The genome of an endosymbiotic methanogen is very similar to those of its free-living relatives.

The methanogenic endosymbionts of anaerobic protists represent the only known intracellular archaea, yet, almost nothing is known about genome structure and content in these lineages. Here, an almost complete genome of an intracellular Methanobacterium species was assembled from a metagenome derived from its host ciliate, a Heterometopus species. Phylogenomic analysis showed that the endosymbiont was closely related to free-living Methanobacterium isolates, and when compared with the genomes of free-living Methanobacterium, the endosymbiont did not show significant reduction in genome size or GC content. Additionally, the Methanobacterium endosymbiont genome shared the majority of its genes with its closest relative, though it did also contain unique genes possibly involved in interactions with the host via membrane-associated proteins, the removal of toxic by-products from host metabolism and the production of small signalling molecules. Though anaerobic ciliates have been shown to transmit their endosymbionts to daughter cells during division, the results presented here could suggest that the endosymbiotic Methanobacterium did not experience significant genetic isolation or drift and/or that this lineage was only recently acquired. Altogether, comparative genomic analysis identified genes potentially involved in the establishment and maintenance of the symbiosis, as well provided insight into the genomic consequences for an intracellular archaeum.

Influence of commercial laying hen housing systems on the incidence and identification of Salmonella and Campylobacter.

The housing of laying hens is important for social, industrial, and regulatory aspects. Many studies have compared hen housing systems on the research farm, but few have fully examined commercial housing systems and management strategies. The current study compared hens housed in commercial cage-free aviary, conventional cage, and enriched colony cage systems. Environmental and eggshell pool samples were collected from selected cages/segments of the housing systems throughout the production cycle and monitored for Salmonella and Campylobacter prevalence. At 77 wk of age, 120 hens per housing system were examined for Salmonella and Campylobacter colonization in the: adrenal glands, spleen, ceca, follicles, and upper reproductive tract. All isolates detected from environmental swabs, eggshell pools, and tissues were identified for serotype. Two predominant Salmonella were detected in all samples:S.Braenderup andS.Kentucky.Campylobacter coli and C. jejuni were the only Campylobacter detected in the flocks. Across all housing systems, approximately 7% of hens were colonized with Salmonella, whereas >90% were colonized with Campylobacter Salmonella Braenderup was the isolate most frequently detected in environmental swabs (P<0.0001) and housing system impacted Salmonella spp. shedding (P<0.0001).Campylobacter jejuni was the isolate most frequently found in environmental swabs (P<0.01), while housing system impacted the prevalence of C. coli and jejuniin ceca (P<0.0001). The results of this study provide a greater understanding of the impact of hen housing systems on hen health and product safety. Additionally, producers and academia can utilize the findings to make informed decisions on hen housing and management strategies to enhance hen health and food safety.

Microbiological impact of three commercial laying hen housing systems.

Hen housing for commercial egg production continues to be a societal and regulatory concern. Controlled studies have examined various aspects of egg safety, but a comprehensive assessment of commercial hen housing systems in the US has not been conducted. The current study is part of a holistic, multidisciplinary comparison of the diverse aspects of commercial conventional cage, enriched colony cage, and cage-free aviary housing systems and focuses on environmental and egg microbiology. Environmental swabs and eggshell pools were collected from all housing systems during 4 production periods. Total aerobes and coliforms were enumerated, and the prevalence of Salmonella and Campylobacter spp. was determined. Environmental aerobic and coliform counts were highest for aviary drag swabs (7.5 and 4.0 log cfu/mL, respectively) and enriched colony cage scratch pad swabs (6.8 and 3.8 log cfu/mL, respectively). Aviary floor and system wire shell pools had the greatest levels of aerobic contamination for all eggshell pools (4.9 and 4.1 log cfu/mL, respectively). Hens from all housing systems were shedding Salmonella spp. (89-100% of manure belt scraper blade swabs). The dry belt litter removal processes for all housing systems appear to affect Campylobacter spp. detection (0-41% of manure belt scraper blade swabs) considering detection of Campylobacter spp. was much higher for other environmental samples. Aviary forage area drag swabs were 100% contaminated with Campylobacter spp., whereas enriched colony cage scratch pads had a 93% positive rate. There were no differences in pathogen detection in the shell pools from the 3 housing systems. Results indicate egg safety is enhanced when hens in alternative housing systems use nest boxes. Additionally, current outcomes indicate the use of scratch pads in hen housing systems needs to be more thoroughly investigated for effects on hen health and egg safety.

