A Cross-sectional Online Questionnaire-based Survey About the Use of Compounded Medications in Veterinary Ophthalmology.

The aims of this survey were to determine how veterinary ophthalmologists worldwide use compounded ophthalmic drugs to treat ocular diseases, define their attitudes regarding compounding pharmacies, and identify commonly dispensed veterinary ophthalmic formulations as well as the diseases for which those preparations are most often prescribed. Respondents voluntar i ly and anonymously completed a questionnaire that was sent to a total of 1014 veterinary ophthalmologists at universities, specialty colleges, and ophthalmology associations in 24 countries. One hundred thirty (12.83%) veterinary ophthalmologists replied. Of those, 87 (66.92%) had worked in  veterinary ophthalmology for more than 10 years. Ten to 30% of their total prescriptions were compounded ophthalmic drugs, the most common of which were tacrolimus and cyclosporine for the treatment of keratoconjunctivitis sicca. Reported advantages of treatment with a compound included the accessibility of preparations that were not commercially available and the ability to customize formulations; reported disadvantages included brief shelf life, delivery time, and cost.

Bartonella species in dromedaries and ruminants from Lower Shabelle and Benadir regions, Somalia.

BACKGROUND: Bartonellosis, caused by bacteria of the genus Bartonella, is a zoonotic disease with several mammalian reservoir hosts. In Somalia, a country heavily reliant on livestock, zoonotic diseases pose significant public health and economic challenges. To the best of our knowledge, no study has been performed aiming to verify the occurrence of Bartonella spp. in Somalia. This study investigated the occurrence and molecular characterization of Bartonella in dromedary (Camelus dromedarius, Linnaeus, 1758), cattle, sheep, and goats from Somalia. MATERIALS AND METHODS: 530 blood samples were collected from various animals (155 dromedary, 199 goat, 131 cattle, and 45 sheep) in Benadir and Lower Shabelle regions. DNA was extracted for molecular analysis, and a qPCR assay targeting the NADH dehydrogenase gamma subunit (nuoG) gene was used for Bartonella screening. Positive samples were also subjected to PCR assays targeting seven molecular markers including: nuoG, citrate synthase gene (gltA), RNA polymerase beta-subunit gene (rpoB), riboflavin synthase gene (ribC), 60 kDa heat-shock protein gene (groEL), cell division protein gene (ftsZ), and pap31 and qPCR targeting the 16-23S rRNA internal transcribed spacer (ITS) followed by Sanger sequencing, BLASTn and phylogenetic analysis. RESULTS: Out of 530 tested animals, 5.1% were positive for Bartonella spp. by the nuoG qPCR assay. Goats showed the highest Bartonella occurrence (17/199, 8.5%), followed by sheep (6/44, 6.8%), cattle (4/131, 3.1%), and dromedary (1/155, 1.9%). Goats, sheep, and cattle had higher odds of infection compared to dromedary. Among nuoG qPCR-positive samples, 11.1%, 14.8%, 11.1%, and 25.9% were positive in PCR assays based on nuoG, gltA, and pap31 genes, and in the qPCR based on the ITS region, respectively. On the other hand, nuoG qPCR-positive samples were negative in the PCR assays targeting the ribC, rpoB, ftsZ, and groEL genes. While Bartonella bovis sequences were detected in cattle (nuoG and ITS) and goats (gltA), Bartonella henselae ITS sequences were detected in dromedary, goat, and sheep. Phylogenetic analysis placed gltA Bartonella sequence from a goat in the same clade of B. bovis. CONCLUSION: The present study showed, for the first time, molecular evidence of Bartonella spp. in dromedary and ruminants from Somalia and B. henselae in sheep and goats globally. These findings contribute valuable insights into Bartonella spp. occurrence in Somali livestock, highlighting the need for comprehensive surveillance and control measures under the One Health approach.

Micropulse transscleral cyclophotocoagulation in canine glaucoma: A retrospective study evaluating sweep velocity.

