Identification of asinine gamma herpesviruses in a donkey with interstitial pulmonary fibrosis, pleural effusion and thrombocytopenia.

A 23-year-old domestic donkey (Equus asinus) referred for severe respiratory distress due to suspected equine asthma. Ultrasound of the chest revealed bilateral irregular pulmonary consolidation and pleural effusion. Airway endoscopy and tracheal wash cytology showed severe neutrophilic inflammation and bacterial culture was positive for Streptococcus equi subsp. zooepidemicus. Despite aggressive treatment, the donkey died in 48 hours. On post-mortem examination, the lung was whitish, collapsed, and firm, with fibrotic multifocal nodular areas. Pleural effusion and pleuritis were detected. Histologically, the lung architecture was markedly replaced by interstitial fibrosis. The histological features observed were suggestive of a severe chronic fibrosing interstitial pleuropneumonia with type 2 pneumocyte hyperplasia and intralesional syncytial cells. Pulmonary fibrosis was associated with the presence of asinine gammaherpesvirus 2 and 5 infection, confirmed by PCR and sequence analysis. The macroscopic and histological pattern of fibrosis was diffuse and interstitial, and the nodular lesions were consistent with spared lung parenchyma, instead of the canonical nodular distribution of the fibrosis observed in equine multinodular pulmonary fibrosis. Asinine herpesviral pulmonary fibrosis is uncommon, but should be considered by clinicians in the list of differentials in donkeys with chronic respiratory signs.

Intermittent mild cold stimulation alleviates cold stress-induced pulmonary fibrosis by inhibiting the TGF-ß1/Smad signaling pathway in broilers.

To investigate the potential protective effect of intermittent cold stimulation on lung tissues of broilers exposed to acute cold stress (ACS). A total of 384 one-day-old broilers were assigned to 4 experimental groups with 6 replicates of 16 birds each: control (CON) and ACS groups were reared at normal feeding temperature from d 1 to 42; cold treatment groups (CS3+ACS and CS9+ACS) were reared, respectively, at 3°C or 9°C for 5 h on alternate days below the CON group from d 15 to 35. Animals in CS3+ACS, CS9+ACS, and ACS groups were exposed at 10°C for 24 h on d 43. Subsequently, lung tissues were collected to perform histopathological examination and measurement of relevant indexes. The results showed that lung tissues in CS9+ACS and ACS groups exhibited increased inflammatory cell infiltrates and collagen deposition compared to the CON group, while this pathological phenomenon was less pronounced in the CS3+ACS group. Compared to CON group, H2O2 and MDA contents were increased, and the activities of antioxidant enzymes (CAT, SOD, GPx, T-AOC) were reduced in CS9+ACS and ACS group (P < 0.05); mRNA and protein levels of inhibitor of NF-κB, Smad7, matrix metallopeptidase (MMP)-2, MMP9, and antioxidant-related genes were downregulated, whereas mRNA and protein levels of genes related to NF-κB/NLRP3 pathway-regulated inflammation and TGF-ß1/Smad pathway-regulated fibrosis were upregulated in cold-stressed broilers (P < 0.05). mRNA levels of heme oxygenase-1, NAD(P)H quinone oxidoreductase-1, and MMP9 were increased in CS3+ACS group (P < 0.05). Moreover, the expression of most antioxidant-related genes was increased, and that of inflammation- and fibrosis-related genes was reduced in CS3+ACS group (P < 0.05). Therefore, cold stress caused oxidative stress and inflammation, leading to pulmonary fibrosis in broilers, whereas intermittent mild cold stimulation at 3°C below normal rearing temperature alleviated fibrosis by inhibiting the TGF-ß1/Smad pathway modulated by the Nrf2/HO-1 and NF-κB/NLRP3 signaling pathway. This study suggests that intermittent mild cold stimulation can be a potential strategy to reduce ACS-induced lung damage in broilers.

Association of equine gammaherpesvirus-5 with facial lymphohistiocytic interface dermatitis in seven adult horses from the United States.

