A descriptive ultrasonographic and histological evaluation of the eyes of humpback whales (Megaptera novaeangliae).

OBJECTIVE: To describe morphological features of Megaptera novaeangliae eyes using ultrasonography and histology. ANIMAL STUDIED: A total of 21 globes from 19 M. novaeangliae were used for the study, including two animals with bilateral assessment. Nine stranded animals were found alive, 10 dead. PROCEDURES: Carcasses were classified according to decomposition state. Globes were assessed ultrasonographically, biometric measurements were taken and then the tissues were sectioned for histological analysis. RESULTS: Seventeen carcasses were classified as decomposition code 2 and two as code 3. Fifteen animals were calves, three juveniles and one adult, twelve males, and five females. The cornea was thinner in the center of the eye and thicker at the periphery. The lens had an oval shape. Fourteen animals showed a structure in the vitreous body which looked triangular and filamentous by ultrasonography and, histologically, originated in the central part of the optic nerve and extended to the posterior region of the lens, composed of connective tissue. The most common abnormalities found by ultrasonograph were retinal detachment (n = 13) and displacement of the lens into the vitreous (n = 4), along with alterations suggestive of hemorrhage, fibrin deposits, and increased echogenicity in the optic nerve (n = 2). CONCLUSIONS: An intraocular structure not previously described in cetaceans was found in this investigation; its function remains unknown. Circulatory changes that were evident in the histopathological analysis may be due to the stranding process and raise the need to consider ophthalmic examinations before reintroducing stranded mysticetes.

Morphometric descriptive report of scleral ossicle rings, by ultrasound and computed tomography, in three Testudines specimens / Relato descritivo morfobiométrico dos anéis de ossículos esclerais, por ultrassonografia e tomografia computadorizada, em três espécimes de Testudines

The scleral ossicle rings function has been related to mechanical protection, muscle fixation, support for eyeball shape and visual accommodation. There are few morphobiometric reports on these rings in different Testudines species, and we performed ultrasound (US) and computed tomography (CT) of the scleral ossicle rings in one green turtle (Chelonia mydas), one black-bellied slider (Trachemys dorbigni) and one red-footed tortoise (Chelonoidis carbonarius). The US and CT of the ossicle rings were performed for anatomical identification. The thickness, density, width, and diameters of each ring were measured. The US and CT of the scleral ossicle rings of three animals showed single and continuous circular structures, located in the anterior pole. These structures were easily observed in C. mydas, whose rings were the biggest, thickest and widest. The T. dorbigni CT presented decreased dimensions and the ossicles were the most difficult to identify. Bone density in the superior region was greater than in the inferior of each ring in all animals. Non-invasive imaging exams are good tools to study the anatomy of the ocular skeleton. The scleral ossicle rings of the three specimens presented general morphological similarities and CT enabled visualizing a greater number of details of the ring bone morphology.

Os anéis de ossículos esclerais têm sua função relacionada à proteção mecânica, fixação muscular, suporte para o formato do bulbo ocular e acomodação visual, contudo existem poucos relatos morfobiométricos sobre esses anéis em diferentes espécies de Testudines. Desta forma, foi realizada a avaliação morfobiométrica, por ultrassom (US) e tomografia computadorizada (TC), dos anéis de ossículos esclerais em uma tartaruga verde (Chelonia mydas), um tigre-d'água (Trachemys dorbigni) e um jabuti-piranga (Chelonoidis carbonarius). Foram realizadas US e TC dos anéis esclerais dos três animais para identificação anatômica, espessura, densidade, largura e diâmetros. A US e a TC dos três animais mostraram estruturas circulares únicas e contínuas, localizadas no polo anterior. Estas estruturas foram facilmente observadas na C. mydas, cujos anéis eram os maiores, mais espessos e mais largos. A TC da T. dorbigni apresentou dimensões reduzidas e os ossículos foram dificilmente identificados. A densidade óssea na região superior foi maior comparativamente a parte inferior de cada anel, em todos os animais. Exames de imagem não invasivos mostraram-se bons instrumentos para estudo do esqueleto escleral. Os anéis de ossículos esclerais dos três espécimes apresentaram semelhanças morfológicas gerais e a TC permitiu visualizar um maior número de detalhes da morfologia óssea do anel.

