Canine Cancer Cells Activate Platelets via the Platelet P2Y12 Receptor.

In addition to their well-known functions in haemostasis, anucleated platelets have a critical role in cancer biology. Many human and non-human cancer types can directly interact with and activate platelets, promoting cancer malignancy and progression. Although naturally occurring canine neoplastic diseases mimic the biologically complex conditions of human cancers more closely than laboratory-bred mice, studies evaluating the relationship between cancer cells and platelets in dogs are scarce, and the effects of tumour cells on platelets in these animals are unknown. To evaluate whether cancer cells could activate canine platelets, we assessed the response of platelet-rich plasma to cultured canine cancer cells using light transmittance aggregometry. Similar to human and murine cancer cell research, we demonstrated that both canine osteosarcoma and mammary carcinoma cells activated canine platelets in vitro, resulting in platelet aggregation. The degree of aggregation was most pronounced at a cancer cell to platelet ratio of 1:200 for most cell lines. Mechanistic studies revealed that the platelet adenosine diphosphate (ADP) receptor P2Y12 is essential for canine platelet aggregation induced by canine cancer. ADP receptor blockage on platelets inhibited >50% of cancer cell-induced maximum platelet aggregation in all cell lines evaluated. As in other species, our results suggest that canine cancers may activate canine platelets in vivo. This mechanism is likely relevant for the biology and progression of cancer in the dog.

Proteinograma e concentração sérica de IgG em potros, do nascimento aos trinta dias de vida, tratados com plasma / Proteinogram and serum IgG concentration in newborn foals up to thirty days of life treated with plasma

Este trabalho teve por objetivo avaliar o proteinograma e concentrações séricas de IgG (após a padronização de teste ELISA) em potros do nascimento aos trinta dias de idade, antes e depois de mamarem colostro e serem tratados com plasma por via intravenosa. Foram utilizados 20 potros e suas respectivas mães, além de quatro animais doadores de plasma. Foram colhidas amostras de sangue dos potros em cinco momentos, logo após o nascimento e antes de mamar colostro (M1), dez horas após nascimento (M2), 24 horas após nascimento e previamente administração do plasma sanguíneo (M3), 48 horas de vida e 24 horas após administração do plasma sanguíneo (M4), e 30 dias após nascimento (M5). Foram colhidos sangue e colostro das éguas progenitoras no momento do parto. A concentração de proteína total (PT) e albumina foram determinadas em analisador bioquímico, a concentração de PT também foi avaliada em refratômetro manual. O fracionamento proteico foi realizado utilizando eletroforese em gel de agarose. A densidade do colostro foi avaliada com colostrômetros de refração BRIX e de densidade específica. A concentração de IgG total de todas as amostras foi determinada por teste ELISA. Com o sistema de ELISA aqui proposto foi possível determinar concentrações de IgG em amostras de soro, plasma e colostro equino com adequada repetibilidade. A média ± desvio padrão da concentração sérica de IgG dos potros ao nascer, foi de 15±8mg/dL, com dez horas de vida foi de 2.408±608mg/dL, se manteve em níveis semelhantes até 48 horas (2.364±784mg/dL) e diminuíram significativamente aos 30 dias de vida (1.414±586mg/dL). A concentração sérica e colostral de IgG nas éguas foi de 1.746±505mg/dL e 7.714±2.619mg/dL, respectivamente. A concentração plasmática de IgG dos doadores de plasma foi de 2.026±148mg/dL. Houve correlação positiva entre as concentrações séricas de IgG e PT (r=0,69 para refratômetro e r=0,76 para bioquímico), GT (r=0,81) e gamaglobulina (r=0,85). Dez horas após o nascimento foi possível verificar a transferência de imunidade passiva, possibilitando adotar medidas profiláticas e/ou terapêuticas em haras de criação de cavalos. Considerando que a proteína total, globulinas totais e fração γ-globulina apresentam correlação com IgG, estas determinações são úteis para monitorar os potros após mamarem o colostro. Um litro de plasma administrado às 24 horas de vida não foi suficiente para aumentar as concentrações séricas de IgG, 24 horas após transfusão, em potros com adequada transferência de imunidade passiva.(AU)

