The viruses of bacteria – bacteriophages – were discovered 20 years after the discovery of viruses. However, this was mainly the bacteriophage research that, after the first 40 years, yielded the ...modern concept of the virus and to large extent formed the grounds of the emerging molecular genetics and molecular biology. Many specific aspects of the bacteriophage research history have been addressed in the existing publications. The integral outline of the events that led to the formation of the key concepts of modern virology is presented in this review. This includes the opposition of F. d’Herelle and J. Bordet viewpoints over the nature of the bacteriophage, the history of lysogeny discovery and of determination of the mechanisms of underlying this phenomenon, the work of the Phage group led by M. Delbruck in USA, the development of the genetic analysis of bacteriophages and other research that eventually led to emergence of the concept of the virus (bacteriophage) as a transmissive genetic program. The review also covers a brief history of early applications of the bacteriophages such as phage therapy and phage typing.
Bacteriophage (phage) is regarded as an antimicrobial alternative for
in food production. However, the development of phage resistance to the host is a main concern for the phage application. This ...study characterized the phage CP39 and investigated the phage resistance of CP39 in
NCTC12662. We determined that phage CP39 belonged to the
family by the WGS and phylogenetic analysis. Phage CP39 was confirmed as a capsular polysaccharide (CPS)-dependent phage by primary
phage typing. It was further confirmed that the phage could not be adsorbed by the acapsular mutant Δ
but showed the same lytic ability in both the wild-type strain NCTC 12662 and the Δ
mutant lacking motile flagella filaments. We further determined that the
gene encoding CDP-glycerol:poly (glycerophosphate) glycerophosphotransferase (CGPTase) in the CPS loci was related to phage CP39 adsorption by SNP analysis and observed a rapid development of phage resistance in NCTC 12662 during the phage infection. Furthermore, we observed a high mutation frequency of
(32%), which randomly occurred in nine different sites in the gene according to colony PCR sequencing. The mutation of the
gene could cause the phase variable expression of non-functional protein and allow the bacteria against the phage infection by modifying the CPS. Our study confirmed the
gene responsible for the CPS-phage adsorption for the first time and demonstrated the phase variable expression as a main mechanism for the bacteria to defend phage CP39. Our study provided knowledge for the evolutionary adaption of bacteria against the bacteriophage, which could add more information to understand the phage resistance mechanism before applying in the industry.
Listeria monocytogenes is an important food-borne pathogen and is widely tested for in food, environmental and clinical samples. Identification traditionally involved culture methods based on ...selective enrichment and plating followed by the characterization of
Listeria spp. based on colony morphology, sugar fermentation and haemolytic properties. These methods are the gold standard; but they are lengthy and may not be suitable for testing of foods with short shelf lives. As a result more rapid tests were developed based on antibodies (ELISA) or molecular techniques (PCR or DNA hybridization). While these tests possess equal sensitivity, they are rapid and allow testing to be completed within 48 h. More recently, molecular methods were developed that target RNA rather than DNA, such as RT-PCR, real time PCR or nucleic acid based sequence amplification (NASBA). These tests not only provide a measure of cell viability but they can also be used for quantitative analysis. In addition, a variety of tests are available for sub-species characterization, which are particularly useful in epidemiological investigations. Early typing methods differentiated isolates based on phenotypic markers, such as multilocus enzyme electrophoresis, phage typing and serotyping. These phenotypic typing methods are being replaced by molecular tests, which reflect genetic relationships between isolates and are more accurate. These new methods are currently mainly used in research but their considerable potential for routine testing in the future cannot be overlooked.
Surveillance of Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium) is generally considered to benefit from molecular techniques like multiple-locus variable-number of tandem ...repeats analysis (MLVA), which allow earlier detection and confinement of outbreaks. Here, a surveillance study, including phage typing, antimicrobial susceptibility testing and the in Europe most commonly used 5-loci MLVA on 1,420 S. Typhimurium isolates collected between 2010 and 2012 in Belgium, was used to evaluate the added value of MLVA for public health surveillance. Phage types DT193, DT195, DT120, DT104, DT12 and U302 dominate the Belgian S. Typhimurium population. A combined resistance to ampicillin, streptomycin, sulphonamides and tetracycline (ASSuT) with or without additional resistances was observed for 42.5% of the isolates. 414 different MLVA profiles were detected, of which 14 frequent profiles included 44.4% of the S. Typhimurium population. During a serial passage experiment on selected isolates to investigate the in vitro stability of the 5 MLVA loci, variations over time were observed for loci STTR6, STTR10, STTR5 and STTR9. This study demonstrates that MLVA improves public health surveillance of S. Typhimurium. However, the 5-loci MLVA should be complemented with other subtyping methods for investigation of possible outbreaks with frequent MLVA profiles. Also, variability in these MLVA loci should be taken into account when investigating extended outbreaks and studying dynamics over longer periods.
