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Hereditary equine regional dermal asthenia (HERDA) is a genetic disease that alters collagen biosynthesis. Affected horses exhibit fragile, hyperextensible skin, especially over the dorsal region. Although ultraviolet (UV) radiation seems to contribute to the regional distribution of lesions and worsening of clinical signs, the molecular mechanisms involved are largely unknown.To evaluate the effect of solar radiation on matrix metalloproteinase MMP1, MMP8 and MMP13 gene expression in the dorsal and ventral skin of HERDA-affected and HERDA-unaffected horses [wild-type (WT) horses].Six HERDA-affected and six unaffected Quarter horses (WT) were paired according to age, sex and coat colour.Horses were submitted to 30 day sunlight restriction, followed by 15 day sunlight exposure. Dorsal and ventral skin biopsies were obtained at six sampling times over 45 days. The expression of MMP1, MMP8 and MMP13 genes was measured by quantitative PCR.Although solar radiation modulated MMP1, MMP8 and MMP13 expression, the effects were more pronounced on MMP1. Sun exposure for three days significantly upregulated MMP1 in the dorsal region when compared to the ventral skin in both unaffected and HERDA-affected horses.This study shows that solar irradiation leads to upregulation of skin collagenase genes particularly MMP1 in the dorsal, sun-exposed skin of horses. Furthermore, this was more marked in HERDA-affected horses. The increased activity of collagenases on the disorganised collagen present in HERDA affected horses would explain why UV radiation leads to deterioration of clinical signs in affected individuals.L'asthénie cutanée régionale héréditaire équine (HERDA) est une maladie génétique qui altère la biosynthèse du collagène. Les chevaux atteints présentent une peau fragile et hyperextensible, en particulier sur la région dorsale. Bien que les rayonnements ultraviolets (UV) semblent contribuer à la distribution régionale des lésions et à l'aggravation des signes cliniques, les mécanismes moléculaires impliqués sont largement méconnus.Évaluer l'effet du rayonnement solaire sur l'expression des gènes de la métalloprotéinase matricielle MMP1, MMP8 et MMP13 dans la peau dorsale et ventrale des chevaux atteints d’HERDA et non atteints [chevaux de type sauvage (WT)]. Animaux : Six Quarter horses (WT) affectés par HERDA et six Quarter horses (WT) non affectés ont été appariés en fonction de l'âge, du sexe et de la couleur du pelage. Matériels et méthodes : Les chevaux ont été soumis à une restriction solaire de 30 jours, suivie d'une exposition solaire de 15 jours. Des biopsies cutanées dorsales et ventrales ont été obtenues à six moments sur 45 jours. L'expression des gènes MMP1, MMP8 et MMP13 a été mesurée par PCR quantitative. Résultats : Bien que le rayonnement solaire module l'expression de MMP1, MMP8 et MMP13, les effets étaient plus prononcés sur MMP1. L'exposition au soleil pendant trois jours a significativement augmenté la MMP1 dans la région dorsale par rapport à la peau ventrale à la fois chez les chevaux non affectés et affectés par HERDA. Conclusions et pertinence clinique : Cette étude montre que l'irradiation solaire conduit à une régulation positive des gènes de la collagénase cutanée, en particulier MMP1 dans la peau dorsale exposée au soleil des chevaux. De plus, cela était plus marqué chez les chevaux atteints de HERDA. L'activité accrue des collagénases sur le collagène désorganisé présent chez les chevaux atteints de HERDA expliquerait pourquoi le rayonnement UV entraîne une détérioration des signes cliniques chez les individus atteints.Introducción- La astenia dérmica regional hereditaria equina (HERDA) es una enfermedad genética que altera la biosíntesis del colágeno. Los caballos afectados exhiben una piel frágil e hiperextensible, especialmente en la región dorsal. Aunque la radiación ultravioleta (UV) parece contribuir a la distribución regional de las lesiones y al empeoramiento de los signos clínicos, se desconocen en gran medida los mecanismos moleculares implicados. Objetivos- Evaluar el efecto de la radiación solar sobre la expresión génica de las metaloproteinasas de matriz MMP1, MMP8 y MMP13 en la piel dorsal y ventral de caballos afectados y no afectados por HERDA [caballos de tipo sin mutación (Wild type)] Animales- Se emparejaron seis caballos Quarter (WT) con seis caballos afectados por HERDA por edad, sexo y color del pelaje. Materiales y métodos- los caballos se sometieron a una restricción de luz solar de 30 días, seguida de una exposición solar de 15 días. Se obtuvieron biopsias de piel dorsal y ventral en seis momentos de muestreo durante 45 días. La expresión de los genes MMP1, MMP8 y MMP13 se midió mediante PCR cuantitativa. Resultados- Aunque la radiación solar moduló la expresión de MMP1, MMP8 y MMP13, los efectos fueron más pronunciados en MMP1. La exposición al sol durante tres días aumentó significativamente la MMP1 en la región dorsal en comparación con la piel ventral tanto en los caballos no afectados como en los afectados por HERDA. Conclusiones y relevancia clínica- este estudio muestra que la radiación solar conduce a una regulación positiva de los genes de colagenasa de la piel, particularmente MMP1 en la piel dorsal expuesta al sol de los caballos. Además, esto fue más marcado en los caballos afectados por HERDA. El aumento de la actividad de las colagenasas en el colágeno desorganizado presente en los caballos afectados por HERDA explicaría por qué la radiación UV conduce al deterioro de los signos clínicos en los individuos afectados.Hintergrund - Die hereditäre equine regionale dermale Asthenie (HERDA) ist eine genetische Erkrankung, welche die Kollagenbiosynthese verändert. Betroffene Pferde zeigen eine fragile, hyperextensible Haut, vor allem über der dorsalen Region. Obwohl die Ultraviolettstrahlung zur regionalen Verteilung der Veränderungen und einer Verschlimmerung der klinischen Zeichen beitragen dürfte, sind die involvierten Molekularmechanismen weitgehend unbekannt. Ziele - Eine Evaluierung der Auswirkung von Sonnenstrahlung auf die Genexprimierung der Matrix Metalloproteinasen MMP1, MMP8 und MMP13 in der dorsalen und ventralen Haut der HERDA-erkrankten und HERDA-freien Pferde [Wildtyp (WT)] Pferde. Tiere - Sechs HERDA-erkrankte und sechs gesunde Quarter Horses (WT) wurden entsprechend ihrem Alter, dem Geschlecht und ihrer Fellfarbe gepaart. Materialien und Methoden - Die Pferde durften 30 Tage nicht ins Sonnenlicht, gefolgt von einer 15-tägigen Exposition im Sonnenlicht. Es wurden dorsale und ventrale Hautbiopsien zu sechs verschiedenen Probezeiten über einen Verlauf von 45 Tagen genommen. Die Exprimierung von MMP1, MMP8 und MMP13 Genen wurde mittels quantitativem PCR gemessen. Ergebnisse - Obwohl die Sonnenstrahlung die Exprimierung von MMP1, MMP8 und MMP13 modulierte, waren die Auswirkungen auf MMP1 besonders deutlich. Eine Sonnenexposition für drei Tage erhöhte MMP1 in der dorsalen Region signifikant im Vergleich zur ventralen Haut sowohl bei an HERDA-erkrankten wie auch bei gesunden Pferden. Schlussfolgerungen und klinische Relevanz: Diese Studie zeigt, dass die Sonnenstrahlung zu einer Upregulierung der Gene der Hautkollagenasen, vor allem MMP1 in der dorsalen, Sonnen-exponierten Haut von Pferden führt. Weiters war dies bei HERDA-erkrankten Pferden deutlicher. Die erhöhte Aktivität der Kollagenasen auf das unorganisierte Kollagen bei HERDA-erkrankten Pferden würde erklären, warum die UV Strahlung zu einer Verschlechterung der klinischen Zeichen von betroffenen Individuen führt.背景: 遺伝性馬局所皮膚無力症(HERDA)は、コラーゲンの生合成を変化させる遺伝病である。患馬の皮膚は脆弱で、特に背側に過伸展を示す。紫外線は病変の局所的な分布と臨床症状の悪化に寄与していると考えられるが、その分子機構はほとんど分かっていない。 目的: 本研究の目的は、HERDA患馬とHERDA非患馬[野生型(WT)馬]の背側および腹側皮膚におけるマトリックスメタロプロテアーゼMMP1、MMP8およびMMP13遺伝子発現に対する太陽光線の影響を評価することであった。 供試動物: HERDAに罹患した馬6頭および非罹患クォーターホース(WT)6頭を、年齢、性別、毛色によってペアにした。 材料と方法:馬は30日間の日光制限を受け、その後15日間の日光照射を受けた。背側および腹側の皮膚生検は、45日間にわたり6回のサンプリングで行った。MMP1、MMP8およびMMP13遺伝子の発現を定量的PCRで測定した。 結果: 太陽光線はMMP1、MMP8、MMP13の発現を調節したが、その影響はMMP1に対してより顕著であった。3日間の日光照射は、非罹患馬とHERDA罹患馬の両方において、腹側皮膚と比較して背側領域でMMP1を有意に上昇させた。 結論と臨床的関連性: 本研究は、太陽照射が、馬の背側、太陽照射を受けた皮膚において、皮膚コラゲナーゼ遺伝子、特にMMP1のアップレギュレーションを引き起こすことを示す。さらに、このことはHERDAの罹患馬でより顕著であった。HERDA患馬に存在する無秩序なコラーゲンに対するコラゲナーゼ活性の増加は、紫外線照射が患馬の臨床症状の悪化につながる理由を説明するものである。.Introdução: A astenia dérmica regional hereditária equina (HERDA) é uma doença genética que altera a biossíntese de colágeno. Os cavalos afetados apresentam a pele frágil e hiperextensível, principalmente na região dorsal. Embora a radiação ultravioleta (UV) pareça contribuir para a distribuição regional das lesões e piora dos sinais clínicos, os mecanismos moleculares envolvidos são amplamente desconhecidos. Objetivos: Avaliar o efeito da radiação solar na expressão gênica das metaloproteinases da matriz MMP1, MMP8 e MMP13 na pele dorsal e ventral de indivíduos afetados por HERDA e não afetados por HERDA cavalos [cavalos do tipo selvagem (WT)]. Animais: Seis cavalos Quarto de Milha (WT) afetados por HERDA e seis não afetados foram pareados de acordo com idade, sexo e cor da pelagem. Materiais e métodos: Os equinos foram submetidos a 30 dias de restrição solar, seguidos de 15 dias de exposição solar. Biópsias de pele dorsal e ventral foram obtidas em seis tempos de amostragem durante 45 dias. A expressão dos genes MMP1, MMP8 e MMP13 foi medida por PCR quantitativa. Resultados: Embora a radiação solar tenha modulado a expressão de MMP1, MMP8 e MMP13, os efeitos foram mais pronunciados em MMP1. A exposição ao sol por três dias aumentou significativamente a MMP1 na região dorsal quando comparada à pele ventral em cavalos não afetados e afetados por HERDA. Conclusões e relevância clínica: Este estudo mostra que a radiação solar leva à regulação positiva dos genes da colagenase da pele, particularmente MMP1 na pele dorsal exposta ao sol de cavalos. Além disso, isso foi mais acentuado em cavalos afetados por HERDA. O aumento da atividade das colagenases sobre o colágeno desorganizado presente em cavalos afetados por HERDA explicaria porque a radiação UV leva à deterioração dos sinais clínicos nos indivíduos afetados.

