Eggshell characteristics and penetration by Salmonella enterica serovar Enteritidis through the production period of a layer flock

1. Egg weight, shell thickness, number of pores, cuticle deposition and ability of Salmonella enterica serovar Enteritidis (SE) to penetrate the shell were determined for eggs from one layer flock through the entire production period. 2. Penetration was assessed by filling the eggs with a selective medium that allowed visualising Salmonella growth on the inside of the shell and membrane complex. After inoculation of each shell with on average 2.59 log cfu, the eggs were stored for up to 20 d at 20 degrees C and 60% relative humidity (RH). 3. On average 38.7% of the eggshells became penetrated. Mostly penetration occurred on d 3. Although it affected all shell characteristics studied, hen age did not significantly influence eggshell penetration. 4. No correlations were observed between any of the shell characteristics studied and the ability of SE to penetrate the shell. The growth of SE on the shell is of major importance because shell contamination at 20 d of storage and SE penetration were highly correlated.     

储存环境对鸡蛋壳表面3种沙门氏菌存活与迁移的影响

为探明不同储存环境对蛋壳表面沙门氏菌存活和向蛋内迁移的影响,在鸡蛋壳表面人工接种肠炎沙门氏菌、鼠伤寒沙门氏菌和鸡白痢沙门氏菌,试验环境设定温度4、25和37℃,环境相对湿度（RH）设定为50%和90%,在不同的储存环境下,各个时间点检测蛋壳表面沙门氏菌数量与蛋内容物中沙门氏菌污染情况。结果显示,4℃环境下可显著减少蛋壳表面3种沙门氏菌的数量;50% RH、25和37℃环境下蛋壳表面3种沙门氏菌的数量均逐渐下降;90% RH、25℃环境下蛋壳表面肠炎沙门氏菌、鼠伤寒沙门氏菌在个别时间点出现数量反弹,其总体数量呈现下降趋势,37℃环境下鸡白痢沙门氏菌的数量快速下降,而肠炎沙门氏菌和鼠伤寒沙门氏菌的数量先下降后上升,并保持在接种的数量级水平。鸡蛋内容物的检测结果显示,肠炎沙门氏菌和鼠伤寒沙门氏菌在高温、高湿环境下更易被检出,而鸡白痢沙门氏菌始终未被检出。综上所述,低温、低湿可减少蛋壳表面沙门氏菌存活,并阻止其菌体向蛋内迁移,高温、高湿环境促进蛋壳表面沙门氏菌的增殖,增加了其向蛋内迁移的几率,不同血清型沙门氏菌在鸡蛋壳表面的存活和向蛋内迁移的侵袭力存在一定差异。     

Effects of Storage Conditions on Survival and Migration of Three Kinds of Salmonella on Eggshell

In order to study the effects of storage conditions on survival and migration of three kinds of Salmonella,S.enteritidis,S.typhimurium and S.pullorum were artificially contaminated on eggshell.Different environment temperatures（4,25 and 37℃）and relative humidity（50% and 90%）were seted,and the number of Salmonellas on the surface of eggshell and egg contents were counted and detected on various time points.The results showed that 4℃ obviously reduced the number of three kinds of Salmonellas.At 50% RH,the number of three kinds of Salmonellas were reduced on eggshell surface at 25 and 37℃ environment.At 90% RH,despite the individual time points,the number of S.enteritidis and S.typhimurium presented downward trend at 25℃.And at the 37℃,the number of S.pullorum rapidly reduced,and the number of S.enteritidis and S.typhimurium firstly decreased and then increased,and finally maintained at the level of inoculated magnitude.The results of egg content detection showed that S.enteritidis and S.typhimurium were more likely to be detected in high temperature and high humidity environment,while S.pullorum was not detected in all kinds of experimental environment.The results suggested that low temperature and low humidity reduced Salmonellas survival on eggshell and inhibited it migration into the egg,and high temperature and high humidity could promote Salmonellas proliferation,increase the risk of its penetration into egg.There were some differences in the survival on eggshell and invasion between the different serotypes Salmonellas.     

