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This study investigated the effect of different prenatal nutrition approaches in 126 pregnant Nellore cows on reproductive and nutrigenetic traits of the male offspring during the finishing phase. For that purpose, three nutritional treatments were used in these cows during pregnancy: PP - protein-energy supplementation in the final third, FP - protein-energy supplementation during the entire pregnancy, and NP - (control) only mineral supplementation. The male progeny (63 bulls; 665 ± 28 days of age) were evaluated for scrotal circumference, seminal traits, number of Sertoli cells and testicular area. We performed a genomic association (700 K SNPs) for scrotal circumference at this age. In addition, a functional enrichment was performed in search of significant metabolic pathways (P 0.05) with inclusion of genes that are expressed in these genomic windows by the MetaCore software. With the exception of major sperm defects (P 0.1), the other phenotypes showed no difference between prenatal treatments. We found genes and metabolic pathways (P 0.05) that are associated with genomic windows (genetic variance explained1%) in different treatments. These molecular findings indicate that there is genotype-environment interaction among the different prenatal treatments and that the FP treatment showed greater major sperm defects compared to the NP treatment.

To address the restrictions caused by the COVID-19 pandemic and to search for assistive learning tools for the subject of Animal Anatomy II and Applied Anatomy, 123 anatomical kits were prepared at the Animal Anatomy Laboratory for students of the Veterinary Medicine course at the University of São Paulo, Faculty of Animal Science and Food Engineering (FZEA/USP) in Pirassununga city, São Paulo, Brazil. The kits contained anatomical pieces for teaching splanchnology and topographic anatomy (two different classes), and they were elaborated based on effective preservation techniques for the preparation of animal anatomical pieces. At the end of each course, we sent an online questionnaire to the students for evaluation of the methodology used. Alternative methods were used to minimize the odour and non-generation of chemical or microbiological contaminants. The acceptance of the kits was unanimous with adherence by all the students, who had the opportunity to experience the Anatomy class in its entirety, without leaving their homes.

Made available in DSpace on 2021-06-25T11:03:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Culture conditions regulate the process of pluripotency acquisition and self-renewal. This study aimed to analyse the influence of the in vitro environment on the induction of porcine induced pluripotent stem cell (piPSCs) differentiation into primordial germ cell-like cells (pPGCLCs). piPSC culture with different supplementation strategies (LIF, bFGF, or LIF plus bFGF) promoted heterogeneous phenotypic profiles. Continuous bFGF supplementation during piPSCs culture was beneficial to support a pluripotent state and the differentiation of piPSCs into pPGCLCs. The pPGCLCs were positive for the gene and protein expression of pluripotent and germinative markers. This study can provide a suitable in vitro model for use in translational studies and to help answer numerous remaining questions about germ cells. Graphical abstract: [Figure not available: see fulltext.] Department of Animal Reproduction School of Veterinary Medicine and Animal Science University of São Paulo Department of Veterinary Medicine School of Animal Sciences and Food Engineering Department of Pharmacology Institute of Biosciences São Paulo State University (UNESP) Department of Surgery Faculty of Veterinary Medicine and Animal Sciences University of Sao Paulo Department of Biochemistry Paulista School of Medicine Federal University of São Paulo (UNIFESP) Department of Preventive Veterinary Medicine and Animal Reproduction Faculty of Agricultural and Veterinary Sciences São Paulo State University (UNESP) Department of Animal Science Faculty of Animal Science and Food Engineering University of São Paulo Department of Pharmacology Institute of Biosciences São Paulo State University (UNESP) Department of Preventive Veterinary Medicine and Animal Reproduction Faculty of Agricultural and Veterinary Sciences São Paulo State University (UNESP)

Encephalic vascular accident, or stroke, is the most common pathology of the central nervous system in humans. It is the second leading cause of death, as well as physical and cognitive disabilities, in developing countries. It is a vascular disorder that may present in an ischemic (more common) or hemorrhagic form. Ozone is a gas capable of oxidizing double bonds of organic molecules, thereby producing lipoperoxides and aldehydes. Stimulation of the immune and antioxidant system, and improvement in tissue vascularization and oxygenation, are few among several effects exerted by ozone. Ozone therapy has previously been shown to be effective in neuromodulation, neuroprotection, and nerve regeneration. The present study aimed to evaluate the effect of targeted mild ozone after inducing cerebral ischemia in vitro. The experiment was divided into two steps; in the first step, neuroblastoma lineage cells (SH-SY5Y) were subjected to 24 hours of hypoxia in an incubator culture chamber. Treatment with different concentrations of ozone (2–10 µg/mL), followed by an 2,5-diphenyl-2H-tetrazolium bromide assay, indicated a possible neuroregenerative effect at low concentrations. The same protocol was applied to canine amniotic membrane stem cells that were evaluated via colorimetric assay spectrophotometry, fluorescence microscopy, and flow cytometry. The metabolic conditions and cellular regeneration in cells at low ozone concentrations (3–8 µg/mL) correlated with lower levels of apoptosis and oxidative stress compared to cells not subjected to hypoxia. High concentrations of ozone (18–30 µg/mL) promoted an increase in rate of apoptosis and cell death. Therefore, we developed a novel protocol that mimics ozone therapy for ischemic stroke, using ozonized culture medium after hypoxia induction. Although more studies are needed to open new avenues for translational medicine, we conclude that ozone has a dose-dependent hormetic effect and can reverse the effect of ischemia in vitro at low concentrations.

Purpose Amniotic membrane stem cells have a high capacity of proliferation, cell expansion, and plasticity, as well as immunomodulatory properties that contribute to maternal-fetal tolerance. Owing to the lack of research on human amniotic membrane at different gestational stages, the canine model is considered ideal because of its genetic and physiological similarities. We aimed to characterize the canine amniotic membrane (CAM) cell lineage in different gestational stages and evaluate the expression of immunomodulatory genes. Materials and Methods Twenty CAMs from early (20-30 days) (n=7), mid- (31-45 days) (n=7), and late gestation (46-63 days) (n=6) stages were studied. The cell features were assessed by cell viability tests, growth curve, colony-forming units, in vitro differentiation, cell labeling for different immunophenotypes, and pluripotent potential markers. The cells were subjected to RT-PCR and qPCR analysis to determine the expression of IDO, HGF, EGF, PGE2, and IL-10 genes. Results CAM cells exhibited a fibroblastoid morphology and adherence to plastic with an average cell viability of 78.5%. The growth curve indicated a growth peak in the second passage and we obtained an average of 138.2 colonies. Osteogenic, chondrogenic, and adipogenic lineages were confirmed by in vitro differentiation assays. Cellular immunophenotyping experiments confirmed the presence of positive mesenchymal markers (CD90 and CD105) and the low or negative expression of hematopoietic markers (CD45 and CD34). Qualitative analysis of the immunomodulatory functions indicated the expression of the IDO, HGF, EGF5, and PGE2 genes. When stimulated by interferon-gamma, CAM cells exhibited higher IDO levels throughout gestation. Conclusion The CAMs from different gestational stages presented features consistent with mesenchymal stem cell lineage; better results were observed during the late gestation stage. Therefore, the gestational stage is a key factor that may influence the functionality of therapies when using fetal membrane tissues from different periods of pregnancy.

