Differentiation of Staphylococcus aureus from freshly slaughtered poultry and strains 'endemic'to processing plants by biochemical and physiological tests

A comparison was made of 27 'endemic' strains of Staphylococcus aureus and 35 strains from freshly slaughtered birds, isolated at five commercial slaughterhouses processing chickens or turkeys. Of 112 biochemical and physiological tests used, 74 gave results which differed among the strains. Cluster analysis revealed several distinct groupings which were influenced by strain type, processing plant and bird origin; these included a single group at the 72% level of similarity containing most of the 'endemic' strains. In comparison with strains from freshly slaughtered birds, a higher proportion of 'endemic' strains produced fibrinolysin, aP-glucosidase and urease and were β-haemolytic on sheep-blood agar. The 'endemic' type also showed a greater tendency to coagulate human but not bovine plasma, and to produce mucoid growth and clumping. The last two properties, relevant to colonization of processing equipment, were less evident in heart infusion broth than in richer media or process water collected during defeathering of the birds.     

Plasmid profiles as indicators of the source of contamination of Staphylococcus aureus endemic within poultry processing plants.

A total of 530 strains of Staphylococcus aureus were isolated from the defeathering machinery of a chicken processing plant and from neck skin samples of carcasses at different stages of processing in two visits 4 weeks apart. Eleven different plasmid profiles were detected in the isolates, eight being common to both visits. The plasmid profiles of the strains forming the majority of the population on the freshly slaughtered birds were rarely present in the strains isolated from the pluckers (except at the entry to the first plucker) and were present in only a small proportion of the strains isolated from carcasses after plucking. However, the profiles from the strains isolated from the pluckers on both visits were different from those forming the majority of the population on the incoming birds but formed the major part of the carcass flora after plucking, suggesting that such strains were endemic. These strains were found as a small proportion of the isolates made from the incoming birds, suggesting that this was the route by which the endemic strains were introduced into the plant. Such endemic strains exhibited a clumping growth, even in liquid shake culture, which may have made it easier for them to become established on the pluckers and to resist cleaning and disinfection. This clumping phenotype was correlated with the presence of a 7.5-megadalton plasmid.     

Plasmid profiles as indicators of the source of contamination of Staphylococcus aureus endemic within poultry processing plants

A total of 530 strains of Staphylococcus aureus were isolated from the defeathering machinery of a chicken processing plant and from neck skin samples of carcasses at different stages of processing in two visits 4 weeks apart. Eleven different plasmid profiles were detected in the isolates, eight being common to both visits. The plasmid profiles of the strains forming the majority of the population on the freshly slaughtered birds were rarely present in the strains isolated from the pluckers (except at the entry to the first plucker) and were present in only a small proportion of the strains isolated from carcasses after plucking. However, the profiles from the strains isolated from the pluckers on both visits were different from those forming the majority of the population on the incoming birds but formed the major part of the carcass flora after plucking, suggesting that such strains were endemic. These strains were found as a small proportion of the isolates made from the incoming birds, suggesting that this was the route by which the endemic strains were introduced into the plant. Such endemic strains exhibited a clumping growth, even in liquid shake culture, which may have made it easier for them to become established on the pluckers and to resist cleaning and disinfection. This clumping phenotype was correlated with the presence of a 7.5-megadalton plasmid.     

Chlorine resistance of Staphylococcus aureus isolated from turkeys and turkey products

When tested in phosphate buffer at pH 7·0, strains of Staphylococcus aureus isolated from turkeys immediately after commercial slaughter were reduced in viability by at least 10000-fold following exposure to 1 mg/1 of free available chlorine for 2 min at 20°C. In contrast, corresponding isolates from samples taken later in the process, and apparently derived from strains capable of colonizing processing equipment ('endemic' strains), were reduced only ca 100-fold; some of these strains survived treatment with 2 mg/1 of free chlorine, when tested under the same conditions. The greater resistance of 'endemic' strains to chlorine may contribute to their persistence in the processing plant.     

Antibiotic Resistance among Escherichia coli O-serotypes from the Gut and Carcasses of Commercially Slaughtered Broiler Chickens: a Potential Public Health Hazard

Presumptive coliform counts and the distribution of Escherichia coli O-serotypes were investigated in chicken rectal contents (175) abdominal cavities (152) and on the carcasses of 44 which had been commercially raised, slaughtered and prepared for sale. Large numbers of E. coli resistant to at least one antibacterial agent were found at each site; comparison of the O-serotypes suggested heavy contamination of the carcass with strains from the gut. The range of O-serotypes was similar to that found in man and some public health implications of cross-infection particularly by handling uncooked birds in the kitchen, are discussed.     