Bacteriology of washed and unwashed eggs. II. Penetration of Salmonella bacteria through the eggshell

The ability of the eggshell to resist penetration of Salmonella bacteria was studied in unwashed and in hand washed eggs, and in eggs, which had been subjected to industrial, large-scale machine washing under strictly controlled conditions.In one part of the investigation, concerning unwashed and machine washed eggs, a defined amount of a broth culture of Salmonella typhi murium was applied on the shell surface, whereafter the eggs were incubated at 4°C for eight weeks or at 30°C for 12 days. At the end of the storage period, the egg contents were examined for the presence of Salmonella. No Salmonella bacteria were detected in the 120 eggs investigated.In another part of the investigation, dealing with 200 unwashed, 200 hand washed and 200 machine washed eggs, a broth culture of Salmonella was applied on the shell surface of eggs, whose contents had been poured out through an opening. In every second egg of each kind, the shell membranes had been scraped off over an area, corresponding to the placement of the bacterial inoculum. The eggshells were filled with enrichment broth and the occurrence of Salmonella bacteria in this broth was examined after incubation.In the eggs with intact shell membranes, the frequencies of Salmonella positive enrichment broths were for unwashed eggs 0.12, for machine washed eggs 0.10, and for hand washed eggs 0.22. The corresponding frequencies in eggs with removed shell membranes were 0.27 (unwashed), 0.42 (machine washed), and 0.60 (hand washed).No statistical difference occurred between unwashed and machine washed eggs when the shell membranes were intact. When the shell membranes were removed this difference was statistically almost significant. The difference between machine washed and hand washed eggs with intact shell membranes was statistically almost significant, while the same difference was highly significant in eggs with removed shell membranes.The author assumes, from the results of the present and a preceding investigation, that the applied washing procedure in no way harms any essential storage properties of uncracked eggs. Considering the known occurrence of hand washing of eggs in Swedish farms — a treatment with documentedly unfavourable influence — it would be desirable if all eggs could be subjected to machine washing under strictly controlled conditions at the grading stations. Such an arrangement ought to result in an increase in the average bacteriologic-hygienic quality of eggs for market.     

Influence of the microstructure on the shell strength of eggs laid by hens of different ages

1. The eggshell is a bioceramic material constructed of columnar calcite crystals preferentially oriented with their c-axis perpendicular to the shell surface. 2. The influence of microstructure (crystal size, shape and crystallographic orientation of crystal grains) on the mechanical properties of eggshells (shell strength) was investigated using eggs from hens of different ages. 3. There was a strong correlation between crystallographic texture and the strength of the eggshell in the case of eggs laid by young hens. The strength of eggshells increased as the preferential orientation of the crystals constituting the eggshell decreased. 4. By comparing two age populations, the effect of hen age on eggshell properties was evaluated. In general, eggshells from aged hens had a lower breaking strength (less than half that of those laid by young hens) and showed a greater variability in their structural properties such as thickness, grain morphology and crystallographic texture. 5. Texture analysis revealed that shells from eggs laid by aged hens have two preferred crystal orientations, after (001) and (104), compared with mainly one, after (001), in eggs laid by young hens. 6. These observed changes in eggshell properties could be due to changes in the organic matrix of the eggshell associated with ageing of the hens.     

Relationship between shell structure and movement of Salmonella enteritidis across the eggshell wall.

1. Ultrastructural analyses of the eggshells of one strain of commercial layers indicated that the cuticular layer is rarely present as an even covering over the shell surface and the paired shell membranes are invariably pitted with holes larger than bacterial dimensions. 2. The above conditions pertain irrespective of the age of the laying bird. 3. Bacterial penetration of the eggshell is independent of pore numbers. 4. In the absence of a functional cuticle and with the shell membranes removed, bacteria are checked in their movement by structural modifications in the mammillary layer.     

The effect of dietary calcium source, concentration and particle size on calcium retention, eggshell quality and overall calcium requirement in laying hens

1. Four different sources of calcium in isonitrogenous and isoenergeric diets were fed to laying hens for 2 weeks when they were 56 and 57 weeks old. The calcium source blends were as follows: 29% fine limestone (LF)+71% large limestone (LG), 32% LF+68% eggshell, 32% LF+68% oyster shell, 50% LF+50% LG. The contents of these blends of calcium in the diets were as follows: 103.3, 93.3, 93.3 and 93.3 g/kg, respectively. 2. The coefficients of calcium retention were significantly higher in 50:50 LF:LG (0.578) and 32:68 LF:eggshell (0.576). The midnight feeding significantly improved the coefficient of calcium retention in all mixtures except 50:50 LF:LG. 3. In the mixtures 29:71 LF:LG and 32:68 LF:oyster shell, there were significantly greater eggshell quality, eggshell weight, eggshell thickness and eggshell strength. Midnight feeding had no significant effect on eggshell quality. 4. In the ration with oyster shells, 96.5% of the retained calcium was deposited in the eggshell, but in rations 32:68 LF:eggshell and 50:50 LF:LG the utilisation was only 73.9 and 78.6%, respectively. 5. To ensure good quality eggshells in the last third of production, the recommendation for calcium is 4.1 g/kg (900 g/kg dry matter, feed intake 110 g/d). As a source of calcium in this stage of production, a feed mixture containing two-thirds large particles should be used (limestone grit or oyster shell).     

