Infectious keratitis following corneal transplantation: A long-term cohort study.

BACKGROUND: To assess the long-term incidence and risk factors for post-keratoplasty infectious keratitis (IK), associated ocular pathogens, and antibiotic resistance profiles. METHODS: Cohort study including 2553 consecutive penetrating, endothelial, and anterior lamellar keratoplasties performed between 1992 and 2020. Medical and microbiological records of patients clinically diagnosed with IK were retrospectively reviewed. MAIN OUTCOME MEASURES: cumulative incidence of IK, infectious agent species, and antibiotics resistance profiles. RESULTS: The average follow-up time after transplantation was 112 ± 96 months. Eighty-nine IK episodes were recorded; microbiological tests were positive in 55/89 (62%). The cumulated incidence of postoperative IK was 5.50%/10.25% at 10/20 years. The occurrence of at least one episode of IK after transplantation was associated with lower graft survival in the long term (p < 0.0001). Rejection risk (adjusted Hazard Ratio, 2.29) and postoperative epithelial complications (HR, 3.44) were significantly and independently associated with a higher incidence of postoperative IK. Infectious agents included 41 bacteria, 10 HSV, 6 fungi, and 1 Acanthamoeba. The rate of antibiotic resistance was 0% for vancomycin, 13% for fluoroquinolones, 20% for rifamycin, 59% for aminoglycosides, and 73% for ticarcillin. In 41% of cases, patients were under prophylactic topical antibiotics before the infectious episode. Topical antibiotics were significantly associated with increased resistance to penicillin, carbapenems, and aminoglycosides. CONCLUSION: IK (mainly bacterial) is a frequent complication of corneal transplantation in the long term. Vancomycin and fluoroquinolones can be considered as first-line treatments. Prolonged postoperative antibiotic preventive treatment is not advisable as it may increase antibiotic resistance.

Hidden in the Eyes-Recurrence of Systemic Hemopathies Reportedly "In Remission": Six Cases and Review of Literature.

Background and Objectives: Secondary ocular localizations of hematological malignancies are blinding conditions with a poor prognosis, and often result in a delay in the diagnosis. Materials and Methods: We describe a series of rare cases of ocular involvement in six patients with hematological malignancies, reportedly in remission, who presented secondary ocular localizations, challenging to diagnose. Two patients had an acute lymphoblastic leukemia (ALL) and developed either a posterior scleritis or a pseudo-panuveitis with ciliary process infiltration. One patient had iris plasmacytoma and developed an anterior uveitis as a secondary presentation. Two patients had a current systemic diffuse large B-cell lymphoma (DLBCL) and were referred either for intermediate uveitis or for papilledema and vitritis with secondary retinitis. Finally, one patient with an acute myeloid leukemia (AML) presented a conjunctival localization of a myeloid sarcoma. We herein summarize the current knowledge of ophthalmologic manifestations of extramedullary hematopathies. Results: Inflammatory signs were associated with symptomatic infiltrative lesions well displayed in either the iris, the retina, the choroid, or the cavernous sinus, from the admission of the patients in the ophthalmological department. These findings suggest that patients with ALL, AML, systemic DLBCL, and myeloma can present with ophthalmic involvement, even after having been reported as in remission following an effective systemic treatment and/or allograft. Conclusions: Early detection of hidden recurrence in the eyes may permit effective treatment. Furthermore, oncologists and ophthalmologists should be aware of those rare ocular malignant locations when monitoring patient's progression after initial treatment, and close ophthalmologic examinations should be recommended when detecting patient's ocular symptoms after treatment.

Cytokines, chemokines and growth factors profile in human aqueous humor in idiopathic uveitis.

To investigate which cytokines, chemokines and growth factors are involved in the immunopathogenesis of idiopathic uveitis, and whether cytokine profiles are associated with. Serum and aqueous humor (AH) samples of 75 patients with idiopathic uveitis were analyzed by multiplex immunoassay. Infectious controls consisted of 16 patients with ocular toxoplasmosis all confirmed by intraocular fluid analyses. Noninfectious controls consisted of 7 patients with Behçet disease related uveitis and 15 patients with sarcoidosis related uveitis. The control group consisted of AH and serum samples from 47 noninflammatory control patients with age-related cataract. In each sample, 27 immune mediators ± IL-21 and IL-23 were measured. In idiopathic uveitis, 13 of the 29 mediators, including most proinflammatory and vascular mediators such as IL-6, IL-8, IL-12, G-CSF, GM-CSF, MCP-1, IP-10, TNF-α and VEGF, were significantly elevated in the aqueous humor when compared to all controls. Moreover, IL-17, IP-10, and IL-21, were significantly elevated in the serum when compared to all controls. We clustered 4 subgroups of idiopathic uveitis using a statistical analysis of hierarchical unsupervised classification, characterized by the order of magnitude of concentrations of intraocular cytokines. The pathogenesis of idiopathic uveitis is characterized by the presence of predominantly proinflammatory cytokines and chemokines and vascular endothelial growth factor with high expression levels as compared to other causes of uveitis. There are indications for obvious Th-1/ IL21-Th17 pathways but also IL9-Th9 and increased IFN-γ-inducing cytokine (IL12) and IFN-γ-inducible CXC chemokine (IP-10). The combined data suggest that immune mediator expression is different among idiopathic uveitis. This study suggests various clusters among the idiopathic uveitis group rather than one specific uveitis entity.