Testicular degeneration by heat is the leading cause of infertility in bulls. Beef cattle are generally farmed under hot and humid conditions, and consequently, the thermotolerance of each breed must be considered in their natural environment. This study aimed to evaluate the reproductive characteristics of Brahman bulls maintained in the grazing system, with or without shadow availability. Ten Brahman bulls aging between 24 and 30 months were allocated in two different paddocks, with or without shadow availability. The heat tolerance test was performed on three non-consecutive typical summer days. The semen samples were collected at four times points in a 14 days interval. The climate conditions were monitored throughout the experiment; and clinical evaluation, testicular consistence and scrotal circumference were measured before every semen collection. In addition, semen was evaluated regarding volume, aspect, turbulence, motility, straight movement, sperm concentration, and morphological exam. The studied Brahman bulls showed a high thermolysis capacity, high heat tolerance, and no differences in semen quality were observed between groups. A degeneração testicular causada pelo calor é a principal causa de infertilidade em touros. Bovinos de corte geralmente são criados em condições de calor e umidade, e, consequentemente, a termotolerância de cada raça deve ser considerada em seu ambiente natural. O presente trabalho teve como objetivo avaliar as características reprodutivas de touros da raça Brahman mantidos em sistema de pastejo, com ou sem disponibilidade de sombra. Dez touros Brahman com idades entre 24 e 30 meses foram alocados em dois piquetes diferentes, com ou sem disponibilidade de sombra. O teste de tolerância ao calor foi realizado em três dias típicos de verão não consecutivos. As amostras de sêmen foram coletadas em quatro momentos em intervalos de 14 dias. As condições climáticas foram monitoradas durante todo o período experimental; e a avaliação clínica, consistência testicular e a circunferência escrotal foram avaliadas antes de cada coleta de sêmen. Ainda, o sêmen foi avaliado quanto ao volume, aspecto, turbulência, motilidade, vigor, concentração espermática e exame morfológico. Os touros estudados da raça Brahman apresentaram alta capacidade de termólise, alta tolerância ao calor, e não foram observadas diferenças na qualidade do sêmen entre os grupos.

Made available in DSpace on 2021-06-25T12:34:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-11-26 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) The cellular reprogramming into pluripotency is influenced by external and internal cellular factors, such as in vitro culture conditions (e.g., environmental oxygen concentration), and the aging process. Herein, we aimed to generate and maintain equine iPSCs (eiPSCs) derived from fibroblasts of a horse older than 20 years and to evaluate the effect of different levels of oxygen tension (atmospheric 20% O-2, 5% O-2, or 20% to 5% O-2) on these cells. Fibroblasts were reprogrammed, and putative eiPSCs were positive for positive alkaline phosphatase detection; they were positive for pluripotency-related genes OCT4, REX1, and NANOG; immunofluorescence-positive staining was presented for OCT4 and NANOG (all groups), SOX2 (groups 5% O-2 and 20% to 5% O-2), and TRA-1-60, TRA-1-81, and SSEA-1 (only in 20% O-2); they formed embryoid bodies; and there is spontaneous differentiation in mesoderm, endoderm, and ectoderm embryonic germ layers. In addition to the differences in immunofluorescence analysis results, the eiPSC colonies generated at 20% O-2 presented a more compact morphology with a well-defined border than cells cultured in 5% O-2 and 20% to 5% O-2. Significant differences were also observed in the expression of genes related to glucose metabolism, mitochondrial fission, and hypoxia (GAPDH, GLUT3, MFN1, HIF1 alpha, and HIF2 alpha), after reprogramming. Our results show that the derivation of eiPSCs was not impaired by aging. Additionally, this study is the first to compare high and low oxygen cultures of eiPSCs, showing the generation of pluripotent cells with different profiles. Under the tested conditions, the lower oxygen tension did not favor the pluripotency of eiPSCs. This study shows that the impact of oxygen atmosphere has to be considered when culturing eiPSCs, as this condition influences the pluripotency characteristics. Sao Paulo State Univ, Fac Agr & Vet Sci, Dept Pathol Reprod & One Hlth, BR-14884900 Sao Paulo, Brazil Univ Sao Paulo FZEA USP, Fac Anim Sci & Food Engn, Dept Vet Med, BR-13635900 Sao Paulo, Brazil Univ Sao Paulo FMVZ USP, Sch Vet Med & Anim Sci, Dept Anim Reprod, BR-13635900 Sao Paulo, Brazil Univ Montreal, Fac Vet Med, Ctr Rech Reprod & Fertilite, Montreal, PQ J2S 2M2, Canada Sao Paulo State Univ, Fac Agr & Vet Sci, Dept Pathol Reprod & One Hlth, BR-14884900 Sao Paulo, Brazil FAPESP: 2015/26818-5 FAPESP: 2018/04009-6 FAPESP: 2013/08135-2-CEPID-CTC

Background Induced pluripotent stem cells (iPSCs) have enormous potential in developmental biology studies and in cellular therapies. Although extensively studied and characterized in human and murine models, iPSCs from animals other than mice lack reproducible results. Methods Herein, we describe the generation of robust iPSCs from equine and bovine cells through lentiviral transduction of murine or human transcription factors Oct4, Sox2, Klf4, and c-Myc and from human and murine cells using similar protocols, even when different supplementations were used. The iPSCs were analyzed regarding morphology, gene and protein expression of pluripotency factors, alkaline phosphatase detection, and spontaneous and induced differentiation. Results Although embryonic-derived stem cells are yet not well characterized in domestic animals, generation of iPS cells from these species is possible through similar protocols used for mouse or human cells, enabling the use of pluripotent cells from large animals for basic or applied purposes. Herein, we also infer that bovine iPS (biPSCs) exhibit similarity to mouse iPSCs (miPSCs), whereas equine iPSs (eiPSCs) to human (hiPSCs). Conclusions The generation of reproducible protocols in different animal species will provide an informative tool for producing in vitro autologous pluripotent cells from domestic animals. These cells will create new opportunities in animal breeding through transgenic technology and will support a new era of translational medicine with large animal models.

