The fate of Salmonella enteritidis PT4 in home-made mayonnaise prepared with citric acid

The fate of Salmonella enteritidis PT4 in home-made mayonnaise prepared with citric acid solution (citric acid concentration of > or = 4.98% (w/v)) was investigated. It was found that pH of mayonnaise is closely related to the ratio of egg yolk to citric acid, and the inactivation rate of the micro-organisms increases as the ratio decreases and/or incubation temperature increases. To achieve Salm. enteritidis PT4-free home-made mayonnaise prepared with pure lemon juice (citric acid concentration > or = 5% (w/v)), it is recommended that the pH should be 3.30 or below, or, in practice, at least 20 ml pure lemon juice per fresh egg yolk should be used. For the use of 20-35 ml pure lemon juice per egg yolk, the product should be held at 22 degrees C or over for at least 72 h and for the use of over 35 ml pure lemon juice per egg yolk, for at least 48 h before consumption or refrigeration.     

An assessment of epidemic outbreaks of salmonellosis connected with poultry plant production]

The causes of the simultaneous rise of salmonellosis morbidity induced by S. enteritidis among the population of three towns in the Perm region were studied. The study revealed the leading role of eggs and chicken meat as factors contributing to the transfer of this infection to the population of different territories, commonly supplied with the products of one poultry plant. The contamination of eggs and chickens with S. enteritidis occurred at the plant due to Salmonella infection of chickens, parallel with the use of nonbalanced mixed fodder, originally intended for feeding swine. Analysis of the epidemic and epizootic processes of Salmonella infection in this epidemic situation made it possible to reliably establish the factors contributing to the transfer of the infective agent and the site of contamination.     

Effects of sexual maturation and Salmonella infection on the expression of Toll-like receptors in the chicken vagina

Toll-like receptors (TLRs) are a critical component of the innate immune response in many vertebrates, including avian species. The recent findings of chicken TLRs (cTLRs) expression in ovarian follicles and in the chicken ovary in vivo, as well as the changes in their expression in response to lipopolysaccharide or Salmonella enteritidis (SE) infection, have broad implications for reproductive physiology and for the prevention of transmission of zoonotic diseases to humans through the consumption of contaminated poultry eggs. Because the main route of egg contamination is from infection of the oviduct and mainly from the vagina, the aim of this study was to investigate the expression of the ten cTLRs identified to date in the chicken oviduct in vivo, to determine whether sexual maturation affects their mRNA abundance and to investigate whether SE infection alters the expression of TLRs in the chicken vagina. RNA was extracted from the vagina of healthy prepubertal, sexually mature and aged birds, and from sexually mature and aged SE infected birds. RT-PCR analysis revealed that all types of cTLRs apart from TLR1-1 were expressed in the vagina of sexually mature birds. Quantitative real-time PCR analysis revealed that the mRNA abundance of TLR2-1, 2-2 and 4 differ with respect to sexual maturation in the chicken vagina. SE infection resulted in a significant induction of TLR5 and 15 in the vagina of sexually mature birds, and in a significant induction of TLR2-1, 4 and 15 in the vagina of aged birds, while a significant down-regulation was observed for TLR7 in the vagina of sexually mature birds. These findings suggest that a TLR mediated immune response mechanism exists in the chicken vagina, playing a crucial role in preventing microbial pathogens from being incorporated into newly forming eggs.     

Large outbreak of food poisoning caused by Salmonella typhimurium definitive type 49 in mayonnaise.

An investigation was conducted to determine the vehicle of infection of an outbreak of food poisoning in a large metropolitan building early in 1988. A questionnaire was distributed to 700 people who had eaten in the building during the week of the outbreak, and attack rates for specific food were calculated. Food and stool samples, environmental samples, and eggs and environmental swabs from the egg suppliers were examined microbiologically. Altogether 474 questionnaires were returned, 120 people reporting gastrointestinal illness. The illness was significantly associated with foods containing mayonnaise. Salmonella typhimurium definitive type 49 was isolated from 76 of the 84 stool samples containing salmonella and from five of the eight samples taken from the chicken house of the main egg supplier. Mayonnaise was probably the vehicle of infection, which was caused by S typhimurium definitive type 49.     

