Differential Viral-Host Immune Interactions Associated with Oseltamivir-Resistant H275Y and Wild-Type H1N1 A(pdm09) Influenza Virus Pathogenicity.

Oseltamivir is a common therapy against influenza A virus (IAV) infections. The acquisition of oseltamivir resistance (OR) mutations, such as H275Y, hampers viral fitness. However, OR H1N1 viruses have demonstrated the ability to spread throughout different populations. The objective of this work was to compare the fitness of two strains of OR (R6 and R7) containing the H275Y mutation, and a wild-type (F) pandemic influenza A (H1N1) 2009 (pdm09) virus both in vitro and in vivo in mice and to select one OR strain for a comparison with F in ferrets. R6 showed faster replication and pathogenicity than R7 in vitro and in mice. Subsequently, R6 was selected for the fitness comparison with the F strain in ferrets. Ferrets infected with the F virus showed more severe clinical signs, histopathological lung lesions, and viral quantification when compared to OR R6-infected animals. More importantly, differential viral kinetics correlated with differential pro-inflammatory host immune responses in the lungs of infected ferrets, where OR-infected animals developed a protective higher expression of type I IFN and Retinoid acid Inducible Gene I (RIG-I) genes early after infection, resulting in the development of milder disease. These results suggest the presence of early specific viral-host immune interactions relevant in the development of influenza-associated lung pathology.

Description and comparison of excretory urography performed during radiography and computed tomography for evaluation of the urinary system in healthy New Zealand White rabbits (Oryctolagus cuniculus).

OBJECTIVE To evaluate the usefulness of excretory urography performed during radiography (REU) and CT (CTEU) in healthy rabbits, determine timings of urogram phases, and compare sensitivities of REU and CTEU for detection of these phases. ANIMALS 13 New Zealand White rabbits (Oryctolagus cuniculus). PROCEDURES Rabbits were screened for signs of systemic and urinary tract disease. An REU examination of each was performed, followed ≥ 5 days later by a CTEU examination. Contrast images from each modality were evaluated for quality of opacification and intervals between initiation of contrast medium administration and detection of various urogram phases. RESULTS Excretory urograms of excellent diagnostic quality were achieved with both imaging modalities. For all rabbits, the nephrographic phase of the urogram appeared in the first postcontrast REU image (obtained between 34 and 40 seconds after initiation of contrast medium administration) and at a median interval of 20 seconds in CTEU images. The pyelographic phase began at a median interval of 1.63 minutes with both imaging modalities. Contrast medium was visible within the urinary bladder at a median interval of 2.20 minutes. Median interval to the point at which the nephrogram and pyelogram were no longer visible in REU images was 8 hours and 2.67 hours, respectively. The CTEU technique was better than the REU technique for evaluating renal parenchyma. CONCLUSIONS AND CLINICAL RELEVANCE Findings suggested that REU and, particularly, CTEU may be valuable tools for the diagnosis of renal and urinary tract disease in rabbits; however, additional evaluation in diseased rabbits is required.

IMAGING DIAGNOSIS-RADIOGRAPHY AND ULTRASONOGRAPHY OF BILATERAL CONGENITAL URETEROVESICAL JUNCTION STENOSIS CAUSING HYDRONEPHROSIS AND HYDROURETER IN A FERRET (MUSTELA PUTORIUS FURO).

An 8-month-old neutered male ferret was presented for vaccination and preventive treatment for adrenal gland disease. Abdominal ultrasound revealed severe bilateral hydronephrosis and hydroureter. Excretory urography and negative contrast cystography showed bilateral ureterovesical junction stenosis. Positive retrograde cystography demonstrated right ureteral reflux. A subcutaneous ureteral bypass was placed in the left kidney; however, it had to be removed 3 months later due to an obstruction and persistent urinary tract infection. Bilateral ureteroneocystotomy was performed. Although congenital strictures are uncommon in veterinary patients, they should be included in the differential diagnosis in ferrets presenting with hydronephrosis and hydroureter of undetermined cause.

Involvement of the different lung compartments in the pathogenesis of pH1N1 influenza virus infection in ferrets.