Morphometric descriptive report of scleral ossicle rings, by ultrasound and computed tomography, in three Testudines specimens / Relato descritivo morfobiométrico dos anéis de ossículos esclerais, por ultrassonografia e tomografia computadorizada, em três espécimes de Testudines

ABSTRACT: The scleral ossicle rings function has been related to mechanical protection, muscle fixation, support for eyeball shape and visual accommodation. There are few morphobiometric reports on these rings in different Testudines species, and we performed ultrasound (US) and computed tomography (CT) of the scleral ossicle rings in one green turtle (Chelonia mydas), one black-bellied slider (Trachemys dorbigni) and one red-footed tortoise (Chelonoidis carbonarius). The US and CT of the ossicle rings were performed for anatomical identification. The thickness, density, width, and diameters of each ring were measured. The US and CT of the scleral ossicle rings of three animals showed single and continuous circular structures, located in the anterior pole. These structures were easily observed in C. mydas, whose rings were the biggest, thickest and widest. The T. dorbigni CT presented decreased dimensions and the ossicles were the most difficult to identify. Bone density in the superior region was greater than in the inferior of each ring in all animals. Non-invasive imaging exams are good tools to study the anatomy of the ocular skeleton. The scleral ossicle rings of the three specimens presented general morphological similarities and CT enabled visualizing a greater number of details of the ring bone morphology.

RESUMO: Os anéis de ossículos esclerais têm sua função relacionada à proteção mecânica, fixação muscular, suporte para o formato do bulbo ocular e acomodação visual, contudo existem poucos relatos morfobiométricos sobre esses anéis em diferentes espécies de Testudines. Desta forma, foi realizada a avaliação morfobiométrica, por ultrassom (US) e tomografia computadorizada (TC), dos anéis de ossículos esclerais em uma tartaruga verde (Chelonia mydas), um tigre-d'água (Trachemys dorbigni) e um jabuti-piranga (Chelonoidis carbonarius). Foram realizadas US e TC dos anéis esclerais dos três animais para identificação anatômica, espessura, densidade, largura e diâmetros. A US e a TC dos três animais mostraram estruturas circulares únicas e contínuas, localizadas no polo anterior. Estas estruturas foram facilmente observadas na C. mydas, cujos anéis eram os maiores, mais espessos e mais largos. A TC da T. dorbigni apresentou dimensões reduzidas e os ossículos foram dificilmente identificados. A densidade óssea na região superior foi maior comparativamente a parte inferior de cada anel, em todos os animais. Exames de imagem não invasivos mostraram-se bons instrumentos para estudo do esqueleto escleral. Os anéis de ossículos esclerais dos três espécimes apresentaram semelhanças morfológicas gerais e a TC permitiu visualizar um maior número de detalhes da morfologia óssea do anel.

Birefringence Analyses Reveal Differences in Supramolecular Characteristics of Corneal Stromal Collagen Fibrils Between Falconiformes and Strigiformes.

This study aimed to assess the birefringent properties of corneal stromal collagen fibrils in birds of the orders Falconiformes (diurnal) and Strigiformes (predominantly nocturnal) to compare their supramolecular organizations. In total, 22 corneas of Falconiformes (Caracara plancus, n = 8; Rupornis magnirostris, n = 10; and Falco sparverius, n = 4) and 28 of Strigiformes (Tyto furcata, n = 16; Pseudoscops clamator, n = 6; and Athene cunicularia, n = 6) were processed histotechnically into 8-µm thick sections. Corneal optical retardation (OR) values related to the form and intrinsic fractions of the total birefringence of collagen fibrils were measured using a polarized light microscope equipped with phase compensators. In addition, the coherence coefficients that inform the local orientation of the fibrils were calculated through video image analysis. All assessments were conducted both in the anterior and posterior stroma of the cornea. Differences were significant when P < 0.05. The results showed supraorganizational differences between fibrils in the anterior stroma of Falconiformes and Strigiformes. The OR values were greater (P < 0.0001) for Falconiformes, indicating that the corneas of these birds contain more collagen fibrils or more aggregated collagen fibrils. In contrast, the coherence coefficients were higher (P = 0.016) for Strigiformes, indicating that the corneal collagen fibers in these birds are highly aligned and have few undulations. A multivariate data matrix constructed for Euclidean distance calculations showed that the dissimilarity between Falconiformes and Strigiformes corneas, in terms of the supraorganization of stromal collagen fibrils, was 4.56%. In conclusion, it is possible that the supraorganizational differences reported in this study may be sources of variation in the visual quality of Falconiformes and Strigiformes. This study provides the necessary evidence to encourage further research associating corneal optical performance to supramolecular characteristics of corneal stromal collagen.

The glycoproteomics of hawk and caiman tears.