The influence of pH and temperature on the behaviour of Salmonella enteritidis phage type 4 in home-made mayonnaise

The behaviour of Salmonella enteritidis phage type 4 in home-made mayonnaise was studied. Samples of mayonnaise were prepared with different pH values using wine vinegar or lemon juice in order to bring down the pH to 5,4.5,4 and 3.6, inoculate and incubated at 4, 24 and 35C for 5 days. The results showed a better bactericidal activity of the vinegar (acetic acid) than the lemon juice (citric acid), both of these acids being more active at higher temperatures. For preventing salmonellosis transmission by home-made mayonnaise the use of vinegar as an acidulant in order to achieve a pH between 3.6 and 4 and storage in a warm place is recommended.     

PCR法对鸡卵黄中Coxiella burnetii菌的测定

[目的]通过用定量PCR加巢式PCR方法,提高了对Coxiella burnetii (C.b)CoMl基因的检出率;通过对鸡卵中病原微生物Coxiella burnetii的基因检测,明确鸡卵的食品安全性;并对明确Coxiella burnetii的流行病学有重要意义.[方法]提取鸡卵DNA,用定量PCR加巢式PCR方法检测上述基因,并对PCR产物进行测序分析,通过间接免疫荧光法观察鸡血白细胞中的微生物.[结果]用定量PCR加巢式PCR方法可检出4个以上的Coxiella burnetii Coml基因,用此方法可测出鸡卵中Coxiella burnetii Coml基因达104-106个,阳性率为5％-22％;对阳性鸡卵Coml基因PCR产物的测序结果显示有变异菌株的存在;免疫荧光法可见鸡卵中含有该微生物.[结论]由此认为鸡卵中存在病原微生物Coxiella burnetii,可能是Q热传染源.     

The influence of egg size and colour on egg consumption by the Algerian hedgehog (<Emphasis Type="Italic">Atelerix algirus</Emphasis>)

We tested the hypothesis that the Algerian hedgehog (Atelerix algirus) consumes smaller eggs in preference to larger ones and that the colour of the eggs (brown or white spotted) does not influence the egg consumption rate. Eggs of three galliform species (the quail, Coturnix japonica; the red legged partridge, Alectoris rufa; the chicken, Gallus domesticus) were offered to 14 individual hedgehogs under experimental conditions. Overall, the hedgehogs consumed 107 of the 225 eggs offered (48%) in 5 days. The colour of the eggs did not influence the rates of egg consumption. However, egg size had a significant influence on the rates of egg consumption. Generalized linear mixed models showed that hedgehogs preferred to consume smaller quail eggs (91%, n = 65) than intermediate partridge eggs (44%, n = 110), whereas the largest chicken eggs (n = 50) were not consumed at all. There was a strong inter-individual variation in egg consumption, particularly of partridge eggs, from 0 to 100%, whereas individual variation in consumption of quail eggs was lower, from 70 to 100%.     

Prolonged incubation period of salmonellosis in an outbreak of Salmonella enteritidis infection

An outbreak of Salmonella enteritidis infections occurred in Otaru, Japan, in September 1997. A total of 143 cases of salmonellosis were reported to the local Public Health Center. In this outbreak, one case had a 214-hr incubation period. We investigated 5 isolates including this case by phage typing and pulsed-field gel electrophoresis (PFGE) to determine the genetic heterogeneity of S. enteritidis. Five isolates were phage typed as reacted, but did not conform (RDNC) with identical reaction patterns and had quite similar PFGE patterns. Thus, the prolonged incubation period may not be attributed to genetic heterogeneity of the organism but rather to other factors.     

Large outbreak of Salmonella enterica serotype paratyphi B infection caused by a goats' milk cheese,France, 1993: a case finding and epidemiological study.

