More Is Not Always Better: Coinfections with Defensive Symbionts Generate Highly Variable Outcomes.

Animal-associated microbes are highly variable, contributing to a diverse set of symbiont-mediated phenotypes. Given that host and symbiont genotypes, and their interactions, can impact symbiont-based phenotypes across environments, there is potential for extensive variation in fitness outcomes. Pea aphids, Acyrthosiphon pisum, host a diverse assemblage of heritable facultative symbionts (HFS) with characterized roles in host defense. Protective phenotypes have been largely studied as single infections, but pea aphids often carry multiple HFS species, and particular combinations may be enriched or depleted compared to expectations based on chance. Here, we examined the consequences of single infection versus coinfection with two common HFS exhibiting variable enrichment, the antiparasitoid Hamiltonella defensa and the antipathogen Regiella insecticola, across three host genotypes and environments. As expected, single infections with either H. defensa or R. insecticola raised defenses against their respective targets. Single infections with protective H. defensa lowered aphid fitness in the absence of enemy challenge, while R. insecticola was comparatively benign. However, as a coinfection, R. insecticola ameliorated H. defensa infection costs. Coinfected aphids continued to receive antiparasitoid protection from H. defensa, but protection was weakened by R. insecticola in two clones. Notably, H. defensa eliminated survival benefits conferred after pathogen exposure by coinfecting R. insecticola Since pathogen sporulation was suppressed by R. insecticola in coinfected aphids, the poor performance likely stemmed from H. defensa-imposed costs rather than weakened defenses. Our results reveal a complex set of coinfection outcomes which may partially explain natural infection patterns and suggest that symbiont-based phenotypes may not be easily predicted based solely on infection status.IMPORTANCE The hyperdiverse arthropods often harbor maternally transmitted bacteria that protect against natural enemies. In many species, low-diversity communities of heritable symbionts are common, providing opportunities for cooperation and conflict among symbionts, which can impact the defensive services rendered. Using the pea aphid, a model for defensive symbiosis, we show that coinfections with two common defensive symbionts, the antipathogen Regiella and the antiparasite Hamiltonella, produce outcomes that are highly variable compared to single infections, which consistently protect against designated enemies. Compared to single infections, coinfections often reduced defensive services during enemy challenge yet improved aphid fitness in the absence of enemies. Thus, infection with multiple symbionts does not necessarily create generalist aphids with "Swiss army knife" defenses against numerous enemies. Instead, particular combinations of symbionts may be favored for a variety of reasons, including their abilities to lessen the costs of other defensive symbionts when enemies are not present.

Phage loss and the breakdown of a defensive symbiosis in aphids.

Terrestrial arthropods are often infected with heritable bacterial symbionts, which may themselves be infected by bacteriophages. However, what role, if any, bacteriophages play in the regulation and maintenance of insect-bacteria symbioses is largely unknown. Infection of the aphid Acyrthosiphon pisum by the bacterial symbiont Hamiltonella defensa confers protection against parasitoid wasps, but only when H. defensa is itself infected by the phage A. pisum secondary endosymbiont (APSE). Here, we use a controlled genetic background and correlation-based assays to show that loss of APSE is associated with up to sevenfold increases in the intra-aphid abundance of H. defensa. APSE loss is also associated with severe deleterious effects on aphid fitness: aphids infected with H. defensa lacking APSE have a significantly delayed onset of reproduction, lower weight at adulthood and half as many total offspring as aphids infected with phage-harbouring H. defensa, indicating that phage loss can rapidly lead to the breakdown of the defensive symbiosis. Our results overall indicate that bacteriophages play critical roles in both aphid defence and the maintenance of heritable symbiosis.

Corneal optical aberrations induced by photorefractive keratectomy.