International audience; Gene editing in large animal models for future applications in translational medicine and food production must be deeply investigated for an increase of knowledge. The mitochondrial transcription factor A (TFAM) is a member of the HMGB subfamily that binds to mtDNA promoters. This gene maintains mtDNA, and it is essential for the initiation of mtDNA transcription. Lately, we generated a new cell line through the disruption of the TFAM gene in bovine fibroblast cells by CRISPR/Cas 9 technology. We showed that the CRISPR/Cas9 design was efficient through the generation of heterozygous mutant clones. In this context, once this gene regulates the mtDNA replication specificity, the study aimed to determine if the post-edited cells are capable of in vitro maintenance and assess if they present changes in mtDNA copies and mitochondrial membrane potential after successive passages in culture. The post-edited cells were expanded in culture, and we performed a growth curve, doubling time, cell viability, mitochondrial DNA copy number, and mitochondrial membrane potential assays. The editing process did not make cell culture unfeasible, even though cell growth rate and viability were decreased compared to control since we observed the cells grow well when cultured in a medium supplemented with uridine and pyruvate. They also exhibited a classical fibroblastoid appearance. The RT-qPCR to determine the mtDNA copy number showed a decrease in the edited clones compared to the non-edited ones (control) in different cell passages. Cell staining with Mitotracker Green and red suggests a reduction in red fluorescence in the edited cells compared to the non-edited cells. Thus, through characterization, we demonstrated that the TFAM gene is critical to mitochondrial maintenance due to its interference in the stability of the mitochondrial DNA copy number in different cell passages and membrane potential confirming the decrease in mitochondrial activity in cells edited in heterozygosis.

The veterinarian has to frequently assess blood tests in equine medicine. The reference values are adapted from international literature, based on examinations of healthy horses, under a standardized climate and physical activity. This study aimed to compare the blood values obtained from sports horses and traction horses, taking into account the type of activity performed. Anamnesis and hematology data from 76 animals were used: 32 members belonged to the Traction Group (TG), corresponding to equines from traction activity (cart horses) and 44 members belonged to the Sport Group (SG), corresponding to equines used in obstacle jumping, both groups were from the Pirassununga region, State of São Paulo. Complete blood counts and plasmatic fibrinogen dosage were evaluated. The animals from the TG had lower erythrocyte count and hematocrit values. The animals from the TG had greater leukocyte values in the leukogram compared to the animals from the SG. Cart horses were found to have lower means of hematological values compared to the jumping horses. Moreover, these values were below the normal physiological range for the species. Keywords: correlation; hematological evaluation; physical activity; horses.

The impact of growth rate (GR) and finishing regime (FR) on growth and meat quality traits of Angus x Nellore crossbred steers, harvested at a constant body weight (530 ± 20 kg) or time on feed (140 days), was evaluated. Treatments were: 1) feedlot, high GR; 2) feedlot, low GR; 3) pasture, high GR and 4) pasture, low GR. Live body composition, carcass and meat quality traits were evaluated. High GR had greater impact on muscle and fat deposition in feedlot-finished, but not in pasture-finished animals. Feedlot animals had higher Longissimus muscle area, backfat thickness, meat luminosity and tenderness when compared to pasture groups. Moreover, pasture- and feedlot-finished animals with similar GR did not differ in the chromatic attributes of non-aged meat, regardless of endpoint. Thus, GR appeared to be the main factor driving beef chromatic parameters, while FR had a major impact on achromatic attributes and tenderness of meat.

Purpose Amniotic membrane stem cells have a high capacity of proliferation, cell expansion, and plasticity, as well as immunomodulatory properties that contribute to maternal-fetal tolerance. Owing to the lack of research on human amniotic membrane at different gestational stages, the canine model is considered ideal because of its genetic and physiological similarities. We aimed to characterize the canine amniotic membrane (CAM) cell lineage in different gestational stages and evaluate the expression of immunomodulatory genes. Materials and Methods Twenty CAMs from early (20–30 days) (n=7), mid- (31–45 days) (n=7), and late gestation (46–63 days) (n=6) stages were studied. The cell features were assessed by cell viability tests, growth curve, colony-forming units, in vitro differentiation, cell labeling for different immunophenotypes, and pluripotent potential markers. The cells were subjected to RT-PCR and qPCR analysis to determine the expression of IDO, HGF, EGF, PGE2, and IL-10 genes. Results CAM cells exhibited a fibroblastoid morphology and adherence to plastic with an average cell viability of 78.5%. The growth curve indicated a growth peak in the second passage and we obtained an average of 138.2 colonies. Osteogenic, chondrogenic, and adipogenic lineages were confirmed by in vitro differentiation assays. Cellular immunophenotyping experiments confirmed the presence of positive mesenchymal markers (CD90 and CD105) and the low or negative expression of hematopoietic markers (CD45 and CD34). Qualitative analysis of the immunomodulatory functions indicated the expression of the IDO, HGF, EGF5, and PGE2 genes. When stimulated by interferon-gamma, CAM cells exhibited higher IDO levels throughout gestation. Conclusion The CAMs from different gestational stages presented features consistent with mesenchymal stem cell lineage; better results were observed during the late gestation stage. Therefore, the gestational stage is a key factor that may influence the functionality of therapies when using fetal membrane tissues from different periods of pregnancy.

Nutritional stimuli during the gestational period in dams have long-term effects on the offspring in terms of health and production rates. This study assessed the effect of fetal programming in 126 pregnant Nellore cows on reproductive and nutrigenetic traits of the progeny during the rearing phase. For that purpose, three nutritional treatments were used in these cows during pregnancy: PP – protein-energy supplementation in the final third, FP – protein-energy supplementation during the entire pregnancy, and NP – (control) only mineral supplementation. The male progeny (64 bulls) was evaluated for scrotal circumference and seminal traits at 12, 15, and 18 months of age. In addition, we performed a genomic association (35 K SNPs) for scrotal circumference at 12 and 18 months of age. Only the total sperm defects showed significant difference between groups (P

The neonatal period is a challenging phase for calves, and during this phase constant adaptations are required. The aim of the present study was to evaluate the invasive hemodynamics with the Swan-Ganz catheter in neonate calves to understand adaptive changes during the first 30 days of life. A prospective and observational study was conducted with 10 Holstein calves. Assessments of the right atrial pressure (RAP), right ventricular pressure (RVP), pulmonary artery pressure (PAP), pulmonary capillary pressure (PW), cardiac output (CO), heart rate (HR), pulmonary vascular resistance (PVR), and blood gas levels were performed. The analyses of PAP, PVR, PW, HR, sO2, and arterial blood gases differed (p &lt, 0.05) between the evaluated periods. Our results indicated transient pulmonary artery hypertension during the process of extrauterine adaptation during the first 30 days of life. This hypertension must be considered as physiological and consequent to the neonatal adaptation process.