The effect of a DNA immunostimulant on inflammatory and immune responses, performance, and health in calves following abrupt weaning and introduction to a concentrate diet was tested. Sixty-four single source Angus crossbred steers were weaned on day 1 and assigned to receive a DNA immunostimulant (TRT) or saline (CON) on days 0, 2, 4, and 6. On day 0, steers received clostridial and respiratory vaccines and anthelmintic; they were then transported 2 h, allocated to pens (n = 8 per pen), and introduced to total mixed ration. Daily intake, ADG, and feed efficiency were measured. Serum haptoglobin, tumor necrosis factor-alpha (TNF-α), and interleukin-1 beta (IL-1β) were assayed by ELISA or AlphaLISA on days 0, 2, 4, 6, 14, and 28; serum-neutralizing antibodies (SNA) to bovine herpesvirus-1 and bovine viral diarrhea virus-1 (BVDV-1) were quantified on days 0, 28, 68, and 135. In a subset of cattle (n = 6 to 8 per treatment group), the percent macrophages and activated gamma delta (γδ) T cells in blood was determined by flow cytometry on days 2 and 6, and expression of mRNA for TNF-α, interferon-gamma (IFN-γ), IL-4, and IL-10 by stimulated blood mononuclear cells was assessed by real-time reverse transcriptase PCR on day 6. After 70 d, cattle were shipped 1,205 km to a feedlot and performance and health were followed. There was a significant effect of time on serum TNF-α, IL-1β, haptoglobin, and SNA (P 0.10). During backgrounding, 1 TRT steer died of enterocolitis. In spite of backgrounding, cattle experienced an outbreak of bovine respiratory disease (BRD) in the feedlot and 1 of 31 TRT cattle and 5 of 32 CON cattle died of BRD. The immunostimulant modified some immune responses during backgrounding. Large variability in inflammatory responses during backgrounding indicated that events around weaning induce systemic inflammation that varies substantially among cattle.

