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1.
Vet Microbiol ; 238: 108429, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31648721

RESUMO

Viral haemorrhagic disease (VHD) and colibacillosis are common diseases in rabbits that cause economic losses worldwide. The effect of colibacillosis on the immune response of vaccinated rabbits against rabbit haemorrhagic disease virus (RHDV) was studied. Four groups (G1-G4) were included. G1 was the negative control group; G2 was the RHDV vaccine group; G3 was the E. coli-infected group; and G4 was the E. coli-infected + RHDV vaccine group. The E. coli infection and RHDV vaccination were simultaneously performed, with another previous infection, 3 days before vaccination. At 28 days post-vaccination (PV), the rabbits (G2-G4) were challenged intramuscularly with 0.5 ml of RHDV at a dose of 103 50% median lethal dose (LD50)/rabbit. The rabbits were observed for clinical signs, body weight gain and mortality rates. Tissue, blood, serum, and faecal samples and rectal swabs were collected at 3, 5, 7, 14, 21 and 28 days PV. Significant clinical signs and mortality and a decrease in BW were observed in the infected + RHDV vaccine group. On the 3rd day post-infection (PI), compared with all the other groups, the vaccinated group (G2) had significantly upregulated hepatic tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels; however, the infected + RHDV vaccine group had significantly higher intestinal levels of TNF-α and IL-6 than the other groups. Furthermore, E. coli infection in vaccinated rabbits led to immunosuppression, as shown by significant decreases (P < 0.05) in heterophil phagocytic activity, the CD4+/CD8+ ratio, and HI antibody responses to RHDV and a significant increase in the heterophil to lymphocyte (H/L) ratio. In conclusion, colibacillosis leads to immunosuppression involving a shift in the equilibrium of cytokines and reduced weight gain and mortality in vaccinated rabbits and could be a contributing factor in RHDV vaccination failure in rabbit farming.


Assuntos
Infecções por Caliciviridae/veterinária , Infecções por Escherichia coli/veterinária , Coelhos/imunologia , Vacinação/veterinária , Vacinas Virais/imunologia , Animais , Infecções por Caliciviridae/imunologia , Infecções por Caliciviridae/mortalidade , Citocinas/genética , Infecções por Escherichia coli/imunologia , Infecções por Escherichia coli/mortalidade , Infecções por Escherichia coli/fisiopatologia , Regulação da Expressão Gênica/imunologia , Vírus da Doença Hemorrágica de Coelhos/imunologia , Coelhos/microbiologia , Coelhos/virologia , Vacinação/normas
2.
J Anim Sci ; 97(10): 4248-4255, 2019 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-31433053

RESUMO

Weanling rabbits frequently exhibit diarrhea or flatulence. Our experiment was conducted to investigate the effect of garlic straw on the performance and intestinal barrier of rabbits. Hyla rabbits (60 d, n = 160) with similar body weight were divided into 4 groups (4 replicates per group and 10 rabbits per replicate): fed a basal diet (control) or fed an experimental diet with 5%, 10%, or 15% garlic straw powder supplement. The results showed that the dietary addition of garlic straw increased significantly the average daily gain and average daily feed intake. Compared with the control, dietary addition of 10% and 15% garlic straw decreased significantly the death rate of rabbit. Rabbits in 10% garlic straw group had a higher secretory immunoglobulins A and immunoglobulins G concentration in jejunum and ileum than control while lower tumor necrosis factor α (TNFα) concentration in jejunum. Compared with the control, dietary addition of 10% garlic straw increased significantly genes expression of zonula occluden protein 2 (ZO2) in jejunum and ileum and mucin4 in ileum while did not alter the genes expression of junctional adhesion molecule 2 (JAM2), JAM3, ZO1, occluding, claudin1, mucin1, mucin6, and toll-like receptor 4 in jejunum and ileum and mucin4 in jejunum. In conclusion, dietary supplement of garlic straw modulates immune responses and enhances intestinal barrier, meanwhile inhibits the synthesis of pro-inflammatory cytokine of TNFα. Besides, our experiment offers positive evidence in improving rabbit health of garlic instead of antibiotics.


Assuntos
Ração Animal/análise , Suplementos Nutricionais/análise , Alho , Coelhos/fisiologia , Animais , Citocinas/metabolismo , Dieta/veterinária , Feminino , Íleo/metabolismo , Mucosa Intestinal/metabolismo , Jejuno/metabolismo , Masculino , Coelhos/imunologia , Fator de Necrose Tumoral alfa/metabolismo
3.
Hum Vaccin Immunother ; 15(10): 2491-2500, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30883271

RESUMO

Invasive meningococcal disease is rare and potentially devastating but often vaccine-preventable. Evaluation of meningococcal vaccine effectiveness is impractical owing to relatively low disease incidence; protection is therefore estimated using serum bactericidal antibody (SBA) assays. Original experiments on natural immunity established a titer of ≥4 as the correlate of protection for SBA assays using human complement (hSBA), but human complement is relatively difficult to obtain and standardize. Use of baby rabbit complement (rSBA assays), per standard guidelines for serogroups A and C, generally results in comparatively higher titers. Postlicensure effectiveness data for serogroup C conjugate vaccines support acceptance of rSBA titers ≥8 as the correlate of protection for this serogroup, but no thresholds have been formally established for serogroups A, W, and Y. Studies evaluating MenACWY-TT (Nimenrix®; Pfizer Inc, Sandwich, UK) immunogenicity have used both hSBA and rSBA assays, and ultimately suggest that rSBA may be more appropriate for these measurements.