Equine herpesvirus-5 (EHV-5) is commonly found in healthy asymptomatic horses worldwide. Although a cause-and-effect relationship has not been thoroughly determined, this virus has been associated with several disease conditions including equine multinodular pulmonary fibrosis (EMPF) and 1 case of interface dermatitis. The authors searched the New York State Animal Health Diagnostic Center database for cases of equine interface dermatitis between 2007 and 2022. Ten cases were identified and scrutinized for viral inclusion bodies which were present in 5 of 10 cases. Two similar cases with interface dermatitis and viral inclusion bodies, which were not part of a retrospective search, were from the Oregon Veterinary Diagnostic Laboratory. The authors describe a total of 7 horses with dermatitis characterized by crusted, alopecic, non-pruritic, non-painful, irregular to annular areas over the face, most commonly the muzzle, for up to several years duration. Histologically, there was a CD3+ T lymphocyte-dominated lymphohistiocytic interface dermatitis with hydropic degeneration, apoptotic keratinocytes, and pigmentary incontinence. Keratinocytes within the upper stratum spinosum and stratum granulosum had glassy pale basophilic intranuclear inclusion bodies consistent with herpesvirus. The presence of EHV-5 was confirmed by quantitative polymerase chain reaction (qPCR) and in situ hybridization in 7 horses and by electron microscopy in 1 horse. One horse later developed EMPF and was euthanized. EHV-5 was not detected with qPCR from 5 control horses and 5 horses with interface dermatitis without histologic evidence of viral inclusion bodies. These are the first cases of facial interface dermatitis associated with EHV-5 reported in the United States.

Amelioration of bleomycin induced pulmonary fibrosis by administration of Salvianolic acid B in mice.

Pulmonary fibrosis is the end­stage manifestation of wide range of respiratory diseases and during pulmonary fibrosis, pulmonary inflammation and epithelial­mesenchymal transition (EMT) play important roles. Salvianolic acid B (SAB) from the herb Salviae miltiorrhiza has been reported to possess an excellent anti­inflammatory, antifibrotic and antioxidant activity. The present study aimed to investigate the ameliorative effect of SAB on bleomycin induced pulmonary fibrosis in mice. Adult albino mice were divided as SHAM/control group (saline alone), BLM group (bleomycin @ 1mg/kg intratracheally once) and SAB groups (BLM challenged once and SAB administration in three dosages @ 5, 10 and 15 mg/kg intraperitoneally daily for 30 days). Lungs wet/dry ratio and protein concentration in bronchoalveolar lavage fluid, MPO activity, oxidative stress markers, hydroxyproline assay, levels of inflammatory cytokines (TNF­α, IL­6 and TGF­ß1), NF­κB activity, histopathology, immunostaining (E­cadherin, vimentin and alpha ­smooth muscle actin) and ultrastructural changes were studied. SAB showed anti­inflammatory and anti­fibrotic effects through inhibition of inflammatory cell infiltration, alveolar structure disruption, and collagen deposition and the expression of several fibrogenic cytokines. SAB also up­regulate E­cadherin and down­regulated vimentin and alpha­smooth muscle actin expression. In conclusion, Salvianolic acid B is effective in alleviating the BLM induced lung fibrosis through suppression of oxidative stress, inflammation, histological, ultrastructural changes and EMT.

Detection of asinine gammaherpesviruses in association with pulmonary fibrosis in free-ranging donkeys.

A mortality event among recently captured feral donkeys (Equus asinus) occurred in south-central Utah in 2016. The deaths were sporadic, and clinical signs were indicative of respiratory disease, likely associated with an infectious etiology. Ten of 13 donkeys autopsied had moderate-to-severe interstitial fibrosing pneumonia, and one had pyogranulomatous pneumonia. Consensus PCRs directed toward the DNA polymerase and DNA packaging terminase subunit 1 for herpesviruses were performed followed by sequencing of the PCR amplicons and phylogenetic analysis. Asinine herpesvirus 4 (AsHV4) and 5 (AsHV5) were consistently identified in lung tissues of affected donkeys. No other herpesviruses were identified, and herpesviral DNA was not detected in lung tissues of 2 donkeys without evidence of respiratory disease. The detection of asinine gammaherpesviruses may have been associated with the lesions described. AsHV4 and AsHV5 have been reported in previous studies as novel gammaherpesviruses based on sequences obtained from donkeys with interstitial pneumonia and marked syncytial cell formation. Our findings suggest that the association of asinine gammaherpesviruses with respiratory conditions in equids deserves further attention.

Reversibility of clinical and computed tomographic lesions mimicking pulmonary fibrosis in a young cat.

