RESUMO
Brucella abortus is a Gram-negative zoonotic pathogen for which there is no 100% effective vaccine. Phagosomes in B. abortus-infected cells fail to mature, allowing the pathogen to survive and proliferate. Interleukin 10 (IL10) promotes B. abortus persistence in macrophages by mechanisms that are not fully understood. In this study, we investigated the regulatory role of IL10 in the immune response to B. abortus infection. B. abortus-infected macrophages were treated with either IL10 siRNA or recombinant IL10 (rIL10), and the expression of phagolysosome- or inflammation-related genes was evaluated by qRT-PCR and Western blotting. Phagolysosome fusion was monitored by fluorescence microscopy. We found that the synthesis of several membrane-trafficking regulators and lysosomal enzymes was suppressed by IL10 during infection, resulting in a significant increase in the recruitment of hydrolytic enzymes by Brucella-containing phagosomes (BCPs) when IL10 signaling was blocked. Moreover, blocking IL10 signaling also enhanced proinflammatory cytokine production. Finally, concomitant treatment with STAT3 siRNA significantly reduced the suppression of proinflammatory brucellacidal activity but not phagolysosome fusion by rIL10. Thus, our data provide the first evidence that clearly indicates the suppressive role of IL10 on phagolysosome fusion and inflammation in response to B. abortus infection through two distinct mechanisms, STAT3-independent and -dependent pathways, respectively, in murine macrophages.
Assuntos
Brucella abortus/fisiologia , Interleucina-10/metabolismo , Lisossomos/metabolismo , Macrófagos/citologia , Macrófagos/microbiologia , Animais , Camundongos , Fagossomos/metabolismo , Células RAW 264.7 , Fator de Transcrição STAT3/metabolismo , Regulação para CimaRESUMO
Lipocalin 2 (Lcn2) is an important innate immunity component against bacterial pathogens. In this study, we report that Lcn2 is induced by Brucella (B.) abortus infection and significantly contributes to the restriction of intracellular survival of Brucella in macrophages. We found that Lcn2 prevented iron uptake by B. abortus through two distinct mechanisms. First, Lcn2 is secreted to capture bacterial siderophore(s) and abrogate iron import by Brucella. Second, Lcn2 decreases the intracellular iron levels during Brucella infection, which probably deprives the invading Brucella of the iron source needed for growth. Suppression of Lcn2 signalling resulted in a marked induction of anti-inflammatory cytokine, interleukin 10, which was shown to play a major role in Lcn2-induced antibrucella immunity. Similarly, interleukin 6 was also found to be increased when Lcn2 signalling is abrogated; however, this induction was thought to be an alternative pathway that rescues the cell from infection when the effective Lnc2 pathway is repressed. Furthermore, Lcn2 deficiency also caused a marked decrease in brucellacidal effectors, such as reactive oxygen species and nitric oxide but not the phagolysosome fusion. Taken together, our results indicate that Lcn2 is required for the efficient restriction of intracellular B. abortus growth that is through limiting iron acquisition and shifting cells to pro-inflammatory brucellacidal activity in murine macrophages.
Assuntos
Brucella abortus/metabolismo , Ferro/metabolismo , Lipocalina-2/metabolismo , Animais , Brucella abortus/imunologia , Brucella abortus/patogenicidade , Proteínas de Transporte de Cátions/metabolismo , Imunidade Inata/fisiologia , Macrófagos/metabolismo , Macrófagos/microbiologia , Camundongos , Células RAW 264.7RESUMO
BACKGROUND: Brucella causes a chronic and debilitating infection that leads to great economic losses and a public health burden. In this study, we demonstrated the brucellacidal effect of heat shock mediated by the induction of pro-inflammatory cytokines, reactive oxygen species (ROS) accumulation and apoptosis in murine macrophages and in mice. RESULTS: RAW264.7 cells were incubated at 43 °C, and BALB/c mice were subjected to whole body hyperthermia. The data showed a reduction in bacterial survival in the mice after daily heat exposure. This was accompanied by increased levels of cytokines TNF, IL-6, IL-1ß and IFN-γ in the sera of the mice. Gene expression of NF-κB and inducible nitric oxide production were also induced in the mouse splenic cells. In parallel with the bacterial reduction in the mouse model, an increased bactericidal effect was observed in RAW264.7 cells after exposure to heat stress. In addition, the heat stress increased both the nuclear translocation of NF-κB and the expression of the heat shock proteins HSP70 and HSP90 in murine macrophages. Furthermore, heat exposure induced the increase of pro-inflammatory cytokines, ROS accumulation and apoptosis but did not affect the production of nitric oxide (NO) in macrophages. CONCLUSION: This study demonstrated the induction of innate immune responses by heat stress that significantly reduced the intracellular survival of B. abortus in vitro and in vivo. Transcriptional factor NF-κB, which is a master regulator, could be termed a key activator of heat-induced immunity against Brucella. The increase in the expression and activation of NF-κB in splenic cells and macrophages was followed by enhanced antimicrobial effectors, including cytokines, ROS and NO that may contribute to the reduction of bacterial survival.