Impact of transport crate reuse and of catching and processing on Campylobacter and Salmonella contamination of broiler chickens

The influence of transport, catching, and processing on contamination of broiler chickens with Salmonella and Campylobacter was investigated. Transport crates were reused with high frequency and were often still contaminated with Salmonella and Campylobacter when they arrived at the farm despite the fact that they were washed at the factory, and thus they were a potential route of infection. These organisms contaminated the feathers of previously Campylobacter- and Salmonella-negative birds going to the processing plant and were isolated from processed carcasses, albeit at a low frequency. The Campylobacter types which were the predominant organisms on the live birds when they arrived at the processing plant were not necessarily the types that were most frequently isolated from processed carcasses. This finding may reflect cross-contamination that occurred during processing or differences in the tolerance of the strains to the hostile environments that the bacteria experienced. The process of catching and putting the birds in crates significantly increased the chance of contamination with Campylobacter (P < 0.001).     

Isolation of Vibrio cholerae from aquatic birds in Colorado and Utah

Vibrio cholerae was isolated from cloacal swabs and freshly voided feces collected from 20 species of aquatic birds in Colorado and Utah during 1986 and 1987. About 17% (198 of 1,131) fecal specimens collected from July 1986 through August 1987 contained the organism. Both O1 and non-O1 V. cholerae strains were isolated from the fecal specimens. Isolates from eight birds (representing five species) agglutinated in O group 1 antiserum. Supernatants of broth cultures from three isolates which typed as V. cholerae O1 serotype Ogawa gave reactions typical of cholera toxin when tested on Y-1 mouse adrenal cell cultures. Several serovars of non-O1 V. cholerae were isolated from the fecal specimens; serovar 22 was the most prevalent type. All non-O1 isolates were cytotoxic to Y-1 mouse adrenal cells. Only non-O1 V. cholerae was detected in water samples collected from the habitat of the birds. The results of this study suggest that aquatic birds serve as carriers and disseminate V. cholerae over a wide area.     

Isolation of Vibrio cholerae from aquatic birds in Colorado and Utah.

Vibrio cholerae was isolated from cloacal swabs and freshly voided feces collected from 20 species of aquatic birds in Colorado and Utah during 1986 and 1987. About 17% (198 of 1,131) fecal specimens collected from July 1986 through August 1987 contained the organism. Both O1 and non-O1 V. cholerae strains were isolated from the fecal specimens. Isolates from eight birds (representing five species) agglutinated in O group 1 antiserum. Supernatants of broth cultures from three isolates which typed as V. cholerae O1 serotype Ogawa gave reactions typical of cholera toxin when tested on Y-1 mouse adrenal cell cultures. Several serovars of non-O1 V. cholerae were isolated from the fecal specimens; serovar 22 was the most prevalent type. All non-O1 isolates were cytotoxic to Y-1 mouse adrenal cells. Only non-O1 V. cholerae was detected in water samples collected from the habitat of the birds. The results of this study suggest that aquatic birds serve as carriers and disseminate V. cholerae over a wide area.     

Effect of avian intestinal microflora possessing adhering and hydrophobic properties on competitive exclusion of Salmonella typhimurium from chicks.

The role of adherence and hydrophobic properties of native gut microflora in competitive exclusion of salmonellas from chicks is reported. Pure bacterial strains were isolated from washed caeca of 3-d-old chicks which had been treated on the day of hatch with a microflora from salmonella-free adult birds. These strains, when added to known mixtures of pure cultures, improved the efficacy of the mixtures in protecting chicks against a challenge of 10(5) cfu of Salmonella typhimurium. Strains from washed caeca and from other sources were also screened for hydrophobic properties. Undefined microflora and strains freshly isolated from washed caeca and which were hydrophobic improved the efficacy of protective mixtures, while none of the combination of hydrophobic strains obtained from stored strains had any beneficial effect.     