OBJECTIVE: To report preliminary results of MP-TSCP in canine patients with glaucoma while evaluating sweep velocity, treatment efficacy, and safety utilizing standardized surgical parameters. ANIMAL STUDIED: Client-owned dogs that underwent MP-TSCP at a veterinary referral hospital. PROCEDURE: Medical records of 19 eyes (14 dogs) treated with MP-TSCP were reviewed. Eleven eyes were treated with a 10 s/sweep (s) velocity and 8 eyes with a 20 s/s. Laser duty cycle was set at 31.3%, mean laser energy was 2766 mW, and mean duration was 120 s per hemisphere. Minimum post-operative follow-up was 6 months. Outcomes of intraocular pressure (IOP), vision status, degree of antiglaucoma medications, and complications were evaluated. RESULTS: There was a significantly lower median (IQR) IOP (<25 mmHg) at each timepoint postoperatively compared to IOP pre-operatively (p < 0.05). The overall success rate for IOP control at 6 months was 84%, 100% for the 10 s/s and 62.5% for the 20 s/s. Vision was preserved in 10/11 (90%) eyes from the 10 s/s group and 4/8 (50%) eyes from the 20 s/s group at 6 months. Average number of glaucoma medications in the 10 s/s group decreased from 4.4 to 1.5 and from 3.3 to 2.5 in the 20 s/s group. Post-operative complications were mild and resolved within 1 week. CONCLUSIONS: MP-TSCP is a non-invasive treatment alternative for canine glaucoma. Employing higher energy levels rendered long-term IOP control while retaining a high safety profile with minimal postoperative complications. There was no statistical difference between a 10 s/s and 20 s/s surgical velocity, both leading to successful surgical outcomes up to 6 months post-operatively.

Supplemental vibrissal extensions as an alternative to improve the tactile sensitivity of blind dogs - a preliminary approach investigation.

OBJECTIVE: This preliminary study suggests a way to artificially extend vibrissae of blind dogs to assist ambulation and avoiding facial contact with obstacles. PROCEDURES: Fourteen irreversibly blind dogs had 5-6 mystacial vibrissae on each side of the face supplementally extended by attaching carefully chosen adult pig hairs to them and were subjected to a maze test before and after the procedure. In three of these dogs the test was repeated one more time after all the extensions had fallen off. Collision counts and course times with and without extensions were analyzed and compared. A p-value > 0.05 was considered significant. RESULTS: Median number of collisions was significantly higher post-extensions (5 IQR 2.25) and after extensions had fallen off (4 IQR 7.50) compared to pre-extensions (1 IQR 1), p = 0.021. Median times were significantly higher pre-extension (25.6 IQR 8.98) and after the extensions had fallen off, compared to the post-extension performance (22.8 IQR 8.55), p = 0.04. CONCLUSION: Vibrissae play an important role in the tactile perception of blind dogs, and our preliminary results suggest that extending this sensory organ possibly improves obstacle location and their quality of life.

A Molecular Phylogenetic Study of the Eye Fluke Philophthalmus lacrymosus (Trematoda: Philophthalmidae) Found in Larus dominicanus (Aves: Laridae) from Brazil.

PURPOSE: Species of the genus Philophthalmus are eye flukes with a complex taxonomy, which began to be improved with the help of molecular data only recently. However, most described species have never been placed into a phylogenetic context. In this study, eye flukes previously found on kelp gulls, Larus dominicanus, from Brazil and identified as Philophthalmus lacrymosus were subjected to molecular analysis. METHODS: For the molecular analyses, we analyzed parasites found in six infected gulls (one worm per bird) collected from different municipalities of the state of Santa Catarina, Brazil. We carried out the amplification and sequencing of the partial region of the 28S and cox1 genes and the data obtained were compared with sequences available to philophthalmid species and subjected to phylogenetic analysis. RESULTS: The isolates of P. lacrymosus from Brazil grouped in well-supported clades with five other species of Philophthalmus with sequences available for comparison. Interspecific divergences of 0.1-1.6% in 28S and 8.2-14.9% in cox1 were found in relation to other isolates of Philophthalmus spp. Two cox1 haplotypes differing in one nucleotide (0.1%) were found between the six eye flukes isolates in gulls from different localities. The Brazilian isolates grouped in a subclade with parasites identified as P. lacrymosus in Portugal; however, the molecular divergences found in cox1 (8.2-8.5%) strongly suggest that these isolates belong to different species. The phylogenetic trees obtained and the intergeneric divergences to species of the genera Cloacitrema and Parorchis did not support the validity of the genus Natterophthalmus, for which P. lacrymosus was proposed as the type species in the past. CONCLUSION: As P. lacrymosus was described from Brazil, we recommend that this name be applied to the South American isolates and that the Portuguese isolates be provisionally considered as Philophthalmus sp., a probable cryptic species. Moreover, data obtained supports the previous morphology-based synonymizing between Natterophthalmus and Philophthalmus. Considering our results and most of previous reports of P. lacrymosus in South America, we suggest this species presents a marine life cycle.