Noninvasive collection of cells used for generation of induced pluripotent stem cells (iPSC) generation would simplify its use for regenerative and reproductive purposes in veterinary medicine, although it is still unpublished in species other than humans. This study aimed to derive urine progenitor cells (UPCs) invitro from urine samples collected from swine, and then to reprogram them into iPSCs. For that, urine samples were collected from three females, and cells were isolated and cultured from each following the human UPCs protocol (Steichen et al. 2017 Curr. Protoc. Hum. Genet. 21, 7.1-21.7.22; https://doi.org/10.1002/cphg.26). Approximately 200mL of urine samples were collected in sterile flasks and centrifuged at 300×g; the pellet was washed in Dulbecco's PBS, resuspended, and cultured in 45% Dulbecco's modified Eagle's medium (DMEM)-high glucose, 5% fetal bovine serum (FBS), 50% Renal Epithelial Cell Growth basal medium (REBM) supplemented with 1% glutamine, 1% nonessential amino acids (NEAA), penicillin/streptomycin (P/S), and REGM supplements hEGF, insulin, hydrocortisone, GA-1000, FBS, transferrin, triiodothyronine, epinephrine (Lonza), and 10ngmL−1 basic fibroblast growth factor (bFGF). The UPCs first colonies were observed approximately 1 week after and resembled epithelial-like cells. At passage 2, one cell line was transduced with murine OCT4, SOX2, KLF4, and C-MYC cDNAs (OSKM) using a lentiviral vector. After 5 days, cells were plated onto mouse embryonic fibroblasts and cultured in knockout DMEM/F12, 20% knockout serum replacement, NEAA, L-glutamine, 2-mercaptoethanol, and P/S supplemented with 10ngmL−1 bFGF. Efficiency of reprogramming was 8.45%, measured by analysing the number of typical iPSC colonies relative to the transduced cells plated, after ~12 days. Three clonal lines (C1, C4, and C6) were maintained invitro and characterised regarding pluripotency markers for at least 30 passages. All three lines were positive for alkaline phosphatase activity in passages 15 and also 22. Immunocytochemistry analysis revealed that C6 (passage 22) was positive for the pluripotency genes OCT4 (1:100, SC), SOX2 (1:500, AB), SSEA1 (1:50, SC), TRA1- 81 (1:50, Millipore), and NANOG (1:100, AB), whereas C1 and C4 (passages 23 and 22, respectively) were positive only for OCT4, SOX2, and SSEA1. The expression of exogenous and endogenous pluripotency factors (OCT4, SOX2, and NANOG) was evaluated by qRT-PCR, comparing the three clonal lines at passages 16/17 and 21/22 and comparing different passages (10, 11, 14, 17, and 22) on C1 line. No statistical difference was observed between cell lines when compared in different passages, perhaps because of the great variation between lines. However, analysis of C1 line over time showed that pluripotency genes increased and exogenous vector expression decreased during early passages (±10 passages); however, after passage 17, OCT4 and NANOG decreased whereas SOX2 and exogenous vector expression increased. In conclusion, it was possible to reprogram UPCs into iPSCs and maintain them in culture for at least 30 passages; however, iPSCS were dependent on exogenous factors. These results represent the partial reprogramming of UPCs to iPSCs in animals for the first time, enabling the generation of invitro disease models using a noninvasive method. Financial support for this study was received from FAPESP (2019/02811-2, 2015/26818-5), CNPq (433133/2018-0), and CAPES.

This review aim to present some clinical problems found in IVP-derived animals focusing on NT procedures and to discuss the possible role of epigenetics in such process. Also, as cell-secreted vesicles have been reported as possible regulators of important physiological reproductive processes such as folliculogenesis and fertilization, it is also presented herein a new perspective of manipulating the pre-implantation period trough effector molecules contained in such vesicles. Nesta revisão, apresentamos alguns problemas clínicos encontrados nos animais derivados de PIV, principalmente derivados de transferência de núcleo, e discutimos o possível papel da epigenética em tais processos. Além disso, uma vez que vesículas secretadas por células têm sido descritas como possíveis reguladores de processos reprodutivos fisiológicos importantes, tais como a foliculogênese e a fertilização, estas são aqui apresentadas como uma possível nova ferramenta para a manipulação do período embrionário pré-implantacional através de moléculas efetoras, contidas em tais vesículas.

RESUMO: As afecções ortopédicas com perda de tecido ósseo são um desafio tanto na medicina veterinária quanto na medicina humana. Portanto, não é raro ortopedistas se depararem com fraturas cominutivas irredutíveis de ossos longos, neoplasias ósseas ou não-uniões, que necessitam de procedimentos cirúrgicos reparadores, por meio da substituição de segmento ou preenchimento de falha óssea com o uso de biomateriais. Pretende-se com esta pesquisa avaliar e comparar a resistência mecânica entre biomateriais naturais, sintéticos e mistos. Foram utilizados sete grupos experimentais compostos por seis corpos de provas cada: Grupo 1 , tecido ósseo cortical de coelho (OSSO - controle); Grupo 2, cimento ósseo (CO); Grupo 3, cimento ósseo autoclavado (COA); Grupo 4, cimento ósseo e macrofragmento ósseo cortical de cão (COMaFO); Grupo 5, cimento ósseo e macrofragmento ósseo autoclavado de cortical de cão (COMaFOA); Grupo 6, cimento ósseo e microfragmento ósseo cortical de cão (COMiFO) e Grupo 7, cimento ósseo e microfragmento ósseo cortical de cão (COMiFOA). Os corpos de prova foram submetidos a ensaios mecânicos de compressão axial controlada em máquina universal de ensaio Emic®. O teste era interrompido quando ocorria queda brusca na curva do gráfico indicando falência da amostra. Em relação à Força máxima, os grupos COA, COMaFOA e COMiFOA não diferiram estatisticamente do grupo controle (OSSO; p>0,01). Já os grupos CO, COMaFO e COMiFO diferiram estatisticamente do grupo controle (OSSO; p0,01). Já os grupos CO, COA, COMaFO e COMiFO diferiram estatisticamente do grupo controle (OSSO; p0,01). Já os grupos CO, COA e COMiFOA diferiram estatisticamente do grupo controle (OSSO; p 0.01). Already the CO groups, COMaFO and COMIFO difeririram statistically the control group (BONE; p 0.01). Already the CO groups, COA, COMaFO and COMIFO differed significantly from the control group (BONE; p0.01). Already the CO groups, COA and COMiFOA differed significantly from the control group (BONE; p