BACKGROUND: Photorefractive keratectomy causes marked alteration to anterior corneal topography, and is likely to induce major changes to the optical aberrations of the eye. METHODS: Six diopters (D) of myopia correction was attempted on one eye of 50 patients, randomly allocated to one of three different treatments: 5-mm or 6-mm single ablation zone, or a double ablation (multizone; -5.00 D correction over 4.6 mm and -1.00 D over 6 mm). Topographic data was used to estimate corneal aberration coefficients. These were compared for effect of ablation zone, before and 1 year after photorefractive keratectomy. The coefficients were used to derive modulation transfer functions for the anterior corneal surface. RESULTS: Corneal spherical aberrations and coma-like aberrations both increased significantly following photorefractive keratectomy (p < 0.001). The mean spherical aberration coefficient increased from 0.36 +/- 0.11 before, to 0.91 +/- 0.37 after treatment, while the mean coma-like aberration coefficient changed from 0.28 +/- 0.16 before, to 0.60 +/- 0.31 after treatment. Ablation zone form had a significant effect on spherical aberration (p = 0.030), but not for coma (p = 0.96). The spherical aberration coefficient increased least for the 6-mm ablation (by 0.38 +/- 0.17), compared with the 5-mm ablation (0.69 +/- 0.45) and the multizone (0.62 +/- 0.38). Corneal modulation transfer functions were reduced significantly following the photorefractive procedure. The effect was greatest for large pupil diameters and for spatial frequencies between 2 and 15 cycles per degree. CONCLUSIONS: Corneal modulation transfer function calculations suggest that a significant loss of visual performance should be anticipated following photorefractive keratectomy, the effect being greatest for large pupil diameters. Results for three ablation zones show that induced aberrations are least for the largest (6 mm) ablation zone.

Effects of ablation diameter, depth, and edge contour on the outcome of photorefractive keratectomy.

PURPOSE: To investigate the effects of the ablation diameter, depth, and edge contour on the outcome of excimer laser photorefractive keratectomy (PRK). METHODS: A prospective study was conducted in which 60 patients (60 eyes) were randomly allocated to 5.00-mm, 6.00-mm, or 5.00 to 6.00-mm multizone treatment groups. All eyes underwent a -6.00 diopter (D) correction using a Summit Omnimed excimer laser. RESULTS: In eyes treated with 6.00-mm diameter zones, the initial hyperopic shift was reduced, with significant differences at 1 and 4 weeks (p < 0.01). At 6 and 12 months, the refractive changes were closer to the intended correction with 6.00-mm diameters. The predictability of PRK was improved with 6.00-mm zones, with a significant reduction in variance of the refractive changes, at all stages postoperatively (p < 0.05 to p < 0.001). Objective measurements of haze were significantly less at 1, 3, and 6 months with 6.00-mm ablations (p < 0.05). There were no differences between the 5.00-mm and the 5.00- to 6.00-mm multizone groups. Computerized measurements of "night" halo were significantly smaller in the 6.00-mm treatment group at 1 week and 1 month (p < 0.05). At 12 months, two patients treated with 5.00-mm zones and three with the 5.00- to 6.00-mm multizone complained of severe night vision disturbances. No 6.00-mm eyes were similarly affected. CONCLUSIONS: Treatment with a 6.00-mm spherical ablation diameter produced less initial overcorrection, improved predictability, and was associated with a reduction in postoperative halos and night vision disturbances. Creating a superficial blend zone with a 5.00- to 6.00-mm multizone treatment had no beneficial effect on the outcome.

Anterior corneal optical aberrations induced by photorefractive keratectomy for hyperopia.

PURPOSE: Photorefractive keratectomy (PRK) for hyperopia requires both a steepening of the central cornea and a flattening of the mid-periphery to achieve its effect and is likely to affect the optical aberrations of the eye. METHODS: Nine patients underwent PRK to correct between +2.00 and +4.00 D of hyperopia (first eye treated for each patient) using the Summit Technology Apex Plus excimer laser. Anterior corneal aberrations for pupil diameters of 3, 5.5 and 7 mm were estimated from corneal topography data (TMS-1), assuming a uni-index, single surface cornea. Refractive error was assessed using retinoscopy and standard subjective tests. RESULTS: Apart from the intended change in refraction (mean spherical equivalent manifest refraction, +4.60 +/- 1.60 D before surgery and +0.70 +/- 1.60 D at 1 year after surgery), the most significant change was in spherical aberration. Anterior corneal spherical aberration was positive (+1.60 +/- 0.60 D for a 5.5-mm pupil) before surgery and became negative after surgery (-1.80 +/- 1.20 D at 1 year). The change in spherical aberration was related to the achieved change in refractive error. CONCLUSIONS: The large change (approximately 3.00 D) in spherical aberration (from positive to negative aberration) has implications for the optical performance of the whole eye, where the effects of lenticular aberration must also be considered.