Environmental sampling methods' influence on detection of pathogens in cage-free aviary housing.

The environmental sampling of layer housing systems is essential to identifying potential pathogens that are of concern to human health. To identify the natural occurrence of pathogens (Listeria, Campylobacter, and Salmonella) at various locations in a cage-free aviary housing system, swabs were collected when hens were 22 to 39 wks of age. Duplicate environmental swabs were taken and inoculated with a low dose (101 cfu) Salmonella Enteritidis (SE) and examined for the recovery of SE from environmental samples. Detection of Listeria (P < 0.0001) and Campylobacter (P < 0.0001) varied between the environmental sample types taken: concrete dust, drag swabs, egg belt dust, manure belt scraper swabs, and wall dust. Detection of Listeria (P < 0.0001) was the highest (70.0%) at the beginning of the study (22 wk) and decreased over time. Detection of Campylobacter (P < 0.001) was also the highest at 22 wk, however the decrease over time was more gradual. Interestingly, detection of Campylobacter (P < 0.0001) was the greatest in concrete dust samples (96.25%), which can be attributed to the presence of rodent excreta in the samples. Drag swabs and manure belt scraper swabs were the best sampling types for high detection of Listeria and Campylobacter. It should be noted that Listeria recovered was not of human health concern. No naturally occurring Salmonella was identified in this study. The recovery of the SE inoculum increased over time, reaching the greatest recovery in drag (81.25%; P < 0.0001), egg belt dust (100.00%; P < 0.0001) and wall dust swabs (100.00%; P < 0.0001) by 39 wk. This high rate of SE recovery occurred just before US mandatory SE environmental monitoring at 40 to 45 wks of age. Based on this study, the use of drag and manure belt scraper swabs are effective in detecting Listeria and Campylobacter in cage-free aviary housing. Along with good pest management, the occurrence of pathogens could be monitored and reduced in laying hen flocks.

The impact of different housing systems on egg safety and quality.

A move from conventional cages to either an enriched cage or a noncage system may affect the safety or quality, or both, of the eggs laid by hens raised in this new environment. The safety of the eggs may be altered either microbiologically through contamination of internal contents with Salmonella enterica serovar Enteritidis (Salmonella Enteritidis) or other pathogens, or both, or chemically due to contamination of internal contents with dioxins, pesticides, or heavy metals. Quality may be affected through changes in the integrity of the shell, yolk, or albumen along with changes in function, composition, or nutrition. Season, hen breed, flock age, and flock disease-vaccination status also interact to affect egg safety and quality and must be taken into account. An understanding of these different effects is prudent before any large-scale move to an alternative housing system is undertaken.

In vitro penetration of Salmonella Enteritidis through yolk membranes of eggs from 6 genetically distinct commercial lines of laying hens.

Although deposition of Salmonella Enteritidis inside yolks is less common than deposition in albumen or on the vitelline (yolk) membrane in naturally contaminated eggs laid by infected hens, bacterial migration into the yolk to reach its nutrient-rich contents could lead to extensive multiplication. The present study used an in vitro egg contamination model to assess the ability of small initial numbers of Salmonella Enteritidis to penetrate the vitelline membrane and multiply inside yolks of eggs laid by 6 genetically distinct commercial lines of hens during 24 h of storage at 30 degrees C. Eggs from each line were tested at 4 different hen ages by inoculation of approximately 100 cfu of Salmonella Enteritidis onto the outside of the vitelline membranes of intact yolks in plastic centrifuge tubes and then adding back the albumen into each tube before incubation. Overall, the frequency of penetration of Salmonella Enteritidis into the yolk contents of eggs from individual lines of hens ranged from 30 to 58% and the mean concentration of Salmonella Enteritidis in yolk contents after incubation ranged from 0.8 to 2.0 log(10) cfu/mL. For both of these parameters, values for one hen line were significantly higher than for 2 other lines, but no other differences were observed. Hen age did not have a significant effect on egg yolk penetration by Salmonella Enteritidis. These results indicate that opportunities for the migration and growth of small initial numbers of Salmonella Enteritidis to attain more dangerous levels inside contaminated eggs during storage at warm temperatures can sometimes vary between different lines of laying hens.

