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This study aimed to verify if the process of artificial insemination (AI) characterized here as animal immobilization, the passage of the semen applicator through the cervix, and deposition of the semen in the uterus, affected cows’ welfare. For this, 18 beef calved cows were selected and divided into two groups: inseminated cows (AIG, n = 9), and not inseminated cows, the control group (CG, n = 9). Body condition score, uterus, and ovary evaluation were performed. Later, both groups were submitted into an estrus synchronization protocol and only the AIG group was inseminated. Blood components of urea, creatinine, AST, GGT, CK, glucose, triglycerides, cholesterol, HDL, LDL, VLDL, NEFA, BHB, cortisol, estradiol, progesterone, albumin, and total protein were measured 30 h before AI, and 4, 24, 48 and 168 h after AI. Statistical differences were considered when P

Em rebanhos leiteiros, a suplementação com ácidos graxos poliinsaturados (PUFAs) é considerada uma ferramenta importante para diminuir o balanço energético negativo de vacas leiteiras durante o periparto e contribuir para a reprodução e sistema imunológico. Os efeitos da suplementação com PUFA sobre estes sistemas têm sido pouco explorados na literatura. Os PUFAs mais comuns na dieta dos ruminantes são Ômega-3 (n-3 PUFA) como ácido linolênico e Ômega-6 (n-6 PUFA) como ácido linoleico. Esta revisão abordará os aspectos gerais do n-3 PUFA, seus efeitos mais relevantes no sistema imune, principalmente seus efeitos nas células imunes, bem como seus efeitos na parte reprodutiva, tanto in vivo como in vitro, enfatizando a ação do n-3 PUFA através de mecanismos moleculares. A incorporação de n-3 PUFAs na dieta de vacas leiteiras exerce efeitos positivos na fertilidade e imunidade. Mais estudos a fim de explorar a função do n-3 PUFA na modulação do sistema imune e parâmetros reprodutivos, desde a investigação da biologia básica até a aplicação a campo de modo clínico e preventivo, devem ser requeridos. In dairy cattle, supplementation with polyunsaturated fatty acids (PUFAs) is considered to be an important tool to decrease the negative energy balance of periparturient dairy cows and improve the reproductive and immune systems. The most common PUFAs added to ruminant diets are omega 3 (n-3 PUFA) as linolenic acid and omega 6 (n-6 PUFA) as linoleic acid. This paper aims to review the potential effects of n-3 PUFA. We consider the effects of n-3 PUFA on the bovine immune system, especially on immune cells, and on in vivo and in vitro reproductive parameters, emphasizing how n-3 PUFAs act as modulators through one or more molecular mechanisms. The incorporation of n-3 PUFA in the dairy cow diet has positive effects on animal fertility and immunity. Future research on n-3 PUFA should be more explored concerning reproduction and immune function, starting from the investigation of basic biology to their potential for application in the clinical and preventive medicine fields.

Submitted by LUISA CUNHA CARNEIRO null (luisacunhacarneiro@hotmail.com) on 2016-02-29T22:54:55Z No. of bitstreams: 1 Tese_Luisa_Cunha_Carneiro.pdf: 2347194 bytes, checksum: 385e788cab291232de3fc02da72024b3 (MD5) Approved for entry into archive by Sandra Manzano de Almeida (smanzano@marilia.unesp.br) on 2016-03-01T14:26:24Z (GMT) No. of bitstreams: 1 carneiro_lc_dr_jabo.pdf: 2347194 bytes, checksum: 385e788cab291232de3fc02da72024b3 (MD5) Made available in DSpace on 2016-03-01T14:26:24Z (GMT). No. of bitstreams: 1 carneiro_lc_dr_jabo.pdf: 2347194 bytes, checksum: 385e788cab291232de3fc02da72024b3 (MD5) Previous issue date: 2016-02-03 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) De modo geral, o objetivo deste estudo foi determinar se as células endometriais bovinas responderam a análogos virais de padrões moleculares associados a patógenos (PAMPs) mediante a produção de citocinas pró-inflamatórias após ativadas pelos receptores “Toll-Like” (TLRs) no endossoma celular e no citoplasma celular pelo genes indutores de ácido retinóico tipo I (RIG-I). No primeiro experimento, amostras uterinas de vacas de corte mestiças pós-púberes foram dissectadas para obtenção de células endometriais epiteliais e estromais. Um controle negativo e quatro PAMPs: LPS, ssRNA, Poly I:C (LMW), Poly (I:C) HMW foram