A comparison between neurological clinical signs, cerebrospinal fluid analysis, cross-sectional CNS imaging, and infectious disease testing in 168 dogs with infectious or immune-mediated meningoencephalomyelitis from Brazil.

This retrospective study evaluated canine patients with presumptively diagnosed meningoencephalomyelitis (ME) based on neurological clinical signs, cerebrospinal fluid (CSF) analysis, cross-sectional imaging, and infectious disease testing with a limited neurological-focused polymerase chain reaction (PCR) panel performed on blood and CSF. The first goal was to determine the proportion of dogs where the condition was caused by an infectious agent versus a probable immune-mediated etiology (i.e., meningoencephalomyelitis of unknown origin; MUO) in our geographic region. The secondary goals of this study were to examine and define associations between abnormal CSF test results and cross-sectional neuroimaging findings, in addition to defining the age and most common neurological clinical signs in each group of ME. A total of 168 dogs matched the inclusion criteria with magnetic resonance imaging (MRI) performed in 130 dogs and computed tomography (CT) performed in 38 dogs. Presumptive MUO was observed in 152/168 (90.5%) of dogs and infectious ME was identified in 16/168 (9.5%) of dogs (p < 0.0001). Canine distemper virus (CDV) was the most common cause of infectious ME in 10/16 dogs (62.5%). Of the total cases with a positive infectious disease result, 3/16 (18.7%) had normal CSF results and 13/16 (81.3%) had abnormal CSF results (p = 0.0078). MRI and CT abnormalities in the brain were detected in 74 and 39% of dogs with inflammatory CSF, respectively. MRI and CT abnormalities in the spinal cord were detected in 90 and 57% of dogs with inflammatory CSF results, respectively. Age was not significantly different between infectious ME and presumptive MUO groups (p = 0.15). Seizures were the most common clinical sign reported for both MUO (36.8% of cases) and infectious ME (31.2% of cases). In conclusion, presumptive MUO is significantly more common than infectious ME in this population of dogs. Furthermore, although normal CSF results were uncommon in dogs with infectious ME, this finding occurred in several patients (3/16), suggesting that infectious disease testing should be considered even in the face of normal CSF results. Finally, MRI was more sensitive than CT in the detection of abnormalities when dogs with ME had inflammatory CSF results but was not 100% sensitive, suggesting CSF analysis should be performed to rule out inflammation even when no abnormalities are detected on MRI or CT.

SELECTED OPHTHALMIC TESTS AND OCULAR DIMENSIONS IN RELATION TO ACTIVITY PATTERN IN THREE NEOTROPICAL NONHUMAN PRIMATES: BLACK-TUFTED MARMOSET (CALLITHRIX PENICILLATA), GUIANAN SQUIRREL MONKEY (SAIMIRI SCIUREUS), AND AZARA'S NIGHT MONKEY (AOTUS AZARAE INFULATUS).

The goals of this study were to compare ocular morphology, determine the reference intervals of selected ophthalmic tests, ocular measurements, intraocular pressure, and tear production, and to establish possible relationships in the visual ecology of three different Neotropical nonhuman primates (NHP). Nineteen black-tufted marmosets (Callithrix penicillate), 24 Guianan squirrel monkeys (Saimiri sciureus), and 24 night monkeys (Aotus azarae infulatus) were included in the study. Schirmer tear test, ocular dimensions, ocular ultrasonography, intraocular pressure, central corneal thickness, and corneal touch threshold were determined. The ratio of the average corneal diameters and axial diameters (CD/AGL) were established. No significant difference was noted between males and females, nor left and right eyes, for all three species for all measurements (P > 0.05). CD/AGL ratio was significantly higher (P < 0.0001) in night monkeys (a nocturnal species) as compared to black-tufted marmoset and Guianan squirrel monkeys (two diurnal species). The reference intervals will aid veterinary ophthalmologists to more accurately diagnose pathological changes in the eyes of these species. In addition, ocular dimension comparison will allow other NHP species to be evaluated and examined in relationship to behavioral traits (nocturnal versus diurnal).