Mutations in the CLCN1 gene are the primary cause of non-dystrophic Hereditary Myotonia in several animal species. However, there are no reports of Hereditary Myotonia in pigs to date. Therefore, the objective of the present study was to characterize the clinical and molecular findings of Hereditary Myotonia in an inbred pedigree. The clinical, electromyographic, histopathological, and molecular findings were evaluated. Clinically affected pigs presented non-dystrophic recessive Hereditary Myotonia. Nucleotide sequence analysis of the entire coding region of the CLCN1 gene revealed the absence of the exons 15 and 16 in myotonic animals. Analysis of the genomic region flanking the deletion unveiled a large intragenic deletion of 4,165 nucleotides. Interestingly, non-related, non-myotonic pigs expressed transcriptional levels of an alternate transcript (i.e., X2) that was identical to the deleted X1 transcript of myotonic pigs. All myotonic pigs and their progenitors were homozygous recessive and heterozygous, respectively, for the 4,165-nucleotide deletion. This is the first study reporting Hereditary Myotonia in pigs and characterizing its clinical and molecular findings. Moreover, to the best of our knowledge, Hereditary Myotonia has never been associated with a genomic deletion in the CLCN1 gene in any other species.

Made available in DSpace on 2020-12-12T00:58:22Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-12-01 Mutations in the CLCN1 gene are the primary cause of non-dystrophic Hereditary Myotonia in several animal species. However, there are no reports of Hereditary Myotonia in pigs to date. Therefore, the objective of the present study was to characterize the clinical and molecular findings of Hereditary Myotonia in an inbred pedigree. The clinical, electromyographic, histopathological, and molecular findings were evaluated. Clinically affected pigs presented non-dystrophic recessive Hereditary Myotonia. Nucleotide sequence analysis of the entire coding region of the CLCN1 gene revealed the absence of the exons 15 and 16 in myotonic animals. Analysis of the genomic region flanking the deletion unveiled a large intragenic deletion of 4,165 nucleotides. Interestingly, non-related, non-myotonic pigs expressed transcriptional levels of an alternate transcript (i.e., X2) that was identical to the deleted X1 transcript of myotonic pigs. All myotonic pigs and their progenitors were homozygous recessive and heterozygous, respectively, for the 4,165-nucleotide deletion. This is the first study reporting Hereditary Myotonia in pigs and characterizing its clinical and molecular findings. Moreover, to the best of our knowledge, Hereditary Myotonia has never been associated with a genomic deletion in the CLCN1 gene in any other species. São Paulo State University (UNESP) School of Veterinary Medicine and Animal Science Instituto de Medicina Veterinária Universidade Federal do Pará São Paulo State University (UNESP) Medical School Department of Pathobiology and Population Medicine College of Veterinary Medicine Mississippi State University São Paulo State University (UNESP) Institute of Bioscience College of Veterinary Medicine University of Minnesota São Paulo State University (UNESP) School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) Medical School São Paulo State University (UNESP) Institute of Bioscience

There is a clear need for low-cost, self-applied, long-lasting approaches to prevent human immunodeficiency virus (HIV) infection in both men and women, even with the advent of pre-exposure prophylaxis (PrEP). Broadly neutralizing antibodies represent an option to improve HIV prophylaxis, but intravenous delivery, cold-chain stability requirements, low cervicovaginal concentrations, and cost may preclude their use. Here, we present an approach to express the anti-GP120 broadly neutralizing antibody PGT121 in the primary site of inoculation, the female reproductive tract, using synthetic mRNA. Expression is achieved through aerosol delivery of unformulated mRNA in water. We demonstrated high levels of antibody expression for over 28 days with a single mRNA administration in the reproductive tract of sheep. In rhesus macaques, neutralizing antibody titers in secretions developed within 4 h and simian-HIV (SHIV) infection of ex vivo explants was prevented. Persistence of PGT121 in vaginal secretions and epithelium was achieved through the incorporation of a glycosylphosphatidylinositol (GPI) anchor into the heavy chain of the antibody. Overall, we present a new paradigm to deliver neutralizing antibodies to the female reproductive tract for the prevention of HIV infections., Graphical Abstract, Santangelo and colleagues report on the aerosol delivery of synthetic mRNAs, diluted in water, to the cervicovaginal epithelium, which expresses membrane-anchored broadly neutralizing antibodies against HIV. In two large animal models, anchored PGT121 was widely dispersed in the reproductive tract and was protective against a SHIV explant challenge.

WOS: 000505610200030, PubMed: 31671225, MicroRNAs modulate male fertility by regulating gene expression. In this study, dynamics of sperm miR-15a, miR-29b and miR-34a from high fertility (HF) and low fertility (LF) bulls using RT-qPCR were evaluated. Bioinformatic tools were employed to ascertain genes of interest of the sperm miRNAs. The expression levels of p53, BCL2, BAX and DNMT1 in bull spermatozoa were determined by immunoblotting. MicroRNA levels of miR-15a and miR-29 were higher in LF sires when compared with those present in HF bulls. Expression levels of miR-34a did not differ between the two groups. We found an inverse correlation between miR-15a and bull fertility. MiR29-b was also negatively associated with fertility scores. BCL2 and DNMT1 were higher in HF bulls while BAX was higher in the LF group. Our data showed a positive correlation between BCL2 and bull fertility. In addition, DNMT1 was positively associated with bull fertility. Furthermore, levels of BAX were negatively linked with bull fertility scores. Identification of miRNAs found in the spermatozoa of sires with different in vivo fertility helps understand the alterations in the fertilising capacity from cattle and other mammals. These potential biomarkers can be used in reproductive biotechnology as fertility markers to assess semen quality and predict male fertility., Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq) of BrazilNational Council for Scientific and Technological Development (CNPq); Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES) of BrazilCAPES; Higher Education Commission of Pakistan under International Research Support ProgramHigher Education Commission of Pakistan; Arab Republic of Egypt; The Republic of Turkey, Ministry of National Education, Directorate General for Higher and Foreign Education; Alta Genetics, Inc; Mississippi Agricultural Experiment Station