BACKGROUND: Glycoproteins are important tear components that participate in the stability of the ocular surface. However, the glycopeptides that are present in the tears of wild animals have not yet been described. This work aimed to describe the glycoproteomic profile of roadside hawk (Rupornis magnirostris) and caiman (Caiman latirostris) tears. METHODS: Tears collected from 10 hawks and 70 caimans using Schirmer tear test strips were used in this study. The samples were submitted to trypsin digestion and separated using a reverse-phase column coupled to a mass spectrometer associated to a nanospray ionization source. The glycoproteins were categorized as: cellular components, biological processes and molecular function, according to the UniProt Knowledgebase. RESULTS: As shown by the liquid chromatography-mass spectrometry, all glycopeptides found were classified as N-type. Of the 51 glycoproteins that were identified in the hawk tear film, the most abundant were ovotransferrin, globulins and complement system proteins. In the caiman tear film, 29 glycoproteins were identified. The most abundant caiman glycoproteins were uncharacterized proteins, ATPases, globulins and proteasome components. Ontological characterization revealed that the glycoproteins were extracellular, and the most identified molecular function was endopeptidase activity for both species. CONCLUSION: Glycoproteins are abundant in the tear film of the bird and reptile species studied herein, and all these molecules were shown to have N-type modifications. Location at the extracellular space and an endopeptidase inhibitor activity were the main cell component and molecular function for both species, respectively. These profiles showed differences when compared to human tears, are possibly linked to adaptive processes and can be the basis for further studies on the search of disease biomarkers.

Tear Film Proteome of Healthy Domestic Cats.

The aim of this study was to investigate the proteins found in tear film of healthy domestic cats. Schirmer tear test strips were used to collect tear samples of twelve healthy cats, which were mixed, centrifuged, and placed in a single 1.5 mL microtube that was frozen at -20°C, until analysis by two-dimensional polyacrylamide gel and mass spectrometry associated with high-performance liquid chromatography. The resulting spectra were analyzed and compared with the Swiss-Prot search tool. Forty peptides were detected in the analyzed protein fragments of 90 spots, with 16 proteins identified. Of these, the authors confirmed what has been already found in other studies: lactotransferrin, serum albumin, allergenic lipocalins, and neutrophil gelatinase-associated lipocalin. Others were considered novel in tear film samples of all species: cyclin-dependent protein kinase, serine/arginine repetitive matrix protein, apelin receptor, secretory protein related to C1q/TNF, Wee1, α-1,4 glucan phosphorylase, and WD repeat domain 1. The network was divided into 11 clusters, and a biological function was assigned. Most of the proteins have functions in the defense and maintenance of feline ocular surface homeostasis. Serum albumin is a bottleneck protein, with a high betweenness value. This paper is a pioneer in reporting, in-depth, the tear film proteome of domestic cats.

Atypical free-floating iridociliary pigmented epithelial cysts and secondary glaucoma in a caiman (Caiman latirostris).

We describe a case of chronic ocular trauma that resulted in fixed and free-floating, pigmented epithelial iridociliary cysts, inflammation, and secondary glaucoma in a caiman (Caiman latirostris). A 20- to 25-year-old male caiman was presented with phthisis bulbi in the right eye, and congested episcleral vessels, corneal leukoma, disorganized anterior chamber, multifocal anterior synechia, and elevated intraocular pressure in the left eye. Ocular ultrasound of the left eye revealed round structures dispersed in the anterior and posterior chambers and vitreous cavity. Bilateral enucleation was performed, and gross pathology of the left eye revealed multiple pigmented cysts attached to the iris and posterior corneal surface causing marked distortion of the anterior uvea, and free-floating in the vitreous cavity. Histopathology demonstrated heavily pigmented cystic structures of iridociliary epithelium origin carpeting the anterior segment surfaces and causing obstruction of the iridocorneal angles, leading to secondary glaucoma. To the authors' knowledge, this is the first report of iridociliary cysts in wildlife species.

Rabbit as an Animal Model for Ocular Surface Disease, Tear Osmolarity, Electrolyte, and Tear Ferning Profiles.

