To investigate the clinical characteristics and treatment outcomes of fungal keratitis caused by Beauveria bassiana. The clinical data of 11 consecutive patients with fungal keratitis caused by Beauveria bassiana in the Eye Hospital of Shandong First Medical University from January 2019 to May 2024 were retrospectively analyzed. The clinical features, etiological characteristics, treatments and outcomes were analyzed. Most patients were farmers (10/11), with three cases caused by plant trauma and five cases having a history of corneal transplantation. Slit-lamp examination revealed that the keratopathy was characterized by dry ulcerative lesions, predominantly circular with relatively irregular borders. Several cases exhibited typical keratopathy signs of fungal keratitis, such as mycelial mat, pseudopodia and hypopyon. No significant immune ring structure was observed in any patients. Both the corneal scraping and in vivo confocal microscopy (IVCM) revealed branched and septate hyphae structures. Morphological and ITS sequencing identification was performed after isolation and cultivation. B. bassiana exhibited typical morphological characteristics: hyphae were branched, septate, transparent and slender; sympodial development of conidia was on a geniculate or zig-zag pattern; conidiogenous cells were flask-shaped, rachiform and sympodially proliferating, often aggregating into sporodochia or synnemata; conidia were hyaline and globose or ovoid in shape. B. bassiana had a relatively high susceptibility to the majority of antifungal agents in vitro. One patient was treated with medical therapy alone. The five patients with a history of corneal transplantation underwent penetrating keratoplasty (PKP). The two patients with ulcer depth < 1/2 corneal thickness and ulcer diameter ≤ 3 mm underwent corneal ulcer debridement combined with medical treatment. The three patients with ulcer depth exceeding 1/2 corneal thickness underwent PKP. Following treatment, no patient experienced recurrence during the follow-up period. Plant trauma, immunosuppression and involvement in agricultural production have been identified as significant risk factors for B. bassiana keratitis. Although the strains exhibited low minimal inhibitory concentrations (MICs) to most antifungal agents in vitro, early diagnosis and treatment remain crucial.

To characterize the clinical features and therapeutic outcomes of fungal keratitis caused by Plectosphaerella cucumerina. A retrospective analysis was conducted on 13 patients diagnosed with P. cucumerina keratitis from July 2018 to April 2024. Clinical manifestations, microbiological identification, antifungal susceptibility profiles, treatment regimens, and prognostic indicators were evaluated. All the 13 patients were farmers, and 6 of them had a history of trauma or the presence of a foreign body prior to the onset of symptoms. The main clinical manifestations were chronic progressive shallow corneal and middle stromal layer ulceration with dry ulcer surface, and moss formation was visible in 7 patients (7/13). No obvious immune ring structure and satellite lesions were observed in all patients. In vivo confocal microscopy (IVCM) showed that the mycelia showed medium strong reflection, some of them were changed like bamboo joints, most of them were 2–3 μm in diameter, and the arrangement was relatively neat and fasciculate. The Angle between the mycelia was small and most of them were acute angles. Drug sensitivity test showed that Amphotericin B, voriconazole and itraconazole were the main sensitive antifungal drugs. The main treatment was sensitive antifungal drugs combined with local lesion resection and voriconazole matrix injection. 2 cases with deep ulcers were treated with corneal transplantation. P. cucumerina keratitis is relatively rare but distinct clinical entity. Diagnosis can be aided by corneal scraping, microbiological culture and IVCM. Emphasizing a comprehensive treatment approach involving medication, debridement, and surgical intervention is crucial to promote rapid healing of the ulcer.

To compare the outcomes of corneal collagen cross-linking combined with lamellar keratoplasty (CXL-LK) versus LK alone in treating medically unresponsive acanthamoeba keratitis (AK). This retrospective, non-randomized controlled clinical study included 11 eyes (CXL-LK group) and 16 eyes (LK group) at a tertiary ophthalmology center. Corneal infiltration, density of acanthamoeba cysts (AC) and pathological changes following CXL in AK patients were assessed. One-year post-operation, refractive outcomes, graft survival, corneal endothelial loss rates and AK recurrence were evaluated. The area and depth of the infiltrates were reduced following CXL in AK patients, and the density of the AC decreased. Histopathological results indicated a reduction in AC volume after CXL, with the contents appearing wrinkled and degraded. At one year postoperatively, the corneal endothelial loss rate was 10.12