Severe cases after pH1N1 infection are consequence of interstitial pneumonia triggered by alveolar viral replication and an exacerbated host immune response, characterized by the up-regulation of pro-inflammatory cytokines and the influx of inflammatory leukocytes to the lungs. Different lung cell populations have been suggested as culprits in the unregulated innate immune responses observed in these cases. This study aims to clarify this question by studying the different induction of innate immune molecules by the distinct lung anatomic compartments (vascular, alveolar and bronchiolar) of ferrets intratracheally infected with a human pH1N1 viral isolate, by means of laser microdissection techniques. The obtained results were then analysed in relation to viral quantification in the different anatomic areas and the histopathological lesions observed. More severe lung lesions were observed at 24 h post infection (hpi) correlating with viral antigen detection in bronchiolar and alveolar epithelial cells. However, high levels of viral RNA were detected in all anatomic compartments throughout infection. Bronchiolar areas were the first source of IFN-α and most pro-inflammatory cytokines, through the activation of RIG-I. In contrast, vascular areas contributed with the highest induction of CCL2 and other pro-inflammatory cytokines, through the activation of TLR3.

Hyperplastic stomatitis and esophagitis in a tortoise (Testudo graeca) associated with an adenovirus infection.

A 2-year-old female, spur-thighed tortoise (Testudo graeca) was presented with poor body condition (1/5) and weakness. Fecal analysis revealed large numbers of oxyurid-like eggs, and radiographs were compatible with gastrointestinal obstruction. Despite supportive medical treatment, the animal died. At gross examination, an intestinal obstruction was confirmed. Histopathology revealed severe hyperplastic esophagitis and stomatitis with marked epithelial cytomegaly and enormous basophilic intranuclear inclusion bodies. Electron microscopy examination revealed a large number of 60-80 nm, nonenveloped, icosahedral virions arranged in crystalline arrays within nuclear inclusions of esophageal epithelial cells, morphologically compatible with adenovirus-like particles. PCR for virus identification was performed with DNA extracted from formalin-fixed, paraffin-embedded tissues. A nested, consensus pan-adenovirus PCR and sequencing analysis showed a novel adenovirus. According to phylogenetic calculations, it clustered to genus Atadenovirus in contrast with all other chelonian adenoviruses described to date. The present report details the pathologic findings associated with an adenovirus infection restricted to the upper digestive tract.

Heterogeneous pathological outcomes after experimental pH1N1 influenza infection in ferrets correlate with viral replication and host immune responses in the lung.

The swine-origin pandemic (p) H1N1 influenza A virus causes mild upper-respiratory tract disease in most human patients. However, some patients developed severe lower-respiratory tract infections with fatal consequences, and the cause of these infections remain unknown. Recently, it has been suggested that different populations have different degrees of susceptibility to pH1N1 strains due to host genetic variations that are associated with inappropriate immune responses against viral genetic characteristics. Here, we tested whether the pathologic patterns of influenza strains that produce different disease outcomes in humans could be reproduced in a ferret model. Our results revealed that the severities of infection did not correspond to particular viral isolate and were not associated with the clinical phenotypes of the corresponding patients. Severe pathological outcomes were associated with higher viral replication, especially in alveolar areas, and with an exacerbated innate cellular immune response that was characterised by substantial phagocytic and cytotoxic cell migration into the lungs. Moreover, detrimental innate cellular responses were linked to the up-regulation of several proinflammatory cytokines and chemokines and the down-regulation of IFNα in the lungs. Additionally, severe lung lesions were associated with greater up-regulations of pro-apoptotic markers and higher levels of apoptotic neutrophils and macrophages. In conclusion, this study confirmed that the clinicopathological outcomes of pH1N1 infection in ferrets were not only due to viral replication abilities but also depended on the hosts' capacities to mount efficient immune responses to control viral infection of the lung.

Congenital Peripheral Vestibular Syndrome in a Domestic Ferret (Mustela putorius furo).