Assuntos
Brucella abortus/crescimento & desenvolvimento , Brucelose/imunologia , Resposta ao Choque Térmico/imunologia , Macrófagos/citologia , Animais , Apoptose , Brucella abortus/imunologia , Núcleo Celular/metabolismo , Citocinas/metabolismo , Modelos Animais de Doenças , Proteínas de Choque Térmico HSP70/metabolismo , Proteínas de Choque Térmico HSP90/metabolismo , Macrófagos/imunologia , Macrófagos/microbiologia , Camundongos , Camundongos Endogâmicos BALB C , Viabilidade Microbiana , Células RAW 264.7 , Espécies Reativas de Oxigênio/metabolismo , Regulação para CimaRESUMO
Brucellosis is one of the most important and widespread zoonosis worldwide responsible for serious economic losses and considerable public health burden. In this study, we investigated the modulatory effect of a microtubule-inhibitor, nocodazole, on B. abortus infection in murine macrophages and in a mouse model. Nocodazole activated macrophages and directly inhibited the growth of Brucella in a dose-dependent manner. Nocodazole increased adhesion but reduced invasion and intracellular growth of Brucella in macrophages although it did not affect co-localization of Brucella with LAMP-1. In addition, nocodazole negatively affected actin polymerization, and weakly activated ERK and p38α but significantly activated JNK in non-infected cells. After subsequent infection, nocodazole weakly inhibited activation of ERK and p38α. For the in vivo tests, nocodazole -treated mice displayed elevated levels of IFN-γ, MCP-1 and IL-10 while Brucella-infected nocodazole -treated mice showed high levels of TNF, IFN-γ, MCP-1, IL-10 and IL-6 as compared to controls. Furthermore, nocodazole treatment reduced inflammation and Brucella proliferation in the spleens of mice. These findings highlight the potential use of nocodazole for the control of brucellosis although further investigations are encouraged to validate its therapeutic use in animal hosts.
Assuntos
Antibacterianos/farmacologia , Brucella abortus/efeitos dos fármacos , Brucelose/microbiologia , Nocodazol/farmacologia , Baço/microbiologia , Actinas/metabolismo , Animais , Aderência Bacteriana/efeitos dos fármacos , Carga Bacteriana , Brucella abortus/patogenicidade , Brucelose/tratamento farmacológico , Brucelose/imunologia , Brucelose/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Modelos Animais de Doenças , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/metabolismo , Macrófagos/microbiologia , Camundongos , Viabilidade Microbiana/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fosforilação , Células RAW 264.7 , Baço/imunologia , Baço/metabolismo , Baço/patologiaRESUMO
BACKGROUND: Brucella abortus is an intracellular pathogen which can infect and persist in host cells through multiple interactions. Above all, its interaction to host cell receptor is important to understand the pathogenic mechanisms of B. abortus. Accordingly, we demonstrated that platelet-activating factor receptor (PAFR) affects host cell response against B. abortus infection. RESULTS: First of all, B. abortus infection to macrophage induces secretion of platelet-activating factor (PAF), which is a PAFR agonist. The stimulation of PAFR by PAF remarkably increases B. abortus uptake into macrophages. It induces Janus kinase 2 (JAK2) and p38α phosphorylation, indicating that PAFR-mediated activation of JAK2 signaling leads to enhanced uptake of B. abortus. Moreover, the dynamics of F-actin polymerization revealed that PAFR-mediated B. abortus uptake is related with the reorganization of F-actin and JAK2. Upon B. abortus phagocytosis, reduced PAFR in the membrane and subsequently increased levels of PAFR colocalization with endosomes were observed which indicate that B. abortus uptake into macrophages allowed PAFR trafficking to endosomes. CONCLUSIONS: This study demonstrated that PAFR has a compelling involvement in B. abortus uptake as a promoter of phagocytosis, which is associated with JAK2 activation. Thus, our findings establish a novel insight into a receptor-related phagocytic mechanism of B. abortus.