The aim of this study was to evaluate serum protein and serum IgG concentrations (after a direct enzyme immunoassay test ELISA optimization) in newborns foals from birth to thirty days of life before and after colostrum consumption and intravenous treatment with plasma. Twenty foals and their respective progenitors as well as four plasma donor's horses were used. Blood samples were obtained from newborn foals at five time points, immediately after birth and before colostrum intake (M1), ten hours after birth (M2), 24 hours after birth and prior administration of blood plasma (M3), 48 hours after birth and 24 hours after plasma administration (M4), and 30 days after birth (M5). Blood and colostrum samples were collected from the progenitor mares immediately postpartum. Concentration of total protein (TP) and albumin were determined using a biochemical analyzer. The TP concentration was also measured by refractometer. Fractions of total serum protein were separated using agarose gel electrophoresis. Colostrum density was evaluated using BRIX refractometer and specific density colostrometer. Total IgG concentration was determined by an enzyme-linked immunosorbent assay. With the ELISA system proposed here it was possible to determine IgG concentrations in serum, plasma, and equine colostrum samples with adequate repeatability. Serum IgG concentration in foals at birth was 15±8mg/dL (mean ± standard deviation) raising at ten hours (2,408±608mg/dL) and remaining at similar levels up to 48 hours of life (2,364±784mg/dL), and decreasing significantly at 30 days of age (1,414±586mg/dL). Serum and colostrum IgG concentrations of mares were 1,746±505mg/dL and 7,714±2,619mg/dL, respectively. The plasma IgG concentrations from donor mares were 2,026±148mg/dL. Total protein, total globulins, and γ-globulin fraction showed correlation with IgG. Ten hours post birth was an adequate time to verify the transfer of passive immunity, allowing to adoption prophylactic and/or therapeutic measures in a horse farms. One liter of plasma administered at 24 hours of life was not sufficient to raise serum IgG concentrations in foals without passive immunity transfer failure.(AU)

Applied Protein and Molecular Techniques for Characterization of B Cell Neoplasms in Horses.

Mature B cell neoplasms cover a spectrum of diseases involving lymphoid tissues (lymphoma) or blood (leukemia), with an overlap between these two presentations. Previous studies describing equine lymphoid neoplasias have not included analyses of clonality using molecular techniques. The objective of this study was to use molecular techniques to advance the classification of B cell lymphoproliferative diseases in five adult equine patients with a rare condition of monoclonal gammopathy, B cell leukemia, and concurrent lymphadenopathy (lymphoma/leukemia). The B cell neoplasms were phenotypically characterized by gene and cell surface molecule expression, secreted immunoglobulin (Ig) isotype concentrations, Ig heavy-chain variable (IGHV) region domain sequencing, and spectratyping. All five patients had hyperglobulinemia due to IgG1 or IgG4/7 monoclonal gammopathy. Peripheral blood leukocyte immunophenotyping revealed high proportions of IgG1- or IgG4/7-positive cells and relative T cell lymphopenia. Most leukemic cells lacked the surface B cell markers CD19 and CD21. IGHG1 or IGHG4/7 gene expression was consistent with surface protein expression, and secreted isotype and Ig spectratyping revealed one dominant monoclonal peak. The mRNA expression of the B cell-associated developmental genes EBF1, PAX5, and CD19 was high compared to that of the plasma cell-associated marker CD38. Sequence analysis of the IGHV domain of leukemic cells revealed mutated Igs. In conclusion, the protein and molecular techniques used in this study identified neoplastic cells compatible with a developmental transition between B cell and plasma cell stages, and they can be used for the classification of equine B cell lymphoproliferative disease.

Histopathological, immunohistochemical, and molecular study of BHV-5 infection in the central nervous system of experimentally infected calves / Estudo histopatológico, imuno-histoquímico e molecular da infecção por BHV-5 no sistema nervoso central de bovinos experimentalmente infectados

Bovine meningoencephalitis caused by BHV-5, a double-stranded DNA enveloped virus that belongs to the family Herpesviridae and subfamily Alphaherpesvirinae, is an important differential diagnosis of central nervous diseases. The aim of this study was to describe the histological changes in the central nervous system of calves experimentally infected with BHV-5 and compare these changes with the PCR and IHC results. Formalin-fixed paraffin-embedded central nervous system samples from calves previously inoculated with BHV-5 were microscopically evaluated and tested using IHC and PCR. All the animals presented with nonsuppurative meningoencephalitis. From 18 evaluated areas of each calf, 32.41% and 35.19% were positive by IHC and PCR, respectively. The telencephalon presented more accentuated lesions and positive areas in the PCR than other encephalic areas and was the best sampling area for diagnostic purposes. Positive areas in the IHC and PCR were more injured than IHC and PCR negative areas. The animal with neurological signs showed more PCR- and IHC-positive areas than the other animals.(AU)

