Ocular mucosal homeostasis of teleost fish provides insight into the coevolution between microbiome and mucosal immunity.

BACKGROUND: The visual organ plays a crucial role in sensing environmental information. However, its mucosal surfaces are constantly exposed to selective pressures from aquatic or airborne pathogens and microbial communities. Although few studies have characterized the conjunctival-associated lymphoid tissue (CALT) in the ocular mucosa (OM) of birds and mammals, little is known regarding the evolutionary origins and functions of immune defense and microbiota homeostasis of the OM in the early vertebrates. RESULTS: Our study characterized the structure of the OM microbial ecosystem in rainbow trout (Oncorhynchus mykiss) and confirmed for the first time the presence of a diffuse mucosal-associated lymphoid tissue (MALT) in fish OM. Moreover, the microbial communities residing on the ocular mucosal surface contribute to shaping its immune environment. Interestingly, following IHNV infection, we observed robust immune responses, significant tissue damage, and microbial dysbiosis in the trout OM, particularly in the fornix conjunctiva (FC), which is characterized by the increase of pathobionts and a reduction of beneficial taxa in the relative abundance in OM. Critically, we identified a significant correlation between viral-induced immune responses and microbiome homeostasis in the OM, underscoring its key role in mucosal immunity and microbiota homeostasis. CONCLUSIONS: Our findings suggest that immune defense and microbiota homeostasis in OM occurred concurrently in early vertebrate species, shedding light on the coevolution between microbiota and mucosal immunity. Video Abstract.

Targeting the Gut-Eye Axis: An Emerging Strategy to Face Ocular Diseases.

The human microbiota refers to a large variety of microorganisms (bacteria, viruses, and fungi) that live in different human body sites, including the gut, oral cavity, skin, and eyes. In particular, the presence of an ocular surface microbiota with a crucial role in maintaining ocular surface homeostasis by preventing colonization from pathogen species has been recently demonstrated. Moreover, recent studies underline a potential association between gut microbiota (GM) and ocular health. In this respect, some evidence supports the existence of a gut-eye axis involved in the pathogenesis of several ocular diseases, including age-related macular degeneration, uveitis, diabetic retinopathy, dry eye, and glaucoma. Therefore, understanding the link between the GM and these ocular disorders might be useful for the development of new therapeutic approaches, such as probiotics, prebiotics, symbiotics, or faecal microbiota transplantation through which the GM could be modulated, thus allowing better management of these diseases.

Investigation of the pathogens contributing to naturally occurring outbreaks of infectious bovine keratoconjunctivitis (pinkeye) using Next Generation Sequencing.

Infectious bovine keratoconjunctivitis (IBK), commonly known as pinkeye, has a marked negative impact on the economy of the cattle industry. Moraxella species, including Mor. bovis and Mor. bovoculi, which have been associated with this disease, colonize clinically healthy eyes as well, suggesting that there are intrinsic changes that may occur to the ocular microbiota or the involvement of additional unrecognized organisms that contribute to IBK. To evaluate this, 104 ocular swabs collected from eyes with IBK or clinically healthy eyes from 16 different cattle herds were subjected to 16 S rRNA gene PCR and next generation sequencing (NGS) analysis. Organisms detected were similar across the herds and there was no difference in the total number of bacterial groups detected among IBK cases and controls. However, the percentages of the different organisms detected varied between the two groups, including Moraxella spp., with more Moraxella spp. in eyes with IBK than controls. Further, using culture and whole genome NGS, a new species of Moraxella (suggested name Mor. oculobovii) was detected from the eyes of cattle from two farms. This strain is non-hemolytic on blood agar, is missing the RTX operon, and is likely a non-pathogenic strain of the bovine ocular microbiome. Alteration of the ocular microbiota composition may have a predisposing role, enhancing bacterial infection and the occurrence of clinical IBK. Future studies are required to evaluate if these changes are permanent or if there is a shift in the microbiome following recovery from the infection and how antibiotics might affect the microbiome.

Effect of Liposomal-Lactoferrin-Based Eye Drops on the Conjunctival Microflora of Patients Undergoing Cataract Surgery.