Effect of avian intestinal microflora possessing adhering and hydrophobic properties on competitive exclusion of Salmonella typhimurium from chicks

The role of adherence and hydrophobic properties of native gut microflora in competitive exclusion of salmonellas from chicks is reported. Pure bacterial strains were isolated from washed caeca of 3-d-old chicks which had been treated on the day of hatch with a microflora from salmonella-free adult birds. These strains, when added to known mixtures of pure cultures, improved the efficacy of the mixtures in protecting chicks against a challenge of 10⁵ cfu of Salmonella typhimurium. Strains from washed caeca and from other sources were also screened for hydrophobic properties. Undefined microflora and strains freshly isolated from washed caeca and which were hydrophobic improved the efficacy of protective mixtures, while none of the combination of hydrophobic strains obtained from stored strains had any beneficial effect.     

Plasmids of Staphylococcus aureus associated with live and processed poultry

A series of 23 strains of Staphylococcus aureus originally isolated from processed poultry was screened for the presence of plasmids. Plasmids were more common in strains of Staph. aureus characteristically associated with live poultry than with strains endemic in poultry plants and strains of human origin. Two plasmids with sizes of 1.65 and 18.2 kilobase pairs (kBp) were present in three strains considered typical of Staph. aureus forma specialis 'altilis' and two plasmids with sizes of 1.65 and 17 kBp were present in three of four strains of Staph. aureus var. gallinae. A 1.65 kBp plasmid was present in all seven strains of these poultry biotypes and in three of 14 'endemic' strains. All the 1.65 kBp plasmids were shown by blot hybridization to share sequence homology. There was also some sequence homology between the 18.2 kBp and 17 kBp plasmids. These results were supported by restriction enzyme digest analyses. A study of cured derivatives of strain PS221 f.sp. 'altilis' suggested that the 18.2 kBp plasmid encoded the genetic determinant(s) responsible for caseolysis. Both the 1.65 and the 18.2 kBp plasmids also exerted an effect on the production of acid from lactose. In no other characteristic did cured strains resemble the plasmid-free 'endemic' strains. This was therefore consistent with the notion that the genetic determinants associated with the cultural characteristics of endemic strains are chromosomally located.     

Impact of Transport Crate Reuse and of Catching and Processing on Campylobacter and Salmonella Contamination of Broiler Chickens

The influence of transport, catching, and processing on contamination of broiler chickens with Salmonella and Campylobacter was investigated. Transport crates were reused with high frequency and were often still contaminated with Salmonella and Campylobacter when they arrived at the farm despite the fact that they were washed at the factory, and thus they were a potential route of infection. These organisms contaminated the feathers of previously Campylobacter- and Salmonella-negative birds going to the processing plant and were isolated from processed carcasses, albeit at a low frequency. The Campylobacter types which were the predominant organisms on the live birds when they arrived at the processing plant were not necessarily the types that were most frequently isolated from processed carcasses. This finding may reflect cross-contamination that occurred during processing or differences in the tolerance of the strains to the hostile environments that the bacteria experienced. The process of catching and putting the birds in crates significantly increased the chance of contamination with Campylobacter (P < 0.001).     

Plasmids of Staphylococcus aureus associated with live and processed poultry

A series of 23 strains of Staphylococcus aureus originally isolated from processed poultry was screened for the presence of plasmids. Plasmids were more common in strains of Staph. aureus characteristically associated with live poultry than with strains endemic in poultry plants and strains of human origin. Two plasmids with sizes of 1·65 and 18·2 kilobase pairs (kBp) were present in three strains considered typical of Staph. aureus forma specialis 'altilis' and two plasmids with sizes of 1·65 and 17 kBp were present in three of four strains of Staph. aureus var. gallinae. A 1·65 kBp plasmid was present in all seven strains of these poultry biotypes and in three of 14 'endemic' strains. All the 1·65 kBp plasmids were shown by blot hybridization to share sequence homology. There was also some sequence homology between the 18·2 kBp and 17 kBp plasmids. These results were supported by restriction enzyme digest analyses. A study of cured derivatives of strain PS221 f.sp. 'altilis' suggested that the 18·2 kBp plasmid encoded the genetic determinant(s) responsible for caseolysis. Both the 1·65 and the 18·2 kBp plasmids also exerted an effect on the production of acid from lactose. In no other characteristic did cured strains resemble the plasmid-free 'endemic' strains. This was therefore consistent with the notion that the genetic determinants associated with the cultural characteristics of endemic strains are chromosomally located.     

Production of phospholipase C (alpha-toxin), haemolysins and lethal toxins by Clostridium perfringens types A to D.