A 3-month-old intact female ferret (Mustela putorius furo) was presented with a 2-month history of ataxia. On physical examination, the ferret had difficulty standing upright. During the neurologic examination, the patient had a left head tilt and positional strabismus, circled to the left, and was ataxic. Results of the complete blood count were consistent with a mild normocytic normochromic anemia. Initial treatment was supportive. Magnetic resonance imaging was performed and revealed an asymmetry of the inner ears. A brainstem auditory evoked response test was also performed. History, clinical signs, and diagnostic test results indicated that the ferret was suffering from congenital peripheral vestibular syndrome and left-sided deafness. Congenital disease should be considered in the differential diagnosis of young ferrets with peripheral vestibular syndrome. Supportive care and physiotherapy can improve balance and motor function, leading to an acceptable quality of life.

Immunohistochemical Study of Aquaporins in an African Grey Parrot (Psittacus erithacus) With Hydrocephalus.

A 5-month-old African grey parrot (Psittacus erithacus) was examined after 3 weeks of weakness, ataxia, mental depression, and seizures. Results of a complete blood cell count and plasma biochemical analysis were unremarkable. Magnetic resonance imaging revealed a severe bilateral hydrocephalus. The bird failed to improve with supportive care, and the owner requested euthanasia. Necropsy findings were severe bilateral hydrocephalus with no evidence of cerebrospinal fluid obstruction. Histologic examination of the brain revealed microspongiosis, edema, gliosis, and neuronal chromatolysis of surrounding periventricular tissue. Aquaporins (AQP) and astrocytes were examined to elucidate the participation of these water channel proteins and glial cells in the pathophysiology and resolution of hydrocephalus. Results showed AQP4 and glial fibrillary acidic protein were overexpressed, especially near the ventricles, but expression of AQP1 was decreased. This is the first report, to our knowledge, of AQP immunolabeling in hydrocephalus in avain species.

Lateral approach to nephrotomy in the management of unilateral renal calculi in a rabbit (Oryctolagus cuniculus).

CASE DESCRIPTION: A 5-year-old 2.23-kg (4.91-lb) spayed female Lop rabbit (Oryctolagus cuniculus) was evaluated because of a history of lethargy, polyuria, and polydipsia of 1 month's duration. The referring veterinarian made a presumptive diagnosis of hypercalcemia of unknown etiology on the basis of a plasma total calcium concentration of 14.0 mg/dL. CLINICAL FINDINGS: Physical examination findings were unremarkable. Hematologic and plasma biochemical analysis revealed nonregenerative, normocytic, normochromic anemia (PCV, 28%) with heteropenia (23%; 1,334 heterophils/mL) and a total plasma calcium concentration of 16 mg/dL. Radiographic and ultrasonographic examination revealed radiodense and hyperechoic structures in both renal pelves and right unilateral nephrolithiasis. TREATMENT AND OUTCOME: Nephrotomy was performed via laparotomy with a right lateral approach. A nephrolith composed of calcium carbonate and carbonate apatite was removed from the right renal pelvis. The rabbit's plasma tested positive for antibodies against Encephalitozoon cuniculi by means of an indirect immunofluorescence assay, and fenbendazole (20 mg/kg [9.1 mg/lb], p.o., q 24 h) was administered for 28 days. Although plasma calcium concentration was not substantially decreased after treatment (values were 14.1 and 15.1 mg/dL 1 week and 3 months after surgery, respectively), the patient did not have clinical signs of hypercalcemia and died of unrelated causes approximately 1 year after surgery. CLINICAL RELEVANCE: To the authors' knowledge, this is the first report of a nephrotomy performed in a rabbit via a lateral abdominal approach. This approach allowed easy location of the kidney and also reduced manipulation, thus decreasing the risk of damaging the abdominal viscera and potentially decreasing postoperative pain.

Cryptococcus gattii chorioretinitis in a ferret.