Preliminary characterization of a novel form of progressive retinal atrophy in the German Spitz dog associated with a frameshift mutation in GUCY2D.

OBJECTIVE: To describe the clinical, preliminary electroretinographic and optical coherence tomography features of a newly identified form of progressive retinal atrophy (PRA) in German Spitzes, and identify the causal gene mutation. ANIMALS: Thirty-three client-owned German Spitz dogs were included. PROCEDURES: All animals underwent a full ophthalmic examination, including vision testing. In addition, fundus photography, ERG, and OCT were performed. A DNA-marker-based association analysis was performed to screen potential candidate genes and the whole genomes of four animals were sequenced. RESULTS: Initial fundus changes were pale papilla and mild vascular attenuation. Oscillatory nystagmus was noted in 14 of 16 clinically affected puppies. Vision was impaired under both scotopic and photopic conditions. Rod-mediated ERGs were unrecordable in all affected dogs tested, reduced cone-mediated responses were present in one animal at 3 months of age and unrecordable in the other affected animals tested. Multiple small retinal bullae were observed in three clinically affected animals (two with confirmed genetic diagnosis). OCT showed that despite loss of function, retinal structure was initially well-preserved, although a slight retinal thinning developed in older animals with the ventral retina being more severely affected. Pedigree analysis supported an autosomal recessive inheritance. A mutation was identified in GUCY2D, which segregated with the disease (NM_001003207.1:c.1598_1599insT; p.(Ser534GlufsTer20)). Human subjects with GUCY2D mutations typically show an initial disconnect between loss of function and loss of structure, a feature recapitulated in the affected dogs in this study. CONCLUSION: We identified early-onset PRA in the German Spitz associated with a frameshift mutation in GUCY2D.

The development of the normal Schirmer tear test results during the dog's first year of life.

OBJECTIVE: To evaluate the relationship between lacrimation and age in a homogeneous group of healthy beagle dogs during the first year of life. MATERIALS AND METHODS: Schirmer tear test I (STT I) was performed at an interval of 12-15 days in both eyes of 16 clinically healthy beagle dogs (eight males and eight females) from 94 to 361 days of age. Three different quadratic polynomial regression equations were estimated for the variation in lacrimation: (1) for the entire period (19 observations), (2) for observations 1-4 (days 94-136), and (3) for observations 5-19 (days 150-361). RESULTS: By fitting quadratic regression equations to different phases of tear production during the dog's first year of life, it was possible to see that with each day of life, lacrimation increased 0.08 times (8%). From days 94 to 136, however, lacrimal production fell 1.1 times with each day of life. From day 150 to 361, production increased by 0.02 (2%) each day of life. In addition, there was a positive significant and moderate linear correlation between body weight and STT I values (p = .01). CONCLUSION: In dogs, during the first year of life, STT I data distribution is parabolic in shape. Age significantly affected tear production. STT I decreased at approximately 108-121 days of age and increased thereafter. Body weight was a significant factor for STT I in young dogs. The establishment of this normal pattern of lacrimation is important for both clinical practitioners and laboratory studies.

Comparison of agarose gel and capillary zone electrophoresis methods with preliminary reference interval generation using capillary zone electrophoresis in bald eagles (Haliaeetus leucocephalus).

BACKGROUND: Electrophoresis can be used to aid in the diagnosis of different diseases in avian species. Capillary zone electrophoresis (CZE) is an automated method that is proposed to be superior to the dye methods used in agarose gel electrophoresis (AGE). However, reference intervals (RIs) for CZE in avian species and comparison studies between electrophoretic methods are lacking. OBJECTIVES: The goals of the current study were to compare AGE and CZE methods and determine reference intervals for CZE using plasma from bald eagles (Haliaeetus leucocephalus). METHODS: Heparinized plasma samples from 44 bald eagles (mean age 18.7 years) under managed care were examined by AGE and CZE. Method comparison analyses were completed, as well as the generation of preliminary RIs using the CZE method and ASVCP guidelines. RESULTS: Passing-Bablok regression and Bland-Altman plots demonstrate that these methods are not equivalent. All fractions were significantly correlated between the methods except for alpha 1 globulin. Inter-assay and intra-assay CVs for CZE were lower or comparable to AGE and ranged from 2.4% to 15.4%, and 0.8% to 8.3%, respectively. CZE resolved more fractions than AGE with two fractions observed in the beta and gamma region vs one for AGE in each region. CONCLUSIONS: CZE provided improved resolution and reproducibility for the quantitation of protein fractions in the bald eagle. Although most fraction results correlated with AGE, these methods were judged as not equivalent, necessitating method-specific Rls. Reference intervals generated from a limited number of mostly aged individuals under managed care should be considered preliminary; additional studies will aid in the production of more robust intervals.