Resumo: Na clínica médica de equinos, explora-se o perfil hematológico do animal, geralmente, com a finalidade de encontrar alterações que não foram constatadas ao exame clínico. A pesquisa de hematozoários em equinos, muitas vezes, apresenta resultados conflitantes entre o quadro clínico apresentado pelo animal e o resultado laboratorial, levantando a hipótese de que a técnica de pesquisa de hematozoários seja a responsável por falhas diagnósticas. Este estudo visa comparar os valores obtidos em exames hematológicos de 15 equinos de esporte e 15 equinos de tração (carroceiros), levando-se em consideração diferenças como características nutricionais, estado de higidez e tipo de atividade realizada, e comparar as diferentes técnicas de pesquisa de hematozoários, como esfregaço sanguíneo e PCR. Verificou-se que apenas os equinos de tração apresentaram valores médios de hemácias, hematócrito e hemoglobina abaixo do considerado fisiológico para a espécie, embora 100% dos animais, de ambos os grupos experimentais, tenham sido considerados positivos para hemoparasitoses por PCR. Verifica-se a superioridade do método de pesquisa de hemoparasitas por PCR, em comparação com esfregaço sanguíneo, realizado por diferentes técnicas, visto que apenas 33,3% dos animais foram considerados positivos para Theileria equi por esta técnica, enquanto que o PCR revelou 100% de positividade, para Theileria equi, Babesia caballi e infecção mista. Nenhum dos animais estudados foi diagnosticado com Anaplasma phagocytophilum (Ehrlichia equi) e Ehrlichia risticcii (Neoricketsia risticii). Verifica-se, então, que muitos dos diagnósticos de ausência de hemoparasitose por exame hematológico e ou esfregaço sanguíneo são errôneos, devido à baixa sensibilidade da técnica e podem repercutir em falha no tratamento ou disseminação dos hemoparasitos e das hemoparasitoses. Ressalta-se, então, a importância de exames como o PCR na elaboração de diagnóstico definitivo. Abstract: The blood profile is usually explored in equine medicine to find changes not detected through clinical examination. The haematozoa search in horses, often presents conflicting results between the clinical picture presented by the animal and the laboratory result, raising the hypothesis that the hematozoa search technique is responsible for diagnostic failures. Thus, this study aims to compare the values obtained from blood tests in 15 sports horses and 15 traction horses (cart horses), taking into account differences such as nutritional characteristics, state of health and type of activity performed, and to compare the different techniques with hematozoa search, as blood smear and PCR. It was found that only the traction horses showed mean values of red blood cells, hematocrit and hemoglobin below the considered physiological for the species, although 100% of the animals of both experimental groups were considered positive for hemoparasitoses by PCR. It was verified the superiority of hemoparasites search method by PCR, compared with blood smear performed by different techniques, since only 33.3% of the horses were considered positive for Theileria equi by this technique, while the PCR showed 100% positive for Theileria equi, Babesia caballi and mixed infection. None of the animals studied was diagnosed with Anaplasma phagocytophilum (Ehrlichia equi) and Ehrlichia risticcii (Neoricketsia risticii). It is shown that many cases of hemoparasitosis absence of diagnostic by hematological exams and or blood smears are erroneous due to the low sensitivity of the technique and may impact on treatment failure or spread of blood parasites and hemoparasitoses. It is noteworthy to stres the importance of tests such as PCR for the definitive diagnosis.

Distemper disease is an infectious disease reported in several species of domestic and wild carnivores. The high mortality rate of animals infected with canine distemper virus (CDV) treated with currently available therapies has driven the study of new efficacious treatments. Mesenchymal stem cell (MSC)-based therapy is a promising therapeutic option for many degenerative, hereditary, and inflammatory diseases. Therefore, the aim of this study was to characterize stem cells derived from the canine fetal olfactory epithelium and to assess the systemic response of animals infected with CDV to symptomatic therapy and treatment with MSCs. Eight domestic mongrel dogs (N = 8) were divided into two groups: support group (SG) (N = 5) and support group + cell therapy (SGCT) (N = 3), which were monitored over 15 days. Blood samples were collected on days 0, 6, 9, 12, and 15 to assess blood count and serum biochemistry (urea, creatinine, alanine transferase, alkaline phosphatase, gamma-glutamyl transferase, total protein, albumin, and globulin), and urine samples were obtained on days 0 and 15 for urinary evaluation (urine I). The results showed a high mortality rate (SG = 4 and SGCT = 2), providing inadequate data on the clinical course of CDV infection. MSC therapy resulted in no significant improvement when administered during the acute phase of canine distemper disease, and a prevalence of animals with high mortality rate was found in both groups due to the severity of symptoms.

This study aimed to verify the influence of the transport in open or closed compartments (0h), followed by two resting periods (1 and 3h) for the slaughter process on the levels of cortisol as a indicative of stress level. At the slaughterhouse, blood samples were taken from 86 lambs after the transport and before slaughter for plasma cortisol analysis. The method of transport influenced in the cortisol concentration (0h; P

Background The selection of beef cattle for feed efficiency (FE) traits is very important not only for productive and economic efficiency but also for reduced environmental impact of livestock. Considering that FE is multifactorial and expensive to measure, the aim of this study was to identify biological functions and regulatory genes associated with this phenotype. Results Eight genes were differentially expressed between high and low feed efficient animals (HFE and LFE, respectively). Co-expression analyses identified 34 gene modules of which 4 were strongly associated with FE traits. They were mainly enriched for inflammatory response or inflammation-related terms. We also identified 463 differentially co-expressed genes which were functionally enriched for immune response and lipid metabolism. A total of 8 key regulators of gene expression profiles affecting FE were found. The LFE animals had higher feed intake and increased subcutaneous and visceral fat deposition. In addition, LFE animals showed higher levels of serum cholesterol and liver injury biomarker GGT. Histopathology of the liver showed higher percentage of periportal inflammation with mononuclear infiltrate. Conclusion Liver transcriptomic network analysis coupled with other results demonstrated that LFE animals present altered lipid metabolism and increased hepatic periportal lesions associated with an inflammatory response composed mainly by mononuclear cells. We are now focusing to identify the causes of increased liver lesions in LFE animals. Electronic supplementary material The online version of this article (doi:10.1186/s12864-015-2292-8) contains supplementary material, which is available to authorized users.