Assuntos
Proteínas do Sistema Complemento/análise , Proteínas do Sistema Complemento/imunologia , Vacinas Meningocócicas/imunologia , Ensaios de Anticorpos Bactericidas Séricos/normas , Animais , Anticorpos Antibacterianos/sangue , Humanos , Infecções Meningocócicas/diagnóstico , Infecções Meningocócicas/prevenção & controle , Coelhos/imunologia , Sorogrupo , Ensaios de Anticorpos Bactericidas Séricos/métodos , Fatores de Tempo , Vacinas Conjugadas/imunologia
4.
J Anim Sci ; 97(4): 1693-1700, 2019 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-30726960

RESUMO

The present study was conducted to investigate the effect of Artemisia argyi on the production performance and intestinal barrier of rabbits. Weaned Hyla rabbits (30 d, n = 160) of similar body weight were divided into 4 groups (40 rabbits per treatment), and they were fed a control diet or fed an experimental diet supplemented with 3%, 6%, or 9% A. argyi. The results showed that the dietary supplementation with A. argyi did not affect the rabbits' food intake and body weight gain regardless of the inclusion level but decreased the diarrhea rate and diarrhea index (P < 0.05). Dietary addition of A. argyi increased the small intestine length and villus height/crypt depth, regardless of the inclusion level (P < 0.05). Compared with the control, the A. argyi supplementation increased the gene expression of zonula occludens 1 (ZO-1) and claudin 1 in all segments of the small intestine and regardless of the level of A. argyi (P < 0.05). In the duodenum, a dietary supplementation with 6% and 9% A. argyi increased the immunoglobulins A (IgA) content (P < 0.05). In the jejunum, the A. argyi supplementation decreased interleukin 2 (IL2) and IL6 content regardless of the inclusion level (P < 0.05). In the ileum, a 3% A. argyi addition decreased IL2 content, whereas a 6% A. argyi addition decreased IL6 content (P < 0.05). Furthermore, 6%-9% A. argyi supplementation increased the IgA content in the ileum (P < 0.05). In conclusion, dietary addition of A. argyi reduces diarrhea and modulates the gut immune function without affecting growth performances of rabbits.


Assuntos
Ração Animal/análise , Artemisia , Suplementos Nutricionais , Regulação da Expressão Gênica/efeitos dos fármacos , Coelhos/imunologia , Animais , Diarreia/tratamento farmacológico , Diarreia/veterinária , Dieta/veterinária , Ingestão de Alimentos , Feminino , Imunoglobulina A/imunologia , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/imunologia , Intestino Delgado/efeitos dos fármacos , Intestino Delgado/imunologia , Intestinos/efeitos dos fármacos , Intestinos/imunologia , Masculino , Coelhos/crescimento & desenvolvimento , Ganho de Peso/efeitos dos fármacos
5.
Science ; 363(6433): 1319-1326, 2019 03 22.
Artigo em Inglês | MEDLINE | ID: mdl-30765607

RESUMO

In the 1950s the myxoma virus was released into European rabbit populations in Australia and Europe, decimating populations and resulting in the rapid evolution of resistance. We investigated the genetic basis of resistance by comparing the exomes of rabbits collected before and after the pandemic. We found a strong pattern of parallel evolution, with selection on standing genetic variation favoring the same alleles in Australia, France, and the United Kingdom. Many of these changes occurred in immunity-related genes, supporting a polygenic basis of resistance. We experimentally validated the role of several genes in viral replication and showed that selection acting on an interferon protein has increased the protein's antiviral effect.


Assuntos
Adaptação Biológica/genética , Imunidade Inata/genética , Myxoma virus/imunologia , Mixomatose Infecciosa/imunologia , Coelhos/genética , Coelhos/virologia , Alelos , Animais , Austrália , Evolução Molecular , França , Frequência do Gene , Variação Genética , Interferon alfa-2/genética , Interferon alfa-2/imunologia , Mixomatose Infecciosa/genética , Polimorfismo de Nucleotídeo Único , População , Coelhos/imunologia , Reino Unido
6.
Talanta ; 192: 288-294, 2019 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-30348391

RESUMO

A multiplex Lateral Flow Immunoassay was developed based on the use of a single Test line and multicolour gold nanoparticles (GNPs) as signal reporters. Red and blue GNPs were linked to antibodies directed towards two different analytes and included in a typical lateral flow immunoassay configuration, in which the Test line was formed by the mixture of two antigens. As a result of the immunoreactions occurring at the Test zone, diverse combinations of red and blue GNPs labels were captured. Therefore, the Test line assumed different colours depending on which - and how much - analyte is present in the sample. The multiplexing capability of the 'colour-encoded assay' is illustrated by the simultaneous detection of aflatoxin B1 (AFB1) and type-B fumonisins (FMs) in wheat and food products that made with wheat. Reproducible detection of AFB1 and FMs contamination in raw and processed food was achieved with visual cut-off levels at 1 ng mL-1 and 50 ng mL-1, respectively. The contaminant was identified based on the colour of the label according with a specific colour code. Furthermore, strips images were acquired by means of a common smartphone and analysed through RGB data analysis providing semi-quantitative detection of the two mycotoxins.