Introduction: Age-related Macular Degeneration (AMD) is a predominant cause of blindness in the elderly. The present study is the first to investigate the alteration of lncRNAs and mRNAs in neovascular AMD. Methods: Nine patients with neovascular AMD were included in the study. The control group comprised seven patients with epiretinal membranes. RNA sequencing was performed to obtain the differentially expressed mRNAs (DEmRNAs) and lncRNAs (DElncRNAs). Then, the DElncRNA-DEmRNA co-expression network, ceRNA network, and immune-related ceRNA subnetwork were constructed. Functional annotation of DEmRNAs between the two groups and DEmRNAs in networks was conducted. The immune cell distribution in neovascular AMD was also evaluated. Real-time qPCR (RT-qPCR) was used to validate the expression levels of key markers. Results: A total of 342 DEmRNAs and 157 DElncRNAs were obtained in neovascular AMD. Functional annotation indicated that these DEmRNAs significantly enriched immune systemrelated processes, such as positive regulation of B cell activation, immunoglobulin receptor binding, complement activation, and classical pathway. The DElncRNA-DEmRNA co-expression network, including 185 DElncRNA-DEmRNA co-expression pairs, and the ceRNA (DElncRNA-miRNA-DEmRNA) network, containing 45 lncRNA-miRNA pairs and 73 miRNAmRNA pairs, were constructed. The immune-related ceRNA subnetwork, including 2 lncRNAs, 5 miRNAs, and 3 mRNAs, was constructed. In addition, the distribution of immune cells was slightly different between the neovascular AMD group and the control group. RT-qPCR validation indicated the consistency between the RT-qPCR results and RNA sequencing results. Conclusion: In conclusion, STC1, S100A1, MEG3, MEG3-hsa-miR-608-S100A1, and MEG3- hsa-miR-130b-3p/hsa-miR-149-3p-STC1 may be related to the occurrence and development of neovascular AMD.

Phaeoacremonium is typically found in the environment and can cause diseases in woody plants. It is rarely responsible for infections in humans. In this report, we present a case of corneal infection caused by Phaeoacremonium iranianum. Caused a fan-shaped grayish-white ulcer measuring approximately 4 mm×3 mm inferior nasal cornea. The ulcer appeared relatively dry, with noticeable swelling and cloudiness in the surrounding cornea. The visual acuity after infection was 0.25. We started an aggressive antifungal therapy. Our antifungal susceptibility testing revealed the Minimum Inhibitory Concentration of anifengin was 4 µg/ml, Micafengin and carpofungin exceeding 8 µg/ml, fluorocytosine was 32 µg/ml, posaconazole was 1 µg/ml, voriconazole was 0.5 µg/ml, itraconazole was 2 µg/ml, fluconazole was 64 µg/ml, and amphotericin was 0.05 µg/ml. The patient underwent keratectomy combined with antifungal therapy. A month later, the conjunctiva of the right eye was congested, and a 4*4 mm corneal opacity was visible inferior nasal cornea, with local obvious thinning and a little keratic precipitates on the inner skin, and the infection has been controlled. We detail the progression and treatment of this infection to contribute to the clinical understanding and management of such cases.

Purpose To compare the outcomes of big-bubble deep anterior lamellar keratoplasty (BB-DALK) and penetrating keratoplasty (PKP) in the management of medically unresponsive Acanthamoeba keratitis (AK). Methods This retrospective study included 27 eyes of BB-DALK and 24 eyes of PKP from a tertiary ophthalmology care centre. Glucocorticoid eye drops were subsequently added to the treatment plan 2 months postoperatively based on the evaluation using confocal laser scanning microscopy. The clinical presentations, best-corrected visual acuity (BCVA), postoperative refractive outcomes, graft survival, and Acanthamoeba recurrence were analyzed. Results The AK patients included in the study were in stage 2 or stage 3, and the percentage of patients in stage 3 was higher in the PKP group ( P = 0.003). Clinical presentations were mainly corneal ulcers and ring infiltrates, and endothelial plaques, hypopyon, uveitis and glaucoma were more common in the PKP group ( P = 0.007). The BCVA and the graft survival rate showed no statistically significant differences between the two groups at 1 year after surgery. However, 3 years postoperatively, the BCVA of 0.71 ± 0.64 logMAR, the graft survival rate of 89.5%, and the endothelial cell density of 1899 ± 125 cells per square millimeter in the BB-DALK group were significantly better than those of the PKP group ( P = 0.010, 0.046, and 0.032, respectively). 3 eyes (11.1%) in the BB-DALK group and 2 eyes (8.3%) in the PKP group experienced Acanthamoeba recurrence, but the rates showed no statistically significant difference between the two groups ( P = 1.000). In the PKP group, immune rejection and elevated intraocular pressure were observed in 5 and 6 eyes, respectively. Conclusion Corneal transplantation is recommended for AK patients unresponsive to antiamoebic agents. The visual acuity and graft survival can be maintained after BB-DALK surgery. Acanthamoeba recurrence is not related to the surgical approach performed, whereas complete dissection of the infected corneal stroma and delayed prescribing of glucocorticoid eye drops were important to prevent recurrence.