SIGNIFICANCE: The tear film promotes ocular surface health and protection through its various constituents' functions. The application of methods for ocular surface examination is essential in the research of diseases that affect the tear film. Rabbits have been used as a model to study some human ocular diseases and to test ophthalmic products. PURPOSE: The aim of the present study was to determine the biochemical profile, osmolarity, and tear ferning patterns of rabbit tears. METHODS: Ten rabbits (Oryctolagus cuniculus) were evaluated for tear osmolarity, tear ferning types and grades (using Rolando and Masmali scales), and biochemical analysis of total protein, urea, and electrolytes (chloride, phosphorus, iron, sodium, potassium, and calcium). RESULTS: Median ± semi-interquartile range for tear osmolarity was 283.5 ± 7.5 mOsm/L. Tear ferning test grades were type 1.0 ± 0.5 on the Rolando scale and 1.0 ± 0.5 on the Masmali scale. Type I and grade 1 were the most commonly observed ferning classifications (60 and 50%, respectively) for the rabbits' tears. Results for electrolytes and other biochemical compounds were as follows: total protein 4.40 g/dL, urea 130.60 mg/dL, chloride 196.51 mEq/L, phosphate 7.35 mg/dL, iron 95.76 µg/dL, sodium 202.04 mmol/L, potassium 12.74 mmol/L, and calcium 11.53 mg/dL. CONCLUSIONS: The results of the various tests described herein may serve as a basis for research using rabbits as an ophthalmic disease model and in the development of diagnostic and therapeutic agents used for ocular health.

Ocular ultrasonography of sea turtles.

BACKGROUND: Environmental changes contribute to the development of ophthalmic diseases in sea turtles, but information on their eye biometrics is scarce. The aim of this study was to describe ophthalmic ultrasonographic features of four different sea turtle species; Caretta caretta (Loggerhead turtle; n = 10), Chelonia mydas (Green turtle; n = 8), Eretmochelys imbricata (Hawksbill turtle; n = 8) and Lepidochelys olivacea (Olive ridley; n = 6) under human care. Corneal thickness, scleral ossicle width and thickness, anterior chamber depth, axial length of the lens, vitreous chamber depth and axial globe length were measured by B-mode sonography with a linear transducer. Carapace size and animal weight were recorded. A sonographic description of the eye structures was established. RESULTS: The four species presented an ovate eyeball, a relatively thin cornea, and a small-sized lens positioned rostrally in the eye bulb, near the cornea, resulting in a shallow anterior chamber. The scleral ossicles did not prevent the evaluation of intraocular structures, even with a rotated eye or closed eyelids; image formation beyond the ossicles and measurements of all proposed structures were possible. B-mode sonography was easily performed in all animals studied. The sonographic characteristics of the eye were similar among the four species. Since there was a correlation between the size of the eye structures and the size of the individual, especially its carapace size, the differences found between E. imbricata and Caretta caretta are believed to be due to their overall difference in size. CONCLUSIONS: Sonography is a valuable tool in ophthalmic evaluation of these species. Only minor differences were found between the species in this study, reinforcing their phylogenetic proximity and their similar functions and habitats.

Comparative Analysis of Tear Composition in Humans, Domestic Mammals, Reptiles, and Birds.

Tears are an important component of the ocular surface protection mechanism and are in close contact with the corneal epithelium and the environment. Their composition is well-known in humans; however, there are few investigations on the composition and function of tears in reptiles, birds and others mammals, which would elucidate the mechanisms governing the maintenance of ocular homeostasis. In this work, electrophoretic profiles and an evaluation of total protein, albumin, urea, glucose, and cholesterol concentrations in tears of semi-aquatic, terrestrial, and marine reptiles (Caiman latirostris, Chelonia mydas, Caretta caretta, Eretmochelys imbricata, Lepidochelys olivacea, and Chelonoidis carbonaria), birds (Tyto furcata, Rupornis magnirostris and Ara ararauna), and mammals (Equus caballus and Canis lupus familiaris) were apresented. Human tear components and respective blood serum samples were used as references. The electrophoretic analysis revealed similarities whithin same Classes. The results of the tear-blood serum relationship and the comparison to human tear components showed particularities that are potentially derived from a homeostatic response to the environment. When the tear compositions of animals belonging to different ecological clusters were compared, marked differences were observed in total protein and urea concentrations. Thus, reptile, bird, and mammalian tears are complex fluids with differing concentrations of biochemical components that are potentially a result of the animals' adaptation to different environments.

The use of tear ferning test in cats for evaluation of ocular surface.

