Effect of time and temperature on growth of Salmonella enteritidis in experimentally inoculated eggs

Influence of time and temperature on Salmonella enteritidis multiplication in experimentally injected eggs was examined. There was an increase in the number of S. enteritidis with the increase in temperature of egg storage. There was less increase of S. enteritidis in eggs stored at 4 degrees C than in eggs held at temperatures higher than 4 degrees C (P less than 0.05). These results suggest a possible method for monitoring commercial eggs for the presence of S. enteritidis. It was concluded that the chances of recovery of S. enteritidis can be increased 10(6)-fold or more by holding the eggs at temperatures of 21 or 27 degrees C for more than 20 days and culturing their contents.     

Effect of faecal extract on the growth of Salmonella enteritidis in artificially contaminated hens' eggs.

1. Salmonella enteritidis PT 4 grew in eggs stored at 25 degrees C, but not at 10 degrees C. 2. The incidence of generalised infection of the egg contents (greater than 10(6) salmonellas/ml) was greater in eggs inoculated with cells suspended in faecal extract compared to those with cells in Ringer's solution. 3. The removal of most of the iron did not decrease the growth-promoting effect of the faecal extract.     

Comparison of serological tests for detection of Mycoplasma gallisepticum antibodies in eggs and chicks hatched from experimentally infected hens

Specific pathogen free hens and males were experimentally infected with Mycoplasma gallisepticum. Eggs were then collected, and a part was incubated and set for hatching. Mycoplasma cultures were performed on infected adults and antibodies to MG were analysed by use of slide agglutination (SA) test and commercial ELISA tests on adults and chicks sera and on yolks from non incubated eggs. Both ELISA tests could detect antibodies in yolks from non incubated eggs laid three weeks after infection. SA and the three ELISA tests revealed positive sera in chicks hatched from eggs laid as soon as one week after infection.     

Control of Salmonella enteritidis phage type 4 experimental infection by fosfomycin in newly hatched chicks

One hundred and twenty 1-day-old broiler chickens were divided into four groups: group I unmedicated and orally challenged with 1.5×10⁸ cfu of Salmonella enteritidis phage type 4; group F infected and treated with 300 ppm of fosfomycin in their drinking water; group CF uninfected and treated, and group C maintained as a control group. Their performance, clinical signs, S. enteritidis PT4 reisolation and biochemistry variables were compared. Group F showed fewer symptoms and gross lesions than those from group I. Fosfomycin treatment at 300 ppm improved body weight at 7 days of age by 42.3%. S. enteritidis PT4 reisolation in group I was higher than in the treated group, but total decontamination of challenged birds was not achieved. There was an increase in the levels of total protein and globulins in group I but not in the treated group. Fosfomycin caused no adverse effects on chickens from group CF, assessed by performance and biochemical variables. The results indicate that fosfomycin could be used in the treatment of S. enteritidis PT4 experimental infection.     

Effect of prior serial in vivo passage on the frequency of Salmonella enteritidis contamination in eggs from experimentally infected laying hens

Experimental infection models are valuable tools for understanding and preventing the deposition of Salmonella enteritidis inside eggs. Oral inoculation is believed to closely simulate naturally occurring S. enteritidis infections of chickens, but oral infection studies have often generated relatively low frequencies of egg contamination. The present study assessed whether repeated in vivo passage of an S. enteritidis strain could affect its ability to cause egg contamination in experimentally infected hens. The incidence of egg contamination was determined in groups of hens inoculated orally with either a phage type 13a S. enteritidis strain or derivatives of this parent strain that were obtained by three successive rounds of passage and reisolation from tissues of infected hens. Passaged S. enteritidis isolates recovered from ovaries and oviducts induced a significantly higher incidence of egg contamination (16.97%) than was attributed to the parent strain (8.27%). However, passaged S. enteritidis isolates recovered from livers and spleens were not associated with a significantly increased frequency of deposition in eggs. By either inducing or selecting for the expression of relevant microbial properties, passage of S. enteritidis through reproductive tissues of chickens may be useful for improving the efficiency at which experimental infection models produce egg contamination.     

"聚焦沙门氏菌"专栏四:种蛋的沙门氏菌污染

沙门氏菌污染是种鸡场要面临的一个问题,这是因为如果种蛋及其内容物受到沙门氏菌的污染,种蛋孵出的子代雏鸡、进而引入该雏鸡的鸡场存在受到污染的可能性. 在过去一段时间中,沙门氏菌阳性种鸡群很可能将污染许多子代鸡群或鸡场,而且这种情况不只在一个国家出现. 需要指出的是,如果我们位居育种"金字塔"的顶端,也就是说处于曾祖代种鸡水平,沙门氏菌通过此金字塔向下垂直传播的后果可能会更加显著,因为更多的鸡群最终将受到污染.     