Background: Fungal keratitis (FK) is characterized by corneal inflammation induced by fungal infection. Thymic stromal lymphopoietin (TSLP) is crucial in FK pathogenesis. This study aimed to explore the impact of TSLP on FK and elucidate the mechanism of TSLP signaling on macrophage-mediated inflammation. Methods: Transformed human corneal epithelial cells (THCEs) were stimulated with heat-killed fragments of Aspergillus fumigatus hyphae. TSLP secretion was assessed using quantitative polymerase chain reaction (qPCR), enzyme-linked immunosorbent assay (ELISA), and western blotting (WB). THCEs stimulated with hyphae or recombinant thymic stromal lymphopoietin (rTSLP) were co-cultured with tohoku hospital pediatrics-1 (THP-1) cell-derived macrophages. The inflammatory activity of rTSLP was evaluated in THP-1 macrophages through real-time PCR, ELISA, WB, and flow cytometry. Additionally, high-throughput transcriptome sequencing was employed to elucidate the underlying mechanisms. Results: Secretion of TSLP by A. fumigatus-stimulated THCEs was correlated with increased inflammation in THP-1 macrophage. Treatment with rTSLP resulted in elevated expression of proinflammatory markers associated with macrophage type 1 (M1) phenotype, including tumor necrosis factor-alpha (TNF-alpha) (p < 0.01), interleukin-1 beta (IL-1 beta) (p < 0.01), inducible nitric oxide synthase (iNOS) (p < 0.001), and CD86 (p < 0.001) at 24 hours. Exposure of THP-1 macrophages to rTSLP induced activation of the nuclear factor kappa B (NF-KB) signaling pathway. The improved level of phosphorylated NF-KB and inflammatory factors (iNOS, IL-1 beta, TNF-alpha) were attenuated by a specific NF-KB inhibitor. Conclusions: In A. fumigatus keratitis, TSLP secreted by THCEs promoted TNF-alpha, IL-1 beta, and iNOS secretion in THP-1-derived macrophages by upregulating the NF-KB signaling pathway. This led to dysregulation of the pro-inflammatory (M1)/alternative (M2) macrophage ratio and polarization of THP-1 macrophages towards the pro-inflammatory phenotype.

This study aims to evaluate the clinical outcomes and efficacy of a modified tectonic corneoscleral graft (TCG) in patients suffering from devastating corneoscleral infections. Thirty-eight eyes from 38 patients who underwent the modified TCG were included in this study. The outcomes measured were recurrence rates, best-corrected visual acuity (BCVA), ocular surface stability, postoperative complications, and graft survival. Among the 38 patients, 23 had fungal infections, 9 had bacterial infections and 6 had Pythium insidiosum infections. At the final follow-up, with an average duration of 25.1 ± 8.6 months, the rate of monocular blindness decreased from 100 to 58