BACKGROUND: In humans with idiopathic pulmonary fibrosis (IPF), specific thoracic computed tomographic (CT) features in the correct clinical context may be used in lieu of histologic examination. Cats develop an IPF-like condition with similar features to humans. As few cats have invasive lung biopsies, CT has appeal as a surrogate diagnostic, showing features consistent with architectural remodeling supporting "end-stage lung". CASE PRESENTATION: A 1-year-old female spayed Domestic Shorthair cat presenting with progressive respiratory clinical signs and thoracic CT changes (reticular pattern, parenchymal bands, subpleural interstitial thickening, pleural fissure thickening, subpleural lines and regions of increased attenuation with traction bronchiectasis and architectural distortion) consistent with reports of IPF was given a grave prognosis for long-term survival. The cat was treated with prednisolone, fenbendazole, pradofloxacin and clindamycin. Five months later, while still receiving an anti-inflammatory dose of prednisolone, the cat was re-evaluated with owner-reported absent respiratory clinical signs. Thoracic CT demonstrated resolution of lung patterns consistent with fibrosis. CONCLUSIONS: Fibrotic lung disease is irreversible. Despite this cat having compatible progressive respiratory signs and associated lung patterns on thoracic CT scan, these abnormalities resolved with non-specific therapy and time, negating the possibility of IPF. While the cause of the distinct CT lesions that ultimately resolved was not determined, infection was suspected. Experimental Toxocara cati infection shows overlapping CT features as this cat and is considered a treatable disease. Improvement of CT lesions months after experimental heartworm-associated respiratory disease in cats has been documented. Reversibility of lesions suggests inflammation rather than fibrosis was the cause of the thoracic CT lesions. This cat serves as a lesson that although thoracic CT has been advocated as a surrogate for histopathology in people with IPF, additional studies in cats are needed to integrate CT findings with signalment, other clinicopathologic features and therapeutic response before providing a diagnosis or prognosis of fibrotic lung disease.

A case series highlighting the role of different gamma-herpesviruses in Equine Multinodular Pulmonary Fibrosis.

INTRODUCTION: This case series describes three cases of equine multinodular pulmonary fibrosis (EMPF) diagnosed at the Clinic for Equine Internal Medicine at the University of Zurich between 2012 and 2017. Current information on etiology and treatment options are presented. Two horses showed mild signs of chronic lower respiratory tract disease and one horse was presented with acute signs of disease including recurrent fever spikes and tachypnea. Diagnosis was achieved by physical examination, radiographic findings, and PCR testing for equine herpesviruses (EHV) of bronchoalveolar lavage (BAL) fluid or lung tissue obtained by biopsy. All horses were euthanized due to continuing deterioration after attempted treatment. Post mortem histological examination of lung tissue showed severe multifocal diffuse to confluent fibrosis in two cases and in another horse a discrete nodular fibrosis pattern. Panherpes nested PCR revealed the presence of equine herpesvirus 5 (EHV-5) DNA in lung tissue of one horse whereas in two other horses, asinine herpes virus 5 (AHV-5) was detected. EMPF should be considered as a differential diagnosis in horses with acute and chronic respiratory disease, including horses non-responsive to treatment for equine asthma.

INTRODUCTION: Cette série de cas décrit trois cas de fibrose pulmonaire multinodulaire équine (EMPF) diagnostiqués à la Clinique de médecine interne équine de l'Université de Zurich entre 2012 et 2017. Des informations actuelles sur l'étiologie et les options de traitement sont présentées. Deux chevaux présentaient de légers signes de maladie chronique des voies respiratoires inférieures et un cheval présentait des signes aigus de maladie, notamment des pics de fièvre récurrents et une tachypnée. Le diagnostic a été obtenu grâce à un examen physique, des résultats radiographiques et des tests PCR pour les virus herpès équins (EHV) du liquide de lavage broncho-alvéolaire (BAL) ou du tissu pulmonaire obtenus par biopsie. Tous les chevaux ont été euthanasiés en raison d'une détérioration continue après une tentative de traitement. L'examen histologique post mortem du tissu pulmonaire a montré une fibrose multifocale diffuse à confluente sévère dans deux cas et chez un cheval un type de fibrose nodulaire discret. La PCR par Panherpes a révélé la présence d'ADN de virus herpès équin 5 (EHV-5) dans le tissu pulmonaire d>un cheval alors que chez deux autres chevaux, le virus de l>herpès asinien 5 (AHV-5) a été détecté. L'EMPF doit être considéré comme un diagnostic différentiel chez les chevaux souffrant d'une maladie respiratoire aiguë et chronique, y compris les chevaux ne répondant pas au traitement de l'asthme équin.

Haemostatic, fibrinolytic and inflammatory profiles in West Highland white terriers with canine idiopathic pulmonary fibrosis and controls.