Ocular lesions in cats diagnosed with systemic sporotrichosis.

OBJECTIVES: To describe the most common ocular lesions and demonstrate the frequency of ophthalmic involvement in a group of cats with systemic sporotrichosis. ANIMALS STUDIED: Two hundred seventy-four cats diagnosed with systemic sporotrichosis. The inclusion criteria included previous positive cytopathological examination, histopathological examination, or fungal culture. PROCEDURES: In a prospective case-control study, 274 cats diagnosed with systemic sporotrichosis underwent ophthalmic evaluation and received treatment for systemic sporotrichosis. Of these animals, 63 had ocular abnormalities which were recorded, and conjunctivitis was scored from 0 to 5. Diagnostic techniques utilized included fungal culture, as well as cytopathological (10 eyes; 10 cats), and histopathological examination of the palpebral conjunctiva and eyes (2 eyes). RESULTS: Cytopathological and histopathological examination of the conjunctiva, as well as fungal culture, proved to be important tests for the detection of Sporothrix sp. Five cats without the evidence of ophthalmic abnormalities also had a positive fungal culture. The identified ocular lesions in animals with systemic sporotrichosis included increased serous discharge (79 eyes; 53 cats), blepharoconjunctivitis (33 eyes; 25 cats), conjunctivitis (39 eyes, 20 cats), blepharitis (9 eyes; 8 cats), uveitis (5 eyes; 3 cats), and Florida keratopathy-like lesions (2 eyes; 1 cat). CONCLUSION: Sporotrichosis should be considered a differential diagnosis for conjunctivitis and blepharoconjunctivitis, especially in endemic areas. Fungal culture and cytopathology of ocular discharge and histopathological examinations of the conjunctiva are important for the diagnosis of ophthalmic sporotrichosis, although not all cats underwent laboratory testing in this study. Ocular discharge could be a source of contagion transmission.

'Candidatus Mycoplasma Haemoalbiventris' and Tick-Borne Pathogens in Black-Eared Opossum (Didelphis aurita) from Southeastern Brazil.

The black-eared opossum (Didelphis aurita) is a South American synanthropic marsupial. The presence of opossums in domestic spaces is relevant in the One-Health context since they are hosts of pathogens and ectoparasites that may affect the health of domestic animals and humans. In this study, we aim to determine the occurrence of hemoplasmas and selected tick-borne pathogens in free-ranging black-eared opossums, along with their molecular characterization, hematological and biochemical evaluation and factors associated with infection, in the municipality of Viçosa, State of Minas Gerais, southeastern Brazil. Thirty black-eared opossums were trapped between March 2021 and June 2022. Ectoparasites were collected. Hematological and biochemical analyses were performed. DNA from EDTA-blood samples were analyzed by PCR and qPCR assays. By molecular analyses, 'Candidatus Mycoplasma haemoalbiventris' was the most prevalent hemoparasite (73.3%), followed by Hepatozoon sp. (22.2%). Significant differences were observed in the number of platelets, and in the concentration of protein and globulins in the animals infected by 'Ca. M. haemoalbiventris' when compared with the negative group. This is the first report of 'Ca. M. haemoalbiventris' infection in D. aurita.

Ocular disease caused by the trematode Philophthalmus lachrymosus in free-living kelp gulls (Larus dominicanus) of Brazil.

In this study, clinical, parasitological and histopathological findings of thirteen kelp gulls (Larus dominicanus) found infected with eyeflukes in Brazil are presented. Parasites detected in the ventral conjunctival fornix were identified as Philophthalmus lachrymosus [mean intensity of infection: 16 (5-36) worms/bird]. Eleven birds (85%) presented signs of systemic disease, such as emaciation, dehydration and depressed consciousness. Conjunctival hyperemia was observed in 22 eyes (85%). Keratitis, corneal ulcers, corneal abscess and chemosis were also detected in some eyes (4-8%). Histopathologic lesions, likely due to the parasite attachment to the conjunctiva, were found in the eyes of one infected bird that died from unrelated causes. Philophthalmosis by P. lachrymosus is here reported as a clinically relevant eye disease in kelp gulls.