A meningoencefalite bovina causada pelo BHV-5, um vírus DNA fita dupla envelopado que pertence à família Herpesviridae e subfamília Alphaherpesvirinae, é um importante diagnóstico diferencial das doenças do sistema nervoso central. O objetivo deste estudo foi descrever as alterações histológicas no sistema nervoso central de bovinos experimentalmente infectados com BHV-5 e comparar estas alterações com os resultados de imunoistoquímica (IHQ) e PCR. Amostras do sistema nervoso central de bezerros previamente inoculados com BHV-5 foram microscopicamente avaliadas e submetidas à IHQ e PCR. Todos os animais apresentaram meningoencefalite não-supurativa. Das 18 áreas avaliadas de cada bezerro, 32,41% e 35,13% foram positivas na IHQ e PCR, respectivamente. O telencéfalo apresentou lesões mais acentuadas e foi mais positivo na PCR do que as demais áreas encefálicas e se apresentou como a melhor área para coleta de material para o diagnóstico. As áreas positivas na IHQ e na PCR apresentaram lesões mais acentuadas do que as áreas negativas para as mesmas técnicas. O animal com sinais neurológicos apresentou mais áreas positivas para PCR e IHQ do que os demais animais.(AU)

Ocular dimensions, corneal thickness, and corneal curvature in quarter horses with hereditary equine regional dermal asthenia.

OBJECTIVES: The aim of this study was to compare ocular dimensions, corneal curvature, and corneal thickness between horses affected with hereditary equine regional dermal asthenia (HERDA) and unaffected horses. ANIMALS: Five HERDA-affected quarter horses and five healthy control quarter horses were used. METHODS: Schirmer's tear test, tonometry, and corneal diameter measurements were performed in both eyes of all horses prior to ophthalmologic examinations. Ultrasonic pachymetry was performed to measure the central, temporal, nasal, dorsal, and ventral corneal thicknesses in all horses. B-mode ultrasound scanning was performed on both eyes of each horse to determine the dimensions of the ocular structures and to calculate the corneal curvature. RESULTS: Each corneal region examined in this study was thinner in the affected group compared with the healthy control group. However, significant differences in corneal thickness were only observed for the central and dorsal regions. HERDA-affected horses exhibited significant increases in corneal curvature and corneal diameter compared with unaffected animals. The ophthalmologic examinations revealed mild corneal opacity in one eye of one affected horse and in both eyes of three affected horses. No significant between-group differences were observed for Schirmer's tear test, intraocular pressure, or ocular dimensions. CONCLUSIONS: Hereditary equine regional dermal asthenia-affected horses exhibit decreased corneal thickness in several regions of the cornea, increased corneal curvature, increased corneal diameter, and mild corneal opacity. Additional research is required to determine whether the increased corneal curvature significantly impacts the visual accuracy of horses with HERDA.

Dermatological and morphological findings in quarter horses with hereditary equine regional dermal asthenia.

BACKGROUND: Hereditary equine regional dermal asthenia (HERDA) is an autosomal recessive disorder affecting quarter horses (QHs); affected horses exhibit characteristic skin abnormalities related to abnormal collagen biosynthesis. HYPOTHESIS/OBJECTIVES: To characterize the thickness and morphological abnormalities of the skin of HERDA-affected horses and to determine the interobserver agreement and the diagnostic accuracy of histopathological examination of skin biopsies from horses with HERDA. ANIMALS: Six affected QHs, confirmed by DNA testing, from a research herd and five unaffected QHs from a stud farm. METHODS: The skin thickness in 25 distinct body regions was measured on both sides in all affected and unaffected horses. Histopathological and ultrastructural evaluation of skin biopsies was performed. RESULTS: The average skin thickness in all of the evaluated regions was thinner in the affected horses. A statistically significant difference between skin thickness of the affected and unaffected animals was observed only when the average magnitude of difference was ≥38.7% (P = 0.038). The interobserver agreement for the histopathological evaluation was fair to substantial. The histopathological sensitivity for the diagnosis of HERDA was dependent on the evaluator and ranged from 73 to 88%, whereas the specificity was affected by the region sampled and ranged from 35 to 75%. CONCLUSIONS AND CLINICAL IMPORTANCE: Despite the regional pattern of the cutaneous signs, skin with decreased thickness was not regionally distributed in the HERDA-affected horses. Histopathological evaluation is informative but not conclusive for establishing the diagnosis. Samples of skin from the neck, croup or back are useful for diagnosis of HERDA. However, the final diagnosis must be confirmed using molecular testing.

