RESUMEN
Various studies have attempted to improve the milk yield and composition in dairy animals. However, no study has examined the effects of milking at different times on milk yield and composition. Therefore, this study aimed to compare the yield, composition, and antimicrobial components of milk obtained from milking at different times in lactating goats. Eight goats were milked once daily at different times for three consecutive weeks (first week: 06:00 h; second week: 09:00 h; and third week: 12:00 h). The light ranged from 06:30 to 19:00 h. Milk and blood samples were collected once a day during milking time. Milking at 09:00 h resulted in a significantly higher milk yield than that obtained after milking at 06:00 and 12:00 h. Prolactin levels in plasma and the fat, Na+, ß-defensin, and S100A7 (antimicrobial component) levels in milk were the lowest in the 09:00 h milking. These results indicate that milk yield, composition, and antimicrobial components can be affected by milking time, which may be related to the altered concentration of prolactin in the blood. These findings provide a rational basis for achieving maximal milk production with strong immunity by changing to a more effective milking time.
Asunto(s)
Cabras , Lactancia , Leche , Prolactina , beta-Defensinas , Animales , Cabras/fisiología , Femenino , Leche/química , Prolactina/sangre , Factores de Tiempo , beta-Defensinas/análisis , Industria Lechera/métodos , Sodio/sangre , Sodio/análisis , Antiinfecciosos/análisisRESUMEN
Mastitis causes significant economic losses to the dairy industry due to decreased milk production in infected cows. Identification of mastitis-causing pathogens, such as streptococci, is necessary for selecting an effective antibiotic for treating mastitis. Although bacterial cultivation is widely used for pathogen identification, it requires more than 24 hr to complete. Contrarily, Lateral flow assays are simple, rapid, and inexpensive testing procedures. In this study, the effectiveness of an immunochromatographic test kit for detecting streptococci in milk samples from cows with clinical mastitis was evaluated as an alternative to bacterial cultivation. The performance of the immunochromatographic test kit for detecting mastitis-causing pathogens was compared with that of bacterial cultivation and real-time quantitative polymerase chain reaction (qPCR). The sensitivity and specificity of the immunochromatographic test kit were 0.800 and 0.875, respectively, compared with bacterial cultivation. Additionally, the κ statistic values of the immunochromatographic test kit was 0.667, indicating substantial agreement with the results of bacterial cultivation. Statistically, sensitivity and specificity of the immunochromatographic kit and real-time qPCR did not differ significantly; thus, the immunochromatographic test kit detected mastitis-causing streptococci as effectively as real-time qPCR. Therefore, the immunochromatographic kit is a rapid, inexpensive, and simple method for detecting streptococci and contributes to the timely selection of appropriate antibiotics for treatment and promotes early recovery from mastitis.
Asunto(s)
Cromatografía de Afinidad , Mastitis Bovina , Leche , Sensibilidad y Especificidad , Infecciones Estreptocócicas , Streptococcus , Animales , Bovinos , Mastitis Bovina/microbiología , Mastitis Bovina/diagnóstico , Femenino , Infecciones Estreptocócicas/veterinaria , Infecciones Estreptocócicas/diagnóstico , Infecciones Estreptocócicas/microbiología , Streptococcus/aislamiento & purificación , Leche/microbiología , Cromatografía de Afinidad/veterinaria , Cromatografía de Afinidad/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Juego de Reactivos para Diagnóstico/veterinariaRESUMEN
We propose a novel strategy for quick and easy preparation of suicide live vaccine candidates against bacterial pathogens. This method requires only the transformation of one or more plasmids carrying genes encoding for two types of biological devices, an unnatural amino acid (uAA) incorporation system and toxin-antitoxin systems in which translation of the antitoxins requires the uAA incorporation. Escherichia coli BL21-AI laboratory strains carrying the plasmids were viable in the presence of the uAA, whereas the free toxins killed these strains after the removal of the uAA. The survival time after uAA removal could be controlled by the choice of the uAA incorporation system and toxin-antitoxin systems. Multilayered toxin-antitoxin systems suppressed escape frequency to less than 1 escape per 109 generations in the best case. This conditional suicide system also worked in Salmonella enterica and E. coli clinical isolates. The S. enterica vaccine strains were attenuated with a >105 