用异源蛋白和蛋壳体外培养鹌鹑胚胎的研究

将50个鹌鹑胚胎在鸡蛋壳中用鸡稀蛋白培养,对种蛋产出体外阶段裸黄状态的鹌鹑胚胎进行培养的方法进行了探讨。1～2.5d、2.5～14d和15～17d胚胎培养温度分别是38.0℃、37.8℃和37.5℃,相对湿度分别是60%、55%和70%,1～14d每小时翻蛋1次,翻蛋角度1～2.5d为90°,2.5～14d为50°,15d后静止落盘。2.5d和15d胚胎存活率以及孵化率分别为92%、78%和28%。结果表明裸黄状态的鹌鹑胚胎不排斥鸡稀蛋白,用鸡稀蛋白培养鹌鹑胚胎可行。     

鸡蛋微生物测定与分析

通过对各类型养鸡场和农贸市场鸡蛋进行微生物测定，结果鸡蛋蛋壳表面和蛋内容物含有大量的各种细菌，其中蛋壳表面沙门氏菌带菌率平均为10％，大肠杆菌带菌率为70％；蛋内容物中部分鸡蛋带有沙门氏菌，大肠杆菌带菌率为46．3％。     

Bacterial shell contamination in the egg collection chains of different housing systems for laying hens

The bacterial eggshell contamination of eating eggs in different commercial housing systems; two conventional cages, one organic aviary system and one barn production, were compared. The total counts of aerobic bacteria and the total counts of Gram-negative bacteria on the shell were used to detect key points where contamination occurred and to study the progress of contamination in the egg collection and transportation chains. The key points in the chain were those where eggs accumulated on a short conveyor belt, initial shell contamination in the alternative housing systems and extra nest-boxes placed on the ground. The high bacterial load of floor eggs (>6.3 log CFU total aerobic flora/eggshell) explains why they cannot be used for eating. On average higher initial shell contamination with total counts of aerobic bacteria was found for eggs from the alternative housing systems compared to the conventional systems; respectively 5.46 compared to 5.08 log CFU/eggshell. However, initial contamination with total counts of Gram-negative bacteria on the shells was less in the alternative systems: 3.31 compared to 3.85 log CFU/shell. Initial bacterial shell contamination tended to correlate positively with the concentration of bacteria in the air of the poultry houses. Storing shell eggs, whether temporarily refrigerated or not, for 9 d or more, resulted in a decrease in bacterial eggshell contamination for both bacterial variables.     

Salmonella typhimurium penetration through the eggshell of hatching eggs

This study was undertaken to demonstrate penetration of Salmonella typhimurium through the eggshell of newly laid broiler hatching eggs. Eggs were challenged either by lightly spraying the bacteria over the blunt end of the egg or by contact with contaminated dry nest litter. Exposure time for both groups was 10 minutes; afterward, all eggs were disinfected and incubated 19 days under normal conditions. Chorioallantoic membranes and yolk sacs were cultured in brain-heart infusion broth on day 19 to demonstrate penetration. Isolation of the bacteria from chorioallantoic membranes and yolk sacs, respectively, were as follows: sprayed group: 100% and 83%; contact group: 59% and 29%. These results showed that although water enhanced S. typhimurium penetration, its presence on the eggshell is not essential for penetration to occur.     

蛋壳胶护膜研究进展

蛋产品安全一直是人们重点关注的问题,也是研究的热点。随着福利养殖的推广,鸡蛋被微生物污染的机率增大。除了加强饲养管理之外,强化鸡蛋自身保护屏障从而抵抗细菌入侵也十分重要。胶护膜是蛋壳最外层覆盖的无色透明膜,鸡蛋产出前1.5~2.0 h于子宫部形成,是蛋抵御微生物污染的重要屏障,研究表明胶护膜较厚的蛋不易被细菌入侵。鸡蛋蛋壳胶护膜主要由糖蛋白（90%）、多糖（4%）、脂质（3%）和以羟基磷灰石晶体形式存在的无机磷（3%）组成。胶护膜中含有溶菌酶C、卵转铁蛋白、ovocleidin-32等多种抗菌蛋白,可有效地阻碍细菌跨壳污染。此外,胶护膜能终止蛋壳矿化、帮助蛋产出前在子宫中翻转、调节水气交换以延长鸡蛋的保存时间。本文对胶护膜的结构、生物学功能、胶护膜品质评价方法及胶护膜质量的影响因素等方面进行了阐述,并对尚待解决的问题展开了探讨。     