Background Demodex blepharitis (DB) is a common disease of the ocular surface. The characteristics of the bacterial community in eyelash roots after Demodex infestation are still unknown. Knowledge of the characteristics of the bacterial community of eyelash follicles in patients with DB can provide valuable insights for guiding the diagnosis and treatment of DB. Methods Twenty-five patients with DB (DB group) and 21 non-DB volunteers (control group) were enrolled in the study. Eyelashes from the upper eyelid of the right eye were sampled, and 16S ribosomal DNA (rDNA) sequencing was performed to determine the V3-V4 regions of the microbial 16S rDNA gene within 1 month of infestation. The sequencing data of the two groups were analyzed and compared. The effect of the bacterium Burkholderia on the survival of Demodex mites was evaluated using Demodex obtained from 12 patients with DB other that the patients in the DB group. Results A total of 31 phyla and 862 genera were identified in the DB and control groups. The five most abundant phyla in the two groups were Proteobacteria, Firmicutes, Actinobacteria, Bacteroidetes and Cyanobacteria. The abundance of Actinomycetes was significantly higher in the DB group than in the control group. At the genus level, the five most abundant genera in the two groups were Pseudomonas , Burkholderia-Caballeronia-Paraburkholderia , Rolstonia and Acinetobacter ; Clostridium sensu stricto 1 was abundant in the control group and Corynebacterium_1 was abundant in the DB group. Compared with the control group, the abundance of Burkholderia-Caballeronia-Paraburkholderia was 2.36-fold lower in the DB group. Linear discriminant analysis Effect Size (LEfSe) analysis revealed Burkholderia-Caballeronia-Paraburkholderia , SC_I_84_unclassified , Nonmyxobacteria and Succinvibrio to be the major biomarkers in the control group and Catenibacterium and Lachnospiraceae NK4A136 group to be the major biomarkers in the DB group. To explore the performance of these optimal marker models, receiver operational characteristic curve analysis was performed, and the average area under the curve value of Burkholderia-Caballeronia-Paraburkholderia was 0.7448. Burkholderia cepacia isolated from normal human eyelashes was fermented, and the Demodex mites isolated from patient eyelashes were cultured together with its fermented supernatant. The results showed that the fermentation supernatant could significantly reduce the survival time of the Demodex mites, suggesting the potential therapeutic value of this bacterium against Demodex . Conclusions The composition of the bacterial community in the eyelashes of DB patients differed from that in eyelashes of healthy volunteers, revealing a decrease in bacterial diversity in infested eyelashes. This decrease may be related to the occurrence and development of DB. The supernatant of Burkholderia cepacia culture medium was found to inhibit the growth of Demodex in eyelash hair follicles, providing a new insight with potential applications for the clinical treatment of Demodex infestation. Graphical Abstract

AIM: To evaluate the correlation between Demodex infestation and keratitis, and to assess demodicosis using a simple approach. METHODS: A modified slit lamp illumination (at 40× magnification) was used to observe Demodex tails in 40 patients with refractory keratitis and 80 healthy controls. Bacterial smear and culture of the conjunctival sac and corneal lesion were performed to identify the pathogen. Tea tree oil ointment (TTOO) was added as a Demodex killing agent for lid scrubs to the treatment when Demodex infestation was confirmed. RESULTS: Demodex tails were found in all patients compared to 42/80 of the controls (P<0.01). Seventeen patients presented blepharitis, while 23 were free of scales and inflammation at the lid margin. The demodicosis was mild, moderate, and severe in 8, 19, and 13 patients, respectively, compared to mild in 42 controls (P<0.01). The keratitis was mild, moderate, and severe in 13, 19, and 8 patients, respectively. The severity of Demodex infestation was not correlated to the severity of keratitis (P=0.126). The growth of Staphylococcus was revealed in nine patients who did not react to antibiotic eye drops prior to the TTOO treatment. Patients' signs and symptoms got resolved after the lid scrub with TTOO. CONCLUSION: Ocular Demodex needs to be checked and treated in refractory keratitis patients with or without blepharitis. A slit-lamp illumination under high magnification favors the judgment of the severity of Demodex infestation.