fold lethal dose. Serum IgG response and protection against the parental pathogenic strain were confirmed. In addition, the live E. coli vaccine strain was significantly more immunogenic and provided greater protection than a formalin-inactivated vaccine. The live E. coli vaccine was not detected after inoculation, presumably because the uAA is not present in the host animals or the natural environment. These results suggest that this strategy provides a novel way to rapidly produce safe and highly immunogenic live bacterial vaccine candidates. IMPORTANCE: Live vaccines are the oldest vaccines with a history of more than 200 years. Due to their strong immunogenicity, live vaccines are still an important category of vaccines today. However, the development of live vaccines has been challenging due to the difficulties in achieving a balance between safety and immunogenicity. In recent decades, the frequent emergence of various new and old pathogens at risk of causing pandemics has highlighted the need for rapid vaccine development processes. We have pioneered the use of uAAs to control gene expression and to conditionally kill host bacteria as a biological containment system. This report proposes a quick and easy conversion of bacterial pathogens into live vaccine candidates using this containment system. The balance between safety and immunogenicity can be modulated by the selection of the genetic devices used. Moreover, the uAA-auxotrophy can prevent the vaccine from infecting other individuals or establishing the environment.
Asunto(s)
Escherichia coli , Salmonella enterica , Humanos , Animales , Escherichia coli/metabolismo , Aminoácidos/metabolismo , Vacunas Atenuadas/genética , Salmonella enterica/metabolismo , Vacunas de Productos InactivadosRESUMEN
We investigated the antimicrobial components in cow milk at dry off and postpartum and their contribution in preventing new high SCC at quarter level. Milk samples from 72 quarters of 19 lactating cows were collected at last milking before dry off and at 7 d after parturition. Milk yield of each cow was recorded and SCC, IgG, IgA, lactoferrin, lingual antimicrobial peptide (LAP), and S100A7 concentrations in each quarter milk sample were measured. The postpartum milk yield was significantly higher than that at dry off. The IgG, IgA and lactoferrin concentrations in milk at dry off were significantly higher than those at postpartum, whereas the LAP concentration was lower. Quarters with SCC < 300 000 cells/ml at both dry off and postpartum were classified as persistent low SCC (PL) whereas those that rose above that same threshold postpartum were classified as new high SCC (NH). At dry off, IgG and LAP concentrations in milk were significantly higher in PL than in NH. These results suggest that high LAP concentrations during the dry period may contribute toward the prevention of new high SCC.
Asunto(s)
Inmunoglobulina A , Inmunoglobulina G , Lactancia , Lactoferrina , Leche , Periodo Posparto , Animales , Bovinos , Femenino , Leche/química , Lactoferrina/análisis , Lactancia/fisiología , Recuento de Células/veterinaria , Inmunoglobulina G/análisis , Inmunoglobulina A/análisis , Mastitis Bovina/prevención & control , beta-DefensinasRESUMEN
In mammary glands, the formation of less-permeable tight junctions (TJs) and the production of antimicrobial compounds like lactoferrin and defensins are important for preventing mastitis. Resveratrol, a polyphenol contained in red grapes, is known to protect mammary epithelial cells (MECs) from oxidative stress; however, oral administration of resveratrol causes a decrease in certain biological processes through conjugation and metabolic conversion. In this study, we determined the beneficial effects of resveratrol on TJs and antimicrobial compounds in cultured goat MECs by adding it to the medium, and in lactating goat mammary glands by topical application for percutaneous absorption. TJ barrier function was evaluated by transepithelial resistance and expression or localization pattern of claudins for culture model in vitro and by somatic cell count, Na+, albumin, and IgG in milk for topical application in vivo. Concentrations of antimicrobial compounds and cytokines were measured using ELISA. Activation of STAT3 was evaluated by Western blotting. Resveratrol strengthened TJ barrier function by upregulating claudin-3 in cultured MECs and topical application to udders reduced somatic cell count, Na+, albumin, and IgG in milk. Resveratrol increased ß-defensin and S100A7 levels in cultured MECs and milk. In addition, resveratrol down-regulated cytokine production and STAT3 pathway. These findings suggest that the topical application of resveratrol to udders may be effective in preventing mastitis.