Inflamação induzida por Adjuvante de Freund: achados clínicos e efeitos sobre a expressão do RNAm da hepcidina em equinos / Freund's adjuvant-induced inflammation: clinical findings and its effect on hepcidin mRNA expression in horses

Hypoferremia observed during systemic inflammatory disorders is regulated by hepcidin. Hepcidin up-regulation is particularly important during acute inflammation, as it restricts the availability of iron, which is necessary for pathogenic microorganism growth before adaptive immunity occurs. The aim of this study was to evaluate the clinical findings and hepatic hepcidin mRNA expression in horses using a Freund's complete adjuvant (FCA) model of inflammation. The expression of hepcidin mRNA in the liver was determined in healthy horses following two intramuscular injections of FCA at 0 h and 12 h. Plasma iron and fibrinogen concentrations were measured at multiple time points between 0 h and 240 h post-FCA injection (PI). Hepcidin mRNA expression was determined by RT-qPCR using liver biopsy samples performed at 0 h (control), 6 h and 18 h PI. The mean plasma fibrinogen level was significantly different from the control values only between 120 and 216 h PI. The mean plasma iron level was significantly lower than the control between 16 and 72 h PI, reaching the lowest levels at 30 h PI (33 % of the initial value), and returned to the reference value from 96 h PI to the end of the experiment. Hepcidin mRNA expression increased at 6 h PI and remained high at 18 h PI. The iron plasma concentration was an earlier indicator of inflammatory processes in horses when compared with fibrinogen and might be useful for the early detection of inflammation in the horse. FCA administration caused the rapid onset of hypoferremia, and this effect was likely the result of up-regulated hepatic hepcidin gene expression. This study emphasizes the importance of hepcidin and iron metabolism during inflammation in horses.

A hipoferremia observada durante os processos inflamatórios sistêmicos é mediada pela hepcidina. O aumento da expressão da hepcidina é particularmente importante durante a inflamação aguda, por restringir a disponibilidade de ferro necessária para o crescimento de microrganismos patogênicos antes que a imunidade adaptativa ocorra. O objetivo deste estudo foi avaliar os achados clínicos e a expressão hepática do RNA mensageiro (RNAm) da hepcidina em cavalos após a indução da inflamação com Adjuvante completo de Freund (FCA). A expressão hepática do RNAm da hepcidina foi determinada em cavalos sadios após duas administrações intramusculares de FCA às 0 h (M0) e 12 h (M12). As concentrações plasmáticas de ferro e fibrinogênio foram mensuradas em múltiplos momentos entre 0 h e 240 h (M240) após a primeira administração de FCA (PI). A expressão do RNAm da hepcidina foi determinada por RT-qPCR usando amostras de biopsias hepáticas colhidas as 0 h (controle), 6 h (M6) e 18 h (M18) PI. A concentração plasmática média de fibrinogênio foi estatisticamente diferente do M0 entre 120 h e 216 h PI. A concentração plasmática média de ferro foi significantemente menor que o controle entre 16 h e 72 h PI, alcançou o nível mais baixo às 30 h PI (33% do valor inicial) e retornou aos valores de referência entre 96 h PI e até o final do experimento. A expressão do RNAm da hepcidina aumentou no M6 e permaneceu alta no M18. A concentração plasmática de ferro foi um indicador precoce da inflamação quando comparada com o fibrinogênio e pode ser útil na detecção precoce da inflamação em cavalos. A administração do FCA causou um rápido início da hipoferremia, e isto foi resultante do aumento da expressão hepática da hepcidina. Estes resultados enfatizam a importância da hepcidina e do metabolismo do ferro durante a inflamação em cavalos.

Allele frequency of hereditary equine regional dermal asthenia in American Quarter horses in Brazil determined by quantitative real-time PCR with high resolution melting analysis.

