The porcine corneal surface bacterial microbiome: A distinctive niche within the ocular surface.

PURPOSE: The ocular surface microbiome has been described as paucibacterial. Until now, studies investigating the bacterial community associated with the ocular surface through high-throughput sequencing have focused on the conjunctiva. Conjunctival samples are thought to reflect and be representative of the microbiome residing on the ocular surface, including the cornea. Here, we hypothesized that the bacterial community associated with the corneal surface was different from those of the inferonasal and superotemporal conjunctival fornices, and from the tear film. METHODS: Both eyes from 15 healthy piglets were sampled using swabs (inferonasal fornix, superotemporal fornix, and corneal surface, n = 30 each) and Schirmer tear test strips (STT, n = 30). Negative sampling controls (swabs and STT, n = 2 each) and extraction controls (n = 4) were included. Total DNA was extracted and high-throughput sequencing targeting the 16S rRNA gene was performed. Bioinformatic analyses included multiple contamination-controlling steps. RESULTS: Corneal surface samples had a significantly lower number of taxa detected (P<0.01) and were compositionally different from all other sample types (Bray-Curtis dissimilarity, P<0.04). It also harbored higher levels of Proteobacteria (P<0.05), specifically Brevundimonas spp. (4.1-fold) and Paracoccus spp. (3.4-fold) than other sample types. Negative control STT strip samples yielded the highest amount of 16S rRNA gene copies across all sample types (P<0.05). CONCLUSIONS: Our data suggests that the corneal surface provides a distinct environmental niche within the ocular surface, leading to a bacterial community compositionally different from all other sample types.

Accuracy of ARFI elastography in the differentiation of cataract stages in dogs

ABSTRACT: Objective was to evaluate the accuracy of elastography in the differentiation between normal and cataract lenses One hundred forty-five eyes of 98 dogs were divided into groups according to cataract stage. Forty-twoeyes were submitted to phacoemulsification. Biometric parameters, echogenicity and echotexture patterns of the anterior, posterior and vitreous chambers, lens and retina-choroid-sclera complexes were evaluated by ocular ultrasound in modes A and B. Deformability, and color (blue color = indicated less rigid structures, color red = more rigid structures) of the lenses were evaluated by the elastogram. The shear wave velocity (SWV; m/s) was calculated in three regions of the lens, both in the cortex and in the nucleus. The SWV of nucleus was statistically different between the normal lenses and with cataracts, and between the stages of cataract (P 0.001). Healthy lenses and incipient cataracts had a more rigid nucleus. Mature cataracts presented lowest nuclear rigidity (P 0.001). On cortical region the SWV was significantly higher (P 0.01) in intumescent and incipient cataracts. SWV less than 2.67m/s indicates cataract with a sensitivity of 72% and specificity of 94%. Values lower than 2.23m/s suggest mature cataract, with sensitivity of 71% and specificity of 76%. SWV greater than 2.66 m/s are associated with normal lenses or incipient cataract, presenting sensitivity of 94% and specificity of 84%. Qualitative method allowed differentiation between healthy and affected lenses and the classification of evolutionary stages. There was a correlation between the degree of stiffness of lens in cortical and nuclear regions (p=00165, r=0.37) and between the balanced saline solution quantitative and surgical time (P 0.01, r=0.73). Degree of stiffness of lens did not correlate with parameters of phacoemulsification. Elastographic proved feasible for evaluating the lens of dogs, characterizing the types of cataracts, and demonstrating increased stiffness of the diseased lenses.

RESUMO: O objetivo foi avaliar a precisão da elastografia na diferenciação entre lentes normais e de catarata. Cento e quarenta e cinco olhos de 98 cães foram divididos em grupos de acordo com o estágio de maturação da catarata. Quarenta e dois olhos foram submetidos à facoemulsificação. Parâmetros biométricos, ecogenicidade e padrões de ecotextura das câmaras anterior, posterior e vítrea, lente e complexos retina-coróide-esclera foram avaliados por ultrassonografia ocular nos modos A e B. A deformabilidade e a coloração (cor azul = indicou estruturas menos rígidas, cor vermelha = estruturas mais rígidas) das lentes foram avaliadas pelo elastograma. A velocidade da onda de cisalhamento (SWV; m/s) foi calculada em três regiões da lente, tanto no córtex quanto no núcleo. A SWV do núcleo foi estatisticamente diferente entre as lentes normais e com catarata e entre os estágios da catarata (P 0,001). Lentes saudáveis e cataratas incipientes tinham um núcleo mais rígido. Cataratas maduras apresentaram menor rigidez nuclear (P 0,001). Na região cortical, a SWV foi significativamente maior (P 0,01) nas cataratas intumescentes e incipientes. Uma SWV menor que 2,67m/s indica catarata com sensibilidade de 72% e especificidade de 94%. Valores inferiores a 2,23m/s sugerem catarata madura, com sensibilidade de 71% e especificidade de 76%. Uma SWV superior a 2,66m/s está associada à catarata normal ou incipiente, apresentando sensibilidade de 94% e especificidade de 84%. O método qualitativo permitiu a diferenciação entre lentes normais de olhos saudáveis e afetadas e a classificação dos estágios evolutivos. A elastografia se mostrara uma ferramenta viável para avaliar as lentes de cães, caracterizando os tipos de catarata e demonstrando maior rigidez das lentes doentes.