BACKGROUND: The tear film is a trilaminar fluid composed mainly of lipids, electrolytes, proteins and water. It is responsible for lubrication, nutrition and protection against microbial and toxic agents. Disruption of any these components may weaken the ocular surface, making it more susceptible to disease. Increasing evidence suggests that qualitative tear film deficiencies are an important predisposing factor or cause of some of the most common and challenging ocular diseases in cats, including conjunctivitis, corneal ulcer, spontaneous chronic corneal epithelial defects (SCCED), pigmentary keratitis, corneal sequestrum and dry eye syndrome. The aim of this study was to describe the tear ferning test in healthy cats and to compare the results by using two grading scales for humans. Tear samples were collected using Schirmer tear test (STT) strips from 60 healthy cats, and, after centrifuging the strips to obtain the samples, the aliquot was placed on clean microscope glass until it dried and the tear ferning patterns were observed under a polarized light microscope and classified according to the Rolando and Masmali grading scales. RESULTS: Ferning patterns in the lower grades showed full crystallization with high density, without gaps between the ferns and branches, forming several nuclei that were easily distinguished. According to the Rolando scale, 50% (60/120), 46.6% (56/120) and 3.4% (4/120) of eyes showed type I, II and III patterns, respectively. According to the Masmali scale, 15% (18/120), 56.6% (68/120 eyes) and 28.4% (34/120) of eyes showed grade 0, 1 and 2 patterns, respectively. No difference was observed between the right and left eyes for both Rolando (P = 0.225) and Masmali (P = 0.683) scales. CONCLUSIONS: The tear ferning test is a qualitative test that can be used in cats as a complementary evaluation of the ocular surface. While the Rolando scale showed an increased prevalence of types I and II, the Masmali scale showed an increased prevalence of grades 1 and 2. This can be attributed to the species-specific differences between human and feline tear film. So Masmali grade 2 can be considered a normal tear pattern for the species, because all the cats used in study were clinically healthy. For this reason, future complementary studies are necessary for comparing healthy eyes and eyes with different ocular surface disease in cats. Both scales can be feasible options for grading tear crystallization in cats, but as Rolando scale included 96.6% of the samples in the 2 types that are considered normal for humans, we think that this scale seemed to be more precise to classify crystallization pattern in cats. The crystallization patterns observed in this study can form the basis for standardizing ocular surface parameters in cats.

Detection of Leptospira spp. in Captive Broad-Snouted Caiman (Caiman latirostris).

Leptospira sp. is an important waterborne zoonotic bacterium, known to cause infection in animals and humans worldwide. The role of reptiles in the transmission of this microorganism is poorly understood and historically neglected. This study aimed to investigate the presence of anti-Leptospira spp. antibodies and leptospiral DNA in captive Caiman latirostris (broad-snouted caiman). Of the 23 reptiles studied by microscopic agglutination test (MAT), 22/23 (95.65%) were considered reactive (titers ≥ 100) and 1/23 (4.35%) non-reactive (titer < 100). The serogroup with highest occurrence was Grippotyphosa (68.18%, n = 15/22) followed by serogroup Djasiman (18.18%, n = 4/22). Specific amplification of Leptospira spp. gene lipL32 was observed in six (26.09%, n = 6/23) blood samples. Five of six samples, previously detected as pathogenic leptospira by PCR, were amplified and sequenced. All the samples corresponded to the pathogenic species Leptospira interrogans (presented 100% of identity) using the PCR targeting to secY gene. We demonstrated high detection of DNA of L. interrogans in crocodilians, and the authors suggest that further research is needed to elucidate the impact of Leptospira spp. infection in health broad-snouted caimans as well as the pathophysiology of leptospirosis in crocodilians.

Anatomical, histological and computed tomography comparisons of the eye and adnexa of crab-eating fox (Cerdocyon thous) to domestic dogs.

An understanding of species' morphological and physiological parameters is crucial to developing conservation strategies for wild animals kept in human care. Detailed information is lacking for crab-eating fox (Cerdocyon thous) eyes and adnexa. Therefore, the aim of this study was to describe anatomical, histological and computed tomography (CT) features of the eye and adnexa in crab-eating fox, compared to domestic dogs. CT of the eye and adnexa of one live animal and a frozen specimen was performed for anatomical identification. In addition, the heads of five animals of each species were fixed in 10% buffered formalin for gross anatomical description of the eye and adnexa using topographic dissection and exenteration techniques. All steps were photographed and features such as location, shape, and distances and relationships between structures were described. For histological evaluation, two eyes of each species were fixed in 10% buffered formalin, processed by routine paraffin inclusion technique and stained with hematoxylin and eosin. The CT scan was difficult to evaluate, mainly that of the frozen head, which did not provide good definition of the soft tissues; nevertheless, it demonstrated the potential for structure visualization and description. The gross anatomical and histological evaluations showed the presence of eyelashes on the upper eyelid and of upper and lower lacrimal points, an incomplete orbit with supraorbital ligament, slightly exposed sclera with discretely pigmented limbus and pigmentation throughout the conjunctiva, and a slit-shaped pupil. Hematoxylin and eosin staining demonstrated structural similarities between the crab-eating fox and domestic dog. Thus, the possibility of using the domestic dog as a study model for the preventive and therapeutic management of wild dogs kept in human care is demonstrated.