Hereditary equine regional dermal asthenia (HERDA) is a genetic disorder that occurs in the American Quarter horse (AQH) and is caused by a c.115G>A missense mutation in the peptidylprolyl isomerase B (PPIB) gene. Using a quantitative real-time PCR high resolution melting analysis genotyping assay for the PPIB mutation, the estimated HERDA allele and carrier frequencies in a sample of Brazilian AQHs were 2.9% and 5.8%, respectively.

Lipopolysaccharide infusion up-regulates hepcidin mRNA expression in equine liver.

Hepcidin has been found to be the key regulator of iron metabolism that leads to hypoferremia during inflammation. Recent work has shown that equine hepcidin is predominantly expressed in the liver of horses. In this study, hepcidin gene expression was determined in the liver and bone marrow of six healthy horses after iv infusion of Escherichia coli O55:B5 LPS. The IL-6 gene expression was also determined in liver and bone marrow samples. Clinical and laboratory evaluations were measured at multiple time points between 0 and 240 h post-LPS infusion (PI). Liver and bone marrow biopsies were taken immediately before (baseline) and at 6 and 18 h PI. In response to endotoxin infusion, all horses showed characteristic clinical signs of endotoxemia. Plasma iron concentration was decreased significantly from the pre-infusion level at 8 h PI. Hypoferremia peak was observed at 12 h and returned to normal levels at 30 h PI. Relative real-time RT-PCR analysis showed that liver hepcidin and IL-6 mRNA expression was up-regulated at 6 h PI. Bone marrow hepcidin relative expression was not influenced by LPS infusion. In another experiment, equine monocyte cultures were stimulated with LPS (1 µg/ml). Monocyte hepcidin and IL-6 gene expression was significantly induced after 2 h of LPS stimulus and returned to baseline levels thereafter. The present study describes that, in horses, LPS infusion up-regulates hepatic hepcidin mRNA expression resulting in early observed hypoferremia and suggests that hepcidin may act as an acute-phase protein in horses.

Influence of experimental inflammatory response on hepatic hepcidin gene expression and plasma iron concentration in sheep.

Hepcidin is a highly conserved disulfide-bonded peptide that plays a central role in iron homeostasis. During systemic inflammation, hepcidin up-regulation is responsible for hypoferremia. This study aimed to analyze the influence of the inflammatory process induced by complete Freund's adjuvant (CFA) or lipopolysaccharide (LPS) on the liver expression of hepcidin mRNA transcripts and plasma iron concentration of sheep. The expression levels of hepcidin transcripts were up-regulated after CFA or LPS. Hypoferremic response was observed at 12 h (15.46 ± 6.05 µmol/L) or 6h (14.59 ± 4.38µmol/L) and iron reached its lowest level at 96 h (3.08 ± 1.18 µmol/L) or 16h (4.06 ± 1.58 µmol/L) after CFA administration or LPS infusion, respectively. This study demonstrated that the iron regulatory hormone hepcidin was up-regulated in sheep liver in response to systemic inflammation. These findings extend our knowledge on the relationship between the systemic inflammatory response, hepcidin and iron, and provide a starting point for additional studies on iron metabolism and the inflammatory process in sheep.

Identification, characterization and expression analysis of hepcidin gene in sheep.

Hepcidin is part of the innate immune system, and it plays a central role in the regulation of iron homeostasis. This peptide has been previously characterized in man, non-human primates, rat, mouse, dog, swine, cattle, horse, fishes, reptiles and birds but until now not in sheep. The aim of this study was to sequence, characterize and perform hepcidin expression analysis in different tissues collected from healthy sheep. The resulting open reading frame consisted of 249 bp predicted to encode an 82 aa peptide with a putative 23 aa signal peptide, a 34 aa pro-region and the 25 aa mature hepcidin. The deduced sequence of the sheep hepcidin precursor was most homologous to Bos taurus and Bubalus bubalis. Hepcidin was predominantly expressed in liver, although high expression was present in abomasum and lower level expression occurred in other tissues. These findings extend our comparative knowledge showing the relationship of sheep hepcidin to other mammalian hepcidins and will be helpful for additional studies on iron metabolism and inflammatory processes in sheep.