Embryonic pluripotent stem cells (ESC) and induced pluripotent stem cells (iPSC) were reported capable of differentiating into primordial germ cell-like (PGCL) and functional gametes in vitro in the murine model (Hikabe et al. 2016 Nature 539, 299-303). The in vitro generation of primordial germ cells (PGC) and gametes from farm animals would greatly contribute to enhance animal production technologies and to the creation of adequate models for several disorders. The present study aimed at the generation of PGC in vitro (iPGC) from iPSC in cattle and their characterisation through pluripotency and germ cell markers. For that, bovine iPSC previously generated and characterised (Bressan et al. 2015 Reprod. Fertil. Dev. 27, 254) were submitted to in vitro differentiation into epiblast-like cells (EpiLC) and iPGC by the protocol adapted from mice (Hayashi et al. 2011 Cell 146, 519-532). The biPS cells were induced into EpiLC by culture in fibronectin-coated (16.7 µg mL−1) 6-well plates in N2B27 culture medium supplemented with 20 ng mL−1 activin A, 12 ng mL−1 basic fibroblast growth factor (bFGF), and 1% knockout serum replacement (KSR) for 48 h and further differentiated into iPGC by non-adherent culture (Agreewell plates, StemCell Technologies, Vancouver, BC, Canada) with GK15 medium (GMEM supplemented with 15% KSR, 0.1 mM nonessential amino acids, 1 mM sodium pyruvate, 0.1 mM 2-mercaptoethanol, 2 mm l-glutamine, and 1% antibiotics) in the presence of 500 ng mL−1 BMP4, 100 ng mL−1 SCF, 500 ng mL−1 BMP8b, and 50 ng mL−1 epidermal growth factor for 4 days. The cells were then characterised regarding morphology, detection of alkaline phosphatase, immunofluorescence for OCT4, DDX4, VASA, and c-Kit proteins, and transcripts of pluripotency-related genes OCT4 and SOX2, as well as of imprinted genes (H19, SNRPN) and imprinted-related (DNMT1, DNMT3B) genes were analysed through RT-qPCR and compared with constitutive genes GAPDH, NAT1, and ACTB. Alkaline phosphatase and immunofluorescence analysis were positive for all specific markers. Interestingly, although OCT4 and SOX2 expression was present in iPS, EpiLC, and iPGC, this last group presented greater OCT4 and lesser SOX2 transcript amounts compared with other groups, suggesting, as expected, that PGC are still pluripotent but may already be differentiating into germ-cell lineages. The expression of H19 was increased in iPGC, whereas the expression of SNRPN was decreased only in the fibroblast group, potentially indicating epigenetic reprogramming process in these cells. Expression of DNMT1 and DNMT3B was not different between pluripotent groups but subtly increased when compared with that in fibroblasts. The results obtained herein represent an important first step in the in vitro generation of PGC and gametes from domestic farm animals, an unprecedented and desirable tool for enhancing new reproductive technologies and providing new understanding of cellular reprogramming and pluripotent germ cell biology. Financially supported by FAPESP grants 2013/08135-2, 2013/13686-8, 2015/26818-5; CNPq 482163/2013-5.

No Brasil, assim como em outros países tropicais, a criação de bovinos é feita de maneira extensiva e, portanto é suscetível às intempéries climáticas. Esse tipo de criação pode levar ao estresse térmico, mesmo em animais zebuínos. Sob condições de estresse térmico é possível a ocorrência de degeneração testicular, e por isso, há a procura por animais tolerantes ao calor. Desta maneira, este trabalho objetivou a avaliação da tolerância ao calor de touros da raça Brahman utilizando o teste de tolerância ao calor, e também a análise da qualidade seminal relacionada à disponibilidade de sombra nos pastos. Para tal, foram utilizados 10 touros da raça Brahman com idades entre 24 e 30 meses. Duas semanas antes do início das análises, três amostras de sêmen foram colhidas de cada animal para o nivelamento biológico do sêmen, e então os touros foram separados em dois grupos: 5 touros foram alocados em pasto com disponibilidade de sombra enquanto 5 touros foram alocados em pasto sem qualquer tipo de sombra. O teste de tolerância ao calor foi realizado em três dias consecutivos típicos de verão, e as colheitas de sêmen foram realizadas em 14 dias durante 2 meses, totalizando 4 colheitas. Anteriormente a todas as colheitas foram avaliadas a consistência testicular e o perímetro escrotal. As características seminais avaliadas foram volume, aspecto, turbilhonamento, motilidade, vigor e concentração e morfologia espermáticas. Os dados obtidos foram analisados com o pacote estatístico Statistical Analysis System (SAS Institute Inc., 2004). Não houve diferenças estatísticas entre os dois grupos de animais. O resultado do teste de tolerância ao calor mostrou que os animais testados são tolerantes ao calor e apresentam boa capacidade termolítica. In Brazil, similarly to others tropical countries, bovines are bred under environmental conditions of extensive system. This breed system type may lead to heat stress even on zebu animals. Under heat stress conditions, testicular degeneration can occur and, therefore, there is a search for animals that are heat tolerant. Therefore, the aim of this work was to evaluate the heat tolerance of Brahman bulls using the heat tolerance test and to verify the occurrence of any possible effect of shadow availability on pasture on semen quality. Ten Brahman bulls aging between 24 and 30 months were used in this work. For biological semen evaluation, three semen samples were collected from each animal two weeks before beginning. The bulls were then separated into two groups: 5 animals were allocated on pasture with shadow availability and 5 bulls were allocated on pasture without any kind of shadow. The heat tolerance test were performed in three consecutive typical summer days and the semen samples were collected each 14 days during 2 months, in a total of 4 semen samples per animal. Testicular consistence and scrotal circumference were measured just before every semen collection. The semen\'s characteristics evaluated were volume, aspect, mass movement, motility, straight movement, sperm concentration and morphological exam. Data obtained from experimental proceedings were analyzed by Statistical Analysis System program (SAS Institute Inc., 2004). No difference was observed (P>0.05) in any of the characteristics analyzed when Brahman bulls were maintained on pastures with or without shadow availability. The performance of tested animals on the heat tolerance test shows that these animals are heat tolerant and present good thermolitic ability.

Background Treatment for horses with pythiosis of a limb is challenging. This study aims to evaluate the effects of administering amphotericin B in a 10 % solution of dimethylsulfoxide by intravenous regional limb perfusion (IRLP) to treat horses for cutaneous pythiosis of a limb. Results All 15 of the horses treated had complete resolutions of their lesion between 6 to 9 weeks after a single IRLP treatment. No complications were observed at the site of venipuncture for IRLP. Before initiation of treatment, there was anemia and marked leucocytosis which resolved following treatment. Serum biochemistry showed no significant changes. Conclusions IRLP administration of amphotericin B in a 10 % DMSO solution was easily performed, relatively inexpensive and an effective treatment for treating horses for pythiosis of a limb and resolved the infection with no complications.