BACKGROUND: Canine idiopathic pulmonary fibrosis (CIPF) is a progressive interstitial lung disease mainly affecting old West Highland white terriers (WHWTs). The aetiology of CIPF is currently unknown and pathogenesis poorly understood. A genetic basis is strongly suspected based on the breed predisposition. CIPF shares clinical and pathological features with human IPF. In human IPF, coagulation disorders favouring a local and systemic pro-thrombotic state have been demonstrated in association with disease severity and outcome. The aim of this study was to compare the systemic haemostatic, fibrinolytic and inflammatory profiles of WHWTs affected with CIPF with breed-matched controls (CTRLs). Additionally, data collected in both groups were interpreted with regard to the reference intervals (when available) to assess possible pro-thrombotic features of the WHWT breed that may be related to CIPF predisposition. A total of 14 WHWTs affected with CIPF and 20 CTRLs were included. RESULTS: WHWTs affected with CIPF had prolonged activated partial thromboplastine time in comparison with CTRLs (12.2 ± 0.9 s vs. 11.5 ± 0.7 s, P = 0.028), whereas results obtained in both groups were all within reference ranges. There was no significant difference between groups for the other factors assessed including plasmatic concentrations of fibrinogen, D-dimers concentration, antithrombin III activity, protein S and protein C activities, anti-factor Xa activity, activated protein C ratio, serum C-reactive protein concentration, and rotational thromboelastometry indices. Platelet count and plasmatic fibrinogen concentration were found to be above the upper limit of the reference range in almost half of the WHWTs included, independently of the disease status. CONCLUSIONS: Results of this study provide no clear evidence of an altered systemic haemostatic, fibrinolytic or inflammatory state in WHWTs affected with CIPF compared with CTRLs. The higher platelet counts and fibrinogen concentrations found in the WHWT breed may serve as predisposing factors for CIPF or simply reflect biological variation in this breed.

Two Cases of Equine Multinodular Pulmonary Fibrosis in Japan.

Equine multinodular pulmonary fibrosis (EMPF) is a recently described form of interstitial pneumonia associated with equine herpesvirus type 5 (EHV-5). This disease has been reported in North and South America, Europe and Oceania but not, to our knowledge, in horses in Japan. We diagnosed EMPF in two Thoroughbred horses in Japan on the basis of gross and histopathological findings. In both cases, significant gross lesions, restricted to the lungs, consisted of numerous firm and coalescing nodules widely distributed throughout the lung. The nodules were <3 cm in diameter and pale white to tan in colour. Microscopically, they showed severe interstitial fibrosis and infiltration of macrophages, neutrophils, lymphocytes and a few eosinophils. The residual alveoli were lined by cuboidal epithelial cells (type II pneumocytes) and filled with many macrophages, which rarely displayed oval eosinophilic to amphophilic intranuclear inclusion bodies. Polymerase chain reaction and sequence analyses identified the glycoprotein H gene of EHV-5, and in-situ hybridization detected EHV-5 in the alveolar macrophages in the lesions. In one case, electron microscopy revealed herpesvirus-like particles and EHV-5 was isolated from pulmonary lesions.

Mediastinal Ectopic Thyroid Carcinoma and Concurrent Multinodular Pulmonary Fibrosis in a Horse.

A 30-year-old Standardbred gelding was referred for chronic pleural effusion. Thoracic ultrasound revealed marked bilateral pleural effusion and a large heterogeneous mass within the cranial mediastinum, which extended from the right 5th to the 11th intercostal space. Subsequently, on thoracic radiographs, the cranial mediastinal mass was confirmed, and a nodular interstitial pattern was identified in the lungs. Because of progressive clinical deterioration of the patient, the owners elected humane euthanasia and necropsy was performed. The mediastinal mass was consistent with an ectopic thyroid carcinoma, and the pulmonary nodules represented equine multinodular pulmonary fibrosis (EMPF). This case report describes a type of mediastinal tumor not previously described in horses. Moreover, it shows the need of including EMPF as a possible differential diagnosis for a nodular interstitial pulmonary pattern in conjunction with a mediastinal or other masses.

Perspectives in veterinary medicine: Description and classification of bronchiolar disorders in cats.