NEW OPTICAL COHERENCE TOMOGRAPHY ANGIOGRAPHY FINDINGS ON AN INTRARETINAL VASCULAR PROCESS SECONDARY TO TOXOPLASMA RETINOCHOROIDITIS: 2 CASE REPORTS.

BACKGROUND/PURPOSE: To determine which retinal layer is primarily involved in intraretinal vascular processes associated with Toxoplasma retinochoroiditis using multimodal imaging, including optical coherence tomography angiography (OCTA). METHODS: Toxoplasma retinal lesions were analyzed through multimodal imaging, including color fundus photographs, fluorescein angiography, indocyanine green angiography, spectral-domain OCT, and OCTA. RESULTS: Two patients with atypical features of Toxoplasma retinochoroiditis are described in the acute phase. The first patient presented with a primary episode of acute Toxoplasma retinitis associated with an intraretinal abnormal vascular process that was detected at the acute phase by indocyanine green angiography and was better delineated by OCTA. Seven months later, the intraretinal vascular process had fully resolved on OCTA imaging. The second patient presented with a recurrence of active Toxoplasma retinochoroiditis adjacent to a pre-existing pigmented scar. He had a similar abnormal intraretinal vascular process that was identified on both fluorescein angiography at the early phase and indocyanine green angiography, which was also confirmed by OCTA imaging. CONCLUSION: We report a new finding associated with Toxoplasma chorioretinitis: an abnormal intraretinal vascular process that resolved without anti-vascular endothelial growth factor therapy but after toxoplasmosis treatment alone.

Anterior chamber tap cytology in acute postoperative endophthalmitis: a case-control study.

AIMS: To determine anterior chamber tap cytology characteristics in acute postoperative bacterial endophthalmitis. METHODS: 488 eyes of 488 patients were included in this retrospective case-control study. The study group included 93 eyes with bacteriologically documented endophthalmitis and 85 eyes with clinical endophthalmitis. The control group included 33 eyes with non-infectious postoperative inflammation, 116 eyes with acute uveitis and 161 cataract surgery eyes with no ocular inflammation. Cytological analysis, direct examination and microbiological cultures were performed in aqueous humour (AqH) samples. Inclusion criteria for the study group were the following: suspected endophthalmitis within 30 days following cataract surgery by phacoemulsification, secondary lens implantation, pars plana vitrectomy or intravitreal injection; best-corrected visual acuity (BCVA) <20/400; hypopyon or cyclitic membrane; absence of visibility of the retina; vitritis at a slit-lamp examination or in ultrasound B-scan. RESULTS: Cell line counts (mainly polymorphonuclear neutrophils) were significantly higher in the two endophthalmitis study subgroups than in the three control subgroups. The study group showed a predominance of polymorphonuclear neutrophils as opposed to the three control subgroups including uveitis (p<0.00001). The best sensitivity/specificity was obtained using a polymorphonuclear neutrophil threshold of 10 per field (sensitivity, 0.90; specificity, 0.75). The sensitivity of the bacterial culture was 32% in the AqH. High neutrophil count was associated with poorer initial BCVA (rs=0.62; p<0.00001) and higher risk of retinal detachment during (p=0.04) and after (p<0.001) hospitalisation. CONCLUSION: Anterior chamber tap cytology is a quick and accessible tool complementary to culture and PCR for the management of acute postoperative endophthalmitis.

Update on fungal keratitis in France: a case-control study.