Tear meniscometry test in wild animals / Teste de meniscometria lacrimal em animais silvestres

ABSTRACT: This study aimed to describe strip meniscometry test values for Amazona aestiva (turquoise-fronted amazon), Ara ararauna (blue-and-yellow macaw) and Caiman latirostris (broad-snouted caiman). Twenty-one Amazona aestiva and 11 Ara ararauna, healthy, adults of unknown sex, and 37 healthy adult Caiman latirostris (27 males and 10 females), were used in this investigation. All animals were manually restrained and I-Tear® Test strip was inserted for 5 s at the edge of the lower tear meniscus, without touching the eyelid or ocular surface, immediately followed by measurement (millimeter per 5 seconds). No differences were reported between left and right eyes for Amazona aestiva (P = 0.824), Ara ararauna (P = 0.262) and Caiman latirostris (P = 0.679). Median and confidence interval were 2.0 (2.25-3.56) mm/5s for Amazona aestiva, 3.0 (2.93-4.15) mm/5s for Ara ararauna and 3.0 (2.55-4.90) mm/5 s for Caiman latirostris. These values for strip meniscometry tear test can be used as parameters for each species and may be helpful for diagnosis and treatment of tear-production disorders in birds and reptiles.

RESUMO: O objetivo deste estudo foi descrever os valores do teste de meniscometria lacrimal para Amazona aestiva (papagaio-verdadeiro), Ara ararauna (arara-canindé) e Caiman latirostris (jacaré-de-papo-amarelo). Vinte e um Amazona aestiva, 11 Ara ararauna adultos, de sexo desconhecido, e 37 Caiman latirostris adultos (27 machos e 10 fêmeas), foram utilizados nesta investigação. Todos os animais foram contidos manualmente e uma tira do I-Tear® test foi inserida por 5 segundos na borda do menisco inferior, sem tocar a pálpebra ou a superfície ocular, e imediatamente mensurada (milímetro por 5 segundos). Não foram encontradas diferenças entre os olhos esquerdos e direitos para Amazona aestiva (P = 0,824), Ara ararauna (P = 0,262) e Caiman latirostris (P = 0,679). Mediana e intervalo de confiança foram de 2,0 (2,25-3,56) mm/5 s para Amazona aestiva, 3,0 (2,93-4,15) mm/5 s para Ara ararauna e 3,0 (2,55-4,90) mm/5 s para Caiman latirostris. Estes valores para o teste de meniscometria podem ser utilizados como parâmetros para estas espécies e podem ser úteis para diagnosticar e tratar distúrbios de produção lacrimal em aves e répteis.

Tear meniscometry test in wild animals / Teste de meniscometria lacrimal em animais silvestres

This study aimed to describe strip meniscometry test values for Amazona aestiva (turquoise-fronted amazon), Ara ararauna (blue-and-yellow macaw) and Caiman latirostris (broad-snouted caiman). Twenty-one Amazona aestiva and 11 Ara ararauna, healthy, adults of unknown sex, and 37 healthy adult Caiman latirostris (27 males and 10 females), were used in this investigation. All animals were manually restrained and I-Tear® Test strip was inserted for 5 s at the edge of the lower tear meniscus, without touching the eyelid or ocular surface, immediately followed by measurement (millimeter per 5 seconds). No differences were reported between left and right eyes for Amazona aestiva (P = 0.824), Ara ararauna (P = 0.262) and Caiman latirostris (P = 0.679). Median and confidence interval were 2.0 (2.25-3.56) mm/5s for Amazona aestiva, 3.0 (2.93-4.15) mm/5s for Ara ararauna and 3.0 (2.55-4.90) mm/5 s for Caiman latirostris. These values for strip meniscometry tear test can be used as parameters for each species and may be helpful for diagnosis and treatment of tear-production disorders in birds and reptiles.(AU)