utilizados. Dois grupos de tratamentos (transfectados e não transfectados) foram analisados durante 24 horas. Em outro experimento, células endometriais foram tratadas apenas com o PAMP Poly (I:C) LMW e um grupo Controle Negativo. Neste, os grupos foram incubados às 0, 2, 6, 12, 24, 36, 48 e 72 horas. Sobrenadantes foram colhidos para desenvolver o teste de ELISA para IL-6 e IL-8. Células epiteliais produziram IL-6 em resposta ao Poly I:C (HMW) quando comparadas com o Controle (Grupo DOTAP positivo; P< 0.05), enquanto que o LPS induziu produção de IL-6 e IL-8 em células estromais (P< 0.05). O uso de um reagente de transfecção entre as células e tratamentos demonstrou efeito (P> 0.05). Ainda, células estromais tratadas por Poly I:C (LMW) demonstraram uma maior produção de IL-6 às 48 e 72 horas (P< 0.05), e para o IL-8 às 6, 12, 24, 36, 48 e72 horas quando comparadas com o grupo Controle (P< 0.05). No segundo experimento, outras amostras uterinas de vacas de corte pós-púberes foram utilizadas. A obtenção de células endometriais estromais e epiteliais foram isoladas pelo mesmo protocolo do primeiro experimento. O PAMP Poly (I:C) LMW e um controle negativo foram utilizados. Proteínas para o RIG-I e p65 foram colhidas após 12, 24, 48 e 72 horas de tratamento. Em resposta a Poly (I:C) LMW, células estromais ativaram o RIG-I às 48 hours (P< 0.05) quando comparadas com o grupo controle. Enquanto que, as células epiteliais não foram suficientemente estimuladas pelo Poly (I:C) LMW na ativação do RIG-I em nenhum momento testado (P> 0.05). A proteína p65 depois de estimulada pela Poly (I:C) LMW foi ativada às 12 horas pelas células estromais (P< 0.05) e às 24 horas pelas células epiteliais (P< 0.05). Conclui-se que, células endometriais bovinas foram essenciais na ativação das vias exercidas tanto pelos TLR como RIG-I com função de iniciar uma defesa imunológica contra infecções virais. In general, the objective of this study was to determine if bovine endometrial cells replied to virus analogs of pathogen associated molecular pattern (PAMPs) by production of proinflammatory cytokines after Toll-Like Receptor (TLR) activation in the cell endosome and after retinoic acid inducible gene – I (RIG-I) stimulation in the cell cytoplasm. In the first experiment, uterine samples from post pubertal cross-breed beef cows were dissected using a protocol to obtain epithelial and stromal cells. A negative control and four different PAMPs: LPS, ssRNA, Poly I:C (LMW), Poly (I:C) HMW were used. Two treatments (transfected and non-transfected) groups were investigated during 24 hours. In the other experiment, endometrial cells were treated with only Poly (I:C) LMW and a negative Control group. All incubated at 0, 2, 6, 12, 24, 36, 48 and 72 hours. Supernatants were collected to develop Elisa for IL-6 and IL-8. Epithelial cells produced IL-6 in response do Poly I:C (HMW) compared to Control (P< 0.05), otherwise, LPS induced IL-6 and IL-8 in stromal (P< 0.05). The transfection Reagent differ between cells and treatments (P> 0.05). Still, in stromal cells treated by Poly I:C (LMW) the production of IL-6 was higher at 48 and 72 hours (P< 0.05), and for IL-8 at 6, 12, 24, 36, 48 and 72 hours when compared to the Control (P< 0.05). In the second experiment, uterine samples from others post pubertal mixed-breed beef cows were used. To obtain stromal and epithelial cells, uterine samples were dissected with the same protocol as the first experiment. The PAMP Poly (I:C) LMW and a negative control were used. Proteins for RIG-I and p65 were collected after 12, 24, 48 and 72 hours. In response to Poly (I:C) LMW induction, stromal cells activated RIG-I at 48 hours (P< 0.05) were compared to the Control group. On the other hand, epithelial cells were not sufficient stimulated Poly (I:C) LMW to activate RIG-I at any time point evaluated (P> 0.05). The protein p65 after stimulated by Poly (I:C) LMW was activated at 12 hours by stromal (P< 0.05) and at 24 hours by epithelial cells (P< 0.05). In conclusion, bovine endometrial cells were elemental factors in the activation of both TLR and RIG-I pathway in order to start an immune defense against viral infection. CNPq: 140625/2013-5