Ascorbic acid biosynthesis in hens producing strong and weak eggshells

1. An experiment was conducted with two strains of layers to ascertain whether the reduction in eggshell strength occurring at the end of the production cycle is the result of reduced ascorbic acid biosynthesis. 2. Hens producing strong and weak eggshells were identified within each strain and egg production, egg weight, per cent shell, shell surface density, plasma, adrenal and hepatic ascorbic acid and renal L-gulonolactone oxidase activity were measured. 3. The strains differed in ascorbic acid synthesis, as measured by L-gulonolactone oxidase activity, and tissue ascorbic acid concentration. 4. Comparison of results from birds producing eggs of similar weight but markedly different in shell strength detected neither a shell strength group x strain interaction nor an effect of shell strength group on plasma and hepatic ascorbic acid and activity of L-gulonolactone oxidase. 5. The results did not support the hypothesis that tissue ascorbate and ascorbic acid biosynthesis are reduced in old hens producing weak eggshells.     

Comparison of hatchability and some egg quality characteristics in spotted and unspotted partridge (Alectoris chukar) eggs

The current research was conducted to determine the comparison of hatchability and some egg quality characteristics in spotted and unspotted partridge (Alectoris chukar) eggs. The research was carried out on 51 male and 102 female chukar partridges (Alectoris chukar) aged 50 wk, which were raised at the Research and Application Farm of the Faculty of Veterinary Medicine of Selcuk University. In total, 244 spotted and 261 unspotted eggs were used in the research. The hatching eggs were stored at 75% RH and 13°C for 14 d. Thirty spotted and 30 unspotted eggs laid on the same day were used for the evaluation of egg quality characteristics. Egg weight, chick weight, egg weight loss during the hatching period, shape index, hatchability, and some egg quality characteristics were evaluated at the end of the research. Spotted eggs exhibited a higher fertility and hatchability, as well as a lower percentage egg weight loss and embryonic mortality when compared with unspotted eggs. No significant differences were observed between spotted and unspotted eggs for egg weight, chick weight, shape index, specific gravity, albumen height, Haugh unit, yolk color green-red chromaticity (a*), albumen pH, and yolk pH. Spotted eggs had higher eggshell color a*, eggshell color blue-yellow chromaticity (b*), and eggshell strength, as well as a lower eggshell color light-dark chromaticity (L*), yolk color L*, and yolk color b* scores when compared with unspotted eggs. It was demonstrated that spots on the eggshells of chukar partridges (Alectoris chukar) improved their hatchability by affecting various quality characteristics of their eggs. It was concluded that spotted eggs are more suitable for hatching.     

Measuring the eggshell strength of 6 different genetic strains of laying hens: techniques and comparisons

1. Eggshell quality was compared in 6 different strains of laying hens. Three strains were commercial; the three others were experimental. 2. Four different variables describing the strength of eggshells were investigated. Three of them were the classical ones eggshell thickness, shell stiffness measured during quasi-static compression and breaking force. Dynamic stiffness, introduced by Coucke (Ph.D. Thesis, KU Leuven, 1998), was the 4th. The fact that this measurement is dynamic could be helpful in genetic selection for eggshell breakage, because forces applied to the egg in practice are dynamic, rather than static. 3. Hisex White hens produce eggs with the strongest eggshell, in terms of all 4 eggshell variables. However, their shell quality in terms of breaking force did not remain constant over the laying period, unlike 4 other strains. 4. All strains showed a decline in quasi-static stiffness over time. 5. The eggshell thickness of three strains showed a decline over time. 6. Dynamic stiffness remained constant or improved in all strain. 7. All variables describing the mechanical eggshell strength gave different information.     

Effects of commercial enzyme preparations on egg and eggshell quality in laying hens

1. Four different commercial enzyme products were added to standard commercial layer diets, based on barley, wheat or triticale. 2. Diets were fed to 4 different strains of commercial laying hen: ISA Brown, Hy-Line CB, Tegel SB2 and Tegel HiSex. 3. Diets were given for 5 weeks prior to measurements of egg and eggshell quality. 4. The inclusion of commercial enzyme products in the diets had no effect on egg weight. 5. Positive effects of the enzymes were: improved eggshell breaking strength, shell weight, percentage shell and shell thickness for the barley-based diet and increased eggshell breaking strength for two of the enzymes with the wheat-based diets. 6. Negative effects of the enzyme products were slightly lighter coloured eggshells and reduced albumen quality for the barley-based diet and for two enzymes with the wheat-based diet. 7. The addition of commercial enzyme preparations had no main effect on egg and eggshell quality for the triticale-based diet, except for effects on yolk colour. 8. Yolk colour was lower than control for enzyme 3 in all diets and for enzyme 4 with the barley- and wheat-based diets. 9. Further investigation is required to elucidate the specific enzyme components that influence egg quality.     