Polioencefalomalacia experimental em bovinos induzida por toxicose por enxofre / Experimental polioencephalomalacia in cattle induced by sulfur toxicosis

O presente trabalho teve como objetivos avaliar os sinais clínicos, as concentrações do sulfeto de hidrogênio ruminal e as alterações anatomopatológicas associadas à intoxicação experimental por enxofre em bovinos. Foram utilizados dez bezerros mestiços leiteiros, sendo que quatro bovinos ingeriram ração sem sulfato de sódio (G1) e seis consumiram ração com sulfato de sódio (G2). Exames clínicos (temperatura retal, frequência cardíaca e respiratória e motricidade ruminal) e laboratoriais (hemograma, fibrinogênio, proteína plasmática, pH do fluido ruminal, concentração do sulfeto de hidrogênio ruminal, líquido cerebrospinal e histopatológico) foram realizados. A temperatura retal, frequência cardíaca, hemograma, fibrinogênio, proteína plasmática, pH do fluido ruminal e os valores do líquido cerebrospinal estavam dentro dos valores de referência para a espécie. Taquipnéia, hipomotricidade ruminal e elevados valores de sulfeto de hidrogênio ruminal foram observados nos bezerros do grupo G2. Um bezerro do grupo G2 apresentou sinais neurológicos e lesões histopatológicas de PEM. Dois animais de cada grupo foram eutanasiados. Lesões microscópicas foram observadas nos bezerros do G2. Histologicamente as alterações observadas foram necrose neuronal cortical e lesões hemorrágicas nos núcleos basais, tálamo, mesencéfalo, ponte e bulbo. O protocolo experimental constituído por uma dieta rica em carboidrato de alta fermentação, baixa quantidade de fibra efetiva e altos níveis de enxofre (0,52%) ocasionou alterações clinicas e histológicas e elevadas concentrações de sulfeto de hidrogênio ruminal compatíveis com quadro de intoxicação por enxofre.

The aims of this study were to evaluate the clinical signs, the ruminal hydrogen sulfide concentration and the histological lesions induced by sulfur toxicosis in cattle. Ten crossbred calves were fed an experimental diet, four without sodium sulfate (G1) and six with (G2). The calves were submitted to clinical (rectal temperature, cardiac and respiratory rate and ruminal motricity) and laboratorial (hemogram, fibrinogen, total plasma protein, ruminal fluid pH, ruminal hydrogen sulfide concentration, cerebrospinal fluid and histopathological) evaluations. Rectal temperature, cardiac rate, hemogram, fibrinogen, total plasma protein, ruminal fluid pH and cerebrospinal fluid values were within normal reference ranges in animals from both groups. Ruminal hypomotricity and increased respiratory rate and ruminal hydrogen sulfide concentration occurred in G2 animals. One out of six calves in G2 developed neurological signs and lesions of PEM. Two calves of each Group were euthanized. Microscopic lesions of PEM were observed in G2 animals. Histologically there were cortical neuronal necrosis and hemorrhagic lesions in basal nuclei, thalamus, midbrain, pons and medulla oblongata. The experimental model consisting of a diet with high carbohydrate and low in long fiber content with high sulfur concentrations (0.52%) resulted in clinical and histological abnormalities and high ruminal hydrogen sulfide concentration consistent with sulpur toxicosis in cattle.

Cloning, sequencing and expression analysis of the equine hepcidin gene by real-time PCR.

Equine serum or plasma iron concentration drops quickly during inflammation. Accumulation of iron inside macrophages and reduction of the intestinal absorption of this element cause hypoferremia during systemic inflammatory processes. These mechanisms are mediated by hepcidin, a 25 amino acids peptide synthesized mainly in the liver in response to iron stores and inflammation. Hepcidin is an important peptide for systemic iron homeostasis and also has antibacterial and antifungal activities. Hepcidin up-regulation is particularly useful during acute inflammation, especially before adaptive immunity occurs, restricting iron availability necessary for pathogenic microorganism growth. Hepcidin gene products have been previously characterized in man, non-human primates, rat, mouse, dog swine, cattle, fishes, reptiles and birds; but until now not in the horse. We have cloned and sequenced equine hepcidin mRNA and performed hepcidin expression analysis in different tissues collected from four healthy horses. The deduced precursor of equine hepcidin was most homologous to Bos taurus and Sus scrofa. The expressed profile of equine hepcidin in liver was very high. Expression in cervical spinal cord and cerebral cortex was much lower than liver but higher than lung, duodenum, stomach, spleen, kidney, skeletal muscle and bladder. This sequence will be helpful for additional studies on iron metabolism and inflammatory process in horses.