Asunto(s)
Antiinfecciosos , Enfermedades de las Cabras , Mastitis , Femenino , Animales , Uniones Estrechas , Lactancia/metabolismo , Resveratrol/farmacología , Resveratrol/metabolismo , Células Epiteliales/metabolismo , Leche/metabolismo , Glándulas Mamarias Animales/metabolismo , Mastitis/tratamiento farmacológico , Mastitis/prevención & control , Mastitis/veterinaria , Antiinfecciosos/farmacología , Cabras , Albúminas/metabolismo , Albúminas/farmacología , Inmunoglobulina G/metabolismo , Enfermedades de las Cabras/metabolismoRESUMEN
In lactating mammary glands, tight junctions (TJs) prevent blood from mixing with milk and maintain epithelial cell polarity, which is important for milk production. This study aimed to investigate the effect of sodium acetate and sodium butyrate (SB) stimulation direction on the TJ barrier function, which is measured with regard to transepithelial electrical resistance and fluorescein flux, in goat mammary epithelial cells. The expression and localization of the TJ proteins claudin-3 and claudin-4 were examined using Western blotting and immunofluorescence. SB treatment in the lower chamber of cell culture inserts adversely affected the TJ barrier function, whereas sodium acetate barely had any effect, regardless of stimulation direction. In addition, SB treatment in the lower chamber significantly upregulated claudin-3 and claudin-4, whereas TJ proteins showed intermittent localization. Moreover, SB induced endoplasmic reticulum (ER) stress. ARC155858, a monocarboxylate transporter-1 inhibitor, alleviated the adverse impact of SB on TJs and the associated ER stress. Interestingly, sodium ß-hydroxybutyrate, a butyrate metabolite, did not affect the TJ barrier function. Our findings indicate that sodium acetate and SB influence the TJ barrier function differently, and excessive cellular uptake of SB can disrupt TJs and induce ER stress.
Asunto(s)
Cabras , Uniones Estrechas , Animales , Femenino , Ácido Butírico/farmacología , Claudina-3 , Claudina-4/genética , Lactancia , Acetato de Sodio , Células Epiteliales , Proteínas de Transporte de MembranaRESUMEN
In the mammary glands, production of antimicrobial components and formation of less-permeable tight junctions (TJs) are important for safe milk production. Previously, we reported that local heat treatment of udders using disposable heating pad enhances the components of innate immunity in lactating goat mammary glands. Gingerol is a polyphenol present in ginger that can induce heat-like effects. However, oral administration of polyphenols causes a decrease in biological activity through conjugation and metabolic conversion. Here, we investigated the effects of gingerol on antimicrobial components and TJs by topically applying it to lactating goat udders. Gingerol application increased the somatic cell count, cathelicidin-2 concentration, and proportion of polymorphonuclear cells in the milk and interleukin-8 production. Moreover, gingerol treatment enhanced ß-defensin-1 production in milk, cultured mammary epithelial cells, and cultured somatic cells. Contrastingly, gingerol treatment did not affect the concentrations of blood-derived components (Na+, albumin, and IgG) in the milk or the TJ barrier function of cultured mammary epithelial cells. These findings suggest that the topical application of gingerol, similar to local heat treatment, to udders enhances the components of innate immunity in mammary glands. These findings may be useful for the prevention of mastitis in milk-producing animals and, hence, safe and stable dairy production.