Non-invasive determination of heart rate in newly hatched chicks.

1. Using a flexible piezoelectric probe, we detected non-invasively the cardiogenic precordial movements of domestic fowl chicks and measured heart rate (HR) daily over the first week after hatching (day 0) for comparison with previously measured embryonic HR. 2. HR of the hatchlings was much more variable than embryonic HR. The mean HR at a given age also varied among the 5 chicks measured. 3. The overall mean HR was 280 +/- 20 (SD, n = 5) bpm on day 0. This value was similar to the prepipping HR and lower than that of externally pipped embryos. It increased to 342 +/- 39 bpm 3 days later.     

Salmonella enteritidis and other Salmonella in laying hens and eggs from flocks with Salmonella in their environment.

Seven Canadian layer flocks with Salmonella enteritidis in their environment were investigated to determine the numbers of hens infected with S. enteritidis, the localization of S. enteritidis in organs of infected hens and the numbers of S. enteritidis-infected eggs produced by two affected flocks. By a microagglutination test (MAT) using S. pullorum antigens, these flocks had more seropositive hens (mean 51.9 +/- 16.9%) than two Salmonella-free flocks (mean 13.0 +/- 4.2%). Culture of tissues of 580 hens (433 seropositive) from the seven flocks detected 26 (4.5%) S. enteritidis-infected hens from two flocks. In one flock, 2/150 hens were infected with S. enteritidis phage type (PT) 8, which was confined to the ceca, and no Salmonella spp. were isolated from 2520 eggs (one day's lay). In the second flock, where 24/150 hens were infected with S. enteritidis PT13, extraintestinal infection was found in nine hens and involved the ovaries and/or oviduct in two hens. Salmonella enteritidis PT13 was isolated from one sample of egg contents and from one sample of cracked shells from among 14,040 eggs (one day's lay) from this flock. The overall prevalence of S. enteritidis-contaminated eggs from the two flocks with infected hens was less than 0.06%. Other Salmonella spp. isolated were S. heidelberg from 58 hens (10%), and S. hadar, S. mbandaka and S. typhimurium from one hen (0.2%) each. The MAT with antigens of S. pullorum had a sensitivity of 81% and a specificity of 24% for detecting S. enteritidis-infected hens.(ABSTRACT TRUNCATED AT 250 WORDS)     

Isolation of Salmonella enteritidis from internal organs of experimentally infected hens

Tissues from experimentally infected hens were examined for the presence of Salmonella enteritidis (SE). SE was recovered from internal organs of both orally inoculated hens and hens infected by horizontal contact transmission. SE was isolated from 58% of the ceca, 51% of the livers, 47% of the spleens, 17% of the ovaries, and 17% of the oviducts of hens sampled during the first 5 weeks after exposure. SE was recovered at a low frequency from all internal organs sampled for as long as 22 weeks after exposure.     

Salmonella contamination of hatching and table eggs: a comparison.

This study determined and compared Salmonella contamination rates of pools of surplus, early and culled hatching eggs from layer and broiler breeder flocks, and of pools of early and regular table eggs from layer flocks. Each pool contained 6 eggs. Five methods were used for the isolation of Salmonella. Nine of 126 pools of culled layer hatching eggs, 2 of 126 pools of surplus layer hatching eggs, and one of 126 pools of early layer hatching eggs were contaminated with Salmonella. All 126 pools of broiler breeder surplus, and early and culled hatching eggs tested negative for Salmonella. All 168 pools of regular table eggs tested negative for Salmonella, whilst one of 84 pools of early table eggs contained Salmonella agona. The pools of culled layer hatching eggs and surplus layer hatching eggs that contained S. typhimurium were derived from the same breeder operation. Similarly, the pools of culled and early layer hatching eggs that contained S. heidelberg were derived from one breeder operation. Pools of culled hatching eggs were more frequently contaminated with Salmonella than other hatching or table eggs. Pools containing eggs that were both cracked and dirty were more frequently contaminated with Salmonella than all other pools of eggs. The overall Salmonella contamination rate of the table eggs was 0.07 to 0.4%. Critical control points (macroscopic classification of the eggs as cracked and dirty) were validated microbiologically.     