Background Orthokeratology (OK) lens wear increases the risk of bacterial infection, but little is known about the microbiota of the conjunctival sac in myopic children wearing OK lenses. This study aimed to investigate the changes of conjunctival microbiota in children after treatment with OK lenses using 16 S rDNA sequencing.Methods Twenty-eight myopic children who had been continuously wearing OK lenses for 12 to 13 months were enrolled in this prospective study. Twenty-two gender- and age-matched myopic children who had not worn OK lenses or discontinued OK lens wear at least 1 year ago were recruited as controls. Conjunctival swabs from each participant were collected for exploration of the microbiota profiles, targeting the V3-V4 regions of the 16 S rRNA gene by MiSeq sequencing. The differences in the microbial community structure and diversity were also compared between groups.Results The bacterial alpha diversity indices in the OK lens group were not different from those in the non-wearer group (P > 0.05, Wilcoxon test), while beta diversity examined using principle coordinate analysis of unweighted UniFrac divided the two groups into different clusters. Proteobacteria, Bacteroidetes, and Firmicutes were the abundant phyla in the conjunctival sac microbiota in both groups (P < 0.05, Mann-Whitney U test). Among children in the OK lens group, the Linear discriminant analysis Effect Size identified the compositional changes in OK lens-associated bacteria. Key functional genera such as Blautia, Parasutterella, and Muribaculum were enriched, whereas Brevundimonas, Acinetobacter, Proteus, and Agathobacter decreased significantly (P < 0.05, Mann-Whitney U test). Phylogenetic investigation of communities by reconstruction of unobserved states also showed altered bacterial metabolic pathways in OK lens-associated microbiota. Moreover, using receiver operating characteristic curves, Brevundimonas, Acinetobacter, Proteus, and Agathobacter alone (the area under the curve was all > 0.7500) or in combination (the area under the curve was 0.9058) were revealed to discriminate OK lens wearers from controls.Conclusions The relative abundance of the microbial community in the conjunctival sac of myopic children can alter after OK lens wear. Brevundimonas, Acinetobacter, Proteus, and Agathobacter may be candidate biomarkers to distinguish between OK lens wearers and non-wearers.

Objective::To analyze the clinical and etiological characteristics of patients with fungal keratitis secondary to infectious endophthalmitis who required evisceration.Methods::In this retrospective case series study, a total of 44 patients (44 eyes) who were diagnosed with fungal keratitis secondary to infectious endophthalmitis and underwent evisceration from January 2012 to December 2021 were collected in Shandong Eye Hospital. The course of the disease and its predisposing factors, ocular and systemic history, clinical manifestations, microbial culture and drug sensitivity test results were analyzed.Results::The 44 patients included accounted for 2.15% of the total cases of fungal keratitis diagnosed by Shandong Eye Hospital in the same period. The median course of the disease was 18.5 days. At admission, 44 eyes had no light perception, and 20 eyes (45%) were complicated with secondary glaucoma. All 44 eyes showed corneal ulcer, involving the whole corneal layer and limbus, and 27 eyes (61%) complicated with anterior chamber empyema. A total of 39 eyes (89%) of microbial culture had fungal growth, including 20 eyes (51%) of Fusarium, 8 eyes (21%) of Aspergillus, 2 eyes (5%) of Alternaria, others including 4 eyes (10%) of Nonsporulating molds, 2 eyes (5%) of Pythium insidiosum, 1 eye (3%) of Scedosporium apiospemum, 1 eye (3%) of Sarocladium, 1 eye (3%) of Phialemoniopsis pluriloculosa. The geometric mean of the minimum inhibitory concentration values of amphotericin B, posaconazole, voriconazole, itraconazole, 5-fluorocytosine, fluconazole, anifensin, micafungin and caspofungin cultivated in this study were 1.597, 1.338, 1.926, 3.291, 64, 189.699, 1.36, 1.16, 1.23 μg/ml.Conclusions::The patients with infectious endophthalmitis secondary to fungal keratitis often present with total corneal ulceration. In addition to the common pathogenic bacteria of Fusarium, Aspergillus and Alternaria, other bacteria with strong pathogenicity and invasiveness such as Nonsporulating molds, Pythium insidiosum and Scedosporium apiospemum are also common.

Purpose To investigate the differences in microbiological characteristics, risk factors, drug resistance, and visual outcomes in three infections: fungal keratitis with hypopyon (FKH), keratitis-related fungal endophthalmitis (FKE), and fungal endophthalmitis without keratitis (FE). Methods An analytical cross-sectional study. Results In total, 14.57% of eyes with FKH progressed to endophthalmitis. Hypopyon, pre-existence of lens problems, topical steroid use and sever keratitis were significantly associated with the development of FKE. The risk factors of the FKH and FE group were mainly plant trauma and open globe trauma, respectively. Keratitis-related endophthalmitis (FKE) showed a significantly higher resistance than the other two groups. The FKH group had the best final visual acuity, while the FKE group had the worst. Conclusion Hypopyon height, pre-existing lens problems, topical steroid use and sever keratitis are risk factors for progression to endophthalmitis in eyes with fungal keratitis, and its progression is not affected by a single fungus. The antifungal drugs resistance in patients with endophthalmitis related to keratitis was significantly higher than that associated with other reasons. Timely diagnosis and risk factor assessment are essential for ensuring early treatment of FKE.