Assuntos
Aflatoxina B1/análise , Grão Comestível/química , Contaminação de Alimentos/análise , Fumonisinas/análise , Imunoensaio/métodos , Aflatoxina B1/imunologia , Animais , Anticorpos/imunologia , Cor , Colorimetria/métodos , Fumonisinas/imunologia , Ouro/química , Limite de Detecção , Nanopartículas Metálicas/química , Coelhos/imunologia , Triticum/química
7.
J Anim Physiol Anim Nutr (Berl) ; 103(1): 363-369, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30353590

RESUMO

Sorghum dried distiller's grains with solubles (S-DDGS) are distillation extract residues from the ethanol fuel industry. In this experiment, two hundred 42-day-old rabbits were randomly allocated to five experimental diets containing 0 g/kg (control), 75, 150, 225 and 300 g/kg S-DDGS. The experiment lasted for 4 weeks. No difference was found in the average daily feed intake (ADFI; p > 0.05). With increasing sorghum inclusion, the average daily gain (ADG) was linearly (p < 0.001) and quadratically (p = 0.039) reduced, while, conversely, the feed conversion ratio (FCR) linearly (p < 0.001) increased. Increasing the amount of S-DDGS in the diet linearly decreased (p < 0.001) the apparent total tract digestibility (ATTD) of dry matter (DM), organic matter (OM), crude protein (CP) and ash. Carcass weight, carcass yield, heart and liver weights were linearly decreased by an increase in the amount of S-DDGS added to diets (p < 0.001), but no difference was observed between the 0, 75 and 150 g/kg S-DDGS groups (p > 0.05). Serum IL-6, IL-10 and SIgA linearly increased (p = 0.008) with increasing levels of S-DDGS in the diet. Rabbits fed 0, 75 and 150 g/kg of S-DDGS had similar IL-6 and IL-10 levels. Statistically significant differences in SIgA were observed between rabbits fed control diets and feed mixtures containing S-DDGS (p < 0.01). To conclude, S-DDGS can safely be added up to 75 g/kg, to the diet of rabbits. Increasing dietary S-DDGS inclusion may decrease the growth performance, nutrient digestibility and carcass traits, and activate immune responses.


Assuntos
Ração Animal/análise , Composição Corporal/efeitos dos fármacos , Dieta/veterinária , Nutrientes/metabolismo , Coelhos/crescimento & desenvolvimento , Sorghum , Fenômenos Fisiológicos da Nutrição Animal , Animais , Digestão/fisiologia , Masculino , Coelhos/imunologia , Distribuição Aleatória
8.
Vet Immunol Immunopathol ; 207: 36-45, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30593349

RESUMO

Rhipicephalus sanguineus s. l. is popularly known as the "brown dog tick" since dogs are its preferential hosts, but the species has been reported to parasitize other mammals, including humans, with significant medical-veterinary importance since it transmits several important pathogenic agents during the feeding period. The tick saliva is a complex mixture that has several functions, including the capability to modulate the hemostatic, inflammatory and immunologic systems of the host, allowing pathogens to settle. Despite knowledge about the immunosuppressive action of tick saliva, little is known about the mechanisms involved in this process and the morphophysiological effects caused by exposure to the salivary gland extract, taking into consideration the different periods of the glandular cycle. Thus, the objective of this study was to analyze the in vitro effects of salivary gland extracts obtained from R. sanguineus s. l. females fed on host rabbits for two (SGE2 - Salivary Gland Extracts of 2 days) and four days (SGE4 - Salivary Gland Extracts of 4 days) on J774 cells (monocyte macrophage cell line) and verify the occurrence of morphological and immunomodulatory alterations in these cells when exposed to different concentrations of these extracts. The results showed that: (i) SGE2 and SGE4 at the concentration of 4 µg/mL presented cytotoxicity to the J774 cells exposed for 24 and 48 hours; (ii) SGE2 at the concentrations of 2 µg/mL(48-hour exposure) and 1 µg/mL (24-hour exposure) and SGE4 at the concentrations of 2 and 1 µg/mL (48-hour exposure) showed proinflammatory activity, confirmed by the increased secretion of NO and proinflammatory cytokine (IL-2), and the presence of morphological characteristics detected by microscopy; and (iii) SGE2 and SGE4 at the concentrations of 0.5 and 0.1 µg/mL had immunomodulatory activity, demonstrated by decreases in the secretion of NO and proinflammatory cytokines (IL2, IL-6 and TNF-α) and increase in the synthesis of IL-10, confirmed by the morphophysiological analysis. These unprecedented data are extremely relevant for future research to identify the processes involved in the ectoparasite-host relationship, as well to develop more efficient tick control strategies.