PURPOSE: To report an epidemiological update of documented fungal keratitis (FK) in a French tertiary ophthalmological centre from 2014 to 2018 in comparison with a previous period from 1993 to 2008. METHODS: Sixty-two consecutive FK documented by microbiological corneal scrapings were compared with the 64 FK of the previous study. Amphotericin B and voriconazole eye drops were administered hourly. Population characteristics, clinical findings, aetiological organisms and treatments were analysed. RESULTS: The most frequently identified fungi were Fusarium (61%), Aspergillus (6.5%) and Candida (5%). Thirty out of 44 cases examined with in vivo confocal microscopy (IVCM) presented filaments. Ten required conventional cross-linking, 9 therapeutic penetrating keratoplasty, and 2 enucleation. Risk factors significantly associated with the absence of response to medical treatment were patient age (p = 0.01), presence of a deep stromal infiltrate at presentation (p = 0.04) and high numbers of filaments in IVCM images (p = 0.01). The two populations were comparable in age, but not in sex ratio males/females (18/44 versus 37/26 in the previous study; p = 0.001). The frequency of contact lens-associated infection increased from 35.5% to 71% (p = 0.0001) between the two periods. Since then, filamentous FK increased from 69% (44/64) to 95% (59/62) (p = 0.0001). A history of keratoplasty was less frequently reported during the last period (3.2% (2/62) versus 17% (11/64) of cases (p = 0.01)). A clear decrease in the frequency of therapeutic keratoplasty was noted from 39% (25/64) to 14% (9/62) (p = 0.02). CONCLUSION: The frequency of filamentous keratomycosis is currently increasing. Elderly patients and the presence of numerous filaments in IVCM are associated with poor clinical outcomes.

Clinical Features and Diagnosis of Anterior Segment Inflammation Related to Cytomegalovirus in Immunocompetent African, Asian, and Caucasian Patients.

Objective: To report the clinical features and treatment outcomes in immunocompetent patients with anterior segment inflammation (ASI) related to human cytomegalovirus (HCMV) depending on their ethnic origin.Material and Methods: Multicenter retrospective study of 38 patients with at least one test, either HCMV-positive PCR or GWc.Results: Features of Posner-Schlossman syndrome were observed in 50% of the eyes, Fuchs heterochromic iridocyclitis in 13% of the eyes, chronic nonspecific anterior uveitis in 21% of the eyes, and corneal endotheliitis in 18% of the eyes. PCR and GWc were positive for HCMV in 50% and 96.2% of the eyes, respectively. Glaucoma was diagnosed in 50% of eyes. Treatment was oral valganciclovir in about half of the patients. Other treatments were intravenous ganciclovir and/or ganciclovir topical ointment and/or intravitreal ganciclovir.Conclusions: No obvious association of specific clinical features with individual ethnicity could be identified. We found a high rate of glaucoma in all ethnic groups. There was a delay in diagnosis and specific treatment of HCMV in most patients.

Moraxella keratitis: epidemiology and outcomes.

We aimed to assess the prevalence, risk factors, and visual outcome of Moraxella keratitis. We retrospectively reviewed the medical charts of patients diagnosed with Moraxella spp. keratitis at the Quinze-Vingts National Ophthalmology Hospital, Paris, France, between January 2016 and December 2018. Definitive microbiological identification was performed on archival strains using matrix-assisted laser desorption ionization time of flight coupled to mass spectrometry. One hundred one culture-proven cases of Moraxella keratitis were identified. The most common isolates were Moraxella lacunata (50%) and Moraxella nonliquefasciens (38%). Systemic predisposing factors, principally diabetes mellitus (13%) were identified in 28% of patients, and 87% of patients had ocular surface conditions, including blepharitis (25%), prior ocular surgery (21%), glaucoma (17%), exposure keratopathy (16%), and trauma (16%). Severely affected inpatients were treated empirically with fortified antibiotics including vancomycin, piperacillin, and gentamicin. The presence of hypopyon and being over the age of 60 years were associated with a poorer final visual acuity (p < 0.05). Adjuvant treatment, mostly amniotic membrane transplantation, was required for 31 eyes. The prognostic factors significantly associated with the need for adjuvant treatment were a larger infiltrate and hypoesthesia. The clinical features including ulcer healing, treatment duration, and infiltrate size were not different between Moraxella species. Keratitis caused by Moraxella spp. are rare in France but may threaten sight. The early identification of patients with a poor ocular surface, particularly those with neurotrophic keratopathy and anesthetic cornea, is crucial to prevent delayed healing of ulcers and the need for adjuvant treatment.

Increase in Ocular Syphilis Cases at Ophthalmologic Reference Center, France, 2012-2015.