RESUMEN
Mammary glands with mastitis are usually treated with antibiotics in combination with anti-inflammatory drug application on the udder skin. Menthol is an anti-inflammatory drug. The aim of the present study was to investigate the effect of surface application of menthol on goat udders on the production of antimicrobial components in milk. Goats (5 Shiba and 11 Tokara goats) were subjected to menthol application to the udder under both healthy and inflammatory conditions. An intramammary infusion of lipopolysaccharides was carried out to induce inflammatory conditions in the udder. Milk samples were collected to determine somatic cell count (SCC) and sodium ion (Na+ ), antimicrobial component and cytokine concentrations. In healthy udders, menthol application increased the concentration of antimicrobial components (S100A7 and S100A8), but not in the control. In the inflamed udder, antimicrobial component (lactoferrin, S100A7, and S100A8) and inflammatory cytokine (IL-1ß) concentrations were higher in the menthol group than in the control group. These results suggest that menthol application on udders augments the antimicrobial component concentration in the mammary gland under both healthy and inflammatory conditions.
Asunto(s)
Antiinfecciosos , Leche , Femenino , Animales , Glándulas Mamarias Animales , Mentol/farmacología , Cabras , Antiinfecciosos/farmacología , CitocinasRESUMEN
The production of antimicrobial components and the formation of less-permeable tight junctions (TJs) are important in the defense system of lactating mammary glands and for safe dairy production. Valine is a branched-chain amino acid that is actively consumed in the mammary glands and promotes the production of major milk components like ß-casein; additionally, branched-chain amino acids stimulate antimicrobial component production in the intestines. Therefore, we hypothesized that valine strengthens the mammary gland defense system without influencing milk production. We investigated the effects of valine in vitro using cultured mammary epithelial cells (MECs) and in vivo using the mammary glands of lactating Tokara goats. Valine treatment at 4 mM increased the secretion of S100A7 and lactoferrin as well as the intracellular concentration of ß-defensin 1 and cathelicidin 7 in cultured MECs. In addition, an intravenous injection of valine increased S100A7 levels in the milk of Tokara goats without influencing milk yield and milk components (i.e., fat, protein, lactose, and solids). In contrast, valine treatment did not affect TJ barrier function either in vitro or in vivo. These findings indicate that valine enhances antimicrobial component production without influencing milk production and TJ barrier function in lactating mammary glands; thus, valine contributes to safe dairy production.
Asunto(s)
Antiinfecciosos , Leche , Femenino , Animales , Leche/metabolismo , Uniones Estrechas/metabolismo , Lactancia/metabolismo , Valina/farmacología , Valina/análisis , Valina/metabolismo , Glándulas Mamarias Animales/metabolismo , Células Epiteliales/metabolismo , Antiinfecciosos/farmacología , Antiinfecciosos/análisis , Antiinfecciosos/metabolismo , CabrasRESUMEN
A bacterial culture of milk is the most common test to determine the presence of mastitis-causing pathogens, which informs appropriate treatment. However, a certain proportion of clinical mastitis milk shows no growth of any mastitis-causing pathogens. We hypothesized that bacterial culture-negative clinical mastitis milk is associated with the activity of antimicrobial components contained in the milk. In this study, the differences in antimicrobial components (lactoferrin, transferrin, lysozyme, lactoperoxidase, and lingual antimicrobial peptide [LAP]) between bacterial culture-positive and culture-negative bovine clinical mastitis milk were investigated using Holstein cows. Our results showed that 37 out of 71 samples of clinical mastitis milk had negative bacterial cultures. The LAP concentration in bacterial culture-negative milk was lower than that in positive milk (31.95 ± 1.64 nM vs. 42.85 ± 4.01 nM). In contrast, the lysozyme concentration in bacterial culture-negative milk was higher than that in positive milk (0.76 ± 0.15 µg/ml vs. 0.42 ± 0.06 µg/ml). In conclusion, the concentration of antimicrobial components was different between bacterial culture-positive and culture-negative bovine clinical mastitis milk, which suggests that antimicrobial components are related to bacterial culture results.