Effect of ablation profile on wound healing and visual performance 1 year after excimer laser photorefractive keratectomy.

BACKGROUND: Early photorefractive keratectomy ablations were of limited diameter and depth to maintain the integrity of the globe and to minimise postoperative haze. This study evaluated the effects of deeper, larger diameter wounds on refractive stability and corneal haze, and investigated the effects of ablation profile on wound healing and visual performance. METHODS: One hundred patients undergoing -3.00D and -6.00D corrections were randomised to receive 5 mm, 6 mm, or multizone treatments. The multizone treatment was 6 mm in diameter, but only the depth of the 5 mm treatment. Outcome was measured by Snellen visual acuity, residual refractive error, objective techniques for haze and halos, pupil diameter, subjective night vision, and requirement for retreatment. RESULTS: Overall, the results of 6 mm treatments were superior to those of 5 mm and multizone treatments: they had a smaller hyperopic shift (p < 0.01), a more predictable (p < 0.001) and stable refractive outcome, less haze (p < 0.05), smaller halos (p < 0.05), fewer subjective night vision problems, and fewer patients required retreatment. CONCLUSIONS: Analysis of these data and a literature review of corneal wound healing demonstrated that the improved outcome associated with the 6 mm beam did not relate to the depth of ablation. The factor with greatest apparent influence on the development of haze and regression was the slope of the wound surface over the entire area of the ablation. Tapering the wound edge provided no additional benefit, and contributed to night vision problems. It is, therefore, recommended that small diameter or multizone treatments should not be used in low and moderate myopia.

Effect of the menstrual cycle on corneal curvature.

The purpose of this research was to determine if a clinically significant variation in corneal curvature occurs with the menstrual cycle. The EyeSys Corneal Analysis System (CAS) was used to measure corneal curvature of a test group of three normally menstruating female subjects over three cycles and compared with a control group of three males, for three pseudocycles of 28 days. Measurements were taken on days 1, 4, 8, 15 and 22 of each of three menstrual cycles, with four CAS images taken on each day. Four points (along 45, 135, 225 and 315 degrees) were selected for each of rings 1, 3, 6 and 8, and the average curvature for each of these rings was calculated. The results showed that no detectable temporal effect occurred with the menstrual cycle, thus demonstrating that either cyclic variation in corneal topography was too small to be identified by this system or it did not take place.

An in vivo investigation of the structures responsible for corneal haze after photorefractive keratectomy and their effect on visual function.

PURPOSE: To make serial measurements of corneal haze and microscopic anatomy after photorefractive keratectomy (PRK) and compare the results with visual function measured at the same time points in the same single group of human subjects. METHODS: Ten patients underwent -6.00-diopter, 6-mm PRK. The patients were reviewed frequently for 12 months. Corneal haze was measured objectively in two ways: (1) an opacification index was determined from the variance in digitized retroillumination images; and (2) light reflected and scattered back from the cornea was assessed by gray-scale analysis of video slit images. In vivo confocal microscopy recorded the anatomic changes occurring in the cornea, and computer analysis of the images quantified the keratocytes and subepithelial deposit. Visual performance was assessed by Snellen visual acuity, contrast sensitivity, and glare-induced visual dysfunction. RESULTS: In the first week, epithelial irregularity resulted in a transient reduction in all aspects of visual function. In the first month, keratocyte disturbances reduced contrast sensitivity at high frequencies and produced glare. Over the next couple of months, the subepithelial deposit resulted in a more prolonged loss of contrast sensitivity at low frequencies and glare-induced visual dysfunction due to the scattering of light. In several patients, these visual defects persisted after 1 year. CONCLUSIONS: Epithelial and keratocyte disturbances only transiently affect visual function. The subepithelial deposit is more persistent and can have a lasting effect on visual performance. Therefore, attempts to improve the visual outcome of PRK must be aimed at controlling the synthesis of subepithelial material.