We describe the frequency, demographic and clinical features, and visual outcomes of ocular syphilis infections observed during 2012-2015 at a tertiary reference center in Paris, France. Twenty-one cases (29 eyes) were identified. The occurrence of ocular syphilis increased from 1 case in 2012 to 5 cases in 2013, 6 cases in 2014, and 9 cases in 2015 (2.22-25.21/1,000 individual patients/year for the period). Among case-patients, an annual 20%-33% were co-infected with HIV. Seventy-six percent of ocular syphilis infections occurred in men who have sex with men. Seventy-five percent of case-patients had a good final visual outcome (best-corrected visual acuity >0.3 logMAR score). Visual outcome was worse for HIV-positive patients than for HIV-negative patients (p = 0.0139). At follow-up, the best visual outcomes were observed in patients whose mean time from first ocular symptom to consultation was 15 days (SD +19 days).

Enzymatic treatment of specimens before DNA extraction directly influences molecular detection of infectious agents.

INTRODUCTION: Biological samples, pharmaceuticals or food contain proteins, lipids, polymers, ammoniums and macromolecules that alter the detection of infectious agents by DNA amplification techniques (PCR). Moreover the targeted DNA has to be released from the complex cell walls and the compact nucleoprotein matrixes and cleared from potential inhibitors. The goal of the present work was to assess the efficiency of enzymatic pretreatments on infectious agents to make DNA available for further extraction and amplification. METHODS: Staphylococcus epidermidis, Streptococcus mitis, Propionibacterium acnes, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Aspergillus niger and Fusarium solani were mixed with an internal control virus and treated with: 1) proteinase K; 2) lyticase and 3) lyticase followed by proteinase K. DNAs was manually extracted using the QIAmp DNA Mini kit or the MagNA Pure Compact automate. DNA extraction yields and the inhibitors were assessed with a phocid Herpesvirus. Bacterial detection was performed using TaqMan real-time PCR and yeasts and filamentous Fungi with HRM (real-time PCR followed by high-resolution melting analysis). RESULTS: Viral DNA was released, extracted and detected using manual and automatic methods without pre enzymatic treatments. Either the manual or the automatic DNA extraction systems did not meet the sensitivity expectations if enzymatic treatments were not performed before: lyticase for Fungi and Proteinase K for Bacteria. The addition of lyticase and proteinase K did not improve results. For Fungi the detection after lyticase was higher than for Proteinase K, for which melting analysis did not allow fungal specification. DISCUSSION: Columns and magnetic beads allowed collecting DNA and separate PCR inhibitors. Detection rates cannot be related to DNA-avidity of beads or to elution but to the lack of proteolysis.

A new workflow for the microbiological diagnosis of febrile neutropenia in patients with a central venous catheter.

OBJECTIVES: We aimed to improve the microbial diagnosis of first episodes of febrile neutropenia (FEFNs) since <30% of episodes are microbiologically documented. Consequently patients are usually treated by empirical antibiotic therapy. METHODS: A prospective study evaluated a new workflow combining: (i) one 40 mL blood culture (BC) sampled from the central venous catheter; (ii) immediate incubation in an automated BC system on the ward; (iii) direct detection of microbial DNA in blood; and (iv) identification and susceptibility testing using rapid methods performed directly on positive BC bottles. Patients were also sampled for the standard workflow with two BC sets incubated in the central laboratory and assessed by classical procedures. RESULTS: One hundred and twenty consecutive FEFNs were included (February 2008-March 2009). The new workflow was as sensitive as the standard workflow, with BC positivity rates of 30% (36/120) and 28% (34/120), respectively (McNemar's χ(2) =0.67, P=0.41). Direct DNA detection was positive in nine episodes (7.5%) that were also positive in BC. The new workflow provided microbiological results significantly earlier than the standard workflow, with a shorter time to BC positivity (median 12 h 31 min, range 7 h 55 min-25 h 37 min versus median 13 h 01 min, range 9 h 31 min-43 h 33 min, P=0.004) and shorter turnaround times for identification and susceptibility testing, with most of the results obtained <24 h after BC sampling. We retrospectively estimated that the new workflow would lead to earlier adequacy of antimicrobial therapy in 30% of documented cases. CONCLUSIONS: Our new process improved the microbiological diagnosis in FEFNs. Cost effectiveness needs to be tested.

Improving antibiotic use in the hospital: Focusing on positive blood cultures is an effective option.