First Report of 'Candidatus Mycoplasma haematomacacae' in Laboratory-Kept Rhesus Monkeys (Macaca mulatta) Maintained in Rio de Janeiro, Brazil.

Health monitoring programs in animals used as experimental models are essential, since only disease-free subjects are considered suitable for research purposes. In laboratory-kept animals, hemoplasmas have been described as an important confounding variable. Different hemoplasma species have been detected infecting non-human primates (NHP) from Brazil. However, the occurrence of hemoplasma species in laboratory-kept NHP in Brazil has not-yet been assessed. Accordingly, this study aimed (i) to screen laboratory-kept rhesus monkeys for hemoplasmas, (ii) to verify if any of the hemoplasma-positive animals demonstrate hematological abnormalities, and (iii) to assess the genotype diversity of hemoplasma species in NHP from Brazil. Five out of eight (62.5%; 95% CI: 3.05-8.63) rhesus monkeys tested positive for hemotropic Mycoplasma spp. by PCR. Sequencing, phylogenetic, distance, and genotype diversity analyses of partial 16S rRNA gene demonstrate that rhesus monkeys were infected by 'Candidatus Mycoplasma haematomacacae' (formerly 'Candidatus Mycoplasma haemomacaque'). Assessments of partial 16S rRNA diversity of hemoplasma species in NHP suggest that at least four genetically diverse groups may occur in Brazil. Although no hematological abnormalities were demonstrated in rhesus monkeys evaluated herein, future studies are needed to elucidate the influence of 'Ca. M. haematomacacae' as a confounding variable on research studies.

Canine and feline uveal melanocytic tumours: Histologic and immunohistochemical characteristics of 32 cases.

OBJECTIVE: Gross, histopathological, and immunohistochemical characteristics of uveal melanocytic neoplasms in dogs and cats were investigated. SAMPLES: Thirty-two enucleated globes with uveal melanocytic neoplasms, 27 from dogs and 5 from cats, were examined. PROCEDURES: Morphological characteristics of uveal melanocytic neoplasms in dogs and cats were evaluated with anti-PNL2, anti-Melan-A, anti-Ki-67, anti-caspase-3, and anti-BAP1 immunomarkers. Statistical analysis was performed to compare canine melanocytomas and melanomas. RESULTS: The 32 uveal neoplasms were classified as melanocytomas (19/27 in dogs) or melanomas (8/27 in dogs, 5/5 in cats). Most tumours (84%) were located in the anterior uvea. Neoplastic cells were classified as epithelioid, spindle-shaped, mixed, or special type (balloon and signet ring cells). The percentage of cells with melanin, melanin concentration within cells, anisocytosis and anisokaryosis, mitotic count, lymphocytic inflammation, necrosis, vascular invasion, and glaucoma were also characterized. Anisocytosis, percentage of neoplastic cells with melanin, mitotic count, and indices (proliferation and apoptotic) varied significantly between canine uveal melanomas and melanocytomas; in general, melanomas had greater cell variability, were less pigmented, and had a higher mitotic count. The melanocytic origin of the neoplasms was confirmed by positive anti-PNL2 immunolabelling (29/32) and positive anti-Melan-A immunolabelling (3/32). In canine uveal melanomas, anisocytosis and anisokaryosis correlated with less pigmentation and minimal pigmentation correlated with a high percentage of immunolabelling for caspase-3. CONCLUSIONS: Uveal melanocytomas were more common in dogs, and uveal melanomas were more frequent in cats. Anisocytosis, percentage of neoplastic cells with melanin, and mitotic count are important histologic characteristics of malignancy to evaluate in uveal melanocytic neoplasms. The proliferation and apoptotic indices are relevant when comparing malignant tumours with benign tumours.

Potential of Human Neural Precursor Cells in Diabetic Retinopathy Therapeutics - Preclinical Model.