The interaction between platelets and tumour cells is important for tumour growth and metastasis. Thrombocytopenia or antiplatelet treatment negatively impact on cancer metastasis, demonstrating potentially important roles for platelets in tumour progression. To our knowledge, there is no information regarding the role of platelets in cancer progression in dogs. This study was designed to test whether canine platelets affected the migratory behaviour of three canine osteosarcoma cell lines and to give insights of molecular mechanisms. Intact platelets, platelet lysate and platelet releasate inhibited the migration of canine osteosarcoma cell lines. Addition of blood leucocytes to the platelet samples did not alter the inhibitory effect on migration. Platelet treatment also significantly downregulated the transcriptional levels of SNAI2 and TWIST1 genes. The interaction between canine platelets or molecules released during platelet activation and these tumour cell lines inhibits their migration, which suggests that canine platelets might antagonize metastasis of canine osteosarcoma. This effect is probably due to, at least in part, downregulation of genes related to epithelial-mesenchymal transition.

Herpesvirus bovino 5 é um alfaherpesvírus causador de meningoencefalite não supurativa em bovinos. Esta doença possui ocorrência natural em surtos ou casos isolados, associadas a baixa morbidade e alta letalidade. Embora estudos anteriores tenham elucidado aspectos relacionados a patogenia da doença, há uma lacuna de conhecimento relacionado aos eventos moleculares que contribuem para a infecção e replicação do BoHV-5. O objetivo do presente estudo foi determinar a expressão gênica in vitro de genes virais (i.e., alfa, beta e gama) e das células hospedeiras (GAPDH e 18S) durante a infecção considerando diferentes momentos de infecção e quantidade de vírus utilizado. Três genes do BoHV-5 (bICP0, UL9, US4), um gene estrutural (GAPDH) e um gene constitutivo (18S) da célula bovina tiveram suas expressões avaliadas por PCR quantitativa (qPCR). Enquanto os genes virais tiveram sua expressão aumentada ao longo do tempo de infecção, o gene hospedeiro teve sua expressão diminuída, demonstrando a ação do vírus na expressão gênica de células bovinas in vitro. O gene constitutivo 18S teve sua expressão mantida durante todos os momentos do experimento. Nossos resultados claramente demonstraram que o GAPDH não deve ser usado como gene de referência em estudos com infecção por BoHV-5 pois é influenciado pela infecção viral. Entretanto, o 18S rRNA foi constitutivamente expresso e pode ser recomendado para normalização em células bovinas infectadas pelo vírus. A expressão de mRNAviral não foi alterada pela quantidade de vírus usada. Todos os genes virais demonstraram o mesmo padrão de expressão ao longo do tempo de infecção. Nossos resultados trazem importantes diferenças comparando aos estudos clássicos que avaliaram a expressão de genes alfa, beta e gama. Mais estudos são necessários para aumentar o conhecimento da biologia molecular do BoHV-5. Estudo utilizando sequenciamento de última geração (i.e., RNA-seq), usando modelos in vitro e in vivo, aparentam ser o próximo passo lógico para acessar as alterações do transcriptoma do hospedeiro e viral ao longo do curso da infecção. Bovine herpesvirus 5 is an alphaherpesvirus that causes nonsuppurative meningoencephalitis in cattle. This disease occurs naturally in either outbreaks or isolated cases, and exhibits low morbidity and high lethality. Although previous studies elucidated crucial aspects involved in the pathogenesis of the disease, there is a paucity of information regarding the molecular events contributing to infection and replication of BoHV-5. The objective of the present study was to determine the in vitro gene expression pattern of BoHV-5 (e.g., alpha, beta, and gamma genes) and host cells genes (GAPDH and 18S) over time utilizing different quantities of inoculated virus. Three BoHV-5 genes (bICP0, UL9, US4) and one structural bovine cell gene had their expression accessed by real-time PCR. While the expression of BoHV-5 genes increased during the course of infection, GAPDH gene expression decreased in the host cells, evidencing the effect of viral infection on the expression of bovine cell genes. The 18S ribosomal RNA (rRNA) gene was constitutively expressed throughout BoHV-5 infection. Our data clearly demonstrates that GAPDH gene should not be used as a reference gene in studies of BoHV-5 infection because it was influenced by viral infection. However, 18S rRNA was constitutively expressed and, therefore, is recommended for normalization of BoHV-5 infection studies in bovine cells. The expression of viral genes transcripts was not altered by increasing number of viral particles added to the culture. All viral genes included here demonstrated the same expression pattern over time and there was no difference in the expression of viral genes among the various time points. Our data show important differences comparing to classical studies regarding herpesvirus alpha, beta, and gamma genes expression. More research is necessary to improve our understanding about the BoHV-5 biology during infection. Studies employing next-generation sequencing (i.e., RNA-seq), using both in vitro and in vivo models, would be the next logical step to grasp the virus and host cell’s transcriptome changes over the course of infection.

Made available in DSpace on 2019-10-04T12:31:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-11-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Type 1 polysaccharide storage myopathy (PSSM1) and malignant hyperthermia (MH) are autosomal dominant genetic diseases caused by a point mutation in equine GYS1 and RYR1, respectively. Although the prevalence of PSSM1 and MH have been described in other countries as causes of myopathy in horses, there are no studies evaluating the allele frequency of those mutations in Brazil. The aim of the present study was to determine the allele frequency of the mutated alleles causing susceptibility for PSSM1 and MH mutations in Quarter Horses (QHs) in Brazil. Deoxyribonucleic acid was purified from blood samples of 741 QHs used in five competitive disciplines (i.e., barrel racing, cutting, halter, racing, and reining). The PCR was optimized to identify the mutations using primers designed from the sequence of equine GYS1 and RYR1. The allele frequencies and the prevalence of PSSM1 were 0.034 and 6.7%, respectively. The highest allele frequency for the GYS1 mutated gene was identified in halter horses (0.163), and the gene mutation was not observed in racing animals. The mutation in the RYR1 was not identified in any of the sampled horses. The PSSM1 allele frequency observed in the present study indicates the relevance of the disease in the differential diagnosis of myopathies in horses. In addition, the absence of the RYR1 causative mutation suggests little importance of MH as a genetic muscle disorder in the Brazilian QHs. (C) 2018 Elsevier Inc. All rights reserved. Univ Fed Uberlandia, Sch Vet Med, Uberlandia, MG, Brazil Univ Estadual Paulista, Sch Vet Med & Anim Sci, Dept Vet Clin Sci, Botucatu, SP, Brazil Mississippi State Univ, Coll Vet Med, Dept Pathobiol & Populat Med, Starkville, MS USA Univ Estadual Paulista, Inst Biosci, Dept Microbiol, Botucatu, SP, Brazil Univ Estadual Paulista, Sch Vet Med & Anim Sci, Dept Vet Clin Sci, Botucatu, SP, Brazil Univ Estadual Paulista, Inst Biosci, Dept Microbiol, Botucatu, SP, Brazil FAPESP: 11/23729-0