OBJECTIVE--To assess the magnitude of a nationwide outbreak of infection with Salmonella enterica serotype paratyphi B and identify the vehicle and source of infection. DESIGN--A case finding study of S paratyphi B infection between 15 August and 30 November 1993; a pair matched case-control study; an environmental investigation at a processing plant that produced a raw goats'' milk cheese incriminated in the outbreak; phage typing and genotyping of food and human S paratyphi B isolates. SETTING--France, 15 August to 30 November 1993. SUBJECTS--273 patients with S paratyphi B infection; 59 pairs of cases and controls matched for age, sex, and city of residence. MAIN OUTCOME MEASURES--Numbers of cases and incidence rates by region of residence and age; matched odds ratios for dairy food preferences. RESULTS--Among the 273 cases there was one death; 203 (78%) strains belonged to phage type 1 var 3. The incidence of infection was greatest in the region where goats'' milk cheese is commonly produced. Comparison of cases and controls showed a 12-fold greater risk of illness (95% confidence interval 1.6 to 92.3) from eating brand A unpasteurised goats'' milk cheese. S paratyphi B isolates of phage type 1 var 3 were recovered from cheese A, goats'' milk at the plant processing cheese A, and goats'' milk supplied to the plant by a single farm. Genotypic IS 200 typing of food and human 1 var 3 phage type isolates showed a common IS 200 pattern. CONCLUSION--This outbreak emphasises the potential health hazards of widely distributed unpasteurised milk products in France and the need for their close bacterial monitoring.     

Analysis of Salmonella enteritidis strains isolated from food-poisoning cases in Taiwan by pulsed field gel electrophoresis, plasmid profile and phage typing

AIMS: To establish the molecular typing data for Salmonella enteritidis due to its increasing role in Salmonella infections in Taiwan. METHODS AND RESULTS: Sixty-three Salm. enteritidis strains isolated from related and unrelated patients suffering from food-borne poisoning during 1991-97 were collected and subjected to pulsed field gel electrophoresis (PFGE), plasmid analysis and phage typing. For PFGE, XbaI, SpeI and NotI restriction enzymes were used for chromosomal DNA digestion. The results showed that, for these 63 Salmonella strains, 10 PFGE pattern combinations were found. Of these, pattern X3 S3 N3 was the major subtype, since 46 strains isolated from different locations at different times during 1991-97 showed this PFGE pattern. Plasmid analysis showed only three plasmid profiles and phage typing showed that most of the Salmonella strains were of the phage type PT4. CONCLUSION: Most of the Salm. enteritidis strains circulating in Taiwan are of very similar genetic types or are highly related and that strains of PFGE pattern X3 S3 N3 are the prevalent and recirculating strains of Salm. enteritidis which caused food-poisoning cases in Taiwan in 1991-97. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides information that in Salmonella infection, certain subtypes of Salm. enteritidis should be scrutinized.     

Lipopolysaccharide-specific IgG in egg yolk from two chicken flocks infected with Salmonella enteritidis

An indirect enzyme-linked immunosorbent assay (ELISA) was used to determine levels of IgG reacting with lipopolysaccharide from Salmonella enteritidis in samples of egg yolk from broiler breeder and egg-laying flocks, some of which were known to be infected with S. enteritidis. There was positive correlation between infection with salmonella and a high reading in the ELISA.     

Colicinogeny in local isolates of salmonellae and plasmid transfer studies

Colicinogeny was determined in local isolates of S. typhimurium and S. enteritidis. Fourteen out of 35 S. typhimurium isolates of hospital origin were colicin producers whereas only one chicken isolate out of 82 S. enteritidis isolates of various origin (human, chicken or egg) produced colicin. A colicin producing, cephalothin (Cpt)- and piperacillin (Prl)-resistant local isolate of Salmonella havana (H32) harbored 4 plasmids of 54.0, 28.4, 2.7 and 1.9 kb. Upon curing its plasmids, the strain lost the ability to produce colicin and resistance to antibiotics and no longer expressed smooth lipopolysaccharide (LPS). The outer membrane protein (OMP) of 34.6 kDa was also lost. Using nalidixic acid (Nal)-resistant mutant of the cured strain in conjugation experiments, 10 out of 27 transconjugants were found to be resistant to Nal and Prl, 10 were resistant to Nal and Cpt and 7 showed Nal, Prl and Cpt resistance. Cpt and Prl resistance were determined by 54.0 and 28.4 kb plasmids, respectively. There was no direct correlation between plasmid contents and colicinogeny, LPS and OMP profiles.     