Crop immune response post-Salmonella enteritidis challenge in eight commercial egg-layer strains and specific-pathogen-free White Leghorn chickens.

The crop immune response against Salmonella Enteritidis (SE) challenge in eight commercial egg-layer strains (five white-egg layer and three brown-egg layer) and specific-pathogen-free (SPF) White Leghorn (WL) hens was investigated. Pre- and post-SE challenge mucosal immune responses within the crops were evaluated. Commercial layers and SPF WL hens were orally challenged with 10(8) CFU/ml SE PT13a and SE nalR PT13, respectively. Crop lavage samples were collected at weekly intervals from day 0 (pre-challenge) to day 25-27 postinfection (PI), and bacteriological examination was performed to monitor progression of SE infection. Crop lavage samples were analyzed for SE-lipopolysaccharide (LPS)-specific IgA using enzyme-linked immunosorbent assay (ELISA). H&E-stained slides of crop sections from day 34 PI and uninfected controls were assessed for lymphoid tissue via light microscopy. Lymphoid areas were graded based on morphology, size, and cellularity using a score 0 to 5 scale. The 0 to 5 (low to high) numerical values represented progressive increases in size and cellular density of lymphoid tissue. Bacterial culture results showed the highest percentage of SE-positive crop lavage samples from all hen groups at day 5-6 PI and day 11-12 PI. A progressive decline in percentage of SE-positive crop lavage samples did occur as time PI lengthened; however, at day 25-27 PI SE persisted in crop lavage samples from SPF WL hens and three commercial white-egg layer strains. A marked increase in SE-LPS-specific IgA was measured in crop lavage samples between day 0 and day 11-12 PI for all hen groups. Crop SE-LPS-specific IgA response remained elevated above day 0 baseline for the duration of the experiment. Well-defined score 3 to 5 lymphoid tissue aggregates were observed in crop tissue sections harvested at day 34 PI. Comparison of crop sections determined a 1.2-4.0 times increase in ratio of lymphoid tissue in day 34 PI SE-challenged hens vs. uninfected control hens.

Marine vertebrate zoonoses: an overview of the DAO special issue.

The role of marine birds, mammals, turtles and fish as vectors of infectious agents of potential risk to humans can be examined from a variety of perspectives. The studies in this DAO Special include a broad survey of multiple agents and species, a sequencing study of Giardia intestinalis haplotypes known to be pathogenic to humans, an assessment of risks to humans working with marine mammals, a source tracking study using E. coli ribotypes, studies of regional Salmonella and Brucella epizootiology, a serology survey and a case report of a herpes simplex infection in a dolphin. Additionally, a recently published study (Venn-Watson et al. 2008; Dis Aquat Org 79:87-93) classifying pure cultures of bacteria from a captive dolphin colony also pertains to this theme. These studies raise the following questions: whether the presence of zoonotic agents in marine vertebrates represents a risk to other marine vertebrates, humans, or both; what are the routes by which these marine vertebrate zoonotic infections are acquired and circulated in the marine ecosystem; to what degree are such agents subclinical versus causes of overt disease in marine vertebrates; what are the subsets of the human population most likely to be affected by such infections; and which human health preventive measures would seem reasonable?

Effects of different litter substrates and induced molt on production performance and welfare quality parameters of white Leghorn hens housed in multi-tiered aviary system.