INTRODUCTION: Postoperative endophthalmitis is typically caused by the patient's conjunctival bacterial flora. Povidone iodine solution (5%) is used perioperatively to obtain periocular and ocular antisepsis. However, an adjunctive prophylaxis procedure could further help control the conjunctival microbial load. Considering the increase in antibiotic resistance, a progressive shift toward alternative methods would be desirable. Somilux® eye drops (Alfa Intes, lactoferrin-based eye drops) are medical devices containing liposomal lactoferrin (LF). This study evaluates the effects on conjunctival microflora of LF-based eye drops used in the preoperative phase in patients scheduled for cataract surgery. METHODS: LF-based eye drops or a vehicle solution (water solution) were instilled 4 times a day starting 3 days before cataract surgery. Before the therapy (T0) and at the time of surgery (T1), a conjunctival swab was performed in both eyes and processed to detect microbial growth, microbiological isolation, and species identification. The outcome was the quantification and characterization of the local microbial flora before and after using LF-based or vehicle-based eye drops. Safety of the treatments was also evaluated. RESULTS: 88 eyes of 44 patients (mean [± SD] age 75 [± 12.6] years) were enrolled. At baseline, 54 conjunctival swabs showed only saprophytic flora, 27 showed only potential pathogenic flora, and seven showed both of them. LF-based eye drops reduced the proportion of potentially pathogenic bacteria (36% at T0 vs. 9% at T1, p = 0.008) compared with the vehicle (41% at T0 vs. 55% at T1, p = 0.302) without altering the physiological ocular microbial composition. No adverse events have been reported. CONCLUSION: Our findings provide a novel contribution to the scientific knowledge on the role of LF in the ophthalmic field, supporting the use of LF-based eye drops as a safe and selective treatment to improve the ocular surface physiological defenses and control the bacterial ocular surface contamination prior to cataract surgery.

Ocular microbiota and lens contamination following Mel4 peptide-coated antimicrobial contact lens (MACL) extended wear.

PURPOSE: The purpose of this study was to investigate the effect of Mel4 antimicrobial peptide-coated contact lenses (MACL) on the microbiota of the conjunctiva and lenses during three months of extended wear. METHODS: One hundred and seventy-six participants were recruited into a randomised, contralateral, double masked, biweekly extended wear MACL and uncoated control lens trial. At the one month and 3-month visit, the conjunctival microbiota was sampled using sterile cotton swabs and contact lenses were collected aseptically. Standard microbiological procedures were employed for culture of the swabs and contact lenses and identification of the isolated microorganisms. RESULTS: Gram-positive bacteria (predominantly coagulase-negative staphylococci) were the most frequently isolated microbes from both contact lenses and conjunctiva. There was no difference in the frequency of isolation of most bacteria or fungi from the conjunctival swabs of eyes wearing either MACL or control lenses. The only exception was a higher frequency of eyes harbouring Staphylococcus arlettae when wearing control lenses (5%) versus MACL (<1%) (p = 0.002). There was no significant difference in the frequency of microbes isolated from MACL or control contact lenses. There were also no differences between lens types in the frequency of isolation of >1 microbial type per sampling occasion for either conjunctiva swabs or contact lenses. CONCLUSION: MACL wear did not change the conjunctival microbiota during extended wear, and the types of microbes isolated from MACL were similar to those isolated from control lenses.

Ocular Microbiota and Intraocular Inflammation.

The term ocular microbiota refers to all types of commensal and pathogenic microorganisms present on or in the eye. The ocular surface is continuously exposed to the environment and harbors various commensals. Commensal microbes have been demonstrated to regulate host metabolism, development of immune system, and host defense against pathogen invasion. An unbalanced microbiota could lead to pathogenic microbial overgrowth and cause local or systemic inflammation. The specific antigens that irritate the deleterious immune responses in various inflammatory eye diseases remain obscure, while recent evidence implies a microbial etiology of these illnesses. The purpose of this review is to provide an overview of the literature on ocular microbiota and the role of commensal microbes in several eye diseases. In addition, this review will also discuss the interaction between microbial pathogens and host factors involved in intraocular inflammation, and evaluate therapeutic potential of targeting ocular microbiota to treat intraocular inflammation.

Preoperative topical liposomal ozone dispersion to reduce bacterial colonization in conjunctival sac and periocular skin: Preliminary study in dogs.