Bilateral exudative chorioretinitis was diagnosed in an 18-month-old male neutered ferret (Mustela putorius furo) with a generalized Cryptococcus gattii infection confirmed by PCR. The animal was referred to the Ophthalmology Service of the Autonomous University of Barcelona (VTH-UAB) for acute onset blindness. Complete ophthalmic examination revealed absent menace response and dazzle reflex in both eyes (OU), as well as subretinal edema located in the tapetal fundus. At that time, the clinical ophthalmologic diagnosis was bilateral exudative chorioretinitis. Treatment with prednisone (0.5 mg/kg PO q24 h) was instituted in addition to the ongoing treatment with fluconazole (10 mg/kg PO q24 h). The following rechecks revealed secondary cataracts with subsequent lens subluxation and panretinal degeneration OU. Despite being blind and the poor prognosis of disseminate cryptococcosis, the patient remained active and in good body condition during 6 months after the initial diagnosis. At that time, the ferret showed ataxia, incontinence, and generalized pain. A magnetic resonance imaging study revealed a mass affecting the spine. The owners declined further investigations and the ferret was humanely euthanized. The postmortem histopathology confirmed the initial diagnosis of cryptococcosis and the presence of intraretinal Cryptococcus spp. To the authors' knowledge, this is the first report of Cryptococcus spp. induced exudative chorioretinitis in a ferret.

Plasma protein electrophoresis of Trachemys scripta and Iguana iguana.

BACKGROUND: Protein electrophoresis is widely applied in veterinary medicine, but is not used often in reptiles, in part because of lack of reference values. OBJECTIVE: The goals of this study were to compare plasma protein profiles obtained by cellulose acetate electrophoresis (CAE) and agarose gel electrophoresis (AGE), measure precision and examine interference by sample hemolysis, and establish preliminary reference intervals for 2 reptile species. METHODS: Heparinized plasma samples from healthy and diseased adult female Iguana iguana (n=40) and Trachemys scripta (n=60) were analyzed by CAE and AGE. Total protein concentration was measured by the biuret method. Electrophoresis results were compared using Bland-Altman plots and Passing-Bablok regression analysis. Precision and the effects of sample hemolysis were determined. Results from clinically healthy animals were used to determine reference intervals. RESULTS: Five protein fractions were identified in both species, with bisalbuminemia observed in 23/40 iguanas. High correlation was observed between the 2 methods for all fractions, with few proportional and systematic errors. Coefficients of variation were lower using AGE vs CAE and for I. iguana vs T. scripta. Two additional bands were observed in hemolyzed samples from T. scripta; 1 additional band was observed for I. iguana. Minimum and maximum values were reported for healthy I. iguana (n=14) and T. scripta (n=22). CONCLUSIONS: Although both methods are acceptable, the performance of AGE was slightly better than that of CAE for analysis of plasma from reptiles. Furthermore, reptile electrophoretic patterns should be interpreted based on the method used, the species analyzed, and the quality of the plasma sample.

Case report: ovarian teratoma in a Mediterranean tortoise.

A 61-year-old female Mediterranean tortoise (Testudo hermannii) presented after 1 week of tetraparesis. Coelomic ultrasonography revealed free fluid and an ovarian mass with multiple follicles. Blood analysis suggested renal failure. After initial medical treatment for kidney disease, the tortoise seemed to improve clinically, but the animal died 2 months later. Necropsy and histopathology revealed an ovarian teratoma. To our knowledge, this is the first reported case of a teratoma in tortoise gonadal tissue.

Relación de la radiación ultravioleta y el pterigión primario / Relation of ultraviolet radiation and primary pterygium

El pterigión está presente a nivel mundial, pero es más común en climas cálidos y secos, predomina en países comprendidos entre 0º y 30º de latitud norte y sur. Existe evidencia epidemiológica en estudios poblacionales experimentales y observacionales que confirman que la radiación ultravioleta es el desencadenante inicial en la aparición del pterigión y un factor de riesgo significativo en su desarrollo. Los fenómenos físicos y climatológicos que explican la patogenia del pterigión son poco conocidos. La epidemiología del pterigión es una herramienta básica para conocer tanto su incidencia como su prevalencia. A partir de esta revisión nos propusimos reflexionar acerca del pterigión como marcador del grado de insolación del organismo y enfatizar en cuanto a la importancia de protección ante la posible exposición a la radiación ultravioleta en el resultado final de una cirugía de pterigión(AU)