Asunto(s)
Antiinfecciosos , Enfermedades de los Bovinos , Mastitis Bovina , Animales , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Antiinfecciosos/uso terapéutico , Bacterias , Bovinos , Enfermedades de los Bovinos/tratamiento farmacológico , Femenino , Lactoferrina/metabolismo , Lactoperoxidasa/metabolismo , Mastitis Bovina/microbiología , Leche/metabolismo , MuramidasaRESUMEN
A single infusion of lipopolysaccharide (LPSs) into the uterus induces inflammation in the mammary gland. This indicates that LPS can translocate from the uterus to the mammary gland. Natural endometritis is characterized by continuous intrauterine inflammation. The aim of the present study was to determine the effect of repeated intrauterine infusion of two different types of LPSs obtained from Escherichia coli O111:B4 (LPS-O111) and O55:B5 (LPS-O55) on the inflammatory status of the mammary glands of goats. Goats were assigned to three groups: LPS-O111, LPS-O55, and saline (control). Saline with (LPS-O111 and 55 groups) and without (control) 100 µg LPS was infused into the uterus continuously for 7 days. Decreased milk yield was detected in both LPS-O111 and LPS-O55 groups 2 days after the first LPS infusion. While somatic cell count (SCC) was significantly increased in all groups 1 day after the first LPS infusion, both LPS infusions further increased SCC 2 days after the first infusion and showed a significantly higher SCC than that in the control group. Plasma LPS-binding protein (LBP) was significantly higher in both LPS groups than in the control group during the days after infusion. In addition, pro-inflammatory cytokines, interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α), and IL-8, were significantly increased in both LPS infusion groups compared with those in the control group. The LPS-O111 infusion resulted in higher SCC, LBP, TNF-α, and IL-8 concentrations than those in the LPS-O55 group. These results suggest that repeated LPS infusion into the uterus can induce more severe mammary gland inflammation than a single infusion. Interestingly, the mammary tissues recovered from inflammation even though the LPS intrauterine infusion was continued.
Asunto(s)
Enfermedades de las Cabras , Mastitis , Animales , Citocinas/metabolismo , Femenino , Enfermedades de las Cabras/inducido químicamente , Cabras/metabolismo , Inflamación/metabolismo , Inflamación/veterinaria , Interleucina-1beta/metabolismo , Interleucina-8/metabolismo , Lipopolisacáridos/toxicidad , Glándulas Mamarias Animales , Mastitis/inducido químicamente , Mastitis/veterinaria , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Ectopic tertiary lymphoid organs (TLOs) have been identified in many organs, such as the lungs, nasal cavities, and kidneys of both mice and humans. Although lymphocyte aggregates have been observed in the mammary glands of ruminants, the details remain unclear. In this study, we investigated the mammary glands of lactating goats for the presence of TLOs. The localization of CD20 (B cells), CD3 (T cells), MECA79 (high endothelial venules), CD40 (follicular dendritic cells), BCL6 (germinal center), and IgA was examined by immunohistochemistry. The concentrations of IgG, IgA, lactoferrin, ß-defensin-1, cathelicidin-2, cathelicidin-7, S100A7, and S100A8 in milk were measured by ELISA. The localization and amount of tight junction (TJ) proteins (claudin-3 and claudin-4) were examined using immunofluorescence and western blotting. We found that 19 out of 30 udders contained lymphocyte aggregates, which showed positive reactions against CD20, CD3, CD40, and MECA79. In addition, large-sized aggregations showed separate localization of B cells and T cells and a positive reaction against BCL6, although BCL6 was sparsely localized in the aggregations. These results indicate that mammary glands of lactating goats contain TLOs. The IgG and IgA concentrations in the milk of TLO-positive goats and the number of IgA-positive cells were higher than those in negative goats. Furthermore, claudin-4 was localized in the TJ region and the amount was higher in TLO-positive mammary glands than that in the negative group, indicating the presence of leakages at TJs. In conclusion, a majority of lactating goat udders have TLOs, which contribute to local immunity by producing immunoglobulins.