Assuntos
Rhipicephalus sanguineus/imunologia , Glândulas Salivares/imunologia , Animais , Linhagem Celular , Citocinas/metabolismo , Relação Dose-Resposta a Droga , Inflamação/induzido quimicamente , Inflamação/veterinária , Interleucina-2/metabolismo , Coelhos/imunologia , Coelhos/parasitologia , Extratos de Tecidos/imunologia , Extratos de Tecidos/farmacologia
9.
Crit Care ; 22(1): 353, 2018 12 20.
Artigo em Inglês | MEDLINE | ID: mdl-30572913

RESUMO

BACKGROUND: Lung diseases such as acute respiratory distress syndrome (ARDS) have a high incidence worldwide. The current drug therapies for ARDS have supportive effects, making them inefficient. New methods such as stromal cell therapy are needed for this problem. METHODS: This research was performed with ten New Zealand rabbits in two groups. Bone marrow aspiration was performed on the treated group, and mesenchymal stem cells were isolated and cultured. The experimental model of ARDS was induced using LPS from Escherichia coli strain O55:B5. Then, 1010 bone marrow mesenchymal stem cells (BM-MSCs) were autologously transplanted intrapulmonary in the treatment group, and 1-2 ml of PBS in the control group. The clinical signs, computed tomographic (CT) scans, echocardiography, blood gas analysis, complete blood count, and cytokine levels were measured before and at 3, 6, 12, 24, 48, 72, and 168 h after BM-MSC transplant. Finally, the rabbits were killed, and histopathological examination was performed. RESULTS: The results showed that BM-MSCs decreased the severity of clinical symptoms, the number of white blood cells and heterophils in the blood, the total cell count, and number of heterophils and macrophages in bronchoalveolar lavage, and balanced the values of arterial blood gases (increase in partial pressure of oxygen and O2 saturation and decrease in the partial pressure of carbon dioxide). They also downregulated the tumor necrosis factor (TNF)-α and interleukin (IL)-6 concentrations and increased the IL-10 concentrations at different times compared with time 0 and in the control group, significantly. In the CT scan, a significant decrease in the Hounsfield units and total lung volume was found by echocardiography, and in comparing the two groups, a significant difference in the parameters was noticed. The histopathology demonstrated that the BM-MSCs were able to reduce the infiltration of inflammatory cells and pulmonary hemorrhage and edema. CONCLUSIONS: This study indicated that BM-MSCs play a significant role in the repair of lung injury.


Assuntos
Pulmão/cirurgia , Células-Tronco Mesenquimais/imunologia , Animais , Contagem de Células Sanguíneas/métodos , Gasometria/métodos , Medula Óssea/cirurgia , Transplante de Medula Óssea , Citocinas/análise , Citocinas/sangue , Ecocardiografia/métodos , Escherichia coli/patogenicidade , Infecções por Escherichia coli/complicações , Infecções por Escherichia coli/veterinária , Coração/microbiologia , Pulmão/imunologia , Pulmão/microbiologia , Células-Tronco Mesenquimais/patologia , Coelhos/imunologia , Síndrome do Desconforto Respiratório do Adulto , Tomografia Computadorizada por Raios X/métodos , Transplante Autólogo/métodos
10.
J Vasc Res ; 55(6): 350-364, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30544118

RESUMO

Endothelial cells of the vascular system are dynamic cells whose molecular adaptability is decisive for the adjustment of homeostasis and organ perfusion. Advanced microscopic techniques, automation processing, and image analysis software was shown to improve the understanding of vascular biology. In this work, we describe advanced methods that allow investigating the dynamics of endothelial cell contacts. The development of viral vectors has contributed significantly to the genetic manipulation of endothelial cells. We used the Gibson assembly as a quick and cheap cloning system for introducing sequences into the lentiviral-based pFUGW vector. Furthermore, classical fluorescence tags such as mCherry and EGFP were compared with self-labeling tags such as Halo and SNAP for their suitability to study junction dynamics in cultured endothelium, and found the self-labeling tags as useful tools. Using such combinations, we found maintained cell junction integrity during shear stress-induced junction remodeling using VE-cadherin-EGFP. Remodeling was accompanied by VE-cadherin plaque formation, indicating that this process is mediated by the for-mation of the actin-driven junction-associated intermittent lamellipodia, JAIL. The combined methods including the Gibson assembly, lentiviral mediated gene transfer, spinning disk-based live cell imaging, and software for quantification allow analyses of the endothelial cell junction dynamics under static and under shear stress conditions.


Assuntos
Clonagem Molecular/métodos , Células Endoteliais/fisiologia , Células Endoteliais/ultraestrutura , Corantes Fluorescentes , Junções Intercelulares/fisiologia , Animais , Anticorpos , Anticorpos Monoclonais , Caderinas/análise , Caderinas/genética , Expressão Gênica , Vetores Genéticos , Cabras/imunologia , Proteínas de Fluorescência Verde/genética , Células Endoteliais da Veia Umbilical Humana , Humanos , Immunoblotting , Junções Intercelulares/química , Camundongos , Coelhos/imunologia , beta Catenina/análise , gama Catenina/análise
11.
Brain Pathol ; 28(6): 806-821, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30062819