Prophylaxis represents a keystone to reduce periocular skin and ocular conjunctiva bacterial load before surgical procedures. Despite many prophylactic agents are available the preferred perioperative ocular surface antimicrobial is still unknown. The purpose of this study was to assess the effectiveness of preoperative liposomal ozone dispersion in reducing bacterial colonization from the conjunctival sac and periocular skin in dogs, in comparison with povidone-iodine and fluoroquinolone. Twenty-two owned dogs consisting with 44 eyes in total scheduled for ophthalmic surgical procedure were enrolled for the study and divided in four groups receiving either ozone dispersion or povidone iodine in eyelid and conjunctiva, fluoroquinolone or placebo. A swab was taken before and after the antisepsis protocol evaluating total microbial count, coagulase positive and negative staphylococci. Statistical analysis revealed a significant decrease in colony forming units (CFU) for total microbial count, coagulase positive and negative staphylococci both for liposomal ozone dispersion and povidone iodine. No statistical differences were detected in median CFU for both one-day placebo and fluoroquinolone preoperative prophylactic topical therapy. The results of this preliminary study demonstrate that liposomal ozone-dispersion is as effective as povidone iodine to reduce preoperative bacterial load in ocular surface.

Chloramphenicol Resurrected: A Journey from Antibiotic Resistance in Eye Infections to Biofilm and Ocular Microbiota.

The advent of multidrug resistance among pathogenic bacteria is devastating the worth of antibiotics and changing the way of their administration, as well as the approach to use new or old drugs. The crisis of antimicrobial resistance is also due to the unavailability of newer drugs, attributable to exigent regulatory requirements and reduced financial inducements. The emerging resistance to antibiotics worldwide has led to renewed interest in old drugs that have fallen into disuse because of toxic side effects. Thus, comprehensive efforts are needed to minimize the pace of resistance by studying emergent microorganisms and optimize the use of old antimicrobial agents able to maintain their profile of susceptibility. Chloramphenicol is experiencing its renaissance because it is widely used in the treatment and prevention of superficial eye infections due to its broad spectrum of activity and other useful antimicrobial peculiarities, such as the antibiofilm properties. Concerns have been raised in the past for the risk of aplastic anemia when chloramphenicol is given intravenously. Chloramphenicol seems suitable to be used as topical eye formulation for the limited rate of resistance compared to fluoroquinolones, for its scarce induction of bacterial resistance and antibiofilm activity, and for the hypothetical low impact on ocular microbiota disturbance. Further in-vitro and in vivo studies on pharmacodynamics properties of ocular formulation of chloramphenicol, as well as its real impact against biofilm and the ocular microbiota, need to be better addressed in the near future.

Determination of baseline values for routine ophthalmic tests in bearded capuchin (Sapajus libidinosus).

BACKGROUND: Establish baseline values for ophthalmic diagnostic tests in Sapajus libidinosus. METHODS: Ophthalmic diagnostic tests, namely Schirmer tear test 1 (STT-1), intraocular pressure (IOP), B-mode ultrasound, culture of the bacterial conjunctival microbiota, and conjunctival exfoliative cytology, were performed in 15 S. libidinosus. RESULTS: Mean values found were as follows: 2.50 ± 2.94 mm/min for the STT-1; 13.3 ± 3.32 mm Hg for the IOP; 2.47 ± 0.41 mm for the depth of the anterior chamber; 2.86 ± 0.96 mm for the axial length of the lens; 10.97 ± 0.48 mm for the depth of the vitreous chamber; and 16.32 ± 1.24 mm for the axial length of the eyeball. The bacterial genus most frequently found was Staphylococcus spp. Conjunctival cytology showed intermediate epithelial, squamous superficial epithelial, and keratinized cells. CONCLUSIONS: Determination of baseline values for eye measurements and ophthalmic tests will assist in the diagnosis of eye diseases in S. libidinosus monkeys.

Aging Eye Microbiota in Dry Eye Syndrome in Patients Treated with Enterococcus faecium and Saccharomyces boulardii.