Aspergillus species is a widespread environmental mould that can cause aspergillosis. The purpose of this study was to investigate the antifungal susceptibility profile and genotypic characterization of clinical Aspergillus isolates from different provinces in Eastern China. The data included the antifungal susceptibility distributions with eight common antifungal drugs, cyp51A gene mutations of triazole-resistant Aspergillus fumigatus sensu stricto, and the genotypic relationships among the A. fumigatus sensu stricto isolates based on microsatellite typing. A. fumigatus sensu lato was the most common clinical Aspergillus species (n = 252), followed by A. flavus (n = 169), A. terreus (n = 37), A. niger (n = 29), and A. nidulans (n = 4). The modal minimum effective concentration values of micafungin and anidulafungin were lower than those of caspofungin for all Aspergillus species. The in vitro efficacy of isavuconazole was similar to that of voriconazole against most Aspergillus species. Sequencing revealed cyp51A gene mutations TR34/L98H, TR34/L98H/S297T/F495I, and TR46/Y121F/T289A in four triazole-resistant A. fumigatus sensu stricto. Phylogenetic analyses using microsatellite markers of A. fumigatus sensu stricto revealed that 211 unique genotypes clustered into two clades. The data demonstrate the diversity of clinically relevant Aspergillus species in Eastern China. Routine antifungal susceptibility testing should be performed to monitor the antifungal resistance and guide clinical therapy. The 6-year multicenter study collected a total of 491 Aspergillus isolates from Eastern China to investigate the in vitro antifungal susceptibility to eight antifungal drugs, the cyp51A gene mutations of triazole-resistant A. fumigatus sensu stricto, and the genetic relatedness through microsatellite typing.

Objective:The aim of this study was to investigate the etiological spectrum of infectious endophthalmitis and analyze the drug susceptibility characteristics of etiological organism in Shandong Province.Methods:A total of 350 patients (350 eyes) were collected from Eye Hospital of Shandong First Medical University (Shandong Eye Hospital) from 2013 to 2020 who were clinically diagnosed with suspected infectious endophthalmitis and received aqueous fluid and (or) vitreous samples for smear test, bacterial culture and fungal culture before examination and retrospectively analyzed. Isolated bacteria and candida were identified and drug sensitive test by Vitek 2 Compact automatic microbiological analysis system. The drug sensitivity data were analyzed by WHONET5.6 and expressed as cases and percentage.Results:Among 350 cases of suspected infectious endophthalmitis, there were 317 cases with exogenous endophthalmitis and 33 cases with endogenous endophthalmitis, accounting for 90.6% and 9.4%, respectively. There were 269 cases with both aqueous humor and vitreous fluid collected, 81 cases with aqueous humor collected. There were 67 cases with Gram-positive cocci, 8 cases with gram-negative bacteria, 8 cases with Gram-positive bacilli and 16 cases with fungi detected by smear, accounting for 28.3%, 67.7%, 8.1%, 8.1% and 16.1%, respectively. For culture in vitro, 135 cases were positive, a positive rate of 38.6%. There were 144 strains of isolated pathogenic organism, including 87 cases with Gram-positive cocci, 12 cases with Gram-positive bacilli, 15 cases with Gram-negative bacilli, and 30 cases with fungi, accounting for 60.4%, 8.3%, 10.5% and 20.8%, respectively. According to intraocular fluid smear and (or) culture positive as the gold standard for the diagnosis of infectious endophthalmitis, there were 211 patients of them with positive intraocular fluid smear and (or) in vitro culture, who were finally diagnosed as infectious endophthalmitis, an positive rate of 60.3%. Among of pathogens, Staphylococcus with negative coagulase was the top1, followed by Streptococcus. The main fungi were Fusarium and Aspergillus. The sensitivity of the isolated strains to levofloxacin ranged from 39.3% to 84.6%, which was higher than that of other drugs.Conclusions:From 2013 to 2020, the ocular trauma was the main pathogenic factor of infectious endophthalmitis in Shandong. Gram-positive coccus, especially Staphylococcus with negative coagulase, were the main pathogenic bacteria of infectious endophthalmitis.