OBJECTIVES: The unsolicited and systematic evaluation of positive blood cultures (pBC) after laboratory report by a single infectious disease specialist (IDS) was evaluated during one year, using a computer-generated alert by the laboratory. The main objectives of IDS counselling were to improve antibiotic use for bloodstream infection (i.e., initiating or modifying therapy) and to stop unjustified therapy for contaminated pBC. METHODS: During the first part of the study (4 months), all pBC in patients from ICUs, medical and surgical wards were analyzed. After an interim analysis, only pBC from medical and surgical wards were evaluated during the second part (8 months). RESULTS: Overall, 1090 episodes of pBC (representing 866 patients) were evaluated and classified as bloodstream infection (65.5%), contamination (29%) or undetermined (5.5%). Forty-three percent of episodes prompted IDS counselling, including initiation (5%), modification (27.5%), withdrawal (3.5%) and diagnosis workup (5%). Restricting the evaluation to medical and surgical wards increased the rate of counselling (61.2% vs. 27.7%, P<0.0001), notably for de-escalating (20% vs. 8%, P<0.0001), initiating (9% vs. 2%, P<0.0001), oral switch (6% vs. 2%, P<0.0001), withdrawing (5% vs. 2%, P=0.002) or reducing the duration of therapy (5% vs. 2%, P=0.002). DISCUSSION AND CONCLUSIONS: In complement to the laboratory report, a computer-generated alert used by the IDS was useful for the management of pBC in hospital. The impact of IDS counselling was more effective when the evaluation was restricted to medical and surgical wards.

Rapid detection of Pseudomonas aeruginosa from positive blood cultures by quantitative PCR.

BACKGROUND: Pseudomonas aeruginosa is responsible for numerous bloodstream infections associated with severe adverse outcomes in case of inappropriate initial antimicrobial therapy. The present study was aimed to develop a novel quantitative PCR (qPCR) assay, using ecfX as the specific target gene, for the rapid and accurate identification of P. aeruginosa from positive blood cultures (BCs). METHODS: Over the period August 2008 to June 2009, 100 BC bottles positive for gram-negative bacilli were tested in order to evaluate performances of the qPCR technique with conventional methods as gold standard (i.e. culture and phenotypic identification). RESULTS: Thirty-three strains of P. aeruginosa, 53 strains of Enterobactericaeae, nine strains of Stenotrophomonas maltophilia and two other gram-negative species were isolated while 3 BCs were polymicrobial including one mixture containing P. aeruginosa. All P. aeruginosa clinical isolates were detected by qPCR except a single strain in mixed culture. Performances of the qPCR technique were: specificity, 100%; positive predictive value, 100%; negative predictive value, 98.5%; and sensitivity, 97%. CONCLUSIONS: This reliable technique may offer a rapid (<1.5 h) tool that would help clinicians to initiate an appropriate treatment earlier. Further investigations are needed to assess the clinical benefit of this novel strategy as compared to phenotypic methods.

Brachyspira pilosicoli bloodstream infections: case report and review of the literature.

Brachyspira pilosicoli is the etiologic agent of human and animal intestinal spirochetosis and is rarely implicated as a cause of bacteremia. Here, we describe the case of a B. pilosicoli spirochetemia in a 53-year-old male patient suffering from cardiogenic shock. This fastidious bacterium was isolated from blood, likely after translocation from the intestinal tract. Blood cultures were positive after 5 days of incubation (one day after the patient's death), highlighting the problem of the recovery of such type of fastidious bacterium. Identification was achieved by molecular methods (16S rRNA sequencing). A review of the English literature found only 8 cases of bacteremia caused by B. pilosicoli, mostly in immunocompromised or critically ill patients. Finally, difficulties in rapid and accurate diagnosis of B. pilosicoli bloodstream infections, in vitro antimicrobial susceptibility of human clinical isolates, and therapeutic options are discussed.

Emergence of a Streptococcus pneumoniae isolate resistant to streptogramins by mutation in ribosomal protein L22 during pristinamycin therapy of pneumococcal pneumonia.

OBJECTIVES: The aim of this study was to characterize the mechanism of resistance to macrolides and streptogramins of a Streptococcus pneumoniae strain isolated from blood cultures in an 80-year-old patient suffering from severe pneumonia unsuccessfully treated with pristinamycin. METHODS: Resistance genes erm(B) and mef(A) were searched for by PCR. Portions of genes for domains V and II of the 23S rRNA (rrl) and genes for ribosomal proteins L4 (rplD) and L22 (rplV) were amplified by PCR from total genomic DNA and sequenced. RESULTS: Resistance genes erm(B) and mef(A) were not detected. Only mutation in the rplV gene encoding ribosomal protein L22 was detected. The strain contained a six amino acid insertion ((107)KRTAHI(108)) in the C-terminus of the ribosomal protein L22. CONCLUSIONS: This is the first report of emergence of a pneumococcus resistant to streptogramins by mutation in ribosomal protein L22 during treatment with pristinamycin.