Accuracy of ARFI elastography in the differentiation of cataract stages in dogs / Acurácia da elastografia ARFI na diferenciação dos estágios da catarata em cães

Objective was to evaluate the accuracy of elastography in the differentiation between normal and cataract lenses One hundred forty-five eyes of 98 dogs were divided into groups according to cataract stage. Forty-twoeyes were submitted to phacoemulsification. Biometric parameters, echogenicity and echotexture patterns of the anterior, posterior and vitreous chambers, lens and retina-choroid-sclera complexes were evaluated by ocular ultrasound in modes A and B. Deformability, and color (blue color = indicated less rigid structures, color red = more rigid structures) of the lenses were evaluated by the elastogram. The shear wave velocity (SWV; m/s) was calculated in three regions of the lens, both in the cortex and in the nucleus. The SWV of nucleus was statistically different between the normal lenses and with cataracts, and between the stages of cataract (P<0.001). Healthy lenses and incipient cataracts had a more rigid nucleus. Mature cataracts presented lowest nuclear rigidity (P<0.001). On cortical region the SWV was significantly higher (P<0.01) in intumescent and incipient cataracts. SWV less than 2.67m/s indicates cataract with a sensitivity of 72% and specificity of 94%. Values lower than 2.23m/s suggest mature cataract, with sensitivity of 71% and specificity of 76%. SWV greater than 2.66 m/s are associated with normal lenses or incipient cataract, presenting sensitivity of 94% and specificity of 84%. Qualitative method allowed differentiation between healthy and affected lenses and the classification of evolutionary stages. There was a correlation between the degree of stiffness of lens in cortical and nuclear regions (p=00165, r=0.37) and between the balanced saline solution quantitative and surgical time (P<0.01, r=0.73). Degree of stiffness of lens did not correlate with parameters of phacoemulsification. Elastographic proved feasible for evaluating the lens of dogs, characterizing the types of cataracts, and demonstrating increased stiffness of the diseased lenses.(AU)

O objetivo foi avaliar a precisão da elastografia na diferenciação entre lentes normais e de catarata. Cento e quarenta e cinco olhos de 98 cães foram divididos em grupos de acordo com o estágio de maturação da catarata. Quarenta e dois olhos foram submetidos à facoemulsificação. Parâmetros biométricos, ecogenicidade e padrões de ecotextura das câmaras anterior, posterior e vítrea, lente e complexos retina-coróide-esclera foram avaliados por ultrassonografia ocular nos modos A e B. A deformabilidade e a coloração (cor azul = indicou estruturas menos rígidas, cor vermelha = estruturas mais rígidas) das lentes foram avaliadas pelo elastograma. A velocidade da onda de cisalhamento (SWV; m/s) foi calculada em três regiões da lente, tanto no córtex quanto no núcleo. A SWV do núcleo foi estatisticamente diferente entre as lentes normais e com catarata e entre os estágios da catarata (P<0,001). Lentes saudáveis e cataratas incipientes tinham um núcleo mais rígido. Cataratas maduras apresentaram menor rigidez nuclear (P<0,001). Na região cortical, a SWV foi significativamente maior (P<0,01) nas cataratas intumescentes e incipientes. Uma SWV menor que 2,67m/s indica catarata com sensibilidade de 72% e especificidade de 94%. Valores inferiores a 2,23m/s sugerem catarata madura, com sensibilidade de 71% e especificidade de 76%. Uma SWV superior a 2,66m/s está associada à catarata normal ou incipiente, apresentando sensibilidade de 94% e especificidade de 84%. O método qualitativo permitiu a diferenciação entre lentes normais de olhos saudáveis e afetadas e a classificação dos estágios evolutivos. A elastografia se mostrara uma ferramenta viável para avaliar as lentes de cães, caracterizando os tipos de catarata e demonstrando maior rigidez das lentes doentes.(AU)