O objetivo deste estudo foi descrever os valores do teste de meniscometria lacrimal para Amazona aestiva (papagaio-verdadeiro), Ara ararauna (arara-canindé) e Caiman latirostris (jacaré-de-papo-amarelo). Vinte e um Amazona aestiva, 11 Ara ararauna adultos, de sexo desconhecido, e 37 Caiman latirostris adultos (27 machos e 10 fêmeas), foram utilizados nesta investigação. Todos os animais foram contidos manualmente e uma tira do I-Tear® test foi inserida por 5 segundos na borda do menisco inferior, sem tocar a pálpebra ou a superfície ocular, e imediatamente mensurada (milímetro por 5 segundos). Não foram encontradas diferenças entre os olhos esquerdos e direitos para Amazona aestiva (P = 0,824), Ara ararauna (P = 0,262) e Caiman latirostris (P = 0,679). Mediana e intervalo de confiança foram de 2,0 (2,25-3,56) mm/5 s para Amazona aestiva, 3,0 (2,93-4,15) mm/5 s para Ara ararauna e 3,0 (2,55-4,90) mm/5 s para Caiman latirostris. Estes valores para o teste de meniscometria podem ser utilizados como parâmetros para estas espécies e podem ser úteis para diagnosticar e tratar distúrbios de produção lacrimal em aves e répteis.(AU)

Correction to: Photography-based method for assessing fluorescein clearance test in dogs.

The original article [1] contained an error whereby the respective legends of Figs. 2 and 3 were mistakenly interchanged. This error has now been amended.

Photography-based method for assessing fluorescein clearance test in dogs.

BACKGROUND: The fluorescein clearance test (FCT) provides insight into the tear film dynamics. The purpose of this study was to describe an inexpensive and practical method for assessing FCT in dogs, using photography and software analysis, and to assess the retention time of 1 vs. 2 eye drops on the canine ocular surface. METHODS: (i) In vivo - Eight healthy German Shepherd dogs were recruited. Following topical anesthesia with 0.5% proxymetacaine, each eye sequentially received (1 week apart) either 1 drop (35 µL) or 2 drops (70 µL) of 0.5% fluorescein. A Schirmer strip was inserted in the ventral conjunctival fornix for 10 s at the following times: each 10 min for 100 min, 24 h, 48 h and 72 h. (ii) In vitro - Schirmer strips were placed for 10 s in contact with microplate wells containing 1 or 2 drops of 0.5% fluorescein. In both experiments, the fluorescein-impregnated Schirmer strips were immediately imaged, and the area and intensity of fluorescein uptake were analyzed with ImageJ software. For the in vitro experiment, images were evaluated by the same examiner (repeatability) or two examiners (reproducibility). RESULTS: Photography-based FCT was easy to perform and showed high repeatability and reproducibility (coefficients of variation ≤2.75%). In vivo, the area and intensity of fluorescein uptake on Schirmer strips were significantly greater at 30 min and 40 min post- fluorescein instillation in the 2 drops vs. 1 drop groups (p ≤ 0.044). Compared to baseline, the residual fluorescein uptake on Schirmer strips was < 5% at 60 min and 90 min in the 1 drop and 2 drops groups, respectively. CONCLUSIONS: Photography-based FCT is a practical and reliable diagnostic tool with various clinical and research applications in veterinary medicine. Instillation of two drops provided greater amount and longer retention on the anesthetized canine ocular surface than a single drop. Fluorescein clearance time of a single drop in dolichocephalic dogs is 60 min.

Evaluation of lacrimal production, osmolarity, crystallization, proteomic profile, and biochemistry of capuchin monkeys' tear film.

BACKGROUND: To evaluate the composition and characteristics of capuchin monkey (CM) tear film. METHODS: Eleven CM (Sapajus sp.) was evaluated. Strip meniscometry test (SMT), osmolarity, and tear ferning test (TFT) (by Rolando and Masmali scales) were assessed. Tear protein profile (SDS-PAGE), and total protein, albumin, urea, glucose, and cholesterol levels in tear film and blood serum were screened. RESULTS: Median ± semi-interquatil range for SMT and osmolarity values were 8.0 ± 1.625 and 303.0 ± 9.875, respectively. TFT for Rolando was 2.0 ± 0.5, and Masmali was 2.0 ± 0.0. Monkeys's tear obtained type II and III for Rolando, and 1 and 2 for Masmali. Tear components showed protein bands among 23-217 kDa, and presence of albumin, urea, glucose, and cholesterol. CONCLUSIONS: The results of SMT, osmolarity, TFT, SDS-PAGE, and tear biochemistry may serve as a reference baseline for CM, and the data may serve as a basis for future experimental model evaluations.