Asunto(s)
Cabras , Lactancia , Tejido Linfoide , Glándulas Mamarias Animales , Animales , Claudina-4 , Femenino , Inmunoglobulina A , Inmunoglobulina G , Tejido Linfoide/anatomía & histología , Glándulas Mamarias Animales/anatomía & histologíaRESUMEN
Short-chain fatty acids activate antimicrobial component production in the intestine. However, their effects on mammary glands remain unclear. We investigated the effects of acetate and butyrate on antimicrobial component production in mammary epithelial cells (MECs) or leukocytes cultured in vitro and in mammary glands of lactating Tokara goats in vivo. Our results showed that butyrate enhanced the production of ß-defensin-1 and S100A7 in MECs. Additionally, the infusion of butyrate into mammary glands through the teats enhanced ß-defensin-1 and S100A7 concentrations in milk. The infusion of acetate also increased ß-defensin-1 and S100A7 concentrations along with those of cathelicidin-2 and interleukin-8, which are produced by leukocytes. Furthermore, acetate promoted cathelicidin-2 and interleukin-8 secretion in leukocytes in vitro. These findings suggest that acetate and butyrate differentially upregulate antimicrobial component production in mammary glands, which could help to develop appropriate treatment for mastitis, thereby reducing economic losses and improving animal welfare in farming environments.
Asunto(s)
Antiinfecciosos , beta-Defensinas , Acetatos/farmacología , Animales , Antibacterianos , Ácido Butírico/farmacología , Femenino , Cabras , Interleucina-8 , Lactancia , Glándulas Mamarias Animales , Leche , Acetato de Sodio/farmacologíaRESUMEN
Mammary epithelial cells (MECs) are the only cells capable of synthesizing lactose. During lactation, alveolar MECs secrete lactose through the apical membrane into the alveolar lumen, whereas alveolar tight junctions (TJs) block the leakage of lactose into the basolateral sides of the MECs. However, lactose leaks from the alveolar lumen into the blood plasma in the mastitis and after weaning. This exposes the basolateral membrane of MECs to lactose. The relationship between lactose in blood plasma and milk production has been suggested. The present study determined whether lactose exposure on the basolateral membrane of mouse MECs adversely affects milk production in vitro. Restricted exposure to lactose on the basolateral side of the MECs was performed using a culture model, in which MECs on the cell culture insert exhibit milk production and less-permeable TJs. The results indicated that lactose exposure on the basolateral side inhibited casein and lipid production in the MECs. Interestingly, lactose exposure on the apical side did not show detectable effects on milk production in the MECs. Basolateral lactose exposure also caused the inactivation of STAT5, a primary transcriptional factor for milk production. Furthermore, p38 and JNK were activated by basolateral lactose exposure. The activation of p38 and JNK following anisomycin treatment reduced phosphorylated STAT5, and inhibitors of p38 blocked the reduction of phosphorylated STAT5 by basolateral lactose exposure. These findings suggest that lactose functions as a partial inhibitor for milk production but only when it directly makes contact with the basolateral membrane of MECs.