Pterygium is present worldwide but it is more common in dry warm climates and predominates in countries situated 0º and 30º on North and South latitudes. There are epidemiological pieces of evidence in experimental and observational population studies that confirm that ultraviolet radiation is the initial unleashing factor in occurrence of pterygium and a significant risk factor for its development. The physical and climate phenomena accounting for the pathogeny of pterygium are almost unknown. Pterygium epidemiology is a basic tool for identifying its incidence and prevalence. On the basis of this review, we intended to make reflections on pterygium as a marker of degree of insolation of the human body and to emphasize the importance of protection against possible exposure to ultraviolet radiations in the final outcome of pterygium surgery(AU)

Relación de la radiación ultravioleta y el pterigión primario: [revisión] / Relation of ultraviolet radiation and primary pterygium: [review]

El pterigión está presente a nivel mundial, pero es más común en climas cálidos y secos, predomina en países comprendidos entre 0º y 30º de latitud norte y sur. Existe evidencia epidemiológica en estudios poblacionales experimentales y observacionales que confirman que la radiación ultravioleta es el desencadenante inicial en la aparición del pterigión y un factor de riesgo significativo en su desarrollo. Los fenómenos físicos y climatológicos que explican la patogenia del pterigión son poco conocidos. La epidemiología del pterigión es una herramienta básica para conocer tanto su incidencia como su prevalencia. A partir de esta revisión nos propusimos reflexionar acerca del pterigión como marcador del grado de insolación del organismo y enfatizar en cuanto a la importancia de protección ante la posible exposición a la radiación ultravioleta en el resultado final de una cirugía de pterigión.

Pterygium is present worldwide but it is more common in dry warm climates and predominates in countries situated 0º and 30º on North and South latitudes. There are epidemiological pieces of evidence in experimental and observational population studies that confirm that ultraviolet radiation is the initial unleashing factor in occurrence of pterygium and a significant risk factor for its development. The physical and climate phenomena accounting for the pathogeny of pterygium are almost unknown. Pterygium epidemiology is a basic tool for identifying its incidence and prevalence. On the basis of this review, we intended to make reflections on pterygium as a marker of degree of insolation of the human body and to emphasize the importance of protection against possible exposure to ultraviolet radiations in the final outcome of pterygium surgery.

Factores predictivos de recurrencia del pterigium primario / Predictive factors of primary pterygium recurrence

El tratamiento exitoso del pterigium constituye un desafío constante para los oftalmólogos. Este tratamiento es netamente quirúrgico, con la intención de lograr su perfeccionamiento sistemático han surgido innumerables técnicas, que se emplean según las particularidades de cada paciente o las del cirujano. El pterigium es de etiología multifactorial, lo cual genera una gran complejidad, cuando se trata de seleccionar la técnica quirúrgica más adecuada. Con el propósito de analizar factores predictivos de recurrencia del pterigium: genéticos, histológicos, angiográficos y morfológicos, se realizó esta revisión bibliográfica(AU)

The successful treatment of pterygiyum is a permanent challenge to be met by ophthalmologists. This kind of therapy is purely surgical and a number of techniques have emerged to attain its systematic improvement, which are used in line with the particularities of every patient or those of the surgeon. Pterygiyum has a multifactor etiology, so it is very complex to select the most suitable technique to operate it. The present literature review was made to analyze predictive factors of pterygium recurrence such as genetic, histological, angiographic and morphological factors(AU)

Estudio comparativo del uso de mitomicina C en el pterigium recidivante / Comparative study of the use of mitomycin C in recurrent pterygium