Assuntos
Brucella abortus/patogenicidade , Macrófagos/imunologia , Macrófagos/microbiologia , Fagocitose , Glicoproteínas da Membrana de Plaquetas/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Citoesqueleto de Actina/metabolismo , Animais , Brucella abortus/metabolismo , Brucelose/metabolismo , Brucelose/microbiologia , Endossomos/metabolismo , Interações Hospedeiro-Patógeno , Janus Quinase 2/metabolismo , Macrófagos/enzimologia , Camundongos , Células RAW 264.7 , Transdução de SinaisRESUMO
Brucellosis is one of the major zoonoses worldwide that inflicts important health problems in animal and human. Here, we demonstrated that dextran sulfate sodium (DSS) significantly increased adhesion of Brucella (B.) abortus in murine macrophages compared to untreated cells. Even without infection, Brucella uptake into macrophages increased and F-actin reorganization was induced compared with untreated cells. Furthermore, DSS increased the phosphorylation of MAPKs (ERK1/2 and p38α) in Brucella-infected, DSS-treated cells compared with the control cells. Lastly, DSS markedly increased the intracellular survival of Brucella abortus in macrophages by up to 48 h. These results suggest that DSS enhanced the adhesion and phagocytosis of B. abortus into murine macrophages by stimulating the MAPK signaling proteins phospho-ERK1/2 and p38α and that DSS increased the intracellular survival of B. abortus by inhibiting colocalization of Brucella-containing vacuoles (BCVs) with the late endosome marker LAMP-1. This study emphasizes the enhancement of the phagocytic and intracellular modulatory effects of DSS, which may suppress the innate immune system and contribute to prolonged Brucella survival and chronic infection.
Assuntos
Aderência Bacteriana/efeitos dos fármacos , Brucella abortus/efeitos dos fármacos , Brucella abortus/crescimento & desenvolvimento , Brucelose/microbiologia , Sulfato de Dextrana/farmacologia , Macrófagos/microbiologia , Animais , Brucella abortus/fisiologia , Brucelose/genética , Brucelose/metabolismo , Interações Hospedeiro-Patógeno , Humanos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Viabilidade Microbiana/efeitos dos fármacos , Células RAW 264.7 , Regulação para Cima/efeitos dos fármacosRESUMO
Stress adversely affects the wellbeing of commercial chickens, and comes with an economic cost to the industry that cannot be ignored. In this paper, we first develop an inexpensive and non-invasive, automatic online-monitoring prototype that uses sound data to notify producers of a stressful situation in a commercial poultry facility. The proposed system is structured hierarchically with three binary-classifier support vector machines. First, it selects an optimal acoustic feature subset from the sound emitted by the laying hens. The detection and classification module detects the stress from changes in the sound and classifies it into subsidiary sound types, such as physical stress from changes in temperature, and mental stress from fear. Finally, an experimental evaluation was performed using real sound data from an audio-surveillance system. The accuracy in detecting stress approached 96.2%, and the classification model was validated, confirming that the average classification accuracy was 96.7%, and that its recall and precision measures were satisfactory.
RESUMO
The diversity of methanogenic archaea associated with Korean Hanwoo cattle was analyzed using mcrA gene sequences from samples of rumen fluid (RF), rectal dung (RD), and barn floor manure (BFM). The predominant species were Methanobrevibacter ruminantium in RF and BFM(63.6% and 62.4%, respectively) and Methanocorpusculum labreanum in RD (53.2%).