BACKGROUND: Aging and oxidative stress seem to play a key role in the onset and progression of ocular surface diseases. Dry Eye Syndrome (DES) is a multifactorial disease of the tears and ocular surface in which symptoms may interfere with the ability to work and carry out daily functions. METHODS: This clinical trial was a pilot study to evaluate the effects of supplementation with mixture (Saccharomyces boulardii MUCL 53837 and Enterococcus faecium LMG S-28935) on the tear film. Following the run-in period subjects were randomized in two groups: group A (n.30 subjects) and group B (n.30 subjects). Group A (control) treated only with substitute tear and group B treated with substitute tear + mixture (probiotic). RESULTS: The data obtained in the two study groups A and B were, respectively the following: Schirmer I: 9.2±0.2 vs. 12.8±0.4 (p< 0.001); Schirmer II: 3.6±0.1 vs. 4.6±0.2 (p<0.001); BUT 3.8±0.3 vs. 6.2±0.2 (p<0.001). Culture test showed initial bacterial growth in group "A" (placebo) 27 out of 60 samples tested, corresponding to 45.0% and "B" after treatment (probiotic) was found positive culture whit growth of bacteria in 18 tests equal to 30.0%. The total numbers of isolations of aerobic and anaerobic bacteria found group A and B after treatment. A reduction of 23 to 16 strains of aerobic and anaerobic isolates from 13 to 7 has been found. CONCLUSION: The administration of probiotics strains was effective in reducing DES. In light of these results, we have identified our probiotic (Saccharomyces boulardii MUCL 53837 and Enterococcus faecium LMG S-28935) activity integration with the action of tear substitutes, along with standardization of clinical parameters of the tear film and microbiological activity in restoring of the microbiota ocular surface subject with DES.

Demodex spp en pacientes con blefaritis crónica / Demodex spp in chronic blepharitis patients

Background: Blepharitis is a very common disease in the ophthalmologic practice generally taking a chronic course with intermittent exacerbations. Several studies have linked the presence of Demodex folliculorum with chronic blepharitis, since the mite has the capacity to perpetuate the follicular inflammatory process. The prevalence of infection by Demodex spp. is variable depending on the population. In Paraguay, information on the frequency of the infestation in patients with chronic blepharitis is not available. Aim : To determine the frequency of Demodex spp, and the ocular microbiota in patients with chronic blepharitis attending the Department of Ophthalmology at the Teaching Hospital of the National University of Asuncion. Patients and Methods: Consecutively, 28 patients with chronic blepharitis, who agreed to participate in the study, were included. Eyes lashes from the upper and lower eyelids were extracted for immediate mite search by direct observation under a light microscope. Samples from eyelids were taken with Kimura spatula and then cultured on blood agar and in enrichment media and incubated in 5% CO2 at 35° C for 72 hours. Results: Among participants, females were more frequent (64%), the age ranged from 17 to 87 years (mean: 38.0; SD: ±13.5 years). The prevalence of Demodex sp was 54%. Bacteria were isolated 92.9% of cases, most frequently coagulase-negative staphylococci (75%). No association was found between socio-demographic or clinical characteristics and the presence of Demodex sp. Conclusion: The observed high prevalence of infestation by Demodex spp in patients with chronic blepharitis is consistent with other studies.

Introducción: La blefaritis es una enfermedad muy común en la práctica oftalmológica, generalmente de curso crónico con exacerbaciones intermitentes. Varios estudios han relacionado la presencia del Demodex folliculorum con la blefaritis crónica, por su capacidad potencial de perpetuar el proceso inflamatorio a nivel folicular. La frecuencia de D. folliculorum varía de acuerdo a la población. Objetivo: Determinar la presencia de Demodex spp y, caracterizar la microbiota ocular en pacientes con blefaritis crónica y dar a conocer estos resultados. Pacientes y Métodos: Las muestras fueron obtenidas de pacientes que consultaron en la Cátedra de Oftalmología del Hospital de Clínicas de la Universidad Nacional de Asunción (UNA). Fueron incluidos en forma consecutiva 28 pacientes con blefaritis crónica que accedieron a participar en forma voluntaria en el estudio. Se extrajeron las pestañas del párpado superior e inferior de los pacientes para la búsqueda inmediata del ácaro por observación directa bajo el microscopio óptico. Para el cultivo de microorganismos se tomaron muestras del párpado con espátula de Kimura, las que fueron cultivadas en agar sangre y en medios de enriquecimiento e incubadas en CO2 al 5% a 35°C durante 72 h. Resultados: El sexo femenino fue más frecuente (64%), el rango de edad estuvo entre 17 y 87 años con una media de 37,9 ± 13,5. Se observó la presencia de Demodex sp en 54% y aislamiento de bacterias en 92,9%; Staphylococcus coagulasa negativa fue el más frecuente (75%). No se encontraron asociaciones entre las características socio-demográficas o clínicas y la presencia de Demodex sp. Conclusión: La alta infestación por Demodex spp observada en los pacientes con blefaritis crónica coincide con otros estudios.