Heritability and genetic correlation of measurements derived from acoustic resonance frequency analysis; a novel method of determining eggshell quality in domestic hens

Cracked eggshells result in economic loss and provide a route for pathogenic organisms to enter the egg. Genetic factors that contribute to shell strength are likely to decrease the risk that an egg will crack when subject to insult. A novel measurement, the dynamic stiffness of the eggshell (Kdyn) was examined to determine if it might be used in the genetic selection of hens with improved eggshell characteristics. The measurement is determined from acoustic resonance frequency analysis. The estimates of heritability for the novel measurement of Kdyn were moderately high and ranged from 0.33 to 0.53 depending on the model used for the estimation. The estimates of genetic correlation of Kdyn with eggshell breaking strength (0.49) and static stiffness (0.57) were positive and relatively large as expected. There was a small negative genetic correlation between Kdyn and egg production from 26 to 50 weeks of age (-0.19) and a moderate one from 58 to 74 weeks of age (-0.36). The moderate heritability and relative independence of Kdyn indicates that this measurement could be used successfully in a breeding programme to improve shell quality and to reduce the incidence of cracks.     

Constant photoperiods and eggshell quality in broiler breeder pullets

Broiler breeder pullets were exposed to constant 10-, 11-, 12-, 13-, 14- or 16-h photoperiods from 3 d of age. Egg weight, eggshell weight and shell thickness index were determined at 52 weeks of age. Egg weight increased by 0.31 g, shell weight decreased by 30 mg and shell thickness index decreased by 0.57 mg/cm2 for each one-hour increase in photoperiod. Whilst the changes in egg weight and eggshell thickness index might be overstated because eggs were collected at the same chronological time, the effect of time of egg-laying within the day was minimal in comparison, and did not negate the conclusion that egg weight increases, and shell weight and thickness index decrease with lengthening photoperiods. The effect of photoperiod on eggshell quality was not due to differences in the rate of lay between treatments. Shell weight was unaffected by time of lay.     

孕酮对鸡蛋壳品质的影响

本试验旨在通过腿肌注射孕酮来研究孕酮对蛋壳品质的影响。选取72只40周龄的海兰褐蛋鸡,随机分为6组,每组12只蛋鸡,单笼饲养。对照组注射花生油;处理1在排卵后5 h注射1.00 mg/kg体重(BW)的孕酮;处理2和处理3分别在排卵后5 h和2 h注射0.25 mg/kg BW的孕酮;处理4和处理5分别在排卵后2 h和1 h注射0.15 mg/kg BW的孕酮。处理后第2天上午收集各组所产鸡蛋,测定蛋壳强度和厚度。结果表明:处理1所产蛋均为破壳蛋,处理2含部分破壳蛋和部分蛋壳完好的鸡蛋,处理3、处理4和处理5没有破壳蛋;与对照组相比,处理1和处理2的蛋壳厚度降低(P<0.05),处理4蛋壳强度和厚度增加(P<0.05),处理3和处理5的蛋壳强度和厚度无显著差异;与对照组比,处理1和处理4的蛋壳超微结构差异显著。结果显示,孕酮可以影响蛋壳强度和厚度,改变蛋壳超微结构,其影响程度与剂量和注射时间有关。     

The effects of sage extract feed supplementation on biochemical parameters, weight of internal organs and Salmonella counts in chickens

We investigated the effects of dietary addition of sage extract on the biochemical parameters, weight of some body organs and changes in the counts of Salmonella Enteritidis PT4 (SE) in experimentally infected chickens. The following diets were used: basal diet, basal diet with addition of an extract of Salvia officinalis L. (S), basal diet and SE, and basal diet and S and SE (SSE). Compared to the SE group, sage extract in the SSE group decreased activities of ALP and ALT and concentrations of glucose and bilirubin on the 4th day post inoculation (p.i.). However, on the 18th day p.i., lower levels of bilirubin and ALT activity only were detected. Addition of sage extract to the diets decreased the counts of Salmonella in the liver, spleen and caecum at both sampling times, along with lower production of mucus in the chickens' intestines. Our results suggest that the addition of sage extract to the diet could be effective in protecting SE-infected chickens.