This Perspectives in Veterinary Medicine article seeks to define, describe putative causes, and discuss key diagnostic tests for primary and secondary bronchiolar disorders to propose a classification scheme in cats with support from a literature review and case examples. The small airways (bronchioles with inner diameters <2 mm), located at the transitional zone between larger conducting airways and the pulmonary acinus, have been overlooked as major contributors to clinical syndromes of respiratory disease in cats. Because the trigger for many bronchiolar disorders is environmental and humans live in a shared environment with similar susceptibility, understanding these diseases in pet cats has relevance to One Health. Thoracic radiography, the major imaging modality used in the diagnostic evaluation of respiratory disease in cats, has low utility in detection of bronchiolar disease. Computed tomography (CT) with paired inspiratory and expiratory scans can detect pathology centered on small airways. In humans, treatment of bronchiolar disorders is not well established because of heterogeneous presentations and often late definitive diagnosis. A review of the human and veterinary medical literature will serve as the basis for a proposed classification scheme in cats. A case series of cats with CT or histopathologic evidence of bronchiolar lesions or both, either as a primary disorder or secondary to extension from large airway disease or interstitial lung disease, will be presented. Future multi-institutional and multidisciplinary discussions among clinicians, radiologists, and pathologists will help refine and develop this classification scheme to promote early and specific recognition and optimize treatment.

Lung lesions of slaughtered horses in southern Brazil / Lesões pulmonares de equinos de matadouro-frigorífico no Sul do Brasil

Respiratory diseases cause significant veterinary costs, reduce performance and require withdrawal of horses. Yet, studies of the causes of pneumonia in horses are scant. This study aimed to describe the pathological and microbiological features of lung lesions in slaughtered horses in southern Brazil. In this study, 84 samples of lungs were examined, and a conclusive diagnosis was obtained in 74 cases. These were composed of bronchopneumonia in 50 cases, followed by granulomatous eosinophilic pneumonia (9/74), recurrent airway obstruction (7/74), lung fibrosis (4/74), lung hemorrhage (3/74) and pulmonary pythiosis (1/74). Bronchopneumonia had grossly firm focally extensive yellow to dark-red areas, which consisted microscopically of multifocal to coalescing infiltrate of degenerate neutrophils. Streptococcus equi subsp. zooepidemicus was identified in 21 of the 50 cases. Granulomatous eosinophilic pneumonia had multifocal pinpoint firm-hard yellow areas, which microscopically were composed of granulomas with a mineralized center surrounded by collagen fibers and severe infiltrate of eosinophils. Recurrent airway obstruction had mild multifocal pinpoint firm white areas that consisted microscopically of large amounts of mucus inside bronchi and bronchiole. Lung fibrosis had two patterns: focally extensive areas of consolidation and firm nodular areas. Microscopically, the first pattern had interstitial to peribronchial fibrosis, while the second had, in addition to the interstitial fibrosis, a severe pneumocyte hyperplasia and an alveolar infiltrate of neutrophils and macrophages with rare intranuclear inclusion bodies (equine herpesvirus 5, EHV-5). Pulmonary pythiosis presented a focal firm nodular area, with multiple kunkers observed in the cut surface, which corresponded microscopically to areas of necrosis surrounded by a mixed inflammatory infiltrate. At the periphery of the necrotic areas, multiple negatively stained hyphae were observed, which were evidenced through Grocott's stain and immunohistochemistry anti-Pythium insidiosum.(AU)

Doenças respiratórias causam em equinos custos significativos com tratamento veterinário, redução de performance e descarte de animais. No entanto, estudos que abordem as causas de pneumonia em equinos são escassos. O objetivo deste estudo foi descrever os aspectos patológicos e microbiológicos de lesões pulmonares em equinos abatidos em matadouro-frigorífico no Sul do Brasil. Neste estudo, 84 amostras de pulmões foram examinadas, e o diagnóstico conclusivo das condições foi obtido em 74 casos. Esses foram compostos por broncopneumonia em 50 casos, seguido por pneumonia granulomatosa eosinofílica (9/74), obstrução aérea recorrente (7/74), fibrose pulmonar (4/74), hemorragia pulmonar (3/74) e pitiose pulmonar (1/74). A broncopneumonia era caracterizada macroscopicamente por áreas focalmente extensas firmes de coloração amarelada a vermelho-escuras, as quais consistiam microscopicamente em infiltrado multifocal a coalescente de neutrófilos degenerados. Streptococcus equi subsp. zooepidemicus foi identificado em 21 dos 50 casos. A pneumonia eosinofílica granulomatosa era caracterizada por áreas multifocais puntiformes firmes a duras e amareladas, que microscopicamente eram compostas por granulomas com área central mineralizada circundados por fibras de colágeno e infiltrado acentuado de eosinófilos. A obstrução aérea recorrente era caracterizada por discretas áreas puntiformes firmes e brancacentas que consistiam microscopicamente em grande quantidade de muco no interior de brônquios e bronquíolos. A fibrose pulmonar exibia dois padrões: áreas de consolidação focalmente extensas e áreas nodulares firmes. Microscopicamente, o primeiro padrão exibia fibrose intersticial a peribronquial, enquanto no segundo padrão havia, além da fibrose intersticial, intensa hiperplasia de pneumócitos e infiltrado alveolar de neutrófilos e macrófagos com raros corpúsculos de inclusão intranucleares (herpesvírus equino 5, EHV-5). A pitiose pulmonar exibia uma área nodular firme focal com múltiplos kunkers ao corte, os quais correspondiam microscopicamente a áreas de necrose circundadas por infiltrado inflamatório misto. À periferia das áreas necróticas, múltiplas imagens negativas de hifas eram observadas, as quais foram evidenciadas através da coloração de Grocott e imuno-histoquímica anti-Pythium insidiosum.(AU)