As principais causas de morte perinatal em bezerros estão de alguma forma relacionadas a problemas cardiorrespiratórios. A maior parte de morte de bezerros ocorre justamente no período de até 48h após o nascimento, quando ainda estão sendo feitos os ajustes fisiológicos necessários à vida extrauterina. Com o aumento de animais advindos das técnicas de produção in vitro, também aumentaram os problemas de sobrevivência e sobrevida dos recém nascidos. Este estudo teve como objetivo geral padronizar valores de referência de hemogasometria e espirometria de bezerros a termo, nascidos por parto espontâneo, e prematuros, nascidos por cesariana, da raça Nelore concebidos por inseminação artificial (IA), assim como estabelecer protocolos de cuidados intensivos neonatais aos animais prematuros. Objetivamente, o estudo compreendeu: 1. A avaliação dos valores de pressão invasiva de átrio direito, ventrículo direito, artéria pulmonar e artéria pulmonar ocluída, hemogasometria, espirometria e capnografia de bezerros da raça Nelore, a termo, nascidos por parto espontâneo; 2. A avaliação e padronização da hemogasometria arterial de bezerros raça Nelore, a termo, nascidos de partos espontâneos; 3. A avaliação da pressão invasiva da artéria pulmonar e dos efeitos do sildenafil sobre os valores de pressão de artéria pulmonar em bezerros da raça Nelore, prematuros, nascidos por cesariana; 4. Diferenças hemodinâmicas (utilizando o cateter de Swan-Ganz), na função pulmonar e nas trocas gasosas entre bezerros a termo nascidos por parto espontâneo e prematuros nascidos por cesariana; 5. O estabelecimento de protocolos de cuidados neonatais para bezerros advindos de gestações complicadas com risco de morte neonatal prematura, onde foram realizadas 3 cesarianas em bezerros concebidos por IA e 16 concebidos por transferência nuclear por de células somática (clonagem), sendo estabelecidos protocolos de terapia neonatal com sildenafil, surfactante e oxigenoterapia. Bezerros da raça Nelore a termo nascidos por parto espontâneo estabilizam seus parâmetros de hemogasometria a valores de animais adultos em até 48 horas após o nascimento e têm valores de hemogasometria mais próximos dos valores normais para a espécie, enquanto os bezerros prematuros, nascidos por cesariana, têm valores mais deslocados em relação a normalidade. A padronização da hemogasometria de bezerros da raça Nelore nascidos por eutocia, pode fomentar as pesquisas com animais advindos de técnicas de PIV, assim como aumentar a sobrevida e sobrevivência além de diminuir a mortalidades desses animais. Já o sildenafil não apresenta efeitos adversos significativos quanto a oxigenação dos bezerros em que foi usado, mesmo quando houve alterações não desejadas da PAP. Dessa forma, quando não existe a possibilidade de se aferir a PAP e suspeita-se de hipertensão, o sildenafil é pode ser usado sem maiores consequências para a saúde do paciente. The main causes of perinatal death in cattle are frequently related to cardiorespiratory distress. A great number of losses are placed during the first 48h after birth, when important physiological adjustments to the extra-uterine life are still in progress. Recent increase of offspring derived from assisted reproductive in vitro biotechnologies resulted in diminished survival rates of the newborn. The present study aimed to obtain standard values of hemodynamic evaluation, blood gas and spirometry of Nelore calves conceived by artificial insemination (AI) born at term, spontaneously, or from premature calves born by cesarean delivery, as well as to establish protocols of neonatal intensive care focusing on premature animals. Objectively, the study comprised: 1. The evaluation of invasive pressure values from right atrium, right ventricle, pulmonary artery and pulmonary wedge pressure, blood gas, spirometry and capnography of Nelore calves born spontaneously; 2. Evaluation and standardization of arterial blood gas of Nelore calves born at term spontaneously; 3. Evaluation of invasive pressure of pulmonary artery (PAP) and effects of sildenafil administration on its values from Nelore premature calves born by cesarean; 4. The comparison of hemodynamic, blood gas and spirometry of calves born at term spontaneously or premature calves born by cesarean delivery, during the first 30 days of life; 5. The establishment of neonatal care protocols focusing on complicated pregnancies at risk of premature neonatal death, when 3 cesarean sections of calves conceived by AI and 16 conceived by nuclear transfer of somatic cells (cloning) were monitored, and therefore protocols involving neonatal therapy with sildenafil, surfactant and oxygen therapy were established. Nelore calves born at term spontaneously have their blood gas stabilized in values similar to adult cattle until 48 hours after birth, and present a blood gas profile more similar to the expected for the species, whereas premature calves born from cesarean delivery present values different from normality. The standardization of blood gas of Nelore calves born at term from eutocic deliveries may promote better perinatal survival rates in offspring derived from in vitro production. The treatment with sildenafil does not present adverse effects related to the oxygen levels of calves even when adverse alterations were observed in PAP. Therefore, whenever hypertension is suspected and measurement of PAP is impaired, the sildenafil treatment may be used without major consequences to the patient health

Background: Skin lesions are the most common clinical complaints in animals. Thus non-neoplastic skin lesions in horses are under diagnosed, especially when laboratory resources are needed in field. Nevertheless the eosinophilic granulomas in mules are poorly reported in the international literature, including laboratory analysis and diagnosis, differential diagnosis, treatment and the evolution of the case. Case: A 17-year-old mule was brought to our veterinary hospital with hard skin nodular lesions, which were irregular in shape and diameter. Body temperature was normal, and no alopecia, itching, erythema or pains were observed. Cytological examination by fine needle aspiration was done and the treatment was based on systemic dexamethasone for nine days and intralesional triamcinolone. Discussion: The cytological examination by fine needle aspiration revealed a high cellularity of eosinophils, and the histopathology analysis confirmed the diagnosis of eosinophilic granuloma. Complete regression of the lesions was achieved after one week of treatment. Although previously reported in horses, eosinophilic granuloma is still poorly documented in mules. The treatment with systemic dexamethasone and intralesional triamcinolone acetonide was effective and lead to total remission.