RESUMO

BACKGROUND: Mutations in the three-prime repair exonuclease 1 (TREX1) gene have been associated with neurological diseases, including Retinal Vasculopathy with Cerebral Leukoencephalopathy (RVCL). However, the endogenous expression of TREX1 in human brain has not been studied. METHODS: We produced a rabbit polyclonal antibody (pAb) to TREX1 to characterize TREX1 by Western blotting (WB) of cell lysates from normal controls and subjects carrying an RVCL frame-shift mutation. Dual staining was performed to determine cell types expressing TREX1 in human brain tissue. TREX1 distribution in human brain was further evaluated by immunohistochemical analyses of formalin-fixed, paraffin-embedded samples from normal controls and patients with RVCL and ischemic stroke. RESULTS: After validating the specificity of our anti-TREX1 rabbit pAb, WB analysis was utilized to detect the endogenous wild-type and frame-shift mutant of TREX1 in cell lysates. Dual staining in human brain tissues from patients with RVCL and normal controls localized TREX1 to a subset of microglia and macrophages. Quantification of immunohistochemical staining of the cerebral cortex revealed that TREX1+ microglia were primarily in the gray matter of normal controls (22.7 ± 5.1% and 5.5 ± 1.9% of Iba1+ microglia in gray and white matter, respectively) and commonly in association with the microvasculature. In contrast, in subjects with RVCL, the TREX1+ microglia were predominantly located in the white matter of normal appearing cerebral cortex (11.8 ± 3.1% and 38.9 ± 5.8% of Iba1+ microglia in gray and white matter, respectively). The number of TREX1+ microglia was increased in ischemic brain lesions in central nervous system of RVCL and stroke patients. CONCLUSIONS: TREX1 is expressed by a subset of microglia in normal human brain, often in close proximity to the microvasculature, and increases in the setting of ischemic lesions. These findings suggest a role for TREX1+ microglia in vessel homeostasis and response to ischemic injury.


Assuntos
Isquemia Encefálica/metabolismo , Isquemia Encefálica/patologia , Encéfalo/metabolismo , Exodesoxirribonucleases/metabolismo , Exodesoxirribonucleases/fisiologia , Microglia/metabolismo , Fosfoproteínas/metabolismo , Fosfoproteínas/fisiologia , Idoso , Animais , Anticorpos/metabolismo , Encéfalo/patologia , Exodesoxirribonucleases/genética , Feminino , Mutação da Fase de Leitura , Células HEK293 , Células HeLa , Doenças Desmielinizantes Hereditárias do Sistema Nervoso Central/genética , Doenças Desmielinizantes Hereditárias do Sistema Nervoso Central/patologia , Homeostase/fisiologia , Humanos , Macrófagos/metabolismo , Masculino , Pessoa de Meia-Idade , Fosfoproteínas/genética , Coelhos/imunologia , Doenças Retinianas/genética , Doenças Retinianas/patologia , Doenças Vasculares/genética , Doenças Vasculares/patologia
12.
Cell Stress Chaperones ; 23(5): 975-984, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29728855

RESUMO

The current study aimed to test the effect of Moringa oleifera extract (MOE), vitamin (Vit) C, and sodium bicarbonate (NaHCO3) on heat stress (HS)-induced alterations in rabbits. Five groups of rabbits were designed as control, HS, HS + MOE, HS + Vit C, and HS + NaHCO3. HS groups were exposed to high temperatures, while treatments were given in drinking water for 6 weeks. Levels of blood cortisol, leptin, IFN-γ, TNF-α, and IL-10 were assayed using ELISA, while adrenaline was assayed calorimetrically. Expression of HSP70, FOXP3, T cell receptor (TCR) γ, and δ mRNA was tested using real-time (RT)-PCR, while HSP70 protein expression was tested using western blotting in liver and kidney tissues. Infiltration of regulatory T cells (Treg; CD25+) and NK (CD56+) cells were tested using immunohistochemistry (IHC). The levels of liver enzymes (ALT & AST), urea, and creatinine were assayed calorimetrically, while body weight gain (BWG) and feed conversion ratio (FCR) were calculated. The results showed increased levels of cortisol, adrenaline, leptin, IFN-γ, TNF-α, ALT, AST, urea, and creatinine but decreased IL-10 in the HS group. Increased expression of HSP70 on both mRNA and protein levels was associated with increased NK and γδ T cells versus decreased Treg cell infiltration in liver and kidney tissues of the HS group. In the same group, BWG was decreased, while FCR was increased with respect to the control group. All treatments used in this study reversed the effects of HS significantly. In conclusion, MOE, Vit C, and NaHCO3 can be added to rabbit diets for the amelioration of HS-induced symptoms.