Effect of valacyclovir on EHV-5 viral kinetics in horses with equine multinodular pulmonary fibrosis.

BACKGROUND: Equine herpesvirus-5 is commonly isolated from the lungs of horses with EMPF, suggesting an etiological link. Valacyclovir is used empirically to treat EMPF; however, no data is available concerning its impact on EHV-5 viral kinetics. OBJECTIVES: To determine the effect of oral administration of valacyclovir on EHV-5 viral load measured by qPCR in blood, nasal secretions (NS) and BALF in horses with EMPF. ANIMALS: Six horses diagnosed with EMPF. METHODS: A prospective clinical trial was performed. Horses received 10 days of PO administered valacyclovir (loading dose 30 mg/kg, maintenance dose 20 mg/kg). Blood, NS, and BALF were collected for EHV-5 viral kinetics analyses during treatment. Blood and NS were collected every other day. BALF was collected on day 0 and day 10. RESULTS: There was no statistical difference in median EHV-5 viral load between day 0 and day 10 for all samples tested. In blood median EHV-5 viral load was 7676 (range 575-39 781) on day 0 and 6822 (range 1136-18 635) glycoprotein B (gB) gene copies per million cells on day 10. For NS median EHV-5 viral load was 2.944 × 106 (range 184 691-3.394 × 109 ) on day 0 and 8.803 × 106 (range 251 186-9.868 × 108 ) gB gene copies per million cells on day 10. For BALF median EHV-5 viral load was 59,842 (range 61-315 655) on day 0 and 185 083 (range 3562-542 417) gB gene copies per million cells on day 10. CONCLUSIONS AND CLINICAL IMPORTANCE: Valacyclovir might not be an effective short-term antiviral treatment but efficacy in treatment of EMPF is unknown.

Equine multinodular pulmonary fibrosis in southern Brazil: pathology and differential diagnosis / Fibrose pulmonar multinodular em equinos no Sul do Rio Grande do Sul: patologia e diagnóstico diferencial

Equine multinodular pulmonary fibrosis (EMPF) diagnosed at the Laboratório Regional de Diagnóstico of Faculdade de Medicina Veterinária, Universidade Federal de Pelotas (LRD/UFPel), is described. Differential aspects of other pulmonary diseases in horses with pneumonia and interstitial fibrosis were discussed. The disease occurred in a 15-year-old equine that presented with clinical signs of respiratory distress, intermittent fever, anorexia, and dyspnea. Macroscopically, there was enlargement of the lungs with whitish, pale, firm and well-delimited nodules, approximately 7-10 cm in diameter, distributed throughout the parenchyma. Histologically, the lung nodules had alveolar spaces with walls covered by cuboidal epithelium containing macrophages, neutrophils, lymphocytes, hyperplasia of type II pneumocytes and, eventually, multinucleated giant cells. The interstitium was markedly thickened by mature fibrous connective tissue and collagen. There were intranuclear inclusion bodies in the macrophages. The PCR technique for detecting the EHV-5 DNA was positive. In a retrospective study of pneumonia cases in horses with interstitial fibrosis diagnosed in the LRD/UFPel, two animals had macroscopic and histological lesions similar to those with EMPF, but they were negative for EHV-5 in PCR. Four cases diagnosed with pneumonia and interstitial tissue fibrosis had a histological pattern that was different from that observed in the EMPF animal, thus eliminating the possibility of EMPF. It is concluded that EMPF is a sporadic disease that should be considered in cases of respiratory disease in horses. Reports of such cases are important to alert technicians about the occurrence of rare diseases in Brazil. It is also necessary to establish the true role of EHV-5 in the pathogenesis of EMPF. Cases of pulmonary fibrosis such as EMPF, in which the virus is not present, should be studied to establish whether it could be an idiopathic form of the disease.(AU)