Made available in DSpace on 2018-12-11T17:22:08Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-10-01 Overo lethal white foal syndrome (OLWFS) is a genetic disorder caused by a dinucleotide mutation in the endothelin receptor type B (EDNRB) gene leading to the death of affected foals shortly after birth. The use of rapid and reliable genetic testing is imperative for the early diagnosis of the mutation avoiding, therefore, either additional suffering or the production of affected animals. In the present study, we developed and validated a high-resolution melting (HRM) genotyping assay to detect the OLWFS causative mutation, and we also determined the frequency of heterozygotes among American Paint horses in Brazil. The HRM genotyping assay resulted in a high sensitivity, specificity, and positive and negative predictive values. The overall estimated frequency of heterozygotes was 21.6%; however, this frequency increased to 89.5% when considering only overo horses. The HRM assay optimized here was a reliable and suitable method for the detection of the dinucleotide mutation observed in the EDNRB gene resulting in a fast, accurate, and precise diagnostic tool. The causative gene mutation of OLWFS is present in heterozygosity in the American Paint Horse population in Brazil and is highly frequent among overo horses. Department of Veterinary Clinical Science School of Veterinary Medicine and Animal Science UNESP - Univ Estadual Paulista Department of Pathobiology and Population Medicine College of Veterinary Medicine Mississippi State University, 240 Wise Center Drive Veterinary Clinic and Surgery Department (DCCV) UFMG Veterinary College Universidade Federal de Minas Gerais UFMG Faculdade de Medicina Veterinária Universidade Federal de Uberlândia Campus Umuarama Department of Genetics Institute of Biosciences UNESP - Univ Estadual Paulista Department of Veterinary Clinical Science School of Veterinary Medicine and Animal Science UNESP - Univ Estadual Paulista Department of Genetics Institute of Biosciences UNESP - Univ Estadual Paulista

RESUMO: 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.

Made available in DSpace on 2018-11-26T17:48:19Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-03-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Glycogen branching enzyme deficiency (GBED) is a fatal autosomal inherited disease affecting horses and caused by a nonsense mutation (c.102C>A) in the GBE1 gene. This disease strongly impairs the glycogen metabolism of affected animals resulting in abortion or early death of foals. Although understanding the prevalence of heterozygous for GBED is imperative to estimate the impact of the disease, prevalence studies have yet to be performed in Brazil. The aim of this study was to determine the prevalence of heterozygous of the GBED mutation in a population of Quarter Horses in Brazil. Blood samples were obtained from 742 animals competing in distinct disciplines including cutting, halter, race, reining, and barrel racing. All samples were submitted to DNA purification, amplification, and sequencing of the target fragment. The overall GBED prevalence of carriers was 7.95% (59/742). The prevalence of heterozygotes by discipline was considered higher in cutting (19.75% [32/162]) and reining (10% [16/160]) subgroups followed by barrel racing (5% [8/160]) and halter (3% [3/100]) horses. There were none heterozygous in the racing subgroup. These results demonstrate the GBED causative mutation in the Brazilian Quarter Horse herd and suggest the occurrence of the disease. Therefore, GBED is a disease that should be considered in the differential diagnosis of abortion and stillbirths, especially if suspect foals are products of two heterozygous. Besides testing aborted foals for the GBED mutation, the molecular diagnosis should be recommended to owners and breeders, at least for those breeding the at-most-risk subgroups of horses, to guiding their mating selections and, therefore, prevent the transmission of the mutation. (C) 2017 Elsevier Inc. All rights reserved. Univ Estadual Paulista, Sch Vet Med & Anim Sci, Dept Vet Clin Sci, Botucatu, SP, Brazil Univ Fed Uberlandia, Sch Vet Med, Uberlandia, MG, Brazil Mississippi State Univ, Dept Pathobiol & Populat Med, Coll Vet Med, Starkville, MS USA Univ Estadual Paulista, Inst Biosci, Dept Microbiol, Botucatu, SP, Brazil Univ Estadual Paulista, Sch Vet Med & Anim Sci, Dept Vet Clin Sci, Botucatu, SP, Brazil Univ Estadual Paulista, Inst Biosci, Dept Microbiol, Botucatu, SP, Brazil FAPESP: 2011/23729-0

Made available in DSpace on 2018-12-11T16:49:59Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-10-01 The Ehlers-Danlos syndrome in horses is a group of genetic connective tissue disorders clinically characterized by skin fragility and hyperextensibility. To date, only two of those conditions (Hereditary Equine Regional Dermal Asthenia and Warmblood Fragile Foal Syndrome [WFFS]) have been characterized based on the causative genetic mutations. This report describes the dermatological and histological findings observed in a 3.5-year-old Mangalarga and Campolina crossbreed mare with recurrent skin wounds. Upon dermatological examination, the mare presented with hyperextensible, fragile, and thin skin areas, and scars distributed mainly along the dorsal regions. Histopathological evaluation of affected skin biopsies revealed collagen fibers abnormalities within the deep dermis. The complete PPIB coding region was amplified, but no mutations were observed. Moreover, the PLOD1 gene mutation responsible for WFFS was not present in this animal. To our knowledge, this is the first report describing a Brazilian non-Quarter horse mare with dermatological and histopathological findings of Ehlers-Danlos syndrome. School of Veterinary Medicine and Animal Science São Paulo State University (Unesp) Department of Pathobiology and Population Medicine College of Veterinary Medicine Mississippi State University Veterinary Practitioners Departamento de Veterinária Universidade Federal de Viçosa Department of Molecular Medicine Cornell University School of Veterinary Medicine and Animal Science São Paulo State University (Unesp)

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.