Assuntos
Archaea/classificação , Archaea/isolamento & purificação , Biodiversidade , Fezes/microbiologia , Esterco/microbiologia , Metano/metabolismo , Rúmen/microbiologia , Animais , Archaea/genética , Archaea/metabolismo , Bovinos , DNA Arqueal/química , DNA Arqueal/genética , Dados de Sequência Molecular , Oxirredutases/genética , Análise de Sequência de DNARESUMO
Brucella abortus, the causative agent of brucellosis, can survive and replicate within host cells. Understanding bacterial virulence factors and bacteria-host cell interactions is critical for controlling brucellosis. However, little is known regarding the pathogenic mechanisms of brucellosis. A lipoprotein mutant (Gene Bank ID: 3339351) of B. abortus showed a lower rate of intracellular replication than did the wild-type strain in HeLa cells and RAW 264.7 macrophages. The adherent activity of the lipoprotein mutant was slightly increased compared to that of the wild-type strain in HeLa cells. After infection into macrophages, the lipoprotein mutant co-localized with either late endosomes or lysosomes. In mice infected with the lipoprotein mutant, fewer lipoprotein mutants were recovered from the spleen at 8 weeks post-infection compared to the wild-type strain. The ability to protect the lipoprotein mutant against infection by the virulent B. abortus strain 544 was similar to that of strain RB51. Our results indicate that the B. abortus lipoprotein is an important factor for survival within phagocytes and mice, and the B. abortus lipoprotein mutant may help improve live vaccines used to control brucellosis.
Assuntos
Brucella abortus/patogenicidade , Brucelose/microbiologia , Brucelose/patologia , Lipoproteínas/metabolismo , Fatores de Virulência/metabolismo , Animais , Carga Bacteriana , Brucella abortus/crescimento & desenvolvimento , Linhagem Celular , Modelos Animais de Doenças , Células Epiteliais/microbiologia , Feminino , Deleção de Genes , Humanos , Lipoproteínas/genética , Macrófagos/microbiologia , Camundongos , Camundongos Endogâmicos BALB C , Baço/microbiologia , Fatores de Virulência/genéticaRESUMO
Automatic detection of pig wasting diseases is an important issue in the management of group-housed pigs. Further, respiratory diseases are one of the main causes of mortality among pigs and loss of productivity in intensive pig farming. In this study, we propose an efficient data mining solution for the detection and recognition of pig wasting diseases using sound data in audio surveillance systems. In this method, we extract the Mel Frequency Cepstrum Coefficients (MFCC) from sound data with an automatic pig sound acquisition process, and use a hierarchical two-level structure: the Support Vector Data Description (SVDD) and the Sparse Representation Classifier (SRC) as an early anomaly detector and a respiratory disease classifier, respectively. Our experimental results show that this new method can be used to detect pig wasting diseases both economically (even a cheap microphone can be used) and accurately (94% detection and 91% classification accuracy), either as a standalone solution or to complement known methods to obtain a more accurate solution.
Assuntos
Auscultação/métodos , Reconhecimento Automatizado de Padrão/métodos , Vigilância da População/métodos , Síndrome Definhante Multissistêmico de Suínos Desmamados/diagnóstico , Síndrome Definhante Multissistêmico de Suínos Desmamados/fisiopatologia , Sons Respiratórios/fisiopatologia , Espectrografia do Som/métodos , Animais , Auscultação/instrumentação , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Espectrografia do Som/instrumentação , Máquina de Vetores de Suporte , SuínosRESUMO
BACKGROUND: Brucella abortus can proliferate within professional and nonprofessional phagocytic host cells and thereby successfully bypass the bacteriocidal effects of phagocytes. However, the intracellular survival mechanism and factors of virulence are not fully understood. METHODS: We have investigated the role of the regulator of G protein signaling 2 (RGS2), an intracellular calcium ([Ca(2+)](i)) regulator of the host cell, in the intracellular survival of B. abortus within phagocytes. RESULTS: B. abortus infection markedly induced RGS2 messenger RNA expression in early phase and increased the [Ca(2+)](i) level up to 24 hours postinfection within macrophages from wild-type mice. The [Ca(2+)](i) level, however, was not influenced by B. abortus infection within macrophages from RGS2-deficient mice. Furthermore, B. abortus survival was reduced within RGS2-deficient macrophages, and hence bacterial proliferation was inhibited in RGS2-deficient mice. Moreover, treatment with the Ca(2+) chelator ethylenediaminetetraacetic acid (EDTA) or 1,2-bis-(2-amino-phenoxy)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester (BAPTA-AM) and the L-type Ca(2+) channel-blocking agent nifedipine or genistein also showed a reduced intracellular replication of B. abortus within macrophages. CONCLUSION: These results indicate that B. abortus infection induces host RGS2 expression and that up-regulation of [Ca(2+)](i) levels is an essential factor for the intracellular survival of B. abortus within phagocytes.