To obtain high yields of extracellular enzymes and toxins for immunological analysis, type culture collection strains of Clostridium perfringens types A to D and 28 fresh isolates of C. perfringens type A from humans were grown in fermenters under controlled conditions in a pre-reduced proteose peptone medium. The type culture collection strains all showed different characteristics with respect to growth rates and pH optima for growth. Production of phospholipase C (alpha-toxin), haemolysin and lethal activity varied considerably between the different types. Growth and extracellular protein production in fermenters with pH control and static or stirred cultures were compared. Production of all extracellular proteins measured was markedly improved by cultivation in fermenters with pH control. Strain ATCC13124 produced five times more phospholipase C than any of 28 freshly isolated strains of C. perfringens type A, grown under identical conditions. Haemolytic and lethal activities of the ATCC strain were equal or superior to the activities of any of the freshly isolated strains. There were no differences in the bacterial yields and in the production of extracellular toxins between type A strains isolated from clinical cases of gas gangrene and abdominal wounds, and those isolated from faecal samples from healthy persons.     

Histomonas meleagridis after One Thousand in vitro Passages

SYNOPSIS. During a 7-year period Histomonas meleagridis survived 1000 passages in Medium 199 fortified with serum and antibiotic-killed bacteria. The histomonads were originally isolated from a chicken's cecal dropping, and the bacteria were cultivated from the cecal contents of a normal turkey. In many respects, the histomonads remained unchanged during cultivation, but in some other respects they changed considerably, perhaps irreversibly.
Morphologically, the histomonads propagated in vitro showed only slight changes. When returned to birds, they resumed the structure characteristic of lumen-dwelling individuals of this species. However, they had long since lost their ability to produce disease in either chickens or turkeys, and organisms of the tissue-dwelling type were rarely seen. The long-cultivated histomonads are actually as nonpathogenic as Histomonas wenrichi , but they have none of the distinguishing morphologic characteristics of the latter.
As has been reported elsewhere, H. meleagridis long maintained in the tissue culture medium has also lost much of its ability to immunize either chickens or turkeys against infection with virulent strains of this parasite. Also lost in the process of adaptation to its restricted medium has been the ability to multiply satisfactorily in vitro with the complement of bacteria normal to the ceca of the birds. However, in some instances, histomonads which have become adapted to in vitro cultivation are still able to live in birds with this diversified flora.
Activity of the histomonads cultivated in vitro differed but little from that of H. meleagridis freshly isolated from birds, when both were viewed under the same conditions. Histomonads from each of the above sources multiplied most satisfactorily in their accustomed habitat, probably because of a difference in nutritional requirements.     

Second Survey of Market Poultry for Salmonella Infection

On 34 days over a period of 30 months, samplings were made of randomly selected birds slaughtered in randomly chosen processing plants. Methods were designed so that Salmonella and paracolon bacteria isolated were indigenous to the intestinal tract of the selected bird. Incidence rates were 5.25% for turkeys, 0.54% for chicken hens, and 1.05% for chicken fryers. Infected birds, providing the potential for contamination, were processed on 58% (turkeys), 25% (chicken hens), and 43% (chicken fryers) of the days. Combined data from the present and previous surveys represent a total of 9,851 carcasses sampled on 94 separate days over a 4.5-year period; these data provide a more accurate analysis of incidence rates and days that positive birds were being slaughtered. For turkeys, chicken hens, and chicken fryers, incidence rates were 4.16, 0.65, and 1.42%, respectively, and days positive were 56, 14, and 32%.     

Similarities and Differences in the Development of Laboratory Strains and Freshly Isolated Strains of Herpes Simplex Virus in HEp-2 Cells: Electron Microscopy

HEp-2 cells infected with two laboratory strains (mP and MP) and two freshly isolated strains (F and G) of herpes simplex virus were fixed at intervals between 4 and 50 hr postinfection and sectioned, and were then examined with the electron microscope. These studies revealed the following. (i) All four strains caused identical segregation of nucleoli and aggregation of host chromosomes at the nuclear membrane. (ii) The development of MP virus could not be differentiated from that of its parent mP strain. (iii) There were quantitative differences between laboratory (mP) and freshly isolated (F) type 1 strains. Thus, cells infected with F contained numerous nuclear crystals of nucleocapsids and relatively few cytoplasmic structures containing enveloped nucleocapsids. Conversely, cells infected with mP or with MP virus contained numerous cytoplasmic structures with enveloped nucleocapsids and relatively few nuclear crystals of nucleocapsids. (iv) There were qualitative differences between type 2 strain (G) isolated from genital lesions and type 1 strains. Thus, cells infected with the G strain contain numerous filaments in nuclei and unenveloped and partially enveloped nucleocapsids in the cytoplasm. Of particular interest is the finding that cytoplasmic membranes in apposition to nucleocapsids were thickened and bent as if they were enveloping the particle. The significance of the qualitative differences in the development of the four strains is discussed.     