More than 90% of the commercial egg production in the United States is pledged to be in cage-free systems by 2025. Management practices like induced molting and litter area management have come under scrutiny because of the housing system change. The aim of this study was to determine the welfare and production implications of different litter substrates and also evaluate induced molting of hens in a cage-free system. Bovan White hens were housed in a multi-tier aviary system with daily access to open litter area of either Astroturf (AT), wood shavings (SH), or straw (ST) and bare concrete floor (CO) serving as control. At 68 wk of age, molt was induced in half of the hens whereas the other half continued without molting to 116 wk. Production and welfare parameters were measured periodically throughout first and second cycles. Litter substrate did not influence hen-day production and case-weight measurements. However, CO had the lowest total number of eggs produced during the first cycle (P < 0.05). Hen-day percentage was approximately 14% greater in molted hens during the second cycle with egg case weight being heavier in non-molt hens toward the end of second cycle (P < 0.05). The only welfare parameter influenced by litter substrate during the first cycle was a greater crop feather loss in AT than ST at mid-lay (P < 0.05). Keel deformations increased with age irrespective of the litter substrate with 91.5% of palpated hens having keel deformations at the end of first cycle (P < 0.05). Molting did not influence the keel palpation and footpad scores whereas frequency of moderate comb wound was greater in molt hens during molt (P < 0.05). Severe feather loss was seen in non-molt hens during the second cycle (P < 0.05). Litter substrate does not affect production and physical parameters of welfare of hens in a multi-tier aviary system. Additionally, induced molting can be successfully carried out in the multi-tier cage-free system.

Hen genetic strain and extended cold storage influence on physical egg quality from cage-free aviary housing system.

In the United States, there is an increase in need for cage-free eggs in retail and food manufacturing sectors. Understanding the impact of cage-free systems and the corresponding management on egg quality is pertinent as the U.S. industry adapts existing housing and builds new cage-free housing structures. A study was conducted comparing 2 brown shell and 2 white shell hen strains housed in a cage-free aviary system. Each set of eggs were placed in cold storage and assessed at 0, 2, 4, 8, and 12 wk. Eggs were collected at 21, 31, 42, and 60 wk of hen age. A full profile of physical quality measurements was conducted on up to 18 intact eggs for each hen strain/egg storage/hen age combination. Egg weight increased approximately 10 g for brown shell and 14 g for white shell eggs as hens aged. Many of the properties monitored were significantly impacted by all 3 main effects (hen strain, egg storage, and hen age) resulting in 3-way interactions. A brown and a white shell strain had stronger shells (44 N; P < 0.0001) than the remaining brown and white shell strains (42 N and 39 N, respectively). The current study also determined volume of shell, total length, maximum width, and percent length at maximum width to more accurately indicate egg shape than shape index. One brown shell strain produced eggs with the most consistent shape characteristics over the hen ages monitored. White shell eggs from the cage-free aviary housing produced the highest whole-egg total solids between 31 to 60 wk of hen age, whereas brown shell eggs resulted in the most consistent level of whole-egg total solids (22-23.5%). The brown and white shell strains in the current study produce cage-free aviary eggs with distinctive physical quality attributes. The outcomes from this study can be utilized by the U.S. egg industry in planning management strategies and market placement of cage-free eggs.

In vitro penetration of egg yolks by Salmonella Enteritidis and Salmonella Heidelberg strains during thirty-six-hour ambient temperature storage.

Although Salmonella deposition inside yolks is uncommon in naturally contaminated eggs, migration through the vitelline membrane into the nutrient-rich yolk contents could enable rapid bacterial multiplication. Egg refrigeration restricts both penetration and growth, but a recently proposed national Salmonella Enteritidis control program would allow unrefrigerated ambient temperature storage of eggs on farms for up to 36 h. The present study used an in vitro egg contamination model to assess the ability of small numbers of 4 Salmonella Enteritidis strains and 4 Salmonella Heidelberg strains to penetrate the vitelline membrane and multiply inside yolks during 36 h of storage at either 20 or 30 degrees C. After inoculation onto the exterior surface of the vitelline membrane, all 8 Salmonella strains penetrated to the yolk contents (at a mean frequency of 45.1%), and most strains grew to significantly higher levels (with a mean (log)10 bacterial concentration of 2.2 cfu/mL) during incubation at 30 degrees C. Significant differences in penetration frequency and yolk multiplication were observed between individual strains and between serotypes (Salmonella Enteritidis > Salmonella Heidelberg for both parameters). Penetration and multiplication were significantly less frequent during incubation at 20 degrees C. These results demonstrate that controlling ambient temperatures during prerefrigeration storage may be an important adjunct to prompt refrigeration for limiting Salmonella growth in eggs and thereby for preventing egg-transmitted human illness.