Assuntos
Brucella abortus/crescimento & desenvolvimento , Cálcio/metabolismo , Cátions Bivalentes/metabolismo , Citosol/microbiologia , Fagócitos/microbiologia , Proteínas RGS/metabolismo , Animais , Brucella abortus/fisiologia , Sobrevivência Celular , Contagem de Colônia Microbiana , Citosol/química , Deleção de Genes , Perfilação da Expressão Gênica , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fagócitos/química , Fagócitos/metabolismo , Proteínas RGS/genética , Baço/microbiologia , Baço/patologiaRESUMO
N-myc downstream-regulated gene 2 (NDRG2) has been studied for its inhibitory effects against growth and metastasis of many tumor cell types. In this study, we showed NDRG2 expression was correlated with favorable recurrence-free survival of patients with breast cancer and inhibited metastasis of breast cancer cells (4T1). NDRG2 expression was examined in 189 breast carcinoma tissues and paired normal breast tissues using immunohistochemistry. Histological and clinicopathological data were correlated using Pearson's chi-square test of independence. NDRG2 expression in human breast cancer tissues was inversely associated with lymph node metastasis and pTNM stage. Furthermore, patients with breast cancer with a high level of NDRG2 expression showed favorable recurrence-free survival (P = 0.038). To study the effect of NDRG2 on metastasis in vivo, we established an NDRG2-overexpressing mouse breast cancer cell line (4T1-NDRG2) and measured the metastasis and survival of 4T1-NDRG2 tumor-bearing mice. To test whether transforming growth factor ß (TGF-ß)- mediated metastasis of 4T1 was inhibited by NDRG2 expression, TGF-Smad-binding element (SBE)-luciferase activity and/or measurement of active TGF-ß were performed in cell or tumor tissue level. 4T1-NDRG2 cells grew gradually and showed less metastatic activity in vivo and low invasiveness in vitro. 4T1-NDRG2 cells showed lower SBE-luciferase activity and lower level of active autocrine TGF-ß than 4T1-Mock did. Correctly, our data show that NDRG2 significantly suppress tumor metastasis by attenuating active autocrine TGF-ß production, and the attenuation might be typically associated with the favorable recurrence-free survival of patients clinically.
Assuntos
Intervalo Livre de Doença , Proteínas/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Proteínas Supressoras de Tumor/metabolismo , Proteínas Adaptadoras de Transdução de Sinal , Animais , Neoplasias da Mama/genética , Neoplasias da Mama/mortalidade , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Feminino , Humanos , Metástase Linfática , Camundongos , Invasividade Neoplásica , Metástase Neoplásica/genética , RecidivaRESUMO
The anticoccidial effects of Galla Rhois (GR) powder, which contains a major tannin-derived component of 52.7%, were evaluated in chickens following oral infection with Eimeria tenella. One-day-old chickens were assigned to five groups (control, unsupplemented, GR 0.5% supplemented [GRS 0.5%], GRS 1.0% [GRS 1.0%] and salinomycin supplemented [SS]). The chickens were fed a standard diet supplemented or not supplemented with GR or salinomycin for 10 days prior to infection. The birds received the supplemented diets continuously until 10 days post infection. The effects of GR on a E. tenella infection were evaluated by several parameters, including body weight gain, feed intake, oocyst excretion, bloody diarrhoea, and lesion scores. Infected chickens on the GRS and SS diets had a relatively moderate body weight loss (reduction ratio < 15%) and improved feed conversion. GRS and SS chickens produced significantly fewer faecal oocysts (P<0.05) and showed milder bloody diarrhoea compared with the E. tenella-infected control group. Furthermore, the lesion scores of both the GRS 0.5% and GRS 1.0% groups were significantly lower than the scores of the unsupplemented group on day 5 post infection. The lesion scores for the GR groups were similar to the scores for the SS group. In conclusion, this study suggests that GR appears to be as efficacious as salinomycin against E. tenella infection. GR supplementation leads to a reduction in infected chickens, although infected chickens are still affected compared with the uninfected control group. GR-based diets may be beneficial in preventing or treating coccidial infections in poultry.