Effect of anaerobic cecal microflora and dietary lactose on colonization resistance of layer chicks to invasive Salmonella enteritidis.

The effect of oral inoculation with anaerobic cultures of cecal microflora and providing lactose in the feed on colonization resistance to invasive Salmonella enteritidis was evaluated in newly hatched leghorn chicks. Salmonella colonization of the ceca, tissue invasion and organ colonization, horizontal transmission, and seroconversion were significantly decreased (P less than 0.01) in chicks inoculated with cecal flora. The addition of lactose to the feed, in the absence of cecal microflora, failed to provide protection. Dietary lactose enhanced colonization resistance in chicks that were inoculated with anaerobic cultures of cecal flora. The results indicated that establishment of normal cecal flora in layer chicks together with the addition of lactose to the diet markedly increases resistance to cecal colonization and organ invasion, and decreases horizontal transmission of S. enteritidis.     

Effect of cadmium injected in ovo on hatching results and the activity of plasma hydrolytic enzymes in newly hatched chicks

The aim of the study was to determine the toxicity of cadmium ions in chick embryos, using plasma hydrolytic enzyme as its biomarker. Hatching eggs (n = 300) from Ross 308 broilers were incubated under standard conditions. On day 4 of incubation, 50 μl of saline solution, containing Cd ions at a concentration from 0 (control group) to 24 μg, was injected in ovo into the egg albumen. The results indicate that the administration of cadmium at doses exceeding 1 μg/egg caused a gradual decrease in hatchability, with an LD50 of 3.9 μg/egg. The greatest differences between the groups in the enzymatic activities studied were found for N-acetyl-β-D-glucosaminidase (NAG), β-D-mannosidase (β-MAN) and arylsulphatase (ARYL). Compared to the control group, in the blood serum of chicks from the groups receiving 3, 6 and 12 μg Cd/egg the NAG activity increased by 79, 108 and 54% and β-MAN activity by 33, 119 and 108%, respectively. Exposure to cadmium at a dose of 1 to 6 μg per egg caused an about 60% increase in ARYL activity while a dose of 12 μg decreased the activity by about 35% below the level observed in the control group. These findings show that cadmium has a similar toxicity mechanism in mammals and birds, which opens the possibility of using NAG activity as a biomarker of the cytotoxic effect of cadmium in birds.     

Efficacy of maternal Salmonella antibodies and experimental oral infection of chicks with Salmonella enteritidis

Distribution of maternally transmitted Salmonella antibodies and their protective effects were studied in the progeny of broiler breeder birds which had been vaccinated with live S. Typhimurium and inactivated S. Enteritidis vaccines. Vaccination resulted in a significant increase of the antibody concentration in yolk of hatching eggs and in serum and jejunum of the progeny of immunized breeder birds. Higher antibody titres for isotypes IgG and IgA were still seen on day 21 of age. Antibody production of isotypes IgA and IgM by the chickens themselves was found between 14 and 21 days of age. Two challenge models (10(2) cfu/bird on day 1 of age and a seeder bird model, respectively) were used to evaluate the efficacy of maternal antibodies against challenge with S. Enteritidis. Using both models numbers of challenge organisms were lower in the caeca of the progeny of immunized parent birds between day 7 and day 21 of age (maximum about 1.5 log10 units) compared with control chicks. The results indicate the efficacy of maternally transferred antibodies but it remains the question of their practical relevance. The effects of acquired maternal antibodies on an active immunization of the progeny of immunized breeder birds with live Salmonella vaccines are discussed.     

柔嫩艾美耳球虫感染对蛋鸡生产肠炎沙门氏菌污染蛋和感染肠炎沙门氏菌敏感性的影响

为揭示球虫感染对蛋鸡的肠炎沙门氏菌敏感性及鸡蛋的肠炎沙门氏菌污染率的影响而进行了本研究。结果显示,柔嫩艾美耳球虫感染对蛋鸡生产肠炎沙门氏菌污染蛋及感染肠炎沙门氏菌的敏感性均具有增强作用。具体表现为：柔嫩艾美耳球虫感染可使肠炎沙门氏菌污染蛋壳显著增多（Ｐ＜０．０５）;球虫感染鸡所产蛋,无论是新鲜的还是贮存１周后的,其蛋内容物的肠炎沙门氏菌阳性率比仅感染肠炎沙门氏菌鸡所产蛋要高;球虫感染鸡盲肠中的沙门氏菌阳性率和沙门氏菌数显著增加（Ｐ＜０．０１）;球虫感染鸡输卵管中的肠炎沙门氏菌也有增多;球虫感染使泄殖腔拭子的肠炎沙门氏菌阳性率增加。研究结果表明,柔嫩艾美耳球虫感染可以增加鸡群中肠炎沙门氏菌的水平及其垂直传播的危险。     