Asunto(s)
Glándulas Mamarias Animales , Factor de Transcripción STAT5 , Animales , Células Epiteliales/metabolismo , Femenino , Lactancia/metabolismo , Lactosa/metabolismo , Lactosa/farmacología , Ratones , Leche/metabolismo , Factor de Transcripción STAT5/metabolismo , Factor de Transcripción STAT5/farmacologíaRESUMEN
In the mammary glands during pregnancy, the alveolar buds are first branched from the mammary ducts after which they form the alveolar luminal structure for milk production postparturition. Body temperature could increase for several reasons, such as infectious disease and heat stress. We have previously reported that high temperature adversely effects on the lactation capacity of mouse mammary epithelial cells (MECs). However, it remains unclear how high temperature influences mammary morophogenesis during pregnancy. In this study, we investigated the effects of high temperature on this mammary alveolar development process using two types of culture models including embedded organoids of MECs in Matrigel; these models reproduced mammary alveolar bud induction and alveolar luminal formation. Results showed that a culture temperature of 41 °C repressed alveolar bud induction and inhibited alveolar luminal formation. In addition, the treatment at 41 °C decreased the number of proliferating mammary epithelial cells but did not affect cell migration. Levels of phosphorylated Akt, -ERK1/2, -HSP90, and -HSP27 were increased in organoids cultured at 41 °C. The specific inhibitors of HSP90 and HSP27 exacerbated the disruption of organoids at 41 °C but not at 37 °C. Furthermore, the organoids precultured at 37 and 41 °C in the alveolar luminal formation model showed differences in the expression levels of caseins and tight junction proteins, which express in MECs in lactating mammary glands, after induction of MEC differentiation by prolactin and dexamethasone treatment in vitro. These results suggest that elevated temperature directly hinders mammary alveolar development; however, heat shock proteins may mitigate the adverse effects of high temperatures.
Asunto(s)
Lactancia , Glándulas Mamarias Animales , Animales , Células Epiteliales/metabolismo , Femenino , Proteínas de Choque Térmico HSP27/metabolismo , Proteínas de Choque Térmico HSP27/farmacología , Lactancia/metabolismo , Glándulas Mamarias Animales/metabolismo , Ratones , Embarazo , Transducción de Señal , TemperaturaRESUMEN
Various antimicrobial components, such as lactoferrin, S100 calcium-binding protein A7 (S100A7), and IgA, produced by epithelial cells and leukocytes in lactating mammary glands are important for host defense against invading pathogens. Increase in milking frequency enhances milk yield in ruminants and implies an increase in frequency of teat stimulation. However, the influence of frequent teat stimulation on the production of antimicrobial components remains unclear. In this study, we investigated the effect of frequent teat stimulation, with and without milk removal, on the lactoferrin, S100A7, and IgA concentrations in milk of lactating Shiba goats in Japan. The lactoferrin, S100A7, and IgA concentrations in milk were measured using ELISA. We found that lactoferrin concentration decreased by frequent teat stimulation with milk removal, although concentrations of IgA and S100A7 increased. Frequent teat stimulation without milk removal also altered the lactoferrin, IgA, and S100A7 concentrations. Furthermore, frequent teat stimulation increased IL-22 concentration, which has been reported to upregulate S100A7 production in cultured human keratinocytes. Thus, these findings indicate that frequent teat stimulation, with or without milk removal, affects antimicrobial components in milk and may be useful for the prevention and treatment of mastitis in ruminants.
Asunto(s)
Inmunoglobulina A , Lactoferrina , Glándulas Mamarias Animales , Animales , Industria Lechera , Femenino , Cabras , Inmunoglobulina A/análisis , Lactancia , Lactoferrina/análisis , Glándulas Mamarias Animales/metabolismo , Leche/químicaRESUMEN
Dairy cows feed on isoflavones as physiologically active substances present in legumes. However, the influences of isoflavones (biochanin A, genistein, formononetin, and daidzein) and their metabolites (p-ethylphenol and equol) on milk components production, tight junctions (TJs), and their regulatory pathways are unclear in bovine mammary epithelial cells (BMECs). In this study, we investigated the influences of isoflavones and their metabolites in BMECs using an in vitro culture model. The influences of isoflavones on milk components production, TJ proteins, and STAT5/STAT3 signaling pathways were different in a type-specific manner. Biochanin A decreased the mRNA expression and secretion of both ß-casein and lactoferrin while a decrease in activated STAT5 and an increase in activated STAT3. In contrast, equol increased claudin-3, which is the main components for less-permeable TJs in lactation, while an increase in activated STAT5. In addition, a mixture of multiple isoflavones based on the intake of red clover increased secretion of lactoferrin, mRNA expression of ß-casein, and amount of claudin-3, but a mixture based on soy did not affect the BMECs. Thus, these results indicate that isoflavones in legumes and the metabolic activity of isoflavones in dairy cows when feeding legumes may affect the milk production ability in BMECs.