Made available in DSpace on 2014-10-01T13:08:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-01-01Bitstream added on 2014-10-01T14:03:54Z : No. of bitstreams: 1 S0100-736X2014000100009.pdf: 659682 bytes, checksum: 4bf40c1af2c43a561b912e0562e61948 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Pro-Reitoria of the Univ. Estadual Univ 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. 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. Universidade Estadual Paulista Faculdade de Medicina Veterinária e Zootecnia Departamento de Clínica Veterinária Universidade Federal de Goiás Escola de Veterinária Departamento de Medicina Veterinária Unesp Instituto de Biociência de Botucatu Departamento de Microbiologia e Imunologia Cornell University Universidade Estadual Paulista Faculdade de Medicina Veterinária e Zootecnia Departamento de Clínica Veterinária Unesp Instituto de Biociência de Botucatu Departamento de Microbiologia e Imunologia FAPESP: 07/07344-6 FAPESP: 07/05008-9 Pro-Reitoria of the Univ. Estadual Univ1946/009/13-PROPe/CDC

Cassia occidentalis is a bush from the Leguminosae family, subfamily Caesalpinoideae, and is a toxic plant of veterinary interest due to the occasional contamination of animal rations. This report describes the clinical and histopathological findings of an outbreak of C. occidentalis poisoning in horses. Twenty mares were poisoned after consuming ground corn contaminated with 8% of C. occidentalis seeds. Of the 20 animals affected, 12 died: 8 mares were found dead, 2 died 6 h after the onset of clinical signs compatible with hepatic encephalopathy and the 2 other animals were subjected to euthanasia 12 h after the onset of the clinical signs. The remaining 8 mares presented with mild depression and decreased appetite, but improved with treatment and no clinical sequelae were observed. In 6 animals that underwent a necropsy, an enhanced hepatic lobular pattern was noted and within the large intestine, a large number of seeds were consistently observed. Hepatocellular pericentrolobular necrosis and cerebral oedema were the main histological findings. In one mare, there was mild multifocal semimembranosus rhabdomyocytic necrosis and haemorrhage. Seeds collected from intestinal contents and sifted from the culpable feedstuff were planted. Examination of the leaves, flowers, fruits and seeds of the resultant plants identified C. occidentalis. Horses poisoned by C. occidentalis seeds demonstrate clinical signs associated with hepatoencephalopathy and frequently die suddenly. Lesions primarily involve the liver and secondarily, the central nervous system. Cassia occidentalis poisoning should be considered a differential diagnosis in horses with hepatoencephalopathy and special caution should be taken with horse rations to avoid contamination with seeds of this toxic plant.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:56:30Z No. of bitstreams: 1 S0102-09352011000300030.pdf: 140179 bytes, checksum: 9296f71faca90d4433c6fae9c04a24e3 (MD5) Made available in DSpace on 2013-08-22T18:56:30Z (GMT). No. of bitstreams: 1 S0102-09352011000300030.pdf: 140179 bytes, checksum: 9296f71faca90d4433c6fae9c04a24e3 (MD5) Previous issue date: 2011-06-01 Made available in DSpace on 2013-09-30T19:50:30Z (GMT). No. of bitstreams: 2 S0102-09352011000300030.pdf: 140179 bytes, checksum: 9296f71faca90d4433c6fae9c04a24e3 (MD5) S0102-09352011000300030.pdf.txt: 13168 bytes, checksum: 337752d64dc95cc56a7048e773521064 (MD5) Previous issue date: 2011-06-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:41:50Z No. of bitstreams: 2 S0102-09352011000300030.pdf: 140179 bytes, checksum: 9296f71faca90d4433c6fae9c04a24e3 (MD5) S0102-09352011000300030.pdf.txt: 13168 bytes, checksum: 337752d64dc95cc56a7048e773521064 (MD5) Made available in DSpace on 2014-05-20T13:41:50Z (GMT). No. of bitstreams: 2 S0102-09352011000300030.pdf: 140179 bytes, checksum: 9296f71faca90d4433c6fae9c04a24e3 (MD5) S0102-09352011000300030.pdf.txt: 13168 bytes, checksum: 337752d64dc95cc56a7048e773521064 (MD5) Previous issue date: 2011-06-01 É descrito o carcinoma de células escamosas (CCE) no fundo vaginal de uma vaca. O diagnóstico de CCE moderadamente diferenciado foi confirmado através do exame histopatológico. Os testes imunoistoquímicos com os marcadores p53 e Ki67 realizados em amostras do tumor confirmaram a mutação na p53 e aumento da proliferação celular. Faculdade de Medicina Veterinária e Zootecnia - UNESP, Campus de Botucatu Faculdade de Medicina Veterinária e Zootecnia - UNESP, Campus de Botucatu

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 6 h (14.59 ± 4.38 μmol/L) and iron reached its lowest level at 96 h (3.08 ± 1.18 μmol/L) or 16 h (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.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:50:56Z No. of bitstreams: 1 S0100-736X2011000100007.pdf: 3278844 bytes, checksum: f9e09c4b9efeeba682c205596d3fbfcd (MD5) Made available in DSpace on 2013-08-22T18:50:56Z (GMT). No. of bitstreams: 1 S0100-736X2011000100007.pdf: 3278844 bytes, checksum: f9e09c4b9efeeba682c205596d3fbfcd (MD5) Previous issue date: 2011-01-01 Made available in DSpace on 2013-09-30T18:18:03Z (GMT). No. of bitstreams: 2 S0100-736X2011000100007.pdf: 3278844 bytes, checksum: f9e09c4b9efeeba682c205596d3fbfcd (MD5) S0100-736X2011000100007.pdf.txt: 70398 bytes, checksum: ee986a3b54e2b3c4f50d9521b40a7a02 (MD5) Previous issue date: 2011-01-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:33:47Z No. of bitstreams: 2 S0100-736X2011000100007.pdf: 3278844 bytes, checksum: f9e09c4b9efeeba682c205596d3fbfcd (MD5) S0100-736X2011000100007.pdf.txt: 70398 bytes, checksum: ee986a3b54e2b3c4f50d9521b40a7a02 (MD5) Made available in DSpace on 2014-05-20T13:33:47Z (GMT). No. of bitstreams: 2 S0100-736X2011000100007.pdf: 3278844 bytes, checksum: f9e09c4b9efeeba682c205596d3fbfcd (MD5) S0100-736X2011000100007.pdf.txt: 70398 bytes, checksum: ee986a3b54e2b3c4f50d9521b40a7a02 (MD5) Previous issue date: 2011-01-01 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. Universidade Federal de Goiás (UFG) Escola de Veterinária Depto Medicina Veterinária Universidade Estadual Paulista Faculdade de Medicina Veterinária e Zootecnia Departamento de Clínica Veterinária Universidade Estadual Paulista Faculdade de Medicina Veterinária e Zootecnia Departamento de Clínica Veterinária