Enhanced gross visualization of chicken Peyer's patch: novel staining technique applied to fresh tissue specimens.

The ileal Peyer's patches (Pp), secondary gut-associated lymphoid tissue of the mucosal immune system, may serve as an important site for monitoring inflammatory and immunologic responses of the host against enteric pathogens. Chicken Pp are often difficult to observe grossly, and a simple technique to enhance visualization of the Pp is lacking. Therefore, we designed a novel staining method that is quick, easy, and accurate to aid in gross identification and recovery of the chicken Pp from fresh tissue specimens. Lower alimentary tracts were harvested from White Leghorn hens and commercial broilers. The ileocecocolic region was excised intact, flushed with deionized water to remove ingesta, and a dilute eosin-Y solution was infused. After 1 min, the eosin-Y was gently extruded. Modified-crystal violet (mCV) was then injected into the gastrointestinal segment, where on the lymphoid tissue area became apparent at the serosal surface. The distal ileal Pp was visible as a pale whitish pink ovoid-focalized area with surrounding gut tissue stained light purple. The exact Pp site could be delineated at the serosal and mucosal surface by gross assessment. Light microscopy evaluation of hematoxylin and eosin-stained tissue slides prepared from the excised Pp site revealed lymphoid tissue aggregations with multiple follicular units indicative of Pp. The novel eosin-Y + mCV staining technique promotes rapid identification and accurate recovery of chicken Pp lymphoid tissue from fresh tissue specimens.

DGGE-based detection method for Quahog Parasite Unknown (QPX).

Quahog Parasite Unknown (QPX) is a significant cause of hard clam Mercenaria mercenaria mortality along the northeast coast of the United States. It infects both wild and cultured clams, often annually in plots that are heavily farmed. Subclinically infected clams can be identified by histological examination of the mantle tissue, but there is currently no method available to monitor the presence of QPX in the environment. Here, we report on a polymerase chain reaction (PCR)-based method that will facilitate the detection of QPX in natural samples and seed clams. With our method, between 10 and 100 QPX cells can be detected in 1 l of water, 1 g of sediment and 100 mg of clam tissue. Denaturing gradient gel electrophoresis (DGGE) is used to establish whether the PCR products are the same as those in the control QPX culture. We used the method to screen 100 seed clams of 15 mm, and found that 10 to 12% of the clams were positive for the presence of the QPX organism. This method represents a reliable and sensitive procedure for screening both environmental samples and potentially contaminated small clams.

Studies on the binding of FMN by apoflavodoxin from Peptostreptococcus elsdenii, pH and NaCl concentration dependence.

1. The pH and ionic strength dependence of the interaction of FMN with apoflavodoxin has been studied by fluorometry in the pH region 2-5, at 22 degrees C. 2. The rate constant of dissociation and the dissociation constant were experimentally determined; the rate constants of association were claculated at a given pH value. These constants depend on the ionic strength. The plots of these constants against the square root of the ionic strength are straight. 3. Our data have been interpreted in terms of the Brönsted theory, which relates chemical reaction rates to ionic strength. The data indicate that the apoenzyme reaches its maximum net positive charge at pH 2.0-2.6. The calculated net charge in this pH region is between 11 and 12 and is in agreement with the theoretical value of 12 as deduced from the primary structure of the protein. The isoelectric point of the holoenzyme is about 4. 4. The rate constant of association extrapolated to zero ionic strength is 3.2-10(5)M-1-s-1 and is pH-independent. 5. The rate constant of dissociation and the dissociation constant extrapolated to zero ionic strength depend on the pH. The results are explained by assuming that there are two protein ionizations with a pK value of 3.4; these ionizing groups are possibly close to the FMN binding site.