Asunto(s)
Isoflavonas , Factor de Transcripción STAT5 , Animales , Caseínas/metabolismo , Bovinos , Claudina-3/metabolismo , Células Epiteliales/metabolismo , Equol/metabolismo , Femenino , Isoflavonas/farmacología , Lactoferrina/metabolismo , Glándulas Mamarias Animales , Leche/metabolismo , ARN Mensajero/metabolismo , Factor de Transcripción STAT5/genéticaRESUMEN
This study aimed to determine whether the innate immune system in the proventriculus of broiler chicks responds to lipopolysaccharide (LPS) and whether this response is affected by Newcastle disease and infectious bronchitis (ND/IB) vaccination. Chicks were divided into 4 groups: nonvaccinated and injected with PBS or LPS (V-L- and V-L+), and vaccinated and injected with PBS or LPS (V+L- and V+L+). Vaccination was performed on d 1, and LPS was intraperitoneally injected on d 11 of age. The gene expression and protein levels of immune molecules, including toll-like receptors (TLRs), antimicrobial peptides, interleukin-1ß (IL-1B), and immunoglobulin A (IgA) in the proventriculus and serum were analyzed. The results showed that the expression levels of TLR21 were higher in vaccinated (V+L-) group than in nonvaccinated (V-L-) group. Gene expression levels of avian ß-defensin (AvBDs) and cathelicidin1 (Cath1) were not different among the 4 groups. However, the results of LC/MS analysis showed that the levels of AvBD2, 6, and 7 significantly increased after the LPS challenge in nonvaccinated and vaccinated chicks; the levels were higher in V-L+ and V+L+ than in V-L- and V+L-, respectively. Immunohistochemistry analysis revealed the localization of AvBD1 protein in the epithelial cells of the surface glands and AvBD2 and CATH1 in the heterophil-like cells in the lamina propria of surface glands. Although IL-1B gene expression and protein concentration in the proventriculus tissues were not different among the 4 groups, serum IL-1B levels were upregulated by LPS in both the nonvaccinated and vaccinated groups (V-L- vs. V-L+, V+L- vs. V+L+). Moreover, IgA levels in the proventriculus and serum were not affected by vaccination or LPS challenge. Taken together, we conclude that LPS derived from gram-negative bacteria upregulates the innate immune system, including antimicrobial peptide synthesis in the proventriculus. ND/IB vaccination may not significantly affect antimicrobial peptide synthesis in response to LPS; however, TLR21 expression is upregulated by that vaccination. The antimicrobial peptides synthesized in the proventriculus probably prevent pathogenic microbes from entering the intestine.
Asunto(s)
Bronquitis , Enfermedad de Newcastle , Vacunas Virales , Animales , Bronquitis/veterinaria , Pollos , Sistema Inmunológico , Inmunoglobulina A , Lipopolisacáridos , Enfermedad de Newcastle/prevención & control , ProventrículoRESUMEN
In lactating mammary glands, alveolar mammary epithelial cells (MECs) produce milk and form less-permeable tight junctions (TJs). However, alveolar TJs are weakened with a reduction in milk production in mammary glands due to mastitis or weaning in the presence of high levels of IL-1ß, IL-6, or TNF-α. In this study, using in vitro cultured model of MECs with milk-producing ability and lactation-specific TJs, we investigated whether the aforementioned cytokines affect MEC TJs. The results showed that TNF-α, IL-1ß, and IL-6 affected lactation-specific TJs in different ways. In particular, upon activation of p38 and JNK signalling, IL-1ß caused rapid disruption of TJs at tricellular contact points. IL-1ß treatment led to decreased CLDN3, CLDN4, and OCLN levels and a weakened TJ barrier. The adverse effects of IL-1ß on TJs were mimicked by anisomycin, which is an activator of p38 and JNK signalling, and were blocked by MEC pretreatment with a p38 