Lipopolysaccharide expression and virulence in mice of Australian isolates of Salmonella enteritidis

The lipopolysaccharide (LPS) of 54 Australian isolates, nine isolates acquired or isolated overseas, and two reference strains of Salmonella enteritidis was studied to assess its relation to pathogenicity. LPS was extracted by proteinase K digestion of whole cells, and analysed by polyacrylamide gel electrophoresis. All isolates possessed an LPS structure identical to that of a reference strain of Salm. enteritidis phage type 4. Representative strains of the clinically prevalent phage types 4, 14 and 26, which express long chain LPS, were assessed for their pathogenicity in mice. Salmonella enteritidis phage type 4 produced a lethal infection in BALB/c mice, but not in C3H/HeJ or Quackenbush (outbred) strains. Phage types 14 and 26 did not produce an obvious infection in any mice, suggesting Australian strains of phage type 4 are more virulent than phage types 14 and 26.     

Detection of two smooth colony phenotypes in a Salmonella enteritidis isolate which vary in their ability to contaminate eggs.

Salmonella enteritidis isolates were obtained from eggs after infection of Leghorn hens with a parent isolate (SE6) known to only infrequently contaminate eggs. Isolates from eggs exhibited two phenotypes that were subtly different. One phenotype was typically smooth, while the other was transiently rough. Both sets of isolates were phage type 13A and positive for D1 epitopes. Immunoblot analysis of entire colonies and gas chromatographic analysis of purified lipopolysaccharide revealed that the phenotypic difference between isolates was due to a quantitative difference in O antigen and possibly a qualitative difference in the lipid A core region. In addition, the two isolates had different opacity properties when examined at 600 nm. When the two isolates were used to inject egg-laying hens, a significant difference in the ability to contaminate eggs was detected.     

Mitigation of avian reproductive tract function by Salmonella enteritidis producing high-molecular-mass lipopolysaccharide

Hens were infected with a wild-type Salmonella enteritidis and its wzz mutant, which lacked the ability to make high-molecular-mass lipopolysaccharide (LPS), in six experiments paired by dosage and route of exposure. Involution of the reproductive tract occurred in 86% of hens that were injected subcutaneously with 108 cfu of the wild-type strain, but none did so when injected with the wzz mutant. In spite of the lack of a specific effect on the reproductive tract, infection of hens with the mutant produced more contaminated eggs and heterophilic granulomas in developing ova (yolks) than wild type; thus, overall, the mutant appeared to be more virulent except after intravenous injection. The mutant also decreased shell quality more often than wild type, regardless of dosage or route of infection. These results suggest that egg-contaminating Salmonella enteritidis that produces high-molecular-mass LPS mitigates signs of illness in poultry by altering the response of the avian reproductive tract to infection, but without altering the incidence of egg contamination following bacteraemia. Further research is warranted to determine whether analyses of shell quality might aid in identification of flocks at risk of producing contaminated eggs.     

The fatty acid composition of oophagous tadpoles (Chirixalus eiffingeri) fed conspecific or chicken egg yolk

We compared the lipid content and fatty acid composition of (1) the egg yolk of three anuran species (Chirixalus eiffingeri, Rhacophorus moltrechti and Buergeria robustus) and chicken eggs; and (2) C. eiffingeri tadpoles fed conspecific eggs or chicken egg yolk. Anuran and chicken egg yolk contained more non-polar than polar lipids but the proportions varied among species. Chicken egg yolk contained low amounts of 22:5n-3 in the polar lipid fraction, and B. robustus eggs did not contain any n-3 or n-6 non-polar lipids. The specific variation of lipid contents and fatty acid composition may relate to the maternal diet and/or breeding biology. In C. eiffingeri tadpoles that fed chicken yolk or frog egg yolk, the dominant components of polar and non-polar lipids were 16:0, 18:0, 18:1, and 18:2n-6, or 20:4n-6 fatty acids. C. eiffingeri eggs contained more n-3 fatty acids (e.g. 18:3n-3 and 20:5n-3) than chicken egg yolk, and tadpoles fed conspecific eggs contained more of these fatty acids than tadpoles fed chicken egg yolk. The compositional differences in the fatty acids between C. eiffingeri tadpoles that fed frog egg or chicken egg yolk are the reflection of the variation in the dietary sources. Our results suggest a direct incorporation of fatty acids into the body without or minimal modification, which provide an important insight into the physiological aspects of cannibalism.     

JAPANESE QUAIL EGG EMBRYO AS A HOST FOR VIRUSES.