The aim of the study was to investigate the potential spread of the previously described multidrug-resistant (MDR) Hungarian clone of Salmonella Infantis of broiler origin in Europe. Therefore, 76 ...strains isolated between 2004 and 2009 from raw meat and fecal samples of broiler origin in nine European countries – including Hungary – were examined by phage typing, antimicrobial resistance typing, pulsed-field gel electrophoresis (PFGE) profile and plasmid analysis. The strains could be divided into two large PFGE clusters with 92% similarity. Cluster A (n=39) contained 15 German, seven Italian, five British, five Polish isolates, one Austrian and one Hungarian isolate. Five Hungarian isolates that were isolated prior to the appearance of the MDR clone also belonged to this cluster. Strains of this cluster comprised seven pulsotypes and 14 different phage types and were mostly susceptible to the 12 antimicrobials tested. Cluster B (n=33) contained all but one of the recent Austrian (n=14) and of Hungarian (n=9), six Polish, one Italian and one German as well as the single Turkish and the Romanian strains, representing five pulsotypes. The strains of this cluster were mostly PT213, with MDR (nalidixic acid-streptomycin-sulphomamide-tetracycline), and the carriage of the same class 1 integron located on a large plasmid (>168kb) characteristic of the current predominant Hungarian clone. The results suggest that two large related clusters (A and B) of S. Infantis isolates can be found in various European countries, of which Austrian and Polish MDR clones of cluster B are identical with, or closely related to, the dominant Hungarian clone. The emergence of a few dominant MDR S. Infantis clones in broilers reported here, raises the possibility that further dissemination of such clones in broilers and in broiler meat may occur and represent a potential threat to public health in Europe.
► Clonality and spread of multidrug-resistant, and of sensitive strains of S. Infantis is different. ► European Salmonella Infantis multidrug resistant clones of broiler origin are closely related. ► More detailed characterization of Salmonella serovars and clonal lines in poultry and man is needed.
The study aimed to identify sources of campylobacter in 10 housed broiler flocks from three United Kingdom poultry companies. Samples from (i) the breeder flocks, which supplied the broilers, (ii) ...cleaned and disinfected houses prior to chick placement, (iii) the chickens, and (iv) the environments inside and outside the broiler houses during rearing were examined. Samples were collected at frequent intervals and examined for Campylobacter spp. Characterization of the isolates using multilocus sequence typing (MLST), serotyping, phage typing, and flaA restriction fragment length polymorphism typing was performed. Seven flocks became colonized during the growing period. Campylobacter spp. were detected in the environment surrounding the broiler house, prior to as well as during flock colonization, for six of these flocks. On two occasions, isolates detected in a puddle just prior to the birds being placed were indistinguishable from those colonizing the birds. Once flocks were colonized, indistinguishable strains of campylobacter were found in the feed and water and in the air of the broiler house. Campylobacter spp. were also detected in the air up to 30 m downstream of the broiler house, which raises the issue of the role of airborne transmission in the spread of campylobacter. At any time during rearing, broiler flocks were colonized by only one or two types determined by MLST but these changed, with some strains superseding others. In conclusion, the study provided strong evidence for the environment as a source of campylobacters colonizing housed broiler flocks. It also demonstrated colonization by successive campylobacter types determined by MLST during the life of a flock.
To determine the prevalence of Salmonella enterica serovars in chicken carcasses in slaughterhouses in Spain and to examine genotypic relations among these serovars. A total of 336 chicken carcasses ...were collected from six slaughterhouses in Northwestern Spain. Salmonellae were isolated (ISO-6579-1993), serotyped, phage-typed, ribotyped and antibiotyped against 20 antibiotics. Salmonella strains were detected in 60 (17·9%) carcasses. Isolates belonged to nine different serotypes, with Salm. Enteritidis being the most common. Three strains (5%) were resistant to one antibiotic and 24 (40%) were multi-resistant (to more than one antibiotic). The most frequently encountered resistances were to sulphamides, fluoroquinolones and tetracycline. Ribotyping was able to differentiate isolates of the same serotype and phage type. The Salmonella serotypes and phage types detected are among those most frequently associated with human diseases in Spain. The large percentage of antimicrobial resistant strains is a matter for concern. A high genetic relationship between strains from different slaughterhouses was found. This study provides detailed information about Salmonella isolates from poultry in Spain. It emphasizes the importance of controlling this pathogen in poultry products, and suggests the need for more prudent use of antibiotics.
