identified in different strains of S. mutans.

Influenza A Virus Surveillance of Migratory Waterfowl in Barbados, West Indies

Migratory waterfowl and shorebirds are regarded as the primordial reservoir of all influenza A viral subtypes and have been repeatedly implicated in avian influenza outbreaks in domestic poultry and swine. Avian influenza zoonotic transmission events have generated increased public health-related and scientific interest in the prevalence and variability of influenza viruses in feral birds and their potential role in human health. All of the 16 hemagglutinin (HA) and 9 neuraminidase (NA) influenza subtypes have been isolated from approximately 90 species of wild birds with the widest distribution in waterfowl of the Order Anseriformes. Barbados is the most easterly of the Caribbean islands, located at 13◦ 6′ N, 59◦ 37′ W, having a tropical climate with temperatures ranging from 22◦C to 31◦C.