Rauscher, Frank J. (National Cancer Institute, Bethesda, Md.), James A. Reyniers, and Miriam R. Sacksteder. Japanese quail egg embryo as a host for viruses. J. Bacteriol. 84:1134-1139. 1962.-The egg embryo of Japanese quail (Coturnix coturnix japonica, Temminck and Schlegel) was found to support readily the growth of a wide range of viruses known to infect chicken eggs. Methods commonly used in studying viruses in chicken eggs were modified and adapted to the quail egg. The procedures are described in detail. The sensitivity of quail eggs to representative viruses of the myxo and pox groups was found to be equal to that of chicken eggs. Preliminary observations of the different responses of quail eggs to standard chicken tumor and to quail-adapted Rous sarcoma virus are described. Viruses known not to grow or induce an observable response in chicken eggs (e.g., the agents of mouse hepatitis, murine leukemias, and visceral lymphomatosis of chickens) also failed to produce such responses in quail eggs.     

A comparison of cecal colonization of <Emphasis Type="Italic">Salmonella enterica</Emphasis> serotype Typhimurium in white leghorn chicks and <Emphasis Type="Italic">Salmonella</Emphasis>-resistant mice

Background  

Salmonellosis is one of the most important bacterial food borne illnesses worldwide. A major source of infection for humans is consumption of chicken or egg products that have been contaminated withSalmonella entericaserotype Typhimurium, however our knowledge regarding colonization and persistence factors in the chicken is small.     

An invasive stink bug as an evolutionary trap for an indigenous egg parasitoid

Invasive alien species can act as ‘evolutionary traps’ for indigenous parasites and predators when the alien species is accepted as prey or a host but is unsuitable for consumption or development. We tested the relationship between acceptance and suitability of eggs of the invasive alien Halyomorpha halys (Hemiptera: Pentatomidae) in North America relative to eggs of the indigenous Podisus maculiventris (Pentatomidae) for the indigenous generalist egg parasitoid Telenomus podisi (Hymenoptera: Scelionidae). T. podisi accepted 0–24 h old H. halys eggs at a rate similar to P. maculiventris eggs (87.5 ± 6.0 and 70.2 ± 9.1 %, respectively). Successful development of T. podisi occurred in 98.3 % of attacked P. maculiventris eggs, but was not observed in H. halys eggs. Oviposition by T. podisi did, however, reduce the developmental success of H. halys embryos relative to unattacked controls by 24.1 % in 0–24 h old eggs and 29.6 % in 24–48 h old eggs. We suggest that as H. halys spreads and increases in abundance in North America, it could operate as an evolutionary trap for indigenous egg parasitoids, thereby indirectly causing an increase in population levels of indigenous pentatomids. This predicted indirect effect would be a result of H. halys eggs acting as an egg sink for T. podisi. We also introduce the concept of a ‘time sink’, which may be particularly relevant for parasitoids such as T. podisi that spend considerable time protecting their reproductive investments.     

Detection and survival of Campylobacter in chicken eggs

AIMS: Campylobacter jejuni, a food-borne human pathogen, is widespread in poultry; however, the sources of infection and modes of transmission of this organism on chicken farms are not well understood. The objective of this study was to determine if vertical transmission of C. jejuni occurs via eggs. METHODS AND RESULTS: Using a temperature differential method, it was shown that Campylobacter had limited ability to penetrate the eggshell. When C. jejuni was directly inoculated into the egg yolk and the eggs were stored at 18 degrees C, the organism was able to survive for up to 14 days. However, viability of C. jejuni was dramatically shortened when injected into the albumen or the air sac. When freshly laid eggs from Campylobacter-inoculated specific pathogen-free (SPF) layers were tested, C. jejuni-contamination was detected in three of 65 pooled whole eggs (5-10 eggs in each pool) via culture and PCR. However, the organism was not detected from any of the 800 eggs (80 pools), collected from the same SPF flock, but kept at 18 degrees C for 7 days before testing. Likewise, Campylobacter was not recovered from any of 500 fresh eggs obtained from commercial broiler-breeder flocks that were actively shedding Campylobacter in faeces. Also, none of the 1000 eggs from broiler breeders obtained from a commercial hatchery were positive for Campylobacter. CONCLUSIONS: These results suggest that vertical transmission of C. jejuni through the egg is probably a rare event and does not play a major role in the introduction of Campylobacter to chicken flocks. SIGNIFICANCE AND IMPACT OF THE STUDY: Control of Campylobacter transmission to chicken flocks should focus on sources of infection that are not related to eggs.