Assuntos
Galinhas , Coccidiose/veterinária , Coccidiostáticos/farmacologia , Eimeria tenella/efeitos dos fármacos , Extratos Vegetais/farmacologia , Doenças das Aves Domésticas/tratamento farmacológico , Ração Animal , Animais , Peso Corporal , Coccidiose/tratamento farmacológico , Coccidiose/parasitologia , Coccidiose/prevenção & controle , Coccidiostáticos/uso terapêutico , Diarreia/veterinária , Suplementos Nutricionais , Fezes/parasitologia , Feminino , Masculino , Oocistos , Extratos Vegetais/uso terapêutico , Doenças das Aves Domésticas/parasitologia , Doenças das Aves Domésticas/prevenção & controle , Piranos/farmacologia , Piranos/uso terapêutico , Organismos Livres de Patógenos Específicos , Aumento de Peso , Redução de PesoRESUMO
Erysipelothrix rhusiopathiae is pathogenic for humans, many domestic animals and wild birds, but infectious cases with clinical symptoms in cats have not been reported. E. rhusiopathiae was recovered from a 4-month Russian blue breed cat with a very poor body condition score of 1 (BCS: 1/5). The isolate was typed as serotype 2b. Mice experimentally infected with the clinical isolate of E. rhusiopathiae through subcutaneous or intraperitoneal routes survived, and the organism was recovered from the spleen and synovial and pericardial fluids. Cats experimentally inoculated with the isolate either orally or subcutaneously survived but commonly exhibited depression and emaciation together with localized erythemal lesion of the skin accompanied by purulent ocular discharge. On hematological analysis, the number of total white blood cells was high compared with that in normal cats. Histological examination revealed congestion and moderate inflammation with focal necrosis. This observation may provide insight on E. rhusiopathiae infection in cats with the possible epidemiological significance and implications as a potential source of infection to other animals and humans.
Assuntos
Doenças do Gato/microbiologia , Depressão/microbiologia , Infecções por Erysipelothrix/microbiologia , Erysipelothrix/isolamento & purificação , Dermatopatias Bacterianas/veterinária , Animais , Bioensaio , Doenças do Gato/psicologia , Gatos , DNA Bacteriano/química , DNA Bacteriano/genética , Depressão/psicologia , Erysipelothrix/genética , Infecções por Erysipelothrix/psicologia , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Testes de Sensibilidade Microbiana/veterinária , Reação em Cadeia da Polimerase/veterinária , Sorotipagem/veterinária , Dermatopatias Bacterianas/microbiologia , Dermatopatias Bacterianas/psicologia , Organismos Livres de Patógenos EspecíficosRESUMO
Two field experiments were conducted to improve the conception rate of Hanwoo cow. The first experiment aimed to investigate the physiological condition of Hanwoo cows on estrus, including metabolic profiles and body condition score (BCS). The second experiment investigated the effect of a novel estrus detector on the artificial insemination (AI) conception rate for Hanwoo cows. For the first experiment, 80 Hanwoo cows (2.5 ± 0.10 of parity), approximately one month before estrus, were housed in 16 pens and offered the experimental diets twice daily with free water access. The BCS were recorded, and blood was collected from the jugular veins just before AI. The collected blood was used to measure physiological conditions, such as metabolite and hormone levels. For the second experiment, each cow was equipped with a neck-mounted estrus detector collar, which had a sensor connected through the internet. Approximately one month before estrus, three hundred sixty Hanwoo cows (2.4 ± 0.21 of parity) were assigned into groups with or without W-Tag collar treatments. The animals were managed the same as in the first experiment. The pregnancy rate reached 55% in the first experiment. The concentration of luteinizing hormone (LH) was higher (p < 0.012; 1.56 vs. 1.08 ng/mL) in cows that were not pregnant (NPG) than in cows that were pregnant (PG) after AI. The BCS and other concentrations of metabolites and hormones in the blood were not different in both NPG and PG cows. The ranges of estrogen, LH, and follicle-stimulating hormone for PG cows were 11.9 to 39.0 pg/mL, < 0.25 to 1.98 ng/mL, and < 0.50 to 0.82 ng/mL, respectively. In the second experiment, cows with the estrus detector had lower days open (p < 0.001; 78.1 vs. 84.8 d), insemination frequency (p < 0.001; 1.26 vs. 2.52), and return of estrus (p < 0.001; 70.9 vs. 79.1 d) than those in cows without the estrus detector. In conclusion, the present study indicated that lower LH concentration just before AI potentially increased the pregnancy rate of Hanwoo cows. Furthermore, the application of estrus detectors to Hanwoo cows could improve the conception success rate for AI.