Does the chicken genotype ‘Géline de Touraine’ have specific carcass and meat characteristics?

The aim of this study was to determine the specific characteristics of carcass and meat from an old French chicken breed, the ‘Géline de Touraine’ (GT), characterised by a very slow-growing rate and usually slaughtered at 120 days of age. For this purpose, we compared the GT with an experimental crossbreed (EC) exhibiting the same growth rate, and with a ‘Label rouge’ (LR) genotype usually slaughtered at 84 days of age. A total of 250 males and 250 females per genotype were reared by separating sexes and genotypes. The growth performances were recorded. At 84 days of age, 80 birds per sex and per genotype were slaughtered. The frequency of clawing and pecking injuries on the carcass was noted. We also measured the skin colour and the thickness of wing membrane. The relative percentages of carcass, breast, thigh + drumstick, abdominal fat, testis or ovary to body weight were determined. On breast and thigh muscles the ultimate pH (pHu) and colour were measured. The juice loss after 3 days’ storage at +4°C and after cooking at 85°C, and the shear force value of Warner–Bratzler were only measured on breast muscles. At 120 days of age, we repeated the same measurements but only on EC and GT genotypes in order to compare birds at the same age or at the respective slaughter age for each production. Whatever the slaughter age, the body weight of males was always higher than that of the females but the carcass yield was similar for both sexes. The females had higher breast yield and carcass fatness but lower thigh + drumstick yield than the males. The yellowness of skin and meat was higher for the females than for the males while the contrary was observed for the redness of the meat. The breast meat of the females also had higher cooking loss than that of the males. GT and EC birds exhibited a higher occurrence of carcass defects and a higher pHu in meat than LR birds. The GT chickens were characterised by a lower breast yield, a higher fattiness and an earlier sexual maturity than the other genotypes, which could confer typical sensorial attributes to their meat. Finally, the EC chickens exhibited a skin and a meat more coloured than the other genotypes, particularly for yellowness, a character which could be under genetic control.     

The Distribution of Staphylococcus aureus in a Poultry Processing Plant

A set of phages previously isolated from poultry strains of Staphylococcus aureus was used to type such isolates from poultry before, during and after processing in a poultry plant. Certain poultry phage types were found to be associated with the live birds rather than the processed carcases. Strains lysed by phages from this group may represent a specific 'poultry'biotype. A site of cross-contamination within the plant was discovered.     

One group of genetically similar Listeria monocytogenes strains frequently dominates and persists in several fish slaughter- and smokehouses

Contamination of foods with the human pathogen Listeria monocytogenes may occur during processing, and the purpose of this study was to determine whether genetically similar strains colonize different processing plants or whether specific persistent strains are unique to each processing plant. We hypothesized that specific L. monocytogenes strains may be better adapted to specific environmental niches in the processing environment. L. monocytogenes contamination patterns were identified by the collection of 686 and 267 samples from the processing environments: raw fish and products of four fish smokehouses and four fish slaughterhouses, respectively. Samples were collected both during production and after cleaning and disinfection. Typically, these samplings were separated by 1 to 3 months. Sampling sites were targeted toward areas likely to harbor the bacterium. L. monocytogenes was isolated from 213 samples, and one strain from each positive sample was typed by RAPD (random amplified polymorphic DNA) analysis with four different primers. The 213 strains were divided into 37 RAPD types. One RAPD type was predominant; 86 of 213 strains belonged to this type. This type was found in three smokehouses and two slaughterhouses and was predominant in three of these plants. A subset of 35 strains was also analyzed by amplified fragment length polymorphism typing, which confirmed the genetic similarity of the groups. Moreover, strains of the dominant RAPD type were indistinguishable from strains isolated frequently from smoked fish products 10 years ago. One smokehouse was surveyed for a year and a half, and the dominant RAPD type persisted throughout the survey period and accounted for 94 of 118 isolates. Our study indicates that strains of L. monocytogenes that are genetically very closely related may be especially adapted to colonizing the processing equipment or especially resistant to cleaning and disinfection.