Assuntos
Ácido Ascórbico/farmacologia , Proteínas de Choque Térmico HSP70/metabolismo , Resposta ao Choque Térmico , Moringa oleifera , Coelhos/imunologia , Coelhos/metabolismo , Bicarbonato de Sódio/farmacologia , Animais , Citocinas/sangue , Epinefrina/sangue , Expressão Gênica/efeitos dos fármacos , Proteínas de Choque Térmico HSP70/genética , Resposta ao Choque Térmico/genética , Resposta ao Choque Térmico/imunologia , Hidrocortisona/sangue , Linfócitos Intraepiteliais/efeitos dos fármacos , Linfócitos Intraepiteliais/imunologia , Rim/efeitos dos fármacos , Rim/metabolismo , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/imunologia , Leptina/sangue , Fígado/efeitos dos fármacos , Fígado/enzimologia , Fígado/metabolismo , Extratos Vegetais/farmacologia , Coelhos/genética , Coelhos/crescimento & desenvolvimento , Linfócitos T Reguladores/efeitos dos fármacos , Linfócitos T Reguladores/imunologia
13.
J Anat ; 233(2): 266-273, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29736914

RESUMO

Sinus-like dilatations of the mammary duct are recognisable in the mammary gland of pregnant and lactating wild European rabbits. These dilatations exhibit a bilaminar epithelial lining, with luminal epithelial cells expressing basal and lateral E-cadherin. Occasional binucleated mammary epithelial cells are present in the luminal layer. Underlying the luminal epithelial cells is a basal layer of cytokeratin 14-positive cells, supported by a thin layer of fibrous tissue. Multi-segmental epithelial proliferation, as indicated by Ki67 expression, is apparent in the luminal epithelial cells, suggesting a capacity for division during pregnancy and lactation. CD3-positive T lymphocytes are present both intraepithelially, suggesting exocytosis, and in foci subjacent to the ductular epithelium. We consider that sinus-like dilatations of the mammary duct may have the potential to give rise to a subset of the mammary gland neoplasms classified as ductal in origin. Milk accumulation in these sinus-like dilatations is likely to provide a niche for bacterial replication in cases of mastitis in rabbits. These structures are an important component of the innate immune system of the mammary gland, both as a physical barrier and as an interface between the milk and mammary immune cells.


Assuntos
Lactação , Glândulas Mamárias Animais/anatomia & histologia , Prenhez , Coelhos/anatomia & histologia , Animais , Células Epiteliais/metabolismo , Feminino , Antígeno Ki-67/metabolismo , Glândulas Mamárias Animais/imunologia , Gravidez , Coelhos/imunologia , Linfócitos T/fisiologia
14.
Animal ; 12(9): 1877-1885, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29224577

RESUMO

Genetic selection and nutrition management have played a central role in the development of commercial rabbitry industry over the last few decades, being able to affect productive and immunological traits of the animals. However, the implication of different energy sources in animals from diverse genetic lines achieving such evolutionary success remains still unknown. Therefore, in this work, 203 female rabbits housed and bred in the same conditions were used from their first artificial insemination until their fifth weaning. The animals belonged to three different genetic types diverging greatly on breeding goals (H line, hyper-prolific (n=66); LP line, robust (n=67) and R line, selected for growth rate (n=67), and were assigned to two experimental diets, promoting major differences in energy source (cereal starch or animal fat)). The aims of this work were to: (1) characterize and describe blood leucocyte populations of three lines of rabbit does in different physiological stages during their reproductive period: first artificial insemination, first weaning, second parturition and fifth weaning; and (2) study the possible influence of two different experimental diets on the leucocyte populations in peripheral blood. Flow cytometry analyses were performed on blood samples taken from females at each different sampling stade. Lymphocyte populations at both weanings were characterized by significantly lower counts of total, CD5+ and CD8+ lymphocytes (-19.8, -21.7 and -44.6%; P<0.05), and higher counts of monocytes and granulocytes (+49.2 and +26.2%; P<0.05) than in the other stages. Females had higher blood counts of lymphocytes B, CD8+ and CD25+ and lower counts of CD4+ at first than at fifth weaning (+55.6, +85.8, +57.5, -14.5%; P<0.05). G/L ratio was higher at both weanings (P<0.05), and CD4+/CD8+ ratio increased progressively from the 1AI to the 5 W (P<0.001). Regarding the effect of genetic type in blood leucocyte counts, LP animals presented the highest counts for total, B, CD5+ and CD8+ lymphocytes (+16.7, +31.8, +24.5 and +38.7; P<0.05), but R rabbits showed the highest counts for monocytes and granulocytes (+25.3 and +27.6; P<0.05). The type of diet given during the reproductive life did not affect the leucocyte population counts. These results indicate that there are detectable variations in the leucocyte profile depending on the reproductive stage of the animal (parturition, weaning or none of them). Moreover, foundation for reproductive longevity criteria allows animals to be more capable of adapting to the challenges of the reproductive cycle from an immunological viewpoint.


Assuntos
Imunidade Celular , Coelhos , Reprodução , Seleção Genética , Animais , Dieta , Feminino , Contagem de Leucócitos , Coelhos/genética , Coelhos/imunologia , Desmame
15.
Comp Med ; 67(6): 498-503, 2017 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-29212581

RESUMO

Because of their ideal size and temperament, rabbits are commonly used in polyclonal antibody production. Immunostimulatory adjuvants-such as Freund complete and incomplete adjuvants as well as various proprietary products-trigger a robust immune response, which increases antibody concentrations. However, these adjuvants can cause excessive soft tissue reactions, prompting concerns regarding animal wellbeing. This study assessed the safety and efficacy of cationic liposome- oligonucleotide complexes (CLDC) as an alternative adjuvant to conventional adjuvants. On days 0 and 14, 15 female New Zealand white rabbits were vaccinated subcutaneously with 15 µg ovalbumin mixed with either CLDC, Freund adjuvant (day 0, complete; day 14, incomplete), or a proprietary adjuvant (n = 5 per group). Antibody titers were measured by direct ELISA on days 0, 14, and 28. Rabbits were palpated daily for lesion development, and all lesions were measured. Rabbits in all groups developed a significant antibody response to ovalbumin over 28 d. However, the differences between groups were not statistically significant. No rabbits in the CLDC group developed skin lesions, whereas 80% of rabbits that received Freund adjuvant and 100% of those that received the proprietary product developed skin lesions. This study demonstrates that CLDC may be a valuable and effective alternative adjuvant for polyclonal antibody production in rabbits-one that avoids the palpable injection-site lesions often seen with other adjuvants.