Acanthamoeba species are among the most ubiquitous protists that are widespread in soil and water and act as both a replicative niche and vectors for dispersal. They are the most important human intracellular pathogens, causing Acanthamoeba keratitis (AK) and severely damaging the human cornea. The sympatric lifestyle within the host and amoeba-resisting microorganisms (ARMs) promotes horizontal gene transfer (HGT). However, the genomic diversity of only A. castellanii and A. polyphaga has been widely studied, and the pathogenic mechanisms remain unknown. Thus, we examined 7 clinically pathogenic strains by comparative genomic, phylogenetic, and rhizome gene mosaicism analyses to explore amoeba-symbiont interactions that possibly contribute to pathogenesis. Genetic characterization and phylogenetic analysis showed differences in functional characteristics between the "open" state of T3 and T4 isolates, which may contribute to the differences in virulence and pathogenicity. Through comparative genomic analysis, we identified potential genes related to virulence, such as metalloprotease, laminin-binding protein, and HSP, that were specific to the genus Acanthamoeba. Then, analysis of putative sequence trafficking between Acanthamoeba and Pandoraviruses or Acanthamoeba castellanii medusaviruses provided the best hits with viral genes; among bacteria, Pseudomonas had the most significant numbers. The most parsimonious evolutionary scenarios were between Acanthamoeba and endosymbionts; nevertheless, in most cases, the scenarios are more complex. In addition, the differences in exchanged genes were limited to the same family. In brief, this study provided extensive data to suggest the existence of HGT between Acanthamoeba and ARMs, explaining the occurrence of diseases and challenging Darwin's concept of eukaryotic evolution. IMPORTANCEAcanthamoeba has the ability to cause serious blinding keratitis. Although the prevalence of this phenomenon has increased in recent years, our knowledge of the underlying opportunistic pathogenic mechanism maybe remains incomplete. In this study, we highlighted the importance of Pseudomonas in the pathogenesis pathway using comprehensive a whole genomics approach of clinical isolates. The horizontal gene transfer events help to explain how endosymbionts contribute Acanthamoeba to act as an opportunistic pathogen. Our study opens up several potential avenues for future research on the differences in pathogenicity and interactions among clinical strains.

To investigate the clinical characteristics and treatment outcomes of Nocardia infection after ocular surface surgery. This is a retrospective study. Eight cases of culture-proven Nocardia infection, which developed within 1 month after ocular surface surgery were included. Demographics and clinical history of patients were investigated. There were 8 eyes (2 left and 6 right) of 8 patients (5 males and 3 females), aged 27–65, with a median age of 52.9 years. Three cases underwent pterygium excision, three were subjected to conjunctival flap covering, and two were treated with lamellar corneal transplantation. The time interval between previous surgery and the onset of symptoms varied from 7 to 28 days (mean = 20.5 ± 7.13 days). All the cases presented grey-white infiltrates at the surgical incision site while appearing with six corneal ulcers and two conjunctival ulcers. Filaments of Nocardia were founded by confocal microscopy in two of the five cases. All responded poorly to medical therapy. Seven of the eight cases were treated with reoperation. Nocardia infection recurred in three cases after reoperation, and one was eviscerated. Surgical trauma is a risk factor for ocular Nocardia infection. Nocardia infection should be suspected when secondary infection occurs in a surgical incision with an atypical clinical presentation. The use of corticosteroids may influence the efficacy of drugs. Complete removal of lesions may lower the recurrence of Nocardia infection with poor drug treatment effects.