Descreve-se a fibrose multinodular pulmonar equina (EMPF) diagnosticado no Laboratório Regional de Diagnóstico da Faculdade de Veterinária da Universidade Federal de Pelotas. Foram discutidos a patologia da doença e os aspectos diferenciais de outras enfermidades pulmonares de equinos que cursam com pneumonia e fibrose intersticial. A doença ocorreu em um equino sem raça definida de 15 anos de idade que apresentou sinais clínicos de dificuldade respiratória febre intermitente, anorexia e dispneia, com evolução de aproximadamente 10 dias. Macroscopicamente havia aumento de volume dos pulmões e nódulos esbranquiçados, pálidos, firmes e bem delimitados, de aproximadamente 7-10 cm de diâmetro, distribuídos pelo parênquima. Histologicamente, o tecido pulmonar apresentava nódulos caracterizados pela presença de espaços alveolares, com paredes revestidas por epitélio cuboidal achatado, contendo macrófagos e neutrófilos e havia, também, linfócitos e hiperplasia de pneumócitos tipo II e eventualmente células gigantes multinuacleadas. O interstício estava acentuadamente espessado por tecido conjuntivo fibroso maduro e por colágeno. Havia corpúsculos de inclusão intranucleares em macrófagos. A técnica de PCR para detecção do DNA de herpes vírus equino-5 (EHV-5) resultou positiva. Em um estudo retrospectivo de casos de pneumonia com fibrose intersticial diagnosticados no LRD entre 2000 e 2015, dois equinos apresentaram lesões macroscópicas e histológicas similares às de EMPF, porém resultaram negativos na PCR para detecção de EHV-5. Quatro casos de pneumonia com fibrose do tecido intersticial apresentaram padrão histológico diverso da EMPF descartando-se a possibilidade de tratar-se da doença. Conclui-se que EMPF é uma enfermidade esporádica, no entanto deve ser levada em consideração em casos de doença respiratória em equinos. A descrição dos casos diagnosticados é importante para alertar técnicos sobre a ocorrência da mesma no Brasil. É necessário estabelecer o real papel do EHV-5 na patogenia da doença. Casos de fibrose pulmonar semelhantes à EMPF em que não esteja presente o vírus, devem ser estudados a fim de ficar estabelecido se poderia ser uma forma idiopática da mesma doença.(AU)

Efectos del sistema endocannabinoide en el sistema respiratorio. Papel de los receptores TRPV1 en la fibrogénesis pulmonar / Endocannabinoid system effects on the respiratory system. The role of TRPV1 receptors in pulmonary fibrosis

Objetivos: establecer un modelo murino de fibrosis pulmonar inducida por bleomicina, investigando el posible papel protector del sistema endocannabinoide (SE) frente a la fibrosis. Métodos: se emplearon ratones salvajes (C5BL/6 y Balb/c) así como la cepa TRPV1-/-. Tras una única dosis intratraqueal de bleomicina, se analizó la respuesta fibrótica mediante un análisis histológico, la determinación de la expresión de marcadores del proceso profibrótico, el estudio de la actividad mieloperoxidasa y del contenido en hidroxiprolina del pulmón, así como el análisis de la expresión génica de VIP, PACAP, IL-1β, IL-6, TNF-α e IL-11, y el estado de activación de las rutas MAPKs (fosfo-JNK, fosfo-ERK) de la ruta de NF-κB (p-IκBα), la ruta de β-catenina y del TGFβ (GSK-3B), la activación de SMAD (pSMAD2) y pSTAT3, a nivel proteico. Resultados: la fibrosis pulmonar inducida por bleomicina en los ratones de la cepa TRPV- /- fue más severa que en la cepa salvaje C5BL/6. El contenido en hidroxiprolina y la actividad mieloperoxidasa fue mayor en los ratones TRPV1-/-. Se detectó un incremento significativo en la expresión génica de citoquinas proinflamatorias (TNF-a, IL-1b, IL11 e IL-6), pero no de VIP o PACAP, en la cepa TRPV1-/-. A nivel proteico, la expresión de pIKBα, pSTAT3, pSMAD2 y pJNK, pero no la de pERK, se vio incrementada en los ratones TRPV1-/-. Conclusiones: el modelo murino de fibrosis pulmonar inducida por bleomicina sigue siendo clave para continuar profundizando los conocimientos acerca de la patogénesis de la FPI. La modulación del SE podría tener un papel protector frente a la fibrosis pulmonar