RESUMO
This study was conducted to evaluate the effects of temperature-humidity index (THI) on the mortality and the panting rates in hens exposed to varying thermal environments. Hens were challenged with an acute elevation in THI in Experiment 1, where dry-bulb temperature and relative humidity were set at ~27°C and 56% at the beginning of the experiment and changed to 36°C and 45% at its conclusion, respectively. In Experiment 2, different groups of hens were exposed to a progressive increase in THI, with similar ranges to those used in the previous experiment. In Experiment 3, the hens used in Experiment 2 were again challenged by THI conditions, the intensity of which ranged between those used in the previous two experiments. In Experiment 4, panting rates were recorded under varying THI. In the last, plasma biochemical profiles were determined in blood taken from hens subjected to experimental conditions similar to those in Experiment 2. When THI was acutely elevated from 24.2° to 32.1°C within 1 h and then maintained over 4.5 h, no mortality was detected in the first hour, but exceeded 95% after 5 h, and reached 100% at 5.5 h. A gradual increase in THI to 31.2°C over 6 h did not result in mortality during the first 3 h. When THI was set below the conditions in Experiment 1 but above those in Experiment 2, mortality was 29% at 4 h, 75% at 5 h, and 79% at 8 h. However, no mortality was detected in their respective control groups. Panting was not observed under 25.3°C and was largely variable under 30°C. However, all hens exhibited panting exceeding 250 counts/min and 60% mortality at 34°C when heat stress continued for a duration of up to 280 min. In Experiment 5, high ambient THI resulted in significant reductions in plasma albumin, amylase and aspartate aminotransferase, compared with those in control group (P < 0.05). These results suggest that an acute elevation of THI has more severe effects on mortality in hens than gradual changes even when temperature and humidity are similar in both cases.
RESUMO
Brucella as a stealthy intracellular pathogen avoids activation of innate immune response. Here we investigated the contribution of an adenosine receptor, Adora2b, during Brucella infection in professional phagocyte RAW 264.7 cells and in a murine model. Adora2b-deficient cells showed attenuated Brucella internalization and intracellular survival with enhanced release of IL-6, TNF-α, IL-12 and MCP-1. In addition, blockade of Adora2b using MRS 1754 treatment in mice resulted in increased total weight of the spleens but suppressed bacterial burden in these organs accompanied by elevated levels of IL-6, IFN-γ, TNF-α, IL-12 and MCP-1, while reduced IL-10. Overall, we proposed that the Adora2b participates in the successful phagocytic pathway and intracellular survival of Brucella in RAW 264.7 cells, and could be a potential therapeutic target for the treatment of acute brucellosis in animals.