O projeto Carroceiro, coordenado e idealizado pela Profa. Dra. Renata Gebara Sampaio Doria, iniciou suas atividades no ano de 2011 e, desde entao, vem contribuindo com muitas familias de Pirassununga e regiao que utilizam equideos para tracao e nao possuem condicoes financeiras para fornecer a seus animais uma assistencia medica veterinaria. O trabalho objetiva a conscientizacao dos proprietarios dos equideos utilizados em carrocas (carroceiros) sobre: zoonoses, saude publica, manejo, bem-estar e promocao da saude desses animais, que sao utilizados para tracao como fonte de subsistencia por muitas familias. Alem de promover a saude e bem-estar dos animais, bem como a conscientizacao dos carroceiros, o projeto auxilia os estudantes de Medicina Veterinaria da Faculdade de Zootecnia e Engenharia de Alimentos da Universidade de Sao Paulo (FZEA-USP) a desenvolver e aprimorar suas habilidades clinico-cirurgicas no tratamento de equideos, complementando a formacao profissional.

This study aimed to evaluate the possible renal and hepatic toxicity of tepoxalin administered before or after isoflurane-induced hypotension, as well as for five consecutive days. Twelve healthy mixed-breed cats, adult males, weighing 4.0±0.8kg were allocated into two groups. They received 25mgkg-1 of tepoxalin orally, two hours before the anesthetic procedure (PRE) or after the procedure (POST) and daily for five days. Cats were anesthetized with isoflurane and the concentration was increased until mean arterial pressure reached 40-60mmHg and kept at this level for 60 minutes. During hypotension, the physiological variables were measured at time 0 and every 10 minutes until 60 minutes, and bleeding time was measured at time 0, 30 and 60 minutes. Blood samples were drawn for a hemogram and determination of concentrations of alanine aminotransferase, alkaline phosphatase, urea, creatinine and Na+ at baseline, 24 hours, 48 hours and 7 days post-hypotension. Urine was collected at baseline, 24 hours, 48 hours and 7 days post-hypotension for determination of concentrations of creatinine, gamma-glutamyltransferase, urine specific gravity, protein, albumin and Na+. During the anesthetic procedure there were no important variations in physiological variables and bleeding time. There were differences only in fractional excretion of Na+, which was elevated at 7 days of evaluation in PRE and in the urine protein/creatinine ratio in PRE, which was higher than in POST at 24 and 48 hours post-hypotension. We conclude that tepoxalin does not cause alterations in hepatic enzymes but can cause discrete renal injury, resulting in proteinuria, in cats subjected to 60min of hypotension. Este estudo teve como objetivo a avaliação da possível toxicidade renal e hepática da tepoxalina administrada antes ou após hipotensão induzida por isofluorano, assim como a sua administração nos cinco dias seguintes à hipotensão. 12 gatos adultos hígidos, machos, sem raça definida e com peso de 4,0±0,8kg foram alocados em dois grupos (n=6). Os animais receberam 25mgkg-1 de tepoxalina pela via oral, duas horas antes do procedimento anestésico (PRE) ou após o procedimento (POST) e diariamente por cinco dias consecutivos. Os gatos foram anestesiados com isofluorano, aumentando-se a sua concentração até que se atingisse uma pressão arterial média entre 40 e 60mmHg, sendo mantida durante 60 minutos. Durante o procedimento de hipotensão, os parâmetros fisiológicos foram mensurados no tempo 0 e a cada dez minutos até o fim do procedimento. O tempo de sangramento da mucosa oral foi avaliado no tempo 0 e aos 30 e 60 minutos de hipotensão. Amostras sanguíneas foram colhidas para a determinação de hemograma, alanina aminotransferase, fosfatase alcalina, ureia, creatinina e sódio no período basal e às 24 horas, 48 horas e sete dias pós-hipotensão. Amostras de urina foram colhidas por meio de cistocentese para a determinação de creatinina, gammaglutamiltransferase, densidade específica, proteínas, albumina e sódio. Durante o período anestésico, não ocorreram alterações referentes aos parâmetros fisiológicos e ao tempo de sangramento. Ocorreram alterações apenas na excreção fracionada de sódio, a qual demonstrou elevação no PRE aos sete dias, e na razão proteína/creatinina na urina, a qual demonstrou elevação do PRE em relação ao POST às 24 e às 48 horas de avaliação. Concluiu-se que a tepoxalina não causou alterações nas enzimas hepáticas, mas pode causar discreta injúria renal, com a presença de proteinúria, em gatos que foram submetidos à hipotensão.

Este estudo teve como objetivo avaliar a influência de recursos de climatização, ventilação e nebulização, sobre a fisiologia e o comportamento de vacas Holandesas alojadas em free-stall, durante o verão do sudeste brasileiro. Foram utilizadas 20 vacas Holandesas submetidas a dois tratamentos com e sem climatização. Os parâmetros ambientais registrados foram temperatura de bulbo seco, umidade relativa do ar e temperatura de globo negro. As variáveis fisiológicas avaliadas foram temperatura retal e frequência respiratória. As variáveis comportamentais registradas foram postura e suas atividades dentro da instalação. Para análise estatística utilizou-se a metodologia de quadrados mínimos por meio do procedimento PROC MIXED e PROC GLM. Apesar das diferenças estatísticas obtidas para as variáveis fisiológicas, as mesmas não foram biologicamente efetivas e indicaram que os animais se encontravam em conforto térmico. Os animais que dispunham de ventilação e nebulização alimentaram-se mesmo nas horas mais quentes do dia. A climatização é uma estratégia que permite maior conforto térmico aos animais e por consequência pode aperfeiçoar a produção leiteira através do aumento no consumo alimentar.

Recombinant coagulation factor IX must be produced in mammalian cells because FIX synthesis involves translational modifications. Human cell culture-based expression of human coagulation factor IX (hFIX) is expensive, and large-scale production capacity is limited. Transgenic animals may greatly increase the yield of therapeutic proteins and reduce costs. In this study, we used a lentiviral system to obtain transgenic cells and somatic cell nuclear transfer (SCNT) to produce transgenic animals. Lentiviral vectors carrying hFIX driven by 3 bovine β-casein promoters were constructed. Bovine epithelial mammary cells were transduced by lentivirus, selected with blasticidin, plated on extracellular matrix, and induced by lactogenic hormones; promoter activity was evaluated by quantitative PCR. Transcriptional activity of the 5.335-kb promoter was 6-fold higher than the 3.392- and 4.279-kb promoters, which did not significantly differ. Transgenic bovine fibroblasts were transduced with lentivirus carrying the 5.335-kb promoter and used as donor cells for SCNT. Cloned transgenic embryo production yielded development rates of 28.4%, similar to previous reports on cloned non-transgenic embryos. The embryos were transferred to recipient cows (N = 21) and 2 births of cloned transgenic cattle were obtained. These results suggest combination of the lentiviral system and cloning may be a good strategy for production of transgenic cattle.