Tissue fibrosis and its correlation with malignancy in canine mammary tumors / Fibrosis tisular y su correlación con malignidad en neoplasias mamarias caninas / Fibrose tecidual e sua correlação com a malignidade em neoplasias mamárias em cadelas

Abstract Background: Fibrosis is present in several pathologies associated with mammary carcinogenesis. Objective: To evaluate and quantify the fibrosis present in malignant and benign mammary neoplasms in bitches. Methods: Eighty-three samples were divided according to histopathological diagnosis into benign (n= 21) and malignant (n= 62) neoplasms. Haematoxylin-eosin and Masson's trichrome were used to locate the connective tissue, and the extent of fibrosis was assessed with image software. Results: Benign neoplasms were classified into adenomas (cystic, complex, and tubular), benign mixed tumor, and ductal and lobular hyperplasia. Malignant neoplasms were classified as carcinomas (complex, mixed tumor, in situ tubular, tubulopapillary, and solid). Grade I was the most prevalent histopathological class, followed by grade II and III. Fibrosis was classified as severe, moderate, or discrete. No significant (p >0.05) difference was observed for the percentage of fibrosis between malignant and benign group neoplasms. However, difference (p=0.028) was found for fibrosis percentage between histopathological subtypes of tumors. The benign subtype of lobular hyperplasia presented differences between cystic adenoma and benign mixed tumor. The in situ malignant tubular carcinoma subtype presented differences between solid and tubulopapillary carcinoma. Conclusions: Fibrosis in canine mammary tumors can be estimated with Massons's trichrome staining.

Resumen Antecedentes: La fibrosis está presente en diversas patologías asociadas con carcinogénesis mamaria. Objetivo: Analizar si existe una correlación entre fibrosis tisular y malignidad en tumores mamarios caninos. Métodos: 83 muestras de tejido mamario fueron divididas en masas benignas (n=21) y malignas (n=62), de acuerdo con sus características histopatológicas. En estas muestras se utilizaron las coloraciones de hematoxilina-eosina y tricromo de Masson para localizar el tejido conectivo y se analizó la proporción y cuantificación de fibrosis en los mismos con un software de imagen especializado. Resultados: Las masas mamarias benignas se diagnosticaron como adenomas (quístico, complejo y tubular), tumor mixto benigno, e hiperplasia ductal y lobular; las masas malignas, como carcinomas (complejo, tumor mixto, tubular in situ, túbulopapilar y solido). Los tumores grado I fueron los más prevalentes, seguidos por los grados II y III. La fibrosis se clasifico como discreta, moderada o severa. No se observó diferencia (p>0,05) en el porcentaje de fibrosis entre neoplasias benignas y malignas. No obstante, el porcentaje de fibrosis mostro diferencias (p=0,028) entre subtipos tumorales. La hiperplasia lobular fue diferente en relación a los adenomas quísticos y tumor mixto mamario. El subtipo maligno carcinoma tubular in situ fue diferente respecto a los carcinomas complejo, mixto maligno, sólido, y tubulopapilar. Conclusiones: La fibrosis en los tumores mamarios caninos se puede estimar por la tinción con tricromo de Masson.

Resumo Antecedentes: A fibrose participa em diversas patologias e ainda possui função adicional associada à carcinogênese mamária. Objetivo: Objetivou-se avaliar a fibrose e correlacionar com a malignidade nas neoplasias mamárias em cadelas. Métodos: 83 amostras foram divididas em dois grupos baseado no diagnóstico histopatológico: neoplasias benignas (n= 21) e neoplasias malignas (n= 62). Hematoxilina e eosina e Tricômico de Masson foram usadas para vizibilização de tecido conjuntivo e avaliação de fibrose através de programa de imagens. Resultados: As neoplasias benignas foram: adenoma (cístico, complexo e tubular), tumor misto benigno, hiperplasia ductal e lobular. As malignas foram: carcinoma complexo, em tumor misto maligno, tubular in situ, tubulopapilar e sólido. O grau histopatológico prevalente foi grau I, seguido do grau II e III. A fibrose nas neoplasias mamárias malignas foi classificada como severa, moderada e discreta. Não houve diferença significativa (p>0,05) na porcentagem de fibrose entre neoplasias malignas e benignas. A estatística revelou diferença significativa (p=0,028) na porcentagem de fibrose em relação ao diagnóstico histopatológico. O subtipo benigno hiperplasia lobular apresentou diferença entre o adenoma cístico e o tumor misto benigno. O subtipo maligno carcinoma tubular in situ apresentou diferença entre carcinoma complexo, misto maligno, sólido e tubulopapilar. Conclusões: A fibrose nos tumores mamários caninos pode ser estimada através da coloração de tricrômio de Masson.