Assuntos
Adjuvantes Imunológicos/farmacologia , Formação de Anticorpos , Oligonucleotídeos/farmacologia , Coelhos/imunologia , Animais , Ensaio de Imunoadsorção Enzimática , Feminino , Adjuvante de Freund/farmacologia , Injeções , Lipossomos , Oligonucleotídeos/administração & dosagem , Ovalbumina/imunologia
16.
Sci Rep ; 7(1): 12202, 2017 09 22.
Artigo em Inglês | MEDLINE | ID: mdl-28939872

RESUMO

Immunodeficient mice have been used predominantly in biomedical research. Realizing that large animal species may have an enhanced ability to predict clinical outcome relative to mice, we worked to develop immunodeficient rabbits by CRISPR/Cas9. We first demonstrated that multiplex embryo transfer efficiently produced multiple lines of single-gene mutant (SGM) founders. Embryos microinjected with single sgRNA targeting FOXN1, RAG2, IL2RG or PRKDC were pooled for embryo transfer. As few as three recipients were used to produce twenty SGM founders for four genes. We then demonstrated the powerful multiplex targeting capacity of CRISPR/Cas9. First, two genes on the same chromosome were targeted simultaneously, resulting in three RAG1/RAG2 double-gene mutant (DGM) founders. Next we microinjected forty-five embryos each with five sgRNAs targeting FOXN1, RAG1, RAG2, IL2RG and PRKDC, and transferred them to two recipients. Five founders were produced: one SGM, two DGM, one triple-gene mutant and one quadruple-gene mutant. The present work demonstrates that multiplex embryo transfer and multiplex gene targeting can be used to quickly and efficiently generate mutant rabbit founders. Four lines of SGM (e.g. FOXN1, RAG2, IL2RG, and PRKDC) immunodeficient rabbits, as well as multigenic mutant immunodeficient rabbits have been produced. These animals may prove useful for biomedical research.


Assuntos
Animais Geneticamente Modificados/imunologia , Transferência Embrionária/métodos , Marcação de Genes/métodos , Engenharia Genética/métodos , Coelhos/imunologia , Animais , Animais Geneticamente Modificados/genética , Sistemas CRISPR-Cas/genética , Feminino , Masculino , Microinjeções/métodos , Modelos Animais , Coelhos/genética
17.
Anim Sci J ; 88(10): 1644-1650, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28544086

RESUMO

The effect of dietary inclusion of probiotics and genetic groups on rabbit performance under hot environmental conditions was studied. A total of 80 rabbits aged 8 weeks were distributed into a completely randomized design in a 4 × 3 factorial arrangement, including four genetic groups and three concentrations of dietary probiotic (0, 200 and 400 g/t feed). The utilized probiotic contained 4 × 109  colony-forming units/g of Bacillus subtilis. Jabali local breed (J), imported Spanish V-line (V) and their crossbreds (»J¾V and ¾J»V) were included in the current study. Final weight and body weight gain were not significantly affected by dietary probiotic levels or genetic group. The feed conversion ratio was better for purebreds than that of crossbreds. A significant improvement in percentage of dressed carcass, mid and hind parts was recorded for rabbits fed a diet containing 400 g probiotic/t feed compared with those fed a basal diet or low probiotic level. Probiotic supplementation had a significant decrease in serum cholesterol. Rabbits given 400 g probiotic/t feed had higher hemoglobin, red blood cells and platelets. Adding 400 g probiotic/t feed to rabbit's diet significantly (P ≤ 0.05) improved cell-mediated immunity compared to the other treatments 48 h post-injection.


Assuntos
Fenômenos Fisiológicos da Nutrição Animal , Bacillus subtilis , Dieta/veterinária , Suplementos Nutricionais , Exposição Ambiental/efeitos adversos , Genótipo , Temperatura Alta/efeitos adversos , Probióticos/administração & dosagem , Coelhos/crescimento & desenvolvimento , Coelhos/genética , Ração Animal , Animais , Peso Corporal , Colesterol/sangue , Contagem de Eritrócitos , Hemoglobinas/metabolismo , Imunidade Celular , Carne , Contagem de Plaquetas , Coelhos/imunologia , Coelhos/metabolismo
18.
Exp Mol Med ; 49(3): e305, 2017 03 24.
Artigo em Inglês | MEDLINE | ID: mdl-28336958

RESUMO

In this review, we explain why and how rabbit monoclonal antibodies have become outstanding reagents for laboratory research and increasingly for diagnostic and therapeutic applications. Starting with the unique ontogeny of rabbit B cells that affords highly distinctive antibody repertoires rich in in vivo pruned binders of high diversity, affinity and specificity, we describe the generation of rabbit monoclonal antibodies by hybridoma technology, phage display and alternative methods, along with an account of successful humanization strategies.