Molecular diagnosis of Ehrlichia canis infection in dogs with uveitis / Diagnóstico molecular da infecção por Ehrlichia canis em cães com uveíte

Canine monocytic ehrlichiosis (CME) is an infectious disease caused by a gram-negative bacterium Ehrlichia canis that has a high global prevalence that leads to high rates of morbidity and mortality in dogs. Among the clinical changes, ophthalmic diseases can lead to permanent blindness and it can be an important clinical sign. The objective of this study was to perform nested polymerase chain reaction (PCR) to diagnose E. canis infection in dogs with bilateral uveitis from the Veterinary Hospital of the Santa Cruz State University. Blood samples were collected and DNA for the molecular diagnosis was extracted from 66 adult dogs of both genders and mixed breeds diagnosed with bilateral uveitis. Thirty-five (53%) dogs showed positive results and presented with iridocyclitis, posterior uveitis, panuveitis, or uveitis with secondary glaucoma. This study demonstrates that nested PCR is an important tool for the differential diagnosis of dogs with bilateral uveitis, as it provides evidence of the infectious agent in the animal.(AU)

Erliquiose Monocítica Canina (EMC) é uma doença infecciosa causada por uma bactéria gram negativa da espécie Ehrlichia canis, que possui alta prevalência mundial e conduz a elevadas taxas de morbidade e mortalidade em cães. Dentre as alterações clínicas, as oftalmopatias podem levar a cegueira permanente e é ser um importante sinal clínico. Objetivou-se realizar nested-PCR para diagnosticar infecção por E. canis em cães portadores de uveíte bilateral provenientes da casuística do Hospital Veterinário da Universidade Estadual de Santa Cruz. Sessenta e seis cães adultos, de ambos sexos e raças variadas, diagnosticados com uveíte bilateral foram submetidos a coleta de sangue e o DNA para realização do diagnóstico molecular foi extraído destas amostras. Foram positivos no teste 35 (53%) cães, que apresentaram iridociclite, uveíte posterior, panuveíte ou uveíte com glaucoma secundário. Esse estudo demonstra a nested-PCR como ferramenta importante no diagnóstico diferencial de cães com uveíte bilateral, por proporcionar a comprovação do agente infeccioso no organismo do animal.(AU)

Molecular diagnosis of Ehrlichia canis infection in dogs with uveitis / Diagnóstico molecular da infecção por Ehrlichia canis em cães com uveíte

Canine monocytic ehrlichiosis (CME) is an infectious disease caused by a gram-negative bacterium Ehrlichia canis that has a high global prevalence that leads to high rates of morbidity and mortality in dogs. Among the clinical changes, ophthalmic diseases can lead to permanent blindness and it can be an important clinical sign. The objective of this study was to perform nested polymerase chain reaction (PCR) to diagnose E. canis infection in dogs with bilateral uveitis from the Veterinary Hospital of the Santa Cruz State University. Blood samples were collected and DNA for the molecular diagnosis was extracted from 66 adult dogs of both genders and mixed breeds diagnosed with bilateral uveitis. Thirty-five (53%) dogs showed positive results and presented with iridocyclitis, posterior uveitis, panuveitis, or uveitis with secondary glaucoma. This study demonstrates that nested PCR is an important tool for the differential diagnosis of dogs with bilateral uveitis, as it provides evidence of the infectious agent in the animal.

Erliquiose Monocítica Canina (EMC) é uma doença infecciosa causada por uma bactéria gram negativa da espécie Ehrlichia canis, que possui alta prevalência mundial e conduz a elevadas taxas de morbidade e mortalidade em cães. Dentre as alterações clínicas, as oftalmopatias podem levar a cegueira permanente e é ser um importante sinal clínico. Objetivou-se realizar nested-PCR para diagnosticar infecção por E. canis em cães portadores de uveíte bilateral provenientes da casuística do Hospital Veterinário da Universidade Estadual de Santa Cruz. Sessenta e seis cães adultos, de ambos sexos e raças variadas, diagnosticados com uveíte bilateral foram submetidos a coleta de sangue e o DNA para realização do diagnóstico molecular foi extraído destas amostras. Foram positivos no teste 35 (53%) cães, que apresentaram iridociclite, uveíte posterior, panuveíte ou uveíte com glaucoma secundário. Esse estudo demonstra a nested-PCR como ferramenta importante no diagnóstico diferencial de cães com uveíte bilateral, por proporcionar a comprovação do agente infeccioso no organismo do animal.