Effects of probiotics and maternal vaccination on Salmonella enteritidis infection in broiler chicks

The effects of probiotics and maternal vaccination with an inactivated Salmonella Enteritidis (SE) vaccine on day-old chicks challenged with SE were evaluated. A 2 X 3 factorial arrangement was used (with or without probiotics; breeders nonvaccinated, vaccinated intramuscularly, or vaccinated intraperitoneally). Three trials were conducted in isolation cabinets and SE challenge was different between trials. The number of SE organisms per chick and the time interval between housing and introduction of seeder birds (hereafter called challenge) were 1.6 X 10(8) and 1 hr (Trial I), 1.8 X 10(6) and 12 hr (Trial II), and 1.2 X 10(4) and 24 hr (Trial III). SE recovery was assessed in ceca and liver at 3, 5, and 7 days postchallenge, and the number of colony-forming units (CFU) in ceca was evaluated at 5 and 7 days postchallenge. The number of SE (log CFU) in the ceca reduced 0.56 log (from 7.59 to 7.03) and 1.45 log (7.62 to 6.17) because of the treatment with probiotics in Trials II and III, respectively. The greater reduction in Trial III indicates the importance of the early use of probiotics on the prevention of SE infection. Treatment with probiotics resulted in a smaller number of SE-positive livers after 5 days postchallenge on Trial III. Although there was no significant effect of maternal vaccination on the number of SE CFU in the ceca, a significant effect of maternal vaccination on the SE CFU was observed in the liver, but not in the ceca at 5 days after challenge.     

Effects of rearing conditions on the colonization of Salmonella enteritidis in the cecum of chicks.

Salmonella enteritidis is the cause of human salmonellosis associated with contaminated eggs. In this study, we artificially challenged S. enteritidis to chicks just after hatching, and the effects of breeding conditions on the intestinal carriage of S. enteritidis were examined. S. enteritidis was not directly detected from spleen, liver and blood, but were constantly isolated from the cecal contents throughout the experiment. When chicks were reared in the unsanitary conditions and in the high housing density, the numbers of S. enteritidis increased. The subsequent experiment was undertaken to examine whether the antibacterial additive in a feed would have any impact on S. enteritidis colonization in chicks. Some antibiotic effective on the growth promotion had an influence on S. enteritidis colonization.     

Dietary lutein and fat interact to modify macrophage properties in chicks hatched from carotenoid deplete or replete eggs

Two experiments were conducted to study the interaction between dietary lutein and fat levels in broiler chicks hatched from lutein depleted (Experiment I) and repleted (Experiment II) eggs. In both experiments, a 2 x 3 factorial arrangement of treatments resulted in six dietary treatments (fat at 3% and 6% and lutein at 0, 25 and 50 mg/kg feed) that were fed for 23 days to 18 birds per treatment (in three replications). In Experiment I, the anti-dinitrophenyl-keyhole-lympet-hemocyanin (anti-DNP-KLM) serum antibody response at day 22 and macrophage phagocytotic index at day 17 did not differ among treatment groups (p > 0.05). The concavalin A and phytohaemagglutinin-P lymphocyte proliferation index at day 19 was greater in birds fed 50 mg of lutein and 3% fat than in birds fed all other diets (p < 0.05). Independent of the level of dietary fat, dietary lutein increased macrophage (day 23) nitrite production measured 46 h after in vitro stimulation with LPS (p < 0.05). Among the birds fed lutein at 25 or 50 mg/kg feed, birds fed 3% fat had higher LPS-induced nitrite production compared to the birds fed 6% fat after 46 (p = 0.014) or 70 h (p < 0.001). In Experiment II, macrophage nitrite production was measured at 54 h after LPS stimulation on days 11, 15, 19 and 23. An interaction between dietary lutein and fat levels on nitrite production was observed on day 19 (p = 0.012), where macrophages from birds fed 0 mg lutein and 3% fat had the highest nitrite production (p = 0.012). Macrophages from birds fed lutein at 25 and 50 mg/kg diet and 3% fat had higher (p = 0.012) nitrite production than birds fed 6% fat. Thus, in birds hatched from lutein deplete and replete eggs, modulation of macrophage nitrite production by lutein is dependent on the level of dietary fat.