PURPOSE: This study aimed to evaluate a cell therapy strategy with human neural precursor cells (hNPCs) to treat diabetic retinopathy (DR) in Wistar rats induced to diabetes by injecting streptozotocin. MATERIAL AND METHODS: The Wharton's jelly mesenchymal stem cells (WJ-MSCs) were isolated, expanded, and seeded onto a biopolymer substrate to develop neurospheres and obtain the hNPCs. The animals were divided into three groups: non-diabetic (ND) n = four, diabetic without treatment (DM) n = nine, and diabetic with cell therapy (DM + hNPCs) n = nine. After 8 weeks of diabetes induction and DR characteristics installed, intravitreal injection of hNPCs (1 × 106 cell/µL) was performed in the DM + hNPCs group. Optical Coherence Tomography (OCT) and Electroretinography (ERG) evaluations were conducted before and during diabetes and after cell therapy. Four weeks posttreatment, histopathological and immunohistochemistry analyses were performed. RESULTS: The repair of the retinal structures in the treated group (DM + hNPCs) was observed by increased thickness of neuroretinal layers, especially in the ganglion cell and photoreceptor layers, higher ERG oscillatory potentials (OPs) amplitudes, and transplanted hNPCs integration into the Retinal Pigment Epithelium. CONCLUSIONS: The results indicate that hNPCs reduced DR progression by a neuroprotective effect and promoted retinal repair, making them potential candidates for regenerating the neuroretinal tissue.

Development of retinal bullae in dogs with progressive retinal atrophy.

OBJECTIVE: To report the development of focal bullous retinal detachments (bullae) in dogs with different forms of progressive retinal atrophy (PRA). PROCEDURES: Dogs with three distinct forms of PRA (PRA-affected Whippets, German Spitzes and CNGB1-mutant Papillon crosses) were examined by indirect ophthalmoscopy and spectral domain optical coherence tomography (SD-OCT). Retinal bullae were monitored over time. One CNGB1-mutant dog was treated with gene augmentation therapy. The canine BEST1 gene coding region and flanking intronic sequence was sequenced in at least one affected dog of each breed. RESULTS: Multiple focal bullous retinal detachments (bullae) were identified in PRA-affected dogs of all three types. They developed in 4 of 5 PRA-affected Whippets, 3 of 8 PRA-affected Germans Spitzes and 15 of 20 CNGB1-mutant dogs. The bullae appeared prior to marked retinal degeneration and became less apparent as retinal degeneration progressed. Bullae were not seen in any heterozygous animals of any of the types of PRA. Screening of the coding region and flanking intronic regions of the canine BEST1 gene failed to reveal any associated pathogenic variants. Retinal gene augmentation therapy in one of the CNGB1-mutant dogs appeared to prevent formation of bullae. CONCLUSIONS: Retinal bullae were identified in dogs with three distinct forms of progressive retinal atrophy. The lesions develop prior to retinal thinning. This clinical change should be monitored for in dogs with PRA.

Assessment of tear film osmolarity using the IPen® Vet osmometer in Pug and Shih-Tzu dogs with and without keratoconjunctivitis sicca.

OBJECTIVE: To establish tear film osmolarity (TFO) values in Pugs and Shih-Tzus, with and without keratoconjunctivitis sicca (KCS). ANIMALS STUDIED: A total of 82 adult dogs were evaluated. PROCEDURE: The inclusion criteria for the healthy group was a Schirmer tear test (STT-1) ≥15 mm/min with no clinical signs of KCS, whereas those with KCS had clinical signs and a STT-1 ≤10 mm/min. All animals underwent complete ophthalmological evaluation prior to STT-1 and TFO. Student's t tests were used to compare STT-1 and TFO in KCS and healthy eyes as well as possible differences in TFO between breeds. In addition, a linear regression to model the relationship between the two variables (STT-1 and TFO) was performed. A P-value ≤ 0.05 was considered statistically significant. RESULTS: STT-1 results were significantly lower (p = 0.0001) in the KCS group (4.46 ± 1.74) compared with the control group (18.80 ± 2.02). Mean TFO results were significantly higher in the KCS group (353.02 ± 16.58 mOsm/L) (p < 0.0001) compared with the control group (315.27 ± 6.15 mOsm/L). The formula Y = 365.059-2.625 * X significantly predicts (p < 0.001) the value of the variable Y (TFO mOsm/L) as a function of the variable X (STT-1 mm/min), with a coefficient of determination of 0.71. CONCLUSIONS: The results revealed differences in TFO and STT-1 between KCS and healthy dogs. Additionally, STT-1 and TFO values were correlated with the aim to use STT-1 values to predict TFO values in brachycephalic breeds.