To compare the clinical and microbiological characteristics in patients with bacterial keratitis with hypopyon (BKH), bacterial keratitis-related endophthalmitis (BKE), and bacterial endophthalmitis without keratitis (BE). Data from all inpatients who were clinically diagnosed with BKH, BKE, and BE from 2018 to 2020 were collected retrospectively. The demographics, predisposing risk factors, clinical characteristics, microbiological profiles, and antibiotic susceptibility of the patients were evaluated. Approximately 9.46% (28/296) of eyes with BKH progressed to endophthalmitis. The hypopyon (OR = 5.35, 95% CI: 2.17–7.08) and corneal perforation (OR = 2.47, 95% CI: 1.04–4.86) were significantly related to the development of BKE. The odds ratios for hypopyon of less than 1 mm, 1–3 mm, and greater than 3 mm were 1, 2.09 (95% CI: 1.17–3.15), and 4.12 (95% CI:2.59–5.68), respectively. The predominant causative pathogen was Staphylococcus epidermidis (36.43%, 38.89%), followed by Streptococci (14.73%, 16.67%), Staphylococcus aureus (8.53%, 7.79%), and Pseudomonas aeruginosa (9.30%, 7.14%) in eyes with BKH and BE. However, the main pathogens were Pseudomonas aeruginosa (37.50%) and Staphylococcus aureus (31.25%) in eyes with BKE. In the BKH, BKE, and BE groups, almost 100% of Staphylococcus aureus isolates were sensitive to vancomycin (97.70%, 100%, 95.56%), about a half were sensitive to fluoroquinolones (51.85%, 39.90%, 62.34%), and approximately 30% were sensitive to trimethoprim/sulfa (27.77%, 21.56%, 33.56%) and cefazolin (41.47%, 20.31%, 38.81%). The susceptibility of Pseudomonas aeruginosa to fluoroquinolones antibiotics was 55.75%, 66.67%, and 62.58%, respectively, in the three groups. The height of hypopyon and corneal perforation are risk factors for progression to endophthalmitis in eyes with bacterial keratitis. When Staphylococcus aureus and Pseudomonas aeruginosa are identified, vigilance is required for advanced endophthalmitis.

PURPOSE:To analyze the incidence and risk factors of postoperative endophthalmitis after primary surgical repair and intraocular foreign body (IOFB) removal within 24 hours of injury.METHODS:The records of all patients treated surgically for open globe injury and IOFB removal at the Eye Hospital of Shandong First Medical University between January 1, 2015, and June 30, 2020, were retrospectively reviewed. Variables included time from injury to operation, cause of injury, details of surgical repair, and follow-up. The incidence and risk factors of endophthalmitis after IOFB removal were studied.RESULTS:During 5 years, 99 patients with IOFB were reviewed. Of these, 19 patients were diagnosed with endophthalmitis on admission, and 5 were suspected of having endophthalmitis during operation. Fifty-four cases had no clinical signs of endophthalmitis on admission and during operation and were treated with operation within 24 hours after the injury. Two patients (2 of 54; 3.70%) developed endophthalmitis after IOFB removal, and the causative agent in both cases was Bacillus cereus.CONCLUSION:The incidence of infectious endophthalmitis after primary surgical repair combined with IOFB removal (≤24 hours) was 3.70% in patients who received a series of standard treatments, and B. cereus infection might be a risk factor.

Background Scedosporium species have drawn significant interest as inhabitants of polluted soil and water and as cause of high mortality in near-drowning patients. So far, most cases have been reported from Europe and Australia, while knowledge on their prevalence and genotypic diversity from Asia is scant. Objectives To increase the knowledge of the genetic diversity and in vitro antifungal susceptibility of Scedosporium species involved in human infections from China. Methods Here, we applied the ISHAM-MLST consensus scheme for molecular typing of Scedosporium species and revealed both high species diversity and high genotypic diversity among 45 Chinese clinical Scedosporium isolates. Results Among the five species, Scedosporium boydii (n = 22) was the most common, followed by S. apiospermum (n = 18), S. aurantiacum (n = 4) and S. dehoogii (n = 1). S. aurantiacum was reported for the first time from clinical samples in China. The predominant sequence types (STs) were ST17 in S. apiospermum, ST4 in S. boydii and ST92 in S. aurantiacum, including four novel STs (ST40, ST41, ST42 and ST43) in S. apiospermum. Based on the CLSI-M38 A2 criterion, voriconazole was the only antifungal compound with low MIC values (MIC90 <= 1 mu g/ml) for all Scedosporium isolates in our study. Conclusions The genetic diversity of clinical isolates of Scedosporium species from China is extremely high, with S. boydii being predominant and S. aurantiacum being firstly reported here. VOR was the only antifungal compound with low MIC values for all Scedosporium isolates in our study, which should be recommended as the firstline antifungal treatment against scedosporiosis in China.