Assuntos
Antagonistas do Receptor A2 de Adenosina/farmacologia , Brucelose/tratamento farmacológico , Imunidade Inata , Macrófagos/microbiologia , Receptor A2B de Adenosina/imunologia , Acetamidas/farmacologia , Agonistas do Receptor A2 de Adenosina/farmacologia , Aminopiridinas/farmacologia , Animais , Brucella abortus/efeitos dos fármacos , Brucella abortus/fisiologia , Brucelose/microbiologia , Citocinas/imunologia , Feminino , Macrófagos/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Fagocitose , Purinas/farmacologia , Células RAW 264.7 , Receptor A2B de Adenosina/genética , Transdução de SinaisRESUMO
The present study was conducted to evaluate the effects of dietary gamma-aminobutyric acid (GABA) in broiler chickens raised in high stocking density (HSD) on performance and physiological responses. A total of 900 male broiler chicks (Ross 308) at 1 d old were assigned in a 2 × 2 factorial arrangement to 4 treatments (10 replicates per treatment) with stocking density, 7.5 birds/m2 (low stocking density; LSD) or 15 birds/m2 (HSD), and dietary GABA, 0 or 100 mg/kg. Chickens raised in HSD exhibited a decrease in body weight gain in all phases (P < 0.05) and feed intake in starter and whole phases (P < 0.01), and an increase in feed conversion ratio in the finisher phase (P < 0.01) compared with LSD-raised chickens. However, dietary GABA did not affect growth performance nor interacted with stocking density on production variables. The HSD vs. LSD increased relative liver weight on d 35 whereas dietary GABA increased relative liver weight and decreased relative bursa weight on d 21. Both stocking density and dietary GABA affected yield and quality of breast and leg muscles. Dietary GABA increased (P < 0.05) width of tibia on d 35 and interacted (P = 0.054) with stocking density on breaking stocking density on d 35. The HSD vs. LSD group lowered (P < 0.05) feather coverage scores. Significant interaction between stocking density and GABA on surface temperature of shank on d 21 was noted (P = 0.024). Dietary GABA exhibited an opposite effect on the concentrations of cecal short-chain fatty acids depending on stocking density leading to a moderate to significant interaction. Stocking density decreased alpha-1-acid glycoprotein whereas dietary GABA decreased heterophil-to-lymphocyte ratio and corticosterone in blood or serum samples. Serum biochemical parameters were altered by stocking density or dietary GABA. It is concluded that dietary GABA alleviated stress indices including corticosterone and heterophil-to-lymphocyte ratio, but failed to reverse stocking density-induced growth depression.
RESUMO
In this study, we investigated the effects of linoleic acid (LA) treatment on Brucella abortus infection in professional phagocyte RAW264.7 cells, particularly during the pathogens invasion and intracellular growth in these cells, as well as in murine model BALB/c mice focusing on bacterial splenic proliferation and immunoregulatory activities. LA inhibited the growth of Brucella in a doseand time-dependent manner. The ability of the pathogen to enter the phagocytes was inhibited as was its survival within these cells. This was accompanied by increased nitrite accumulation in these cells at 24 h post-infection. The concentration of LA used in the present study did not affect the total body weight or liver function of the mice. During Brucella infection, the total splenic weight of these animals was not changed; rather, resistance to bacterial proliferation was enhanced in the spleen. Furthermore, mice treated with LA displayed elevated levels of IL-12 and IFN-γ but reduced levels of IL-10 during infection. The findings in this study showed the regulatory role of LA against B. abortus infection suggesting its potential use in designing intervention strategy for brucellosis.
Assuntos
Antibacterianos/farmacologia , Brucella abortus/efeitos dos fármacos , Brucelose/microbiologia , Ácido Linoleico/farmacologia , Animais , Antibacterianos/química , Citocinas/metabolismo , Modelos Animais de Doenças , Ácido Linoleico/química , Camundongos , Camundongos Endogâmicos BALB C , Células RAW 264.7 , Baço/efeitos dos fármacosRESUMO
Brucellosis is an emerging infectious disease affecting humans and animals. In this study, we investigated the in vitro and in vivo effects of tannic acid (TA) against Brucella abortus infection. After infection, F-actin polymerization and mitogen-activated protein kinases (MAPKs) (ERK 1/2 and p38α) phosphorylation were reduced in TA-treated cells compared with that in control cells. The mice were infected via an intraperitoneal route and were orally given TA or phosphate-buffered saline for 14 days. Spleen weights of the TA-treated and control mice were not different; however, splenic proliferation of B. abortus was significantly reduced in the TA-treated group. Immune response analysis showed that, compared with the control group, non-infected TA-treated mice displayed increased levels of interferon-γ (IFN-γ), monocyte chemoattractant protein-1 (MCP-1), and interleukin-10 at 3 days post-infection and a further increase in IFN-γ and MCP-1 at 14 days post-infection. In contrast, compared with the control group, infected TA-treated mice displayed elevated levels of IFN-γ at 3 days post-infection, which continued to increase at 14 days post-infection, as was also observed for tumor necrosis factor. Taken together, the results showing TA activation of cytokine production and inhibition of bacterial proliferation in the host highlight a potential use of TA treatment in the control of Brucella infection.