Penetration of Salmonella enteritidis and Salmonella heidelberg into egg yolks in an in vitro contamination model.

Eggs that harbor Salmonella in their edible contents pose a significant risk of transmitting disease to consumers. Although Salmonella deposition inside yolks does not usually occur at a high frequency in naturally contaminated eggs, bacterial penetration through the vitelline membrane could lead to rapid and extensive multiplication in the nutrient-rich yolk contents. The present study used an in vitro egg contamination model to assess the ability of Salmonella strains to penetrate the vitelline membrane and multiply inside yolks. An S. enteritidis strain and 2 Salmonella heidelberg strains, initially inoculated onto the outside of the vitelline membrane, were able to enter the yolk contents (at frequencies ranging from 10 to 25% of experimentally contaminated eggs) during 24 h of incubation at 30 degrees C. Variants of these parent strains, obtained by in vivo passage into eggs laid by infected hens, penetrated the yolk membrane at significantly higher frequencies. These results demonstrate that pathogens such as S. enteritidis and S. heidelberg can penetrate into and begin to multiply inside the yolks of contaminated eggs during the first day of storage at warm temperatures.

A dual-labeling method for identifying differentially expressed genes: use in the identification of cDNA clones that hybridize to RNAs whose abundance in tomato flowers is potentially regulated by gibberellins.

A method for identifying cDNA clones that hybridize to differentially expressed RNAs is described. Briefly, the RNA population in which the RNAs of interest are more abundant is used as a template for the synthesis of 35S-labeled cDNAs and another RNA population in which the RNAs of interest are less abundant is used as a template for the synthesis of 32P-labeled cDNAs. The labeled cDNAs are pooled and hybridized to plaque or colony lifts constructed from a cDNA library. Clones that hybridize to RNAs that are differentially expressed are identified using differential autoradiography/fluorography to discriminate between the 32P and 35S isotopes. We have used this method to identify cDNA clones that hybridize to mRNAs that are more abundant in the flowers of wild-type tomato than in the flowers of mutants that have low endogenous levels of gibberellins.

Closed-system technique for implantation of iris-supported negative-power intraocular lens.

BACKGROUND: The iris-supported claw lens can reversibly correct a wide range of ametropia and aniseikonia with excellent predictability and stability. However, concerns regarding its potential harm to the corneal endothelium have prevented its widespread use. Our experience with the closed-system surgical technique and the spreader device used in 180 cases between 1987 and 1993 has led us to conclude that potential complications are more likely due to the technique of implantation rather than to the properties of the lens itself. METHOD: We conducted a prospective study of 35 eyes of 20 consecutive patients (mean age, 38 years; range, 21 to 55 years) operated on between August 1993 and August 1994. Preoperative spherical equivalent refractions ranged from -6.00 to -21.25 diopters (D) (mean, -12.50 D). Follow up was 6 months (86% of eyes) to 1 year (57% of eyes). Spectacle-corrected visual acuity and endothelial cell density were measured prior to, and 1, 6, and 12 months following lens implantation. All data were analyzed using the paired t-test. RESULTS: Mean endothelial cell density dropped insignificantly (p > .10):by 1.22% at 1 month (n = 34), by 2.25% at 6 months (n = 27), and by 1.21% at 12 months (n = 18). Spectacle-corrected visual acuity increased significantly (p < .001): from a mean of 0.61 preoperatively to 0.77 at 1 month, 0.84 at 6 months, and 0.93 at 12 months postoperatively. At 12 months, 15 eyes (75%) had a refraction within +/-0.50 D, 19 eyes (95%) within +/-1.00 D, and 20 eyes (100%) within +/-2.00 D of emmetropia. CONCLUSIONS: Our results suggest that the closed-system approach using the spreader device allows safe implantation of the Worst iris-supported claw lens.

The suction bridge for radial keratotomy may avoid late hyperopic shift.