Made available in DSpace on 2022-04-28T19:05:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-01-01 The objective of this study was to evaluate the incidence of repeat breeder (RB) in crossbred dairy cows submitted to a timed artificial insemination (TAI) and estrous synchronization protocols. Data from 371 heifers and 997 lactating crossbred dairy cows were analyzed. Cows with more than 30 DPP with good uterine condition, no lameness or clinical mastitis, and body condition score ≥ 2.5 were used. The cows were divided between two groups, as: group 1: TAI – those cows with no CL at ultrasound exam that were enrolled in a TAI protocol and group 2: estrous synchronization – all those cows that had CL. The incidences of RB and factors relative to it were analyzed by logistic regression in SAS. The incidence of RB condition was higher in lactating cows than nonlactating heifers (P < 0.001). There was a tendency of RB condition to higher in cows that calved during spring/summer than those calved at autumn/winter season. The parturition condition had no effect on RB incidence (P > 0.001); however, lactation degree influenced the incidence of RB (P

CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior Contaminação bacteriana que normalmente infecta o endométrio bovino no pós-parto prejudica a defesa imunológica quando há estresse metabólico, levando a endometrite e infertilidade. A resposta do endométrio contra bactérias depende da imunidade inata, com o reconhecimento de padrões moleculares associados a patógenos, estimulando a inflamação, caracterizadas por secreção de interleucinas (IL)-1p, IL-6 e IL-8. Entretanto, animais frequentemente falham em eliminar a infecção microbiana quando apresentam reduzidas concentrações de glicose e glutamina, que são fontes de energia para o endométrio no conhecido balanço energético negativo. A contínua inflamação e resposta imune inata contra infecção bacteriana no útero após o parto compromete o bem-estar animal. Neste estudo testou-se a hipótese de que as vias homeostáticas conferidas pela glicose, glutamina e biotina integram o metabolismo energético e a imunidade inata no tecido endometrial bovino. A ausência de glicose reduziu a secreção de IL-1P, IL-6 e IL-8 em culturas de órgãos endometriais desafiadas com a endotoxina lipopolissacarídeo (LPS). Depleção de glutamina também reduziu a atividade inflamatória do endométrio contra LPS. Entretanto, as atividades de glicose e glutamina dependem da presença de glicólise. A vitamina biotina demonstrou ter caráter anti- inflamatório, reduzindo a produção de citocinas pró-inflamatórias no endométrio. Em conclusão, o estresse energético metabólico afetou as respostas inflamatórias à infecções bacterianas no endométrio. Providenciamos dados que demonstram como o balanço energético negativo pode estar ligado a doença uterina no pós-parto. Bacteria that usually infect the bovine endometrium at postpartum impairs immune defense when exist metabolic stress, leading to endometritis and infertility. The response of the endometrium to bacteria depends on innate immunity, with the recognition of pathogen-associated molecular patterns by stimulating inflammation, characterized by secretion of interleukins (IL) -1p, IL-6 and IL-8. However, animals often fail to eliminate the microbial infection when they have reduced levels of glucose and glutamine, which are sources of energy to the endometrium in the negative energy balance. Continuous inflammation and innate immune response to bacterial infection in the uterus after calving compromise animal welfare. In this study we tested the hypothesis that homeostatic pathways conferred by glucose , glutamine and biotin, link the energy metabolism and innate immunity in the bovine endometrial tissue. The deprivation of glucose reduced the secretion of IL-1P, IL-6 and IL-8 in cultures of endometrial organs challenged with the endotoxin lipopolysaccharide (LPS). Glutamine depletion also reduces the inflammatory activity of the endometrium to LPS. However, glucose and glutamine activities depend on the presence of glycolysis. The vitamin biotin demonstrates anti-inflammatory nature, reducing the production of proinflammatory cytokines in the endometrium. In conclusion, the energy metabolic stress disrupts inflammatory responses to bacterial infections in the endometrium. We provide data on how negative energy balance may be linked to postpartum uterine disease. Tese (Doutorado)