Assuntos
Anticorpos Monoclonais Humanizados/imunologia , Coelhos/imunologia , Animais , Humanos , Técnicas Imunológicas/métodos
19.
Animal ; 11(5): 854-863, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-27745563

RESUMO

Limiting the post-weaning intake of the young rabbit is known to improve its resistance to digestive disorders, whereas a degradation of its housing hygiene is assumed to have a negative impact on its health. This study aims at providing insights into the mechanism of digestive health preservation regarding both host (growth and immune response) and its symbiotic digestive microbiota. A 2×2 factorial design from weaning (day 28) to day 64 was set up: ad libitum intake or restricted intake at 70% of ad libitum, and high v. low hygiene of housing (n=105 per group). At day 36 and day 45, 15 animals/group were subcutaneously immunized with ovalbumin (OVA) to assess their specific immune response. Blood was sampled at 36, 45, 57 and 64 days of age to determine total and anti-OVA immunoglobulin type G (IgG) and haptoglobin levels. The cecal bacterial community was explored (18 per group) by 454 pyrosequencing of genes coding for the 16S ribosomal RNA, whereas cecal pH, NH3 and volatile fatty acid (VFA) concentrations were measured to characterize fermentative activity. A 30% reduction in feed intake reduced the growth by only 17% (P<0.001), and improved the feed conversion ratio by 15% (P<0.001), whereas the degradation of hygiene conditions slightly decreased the feed intake in ad libitum fed rabbits (-3.5%, P<0.02). As poor hygiene conditions did not affect weight gain, feed conversion was improved from day 42 (P<0.05). Restricted feeding led to a lower mortality between day 28 and day 40 (P=0.047), whereas degraded hygiene conditions decreased overall morbidity (7.8% v. 16.6%; P<0.01). Both a reduced intake and low hygiene conditions of housing affected microbiota composition and especially dominant genera belonging to the Ruminococcaceae family (P<0.01). Moreover, low hygiene was associated with a higher Ruminococcaceae/Lachnospiraceae ratio (3.7 v. 2.4; P<0.05). Cecal total VFA and pH were increased (+19%; P<0.001) and decreased (-0.1 pH unit; P<0.05), respectively, in feed-restricted rabbits. Neither specific anti-OVA IgG nor haptoglobin was affected by treatments. Total IgG concentrations were the highest in animals raised in poor hygiene conditions after 8 days of restriction, but decreased after 19 days of restriction in high hygiene conditions (-2.15%; P<0.05). In conclusion, the degradation of hygiene conditions failed to induce a systematic specific and inflammatory response in rabbit, but reduced morbidity instead. Our results suggest that the microbiota composition would be a helpful source of biomarkers of digestive health.


Assuntos
Restrição Calórica , Microbioma Gastrointestinal , Abrigo para Animais , Imunidade Inata , Coelhos/fisiologia , Animais , Comportamento Alimentar , Feminino , Higiene , Masculino , Coelhos/crescimento & desenvolvimento , Coelhos/imunologia , Coelhos/microbiologia
20.
Dev Comp Immunol ; 65: 280-288, 2016 12.
Artigo em Inglês | MEDLINE | ID: mdl-27492646

RESUMO

Infection with rabbit hemorrhagic disease virus (RHDV) can cause acute liver failure (ALF), leading to severe mortality in rabbits. Inflammatory response, especially the expression of inflammatory cytokines such as interleukin (IL)-1ß, tumor necrosis factor (TNF)-α, and IL-6, may play major roles in mediating and amplifying the ALF. Among these cytokines, IL-6 is a multifunctional cytokine with a central role in various physiological inflammatory and immunological processes. In this study, we found that RHDV infection significantly upregulated IL-6 gene expression in vivo. Next, the rabbit IL-6 promoter was cloned and analyzed. Transfection of full-length RHDV cDNA in RK-13 cells upregulated the activity of the IL-6 promoter. A series of 5' deletion constructs demonstrated that AP-1 (activator protein 1), NF-IL6 (nuclear factor interleukin-6), and NF-κB (nuclear factor kappa B) elements were critical for RHDV-induced IL-6 transcription. Besides, the CREB (cAMP-response element binding protein) element may also play an accessory effect on RHDV-induced IL-6 transcription. Collectively, the results elucidate the mechanism of IL-6 induction, and enrich the RHDV pathogenesis in rabbit.


Assuntos
Infecções por Caliciviridae/imunologia , Células Epiteliais/imunologia , Vírus da Doença Hemorrágica de Coelhos/imunologia , Inflamação/imunologia , Interleucina-6/metabolismo , Falência Hepática Aguda/imunologia , Coelhos/imunologia , Animais , Linhagem Celular , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Células Epiteliais/virologia , Interleucina-6/genética , Mutagênese Sítio-Dirigida , NF-kappa B/metabolismo , Regiões Promotoras Genéticas/genética , Fator de Transcrição AP-1/metabolismo , Regulação para Cima
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