Salmonella is an important foodborne pathogen worldwide and is commonly isolated from pigs and pig products in Ireland. Pigs, reared in an environment free of Salmonella spp. or with low levels of ...infection, may acquire infection or become contaminated during transport, lairage or post-slaughter. The main objective of this study was to determine the role of the abattoir as a potential factor that contributes to the dissemination of Salmonella spp. in slaughter pigs from herds with a low Salmonella seroprevalence (≤10%). A total of 128 pigs from eight herds were monitored from farm through the slaughter process in three separate abattoirs. The prevalence of Salmonella spp. was determined in samples collected from trucks, lairage pens and the slaughterline before pigs entered, from pigs after slaughter (caecal contents and ileocaecal lymph nodes) and carcass surfaces post-evisceration. Isolates were characterised by serotype, phage type and pulsed-field gel electrophoresis (PFGE) patterns. Of the swabs taken from the trucks, lairage and slaughterline, before the pigs entered, 4.3% (3/70), 80% (64/80) and 16.7% (4/27) were positive for Salmonella spp., respectively. The proportion of pigs showing serological evidence of infection was 3.1% (4/128). Salmonella spp. were isolated from the ileocaecal lymph nodes and caecal contents of 14.8% (19/128) and 11.7% (15/128) of pigs, respectively, and 13/128 (10.2%), 5/128 (3.9%), 2/111 (1.8%) and 8/111 (7.2%) carcass swabs pre wash, post wash, post chill and belly-strip samples, respectively, were Salmonella-positive. There was only slight agreement between serological and bacteriological data at the pig level. Salmonella isolates from 45% of all positive pig samples and 82% of positive carcass samples were indistinguishable, based on PFGE patterns, from salmonellae isolated from the lairage and slaughterline. Based on these results it is concluded that the lairage and the slaughterline provide a substantial source for Salmonella contamination of pigs and carcasses.
Salmonella enterica subspecies enterica serovar Enteritidis (SE) is one of the most common causes of human salmonellosis and in Canada currently accounts for over 40% of human cases. Reliable ...subtyping of isolates is required for outbreak detection and source attribution. However, Pulsed-Field Gel Electrophoresis (PFGE), the current standard subtyping method for Salmonella spp., is compromised by the high genetic homogeneity of SE. Multiple Locus Variable Number Tandem Repeat Analysis (MLVA) was introduced to supplement PFGE, although there is a lack of data on the ability of MLVA to subtype Canadian isolates of SE. Three subtyping methods, PFGE, MLVA and phage typing were compared for their discriminatory power when applied to three panels of Canadian SE isolates: Panel 1: 70 isolates representing the diversity of phage types (PTs) and PFGE subtypes within these PTs; Panel 2: 214 apparently unrelated SE isolates of the most common PTs; and Panel 3: 27 isolates from 10 groups of epidemiologically related strains.
For Panel 2 isolates, four MLVA subtypes were shared among 74% of unrelated isolates and in Panel 3 isolates, one MLVA subtype accounted for 62% of the isolates. For all panels, combining results from PFGE, MLVA and PT gave the best discrimination, except in Panel 1, where the combination of PT and PFGE was equally as high, due to the selection criteria for this panel. However, none of these methods is sufficiently discriminatory alone for reliable outbreak detection or source attribution, and must be applied together to achieve sufficient discrimination for practical purposes. Even then, some large clusters were not differentiated adequately. More discriminatory methods are required for reliable subtyping of this genetically highly homogeneous serovar. This need will likely be met by whole genome sequence analysis given the recent promising reports and as more laboratories implement this tool for outbreak response and surveillance.
•MLVA, PFGE and phage typing were evaluated for subtyping Salmonella Enteritidis.•MLVA improved isolate discrimination but many isolates shared a common MLVA type.•Combining the three methods gave the best results but was not always adequate for subtyping.
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