El pterigium constituye un problema en la práctica oftalmológica por su alta prevalencia y elevada frecuencia de recidiva. El propósito de este estudio fue el de comparar la técnica quirúrgica de injerto autólogo de conjuntiva con células límbicas más mitomicina C, con la de injerto de membrana amniótica con mitomicina C, en pterigium recidivante, en cuanto a la tasa de recidiva, complicaciones posoperatorias inmediatas, y tiempo libre de recidiva (meses). Se realizó un ensayo clínico controlado prospectivo con n= 57 pacientes portadores de pterigium recidivante. Se conformaron 2 grupos: grupo 1, injerto de conjuntiva antólogo con células límbicas más mitomicina C; y el grupo 2, injerto de membrana amniótica más mitomicina C. Se comparó el comportamiento de las técnicas empleadas a partir de sus resultados. El seguimiento fue ± de 1-6 meses. La agudeza visual mejoró en 60 por ciento de los casos. La recidiva fue de 2 por ciento para el grupo 1 y de 6 por ciento para el grupo 2. El método de injerto conjuntival con limbo resultó más efectivo para el tratamiento del pterigium recidivante. Él método de membrana amniótica constituye una alternativa para el tratamiento de casos en los cuales no sea viable la aplicación de de injerto antólogo(AU)

Pterygium is a problem in ophthalmology practice because of its high prevalence and frequency of recurrence. The objectives of this study was to compare the surgical technique of autologous conjuctival graft with limbal cells plus mitomycin C with that of amniotic membrane graft treated with mitomycin C in recurrent pterygium in terms of recurrence rates, immediate postoperative complications and recurrence-free time (months). A prospective controlled clinical assay was carried out in 57 patients suffering recurrent pterygium. They were divided into two groups; the group 1 that had autologous conjuctival graft with limbal cells plus mitomycin C, and the group 2 with amminotic membrane graft plus mitomycin C. The used techniques were compared on the basis of their results. The follow-up period extended from one to six months. Visual acuity improved in 60 percent of cases. Recurrence occurred in 2 percent of cases in group 1 and 6 percent in group 2. The conjunctival graft with limbal cells method was more effective for the recurrent pterygium treatment. The amniotic membrane method constitutes an alternative therapy for cases in which autologous graft is not applicable(AU)

Estudio comparativo del uso de mitomicina C en el pterigium recidivante / Comparative study of the use of mitomycin C in recurrent pterygium

El pterigium constituye un problema en la práctica oftalmológica por su alta prevalencia y elevada frecuencia de recidiva. El propósito de este estudio fue el de comparar la técnica quirúrgica de injerto autólogo de conjuntiva con células límbicas más mitomicina C, con la de injerto de membrana amniótica con mitomicina C, en pterigium recidivante, en cuanto a la tasa de recidiva, complicaciones posoperatorias inmediatas, y tiempo libre de recidiva (meses). Se realizó un ensayo clínico controlado prospectivo con n= 57 pacientes portadores de pterigium recidivante. Se conformaron 2 grupos: grupo 1, injerto de conjuntiva antólogo con células límbicas más mitomicina C; y el grupo 2, injerto de membrana amniótica más mitomicina C. Se comparó el comportamiento de las técnicas empleadas a partir de sus resultados. El seguimiento fue ± de 1-6 meses. La agudeza visual mejoró en 60 por ciento de los casos. La recidiva fue de 2 por ciento para el grupo 1 y de 6 por ciento para el grupo 2. El método de injerto conjuntival con limbo resultó más efectivo para el tratamiento del pterigium recidivante. Él método de membrana amniótica constituye una alternativa para el tratamiento de casos en los cuales no sea viable la aplicación de de injerto antólogo.

Pterygium is a problem in ophthalmology practice because of its high prevalence and frequency of recurrence. The objectives of this study was to compare the surgical technique of autologous conjuctival graft with limbal cells plus mitomycin C with that of amniotic membrane graft treated with mitomycin C in recurrent pterygium in terms of recurrence rates, immediate postoperative complications and recurrence-free time (months). A prospective controlled clinical assay was carried out in 57 patients suffering recurrent pterygium. They were divided into two groups; the group 1 that had autologous conjuctival graft with limbal cells plus mitomycin C, and the group 2 with amminotic membrane graft plus mitomycin C. The used techniques were compared on the basis of their results. The follow-up period extended from one to six months. Visual acuity improved in 60 percent of cases. Recurrence occurred in 2 percent of cases in group 1 and 6 percent in group 2. The conjunctival graft with limbal cells method was more effective for the recurrent pterygium treatment. The amniotic membrane method constitutes an alternative therapy for cases in which autologous graft is not applicable.