Association between spinal cord compression ratio in magnetic resonance imaging, initial neurological status, and recovery after ventral slot in 57 dogs with cervical disc extrusion.

This retrospective, unblinded, single rater study evaluated images obtained from magnetic resonance imaging (MRI) of dogs with cervical intervertebral disc extrusion before being submitted to ventral slot decompression (VSD). Dogs were re-evaluated systematically at 10 and 30 days after VSD. The objectives of this study were to investigate the associations between the following parameters: (1) The maximal spinal cord compression ratio (SCCR) as seen on transverse MRI and pre-surgical neurological status (NS) grade; we hypothesized that dogs with greater SCCR will have worse pre-surgical NS grade at presentation; (2) Pre-surgical NS grade and postoperative recovery; we hypothesized that worse pre-surgical NS grade will be associated with longer postoperative recovery time; (3) SCCR and postoperative recovery; we hypothesized that dogs with higher SCCR will have longer recovery time; (4) Location of extrusion (cranial vs. caudal) and initial NS grade and outcomes; we hypothesized that caudal cervical extrusion will have worse NS grade and longer time to recovery; (5) Longitudinal extension of ventral CSF signal loss on HASTE pulse sequence and NS grade and time to recovery; we hypothesized that dogs with longer HASTE CSF attenuation will have higher NS grade and longer time to recovery. There was no significant association between SCCR and NS grade, suggesting that this relationship in the cervical region is similar to what is observed in the thoracolumbar region, rejecting our first hypothesis. There was a significant difference between ambulatory tetraparesis dogs versus non-ambulatory tetraparesis dogs regarding complete recovery at 10 days: dogs with NS grade 1, 2, or 3 overall recovered faster than dogs with NS grade 4. However, there was no significant difference between these groups regarding complete recovery at 30 days, thereby accepting our second hypothesis at 10 days and rejecting it at 30 days. There was no correlation between SCCR and recovery time, rejecting our third hypothesis. Caudal cervical extrusion did not show higher NS grade or longer recovery time than cranial extrusion, rejecting our fourth hypothesis. CSF attenuation length ratio on HASTE images was not significantly correlated with NS grade but weakly correlate with post-surgical recovery time, partially accepting our fifth hypothesis.

Subconjunctival use of allogeneic mesenchymal stem cells to treat chronic superficial keratitis in German shepherd dogs: Pilot study.

Background: Chronic superficial keratitis (CSK) is an ocular condition in dogs characterized by corneal opacification leading to visual function impairment. Control of this chronic condition requires the use of topical immunomodulators or corticosteroids daily. Regenerative medicine has shown promising results in several fields of medicine. Aim: The aim of this study was to evaluate the clinical effect of allogeneic mesenchymal stem cells (MSCs) of adipose tissue applied via subconjunctival in dogs with CSK. Methods: A series of cases of eight dogs diagnosed with CSK were divided into two groups, four dogs each; the conventional treatment group received prednisolone 1% as topical eye drops and the experimental group (EG) received allogeneic MSCs transplantation. The dogs had not previously been treated for CSK. Systemic and ophthalmologic examinations were performed to exclude other abnormalities. An administered amount of MSC (1 × 106 cells each time) was injected via subconjunctival in the peri-limbal region at 0 and 30 days. The animals were followed for 110 days for clinical evaluation, and, at the same time, the images of the corneal abnormalities were obtained and analyzed in the ImageJ software. The statistical analysis was performed in the GrandPrism 7.0 software. Results: Initial and final images revealed that areas with neovascularization, inflammatory infiltrate, and opacity regressed in most eyes in both groups (7/8 eyes in each group) at the end of the 110 days, p = 0.0391 and p = 0.0078 respectively, but this response was minor in the EG comparing to conventional group (CG) (p = 0.026). No local or systemic side effects were observed. Conclusions: Despite the small melioration, MSCs treatment suggests clinical improvement in patients with CSK after 110 days without any local or systemic side effects. However, the improvement achieved was significantly less than the observed within CG. Further studies still are needed to evaluate the use and benefits of stem cells as an adjunct treatment for CSK.