BACKGROUND: Radial keratotomy may induce late hyperopic shift. We present data on 140 consecutive eyes with a follow-up of up to 3 years that underwent radial keratotomy with the RK suction bridge. METHODS: We conducted a retrospective study of 140 consecutive eyes that had radial keratotomy between 1987 and 1994. Mean preoperative spherical equivalent was -5.21 D (range -2.00 to -9.75 D). All operations were performed by one surgeon (JHK) with the RK suction bridge. A suction ring maintaining physiological intraocular pressure immobilized the eye and left a peripheral rim of uncut cornea. The ring incorporated an eccentric bridge that guided the radial keratotomy knife. The knife setting was 90% of the central corneal thickness, measured by pachymetry. Spherical equivalent refraction and spectacle corrected visual acuity were measured at 1 week, 1, 3, 6 months, 1 year, and 3 years after radial keratotomy. RESULTS: The mean preoperative spherical equivalent refraction of -5.21 D dropped to -0.43 D at 1 week (n = 136), -0.71D at 1 month (n = 120), -0.85 D at 3 months (n = 95), -0.74 D at 6 months (n = 73), -0.77 D at 12 months (n = 79), and -0.85 D at 3 years (n = 67). Compared to 1 month spherical equivalent, at 3 years three eyes (4.4%) had moved > = or 1.00 D toward hyperopia. One eye (1.4%) shifted by 1.25 D. Paired t-tests of mean spherical equivalent refraction did not reveal significant shifts toward hyperopia. Mean preoperative spectacle-corrected visual acuity was slightly diminished at 1 week and was equal or better thereafter. CONCLUSIONS: Our 3-year data suggest that a late hyperopic shift following radial keratotomy may be prevented if an intact peripheral rim is maintained and cutting depth does not exceed 90% of the lowest corneal thickness.

Understanding Salmonella enteritidis in laying chickens: the contributions of experimental infections.

A significant proportion of human Salmonella enteritidis (SE) outbreaks in recent years has been traced to the consumption of contaminated eggs produced by infected laying flocks. Experimental SE infections in chickens have been used to acquire a considerable amount of basic information about the interaction between SE and the avian host. After oral or parenteral inoculation of chickens, SE can colonize the intestinal tract, invade and disseminate to reach numerous internal organ sites, and elicit the production of specific antibodies in serum and egg yolks. Experimental infection of laying hens can result in the deposition of SE in the contents of eggs before oviposition, although generally in rather small numbers and at a relatively low frequency. The consequences of experimental SE infection have been shown to vary significantly with the strain and dose of the inoculum. Some SE isolates have been shown to produce various clinical effects, including decreased egg production by hens. The information provided by experimental SE infections in chickens has played an important role in the formulation of appropriate strategies for reducing the incidence of SE in commercial laying flocks and thereby also reducing the incidence of transmission of SE to consumers via contaminated eggs.

Multiplication in egg yolk and survival in egg albumen of Salmonella enterica serotype Enteritidis strains of phage types 4, 8, 13a, and 14b.

Refrigeration of eggs is vital for restricting the multiplication of Salmonella enterica serotype Enteritidis contaminants, but differences between Salmonella Enteritidis strains or phage types in their survival and multiplication patterns in egg contents might influence the effectiveness of refrigeration standards. The present study compared the abilities of 12 Salmonella Enteritidis isolates of four phage types (4, 8, 13a, and 14b) to multiply rapidly in egg yolk and to survive for several days in egg albumen. The multiplication of very small numbers of Salmonella Enteritidis inoculated into yolk (approximately 10(1) CFU/ml) was monitored during 24 h of incubation at 25 degrees C, and the survival of much larger numbers of Salmonella Enteritidis inoculated into albumen (approximately 10(5) CFU/ml) was similarly evaluated during the first 3 days of incubation at the same temperature. In yolk, the inoculated Salmonella Enteritidis strains multiplied to mean levels of approximately 10(3) CFU/ml after 6 h of incubation and 10(8) CFU/ml after 24 h. In albumen, mean levels of approximately 10(4) CFU/ml or more of Salmonella Enteritidis were maintained through 72 h. Although a few differences in multiplication and survival were observed between individual isolates, the overall range of values was relatively narrow, and no significant differences (P < 0.05) were evident among phage types.