Made available in DSpace on 2018-11-26T15:28:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-11-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) 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. Cornell Univ, Coll Vet Med, Dept Clin Sci, Ithaca, NY 14853 USA Univ Estadual Paulista, Dept Vet Clin Sci, Sch Vet Med & Anim Sci, Sao Paulo, Brazil Cornell Univ, Coll Vet Med, Dept Populat Med & Diagnost Sci, Ithaca, NY 14853 USA Cornell Univ, Coll Vet Med, Dept Biomed Sci, Ithaca, NY 14853 USA Univ Estadual Paulista, Dept Vet Clin Sci, Sch Vet Med & Anim Sci, Sao Paulo, Brazil FAPESP: 2010/08774-7

Made available in DSpace on 2015-08-26T19:19:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-05-01. Added 1 bitstream(s) on 2015-08-31T13:04:58Z : No. of bitstreams: 1 S0103-84782015000500854.pdf: 250535 bytes, checksum: d8e6c556effb6a40d3095d349c5ccf07 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) A paralisia periódica hipercalêmica (HYPP) é uma das principais enfermidades genéticas de caráter dominante que acometem cavalos da raça Quarto de milha (QM). A HYPP é causada por uma mutação pontual no gene SCN4A e, apesar de estar presente nos cavalos QM no Brasil, dados sobre a prevalência da HYPP são escassos. O objetivo deste trabalho foi verificar a prevalência da mutação responsável pela HYPP em cavalos QM, utilizados nas modalidades esportivas de rédeas (n=160), apartação (n=160), tambor e baliza (n=160), corrida (n=160) e conformação (n=101). Foram utilizados DNA sanguíneo dos 741 equinos; o teste genético para enfermidade foi padronizado e as amostras sequenciadas para identificação da mutação no gene alvo. A prevalência de HYPP na população amostrada foi de 4,2%, sendo que somente na linhagem de conformação foram identificados animais positivos (30,7%). Medidas de controle mais efetivas devem ser adotadas para diminuir a prevalência da HYPP. Hyperkalemic Periodic Paralysis (HYPP) is one of the major dominant genetic diseases which affect Quarter horses (QH). The HYPP is caused by a point mutation in the SCN4A gene and despite the presence of HYPP in Brazilian QH, limited data on the disease prevalence are available. The aim of this study was to investigate the HYPP mutation in QH belonging to reining (n=160), cutting (n=160), barrel racing (n=160), racing (n=160) and halter (n=101) competitive disciplines. Blood DNA from 741 horses were used. Genetic tests were standardized and samples were sequenced to identify the mutation on the target gene. The prevalence of HYPP on the sampled population was 4.2% and the positive animals (30.7%) were only identified in the halter lineage. More effective actions on HYPP control should be done to reduce the disease prevalence. Universidade Federal de Uberlândia Faculdade de Medicina Veterinária Universidade Estadual Paulista Júlio de Mesquita Filho Faculdade de Medicina Veterinária e Zootecnia UNESP Instituto de Biociências de Botucatu UNESP FMVZ Departamento de Clínica Veterinária FAPESP: 2011/23729-0

Made available in DSpace on 2015-10-21T13:09:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-04-01. Added 1 bitstream(s) on 2015-10-22T09:53:11Z : No. of bitstreams: 1 S0100-736X2015000400337.pdf: 409488 bytes, checksum: 55affbb4b281ddd22ce343505f842431 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) 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. 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. Univ Estadual Paulista Unesp, Fac Med Vet &Zootecnia, Dept Clin Vet, BR-18618970 Botucatu, SP, Brazil Univ Fed Goias, Escola Vet Zootecnia, Dept Med Vet, BR-74001970 Goiania, Go, Brazil FMVZ Unesp, Dept Higiene Vet &Saude Publ, Botucatu, SP, Brazil Univ Estadual Londrina, Dept Vet Prevent Med, Ctr Ciencias Agr, Lab Virol Anim, BR-86057970 Londrina, PR, Brazil Univ Estadual Paulista Unesp, Fac Med Vet &Zootecnia, Dept Clin Vet, BR-18618970 Botucatu, SP, Brazil FMVZ Unesp, Dept Higiene Vet & Saude Publ, Botucatu, SP, Brazil FAPESP: 2008/07697-9

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.

Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:29:49Z No. of bitstreams: 0 Made available in DSpace on 2014-05-27T11:29:49Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-07-01 Two Quarter Horse mares with hereditary equine regional dermal asthenia (HERDA) were diagnosed with metastatic squamous cell carcinoma (SCC) associated with chronic nonhealing wounds. The lesions were similar to the development of SCC from chronic nonhealing ulcers, known as Marjolin's ulcers in humans. The horses showed recurrent skin wounds in the saddle and paralumbar regions and were confirmed by molecular techniques as having HERDA. Both horses were maintained as research animals for prolonged periods and received regular veterinary care and wound treatment. Both horses were ultimately euthanized because of their chronic progressive wounds, coupled with declining health. At necropsy, the nonhealing wounds were found to be complicated by infiltrative SCC; both horses had metastasis to lungs. Chronically inflamed, recurrent skin wounds that heal slowly and incompletely as a consequence of HERDA are proposed as a major pathogenetic factor in tumorigenesis. Consistent findings with respect to proliferation index (Ki-67) and mutations of p53 tumor suppressor gene were confirmed by immunohistochemistry in one horse. SCC consistent with Marjolin's ulcer has been previously suggested in association with chronic ulcers or burn scars in horses, but this is the first report of an association with chronic poor healing wounds in HERDA horses. © 2013 Elsevier Inc. Department of Veterinary Clinical Science School of Veterinary Medicine and Animal Science Universidade Estadual Paulista (UNESP), Botucatu, São Paulo Department of Large Animal Clinical Sciences College of Veterinary Medicine Michigan State University, East Lansing, MI Department of Pathobiology and Population Medicine Mississippi State University, Mississippi State, MS Department of Veterinary Medicine School of Veterinary Medicine Universidade Federal de Goiás (UFG), Goiânia, Goiás Department of Small Animal Clinical Sciences College of Veterinary Medicine Michigan State University, East Lansing, MI Department of Veterinary Medicine Universidade Federal de Viçosa (UFV), Viçosa, Minas Gerais Department of Clinical Sciences Mississippi State University, Mississippi, MS Department of Veterinary Clinical Science School of Veterinary Medicine and Animal Science Universidade Estadual Paulista (UNESP), Botucatu, São Paulo

Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:30:03Z No. of bitstreams: 0Bitstream added on 2014-05-27T14:43:25Z : No. of bitstreams: 1 2-s2.0-84880335347.pdf: 2379162 bytes, checksum: db53290b788805d40796a665b355e5a3 (MD5) Made available in DSpace on 2014-05-27T11:30:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-07-25 Introduction. Tendon injury is a major cause of lameness and decreased performance in athletic equines. Various therapies for tendonitis have been described; however, none of these therapies results in complete tissue regeneration, and the injury recurrence rate is high even after long recovery periods involving rest and physiotherapy. Methods. A lesion was induced with collagenase gel in the superficial digital flexor tendon in the center portion of the metacarpal region of eight equines of mixed breed. After two weeks, the lesions of the animals in the treated and control groups were treated through the intralesional administration of mesenchymal stem cells derived from adipose tissue (adMSCs) suspended in platelet concentrate (PC) and with phosphate buffered saline (PBS), respectively. Serial ultrasound analyses were performed every two weeks. After 16 weeks of therapy, a biopsy was performed for histopathological, immunohistochemical and gene expression (type I collagen (COL1A1), type III collagen (COL3A1), tenascin-C (TNC), tenomodulin (TNMD), and scleraxis (SCX)) analyses. Results: Differences in the ultrasound and histopathological analyses were observed between the groups. Improved results were reported in the group treated with adMSCs suspended in PC. There was no difference in the gene expression levels observed after the different treatments. The main results observed from the histopathological evaluation of the treated group were as follows: a prevention of the progression of the lesion, a greater organization of collagen fibers, and a decreased inflammatory infiltrate. A lack of progression of the lesion area and its percentage was observed in the ultrasound image, and increased blood flow was measured by Power Doppler. Conclusions: The use of adMSCs combined with PC for the therapy of experimentally induced tendonitis prevented the progression of the tendon lesion, as observed in the ultrasound examination, and resulted in a greater organization and decreased inflammation, as observed in the histopathological evaluation. These data demonstrate the therapeutic potential of this therapy for the treatment of equine tendonitis. © 2013 Carvalho et al.; licensee BioMed Central Ltd. Department of Veterinary Surgery and Anesthesiology School of Veterinary Medicine and Animal Science São Paulo State University Rubião Junior, 18618-970 Botucatu, São Paulo Department of Veterinary Clinic School of Veterinary Medicine and Animal Science São Paulo State University Rubião Junior, 18618-970 Botucatu, São Paulo Department of Animal Reproduction and Veterinary Radiology School of Veterinary Medicine and Animal Science São Paulo State University Rubião Junior, 18618-970 Botucatu, São Paulo Blood Center Botucatu Medical School São Paulo State University Rubião Junior, 18618-970 Botucatu, São Paulo Department of Veterinary Surgery and Anesthesiology School of Veterinary Medicine and Animal Science São Paulo State University Rubião Junior, 18618-970 Botucatu, São Paulo Department of Veterinary Clinic School of Veterinary Medicine and Animal Science São Paulo State University Rubião Junior, 18618-970 Botucatu, São Paulo Department of Animal Reproduction and Veterinary Radiology School of Veterinary Medicine and Animal Science São Paulo State University Rubião Junior, 18618-970 Botucatu, São Paulo Blood Center Botucatu Medical School São Paulo State University Rubião Junior, 18618-970 Botucatu, São Paulo

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.

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.

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.

RESUMO A análise do líquido cefalorraquidiano (LCR) é uma importante ferramenta no diagnóstico das doenças neurológicas dos bovinos. A coleta do LCR em diferentes momentos facilita o monitoramento do quadro clínico e/ou a avaliação da eficácia de tratamentos estabelecidos. Todavia, os efeitos de coletas consecutivas sobre os parâmetros citológicos e bioquímicos do LCR de bovinos saudáveis ainda não foram analisados. Dessa forma, o objetivo do presente estudo foi avaliar potenciais diferenças citológicas e bioquímicas no LCR de bovinos sadios, obtidos em dois momentos, com intervalo de 96 horas. O LCR foi submetido à análise citológica e à determinação do teor de proteína total. Na segunda coleta, verificou-se aumento significativo da celularidade, porém ainda dentro do intervalo de referência para a espécie, diminuição de linfócitos e elevação de células mononucleares em relação à primeira coleta. No entanto, os números de hemácias (/µL), células nucleadas (/µL), neutrófilos (%), eosinófilos (%) e a concentração da proteína total (mg/dL) continuaram dentro dos valores de referência 96 horas após a realização da primeira punção. Conclui-se que os procedimentos de coleta de LCR consecutivos não provocaram inflamação local detectável 96 horas após a primeira punção, o que sugere que, quando necessário, coletas de LCR seriadas, por si só, não interfeririam nos resultados do exame. ABSTRACT The analysis of cerebrospinal fluid (CSF) is important for the diagnosis of neurological diseases in cattle. However, the effect of consecutive collections on the cytological and biochemical parameters of the CSF has not been evaluated yet. The objective of the present study was to evaluate potential differences on the cytological and biochemical analysis of CSF obtained from healthy cattle at two sampling times with 96 hours of interval in between. Total and differential cell counts, and total protein concentration were analyzed in the CSF. Increased cellularity and significant reduction of lymphocyte and elevated numbers of mononuclear cells were observed on the second collection. However, erythrocyte count (/µl), total leukocyte count (/µl), neutrophils (%), eosinophils (%) and total protein concentration (mg/dl) remained within the reference range 96 hours after the completion of the first puncture. In conclusion, consecutive CSF collections cause no detectable local inflammation 96 hours after the first puncture suggesting that repeated collections do not interfere in the CSF results.