Objectives: to establish a murine model of bleomycin-induced pulmonary fibrosis, analysing the possible protective role of the Endocannabinoid System (ES) against fibrosis. Methods: wild C5BL/6 and Balb/c mice, as well as the genetically modified strain TRPV1- /- were used. After a single dose of intratracheal bleomycin, the fibrotic response was analysed though histologic studies, the assessment of proinflammatory markers, myeloperoxidase activity, hydroxyproline content, genetic expression of VIP, PACAP, IL-1 β, IL-6, TNF-α and IL-11, as well as MAPK route (phospho-JNK, phospho-ERK), NF-κB (p-IκBα), β-cathenin, TGF-β (GSK-3B), SMAD (p-SMAD2) and pSTAT3, at a protein level. Results: pulmonary fibrosis was more severe in TRPV1-/- mice compared to C5BL/6 mice. A significant increase in proinflammatory markers such as TNF-α, IL-1β, IL11 and IL-6, but not VIP or PACAP, was observed. pIKBα, pSTAT3, pSMAD2 and pJNK, but not pERK, were increased at a protein level in TRPV-/- mice. Conclusions: the murine model of bleomycin-induced lung fibrosis remains a keystone to pioneer current investigation in lung fibrosis. Modulation of the ES might have a protective role

Antiherpetic Drugs in Equine Medicine.

Since vaccination may not prevent disease, antiherpetic drugs have been investigated for the therapy of several equine herpesviruses. Drug efficacy has been assessed in horses with disease, but most evidence is in vitro, in other species, or empirical. Oral valacyclovir is most often administered in the therapy of equine herpesvirus type-1 (EHV-1) to protect adult horses from equine herpesvirus myeloencephalopathy, while oral acyclovir is frequently administered for EHV-5 infection in the therapy of equine multinodular pulmonary fibrosis. Other antiherpetic drugs are promising but require further investigation. Several topical drugs are also empirically used in the therapy of equine viral keratitis.

Assessment of quantitative polymerase chain reaction for equine herpesvirus-5 in blood, nasal secretions and bronchoalveolar lavage fluid for the laboratory diagnosis of equine multinodular pulmonary fibrosis.

REASONS FOR PERFORMING STUDY: The ante mortem diagnosis of equine multinodular pulmonary fibrosis (EMPF) relies on histopathological results and polymerase chain reaction (PCR)-positive equine herpesvirus (EHV)-5 testing of lung tissue. Polymerase chain reaction detection of EHV-5 in bronchoalveolar lavage fluid (BALF) is commonly used to support a diagnosis of EMPF. However, the diagnostic power of EHV-5 testing on BALF and other biological samples such as blood and nasal secretions has yet to be shown to support a diagnosis of EMPF. OBJECTIVES: To determine the frequency of detection and the viral loads of EHV-5 by quantitative PCR (qPCR) in blood, nasal secretions and BALF from horses confirmed with EMPF, healthy horses and horses with non-EMPF pulmonary diseases. STUDY DESIGN: Prospective study. METHODS: The study population consisted of 70 adult horses divided into 4 groups based on a combination of clinical findings, cytology of BALF, imaging studies of the thoracic cavity and histopathology of pulmonary tissue: control group (n = 14), EMPF group (n = 11); inflammatory airway disease group (n = 32); and non-EMPF interstitial lung disease group (n = 13). For each horse, whole blood, nasal secretions and BALF were available for EHV-5 qPCR testing. Sensitivities, specificities and their respective 95% confidence intervals were calculated for viral loads from blood, nasal secretions and BALF. In addition, these measures were calculated for combined use of blood and nasal secretions. RESULTS: The detection of EHV-5 in BALF was strongly associated with EMPF (sensitivity 91%, specificity 98.3%). Detection of EHV-5 in blood was, independent of the viral loads, strongly associated with EMPF with a sensitivity of 91% and specificity of 83.1%. The detection of EHV-5 in nasal secretions displayed the highest sensitivity (72.7%) and specificity (83.1%) at a level of >245,890 glycoprotein B target genes/million cells to support a diagnosis of EMPF. Dually positive blood and nasal secretions at any viral loads in support of EMPF yielded a sensitivity and specificity of 90% and 89.8%, respectively. CONCLUSIONS: Although histopathological confirmation (lung biopsy) is considered the gold standard for EMPF diagnosis, results of qPCR testing of BALF or a combination of whole blood and nasal secretions should be regarded as clinically useful in support of this diagnosis. The latter testing may be relevant when dealing with horses in respiratory distress, for which invasive procedures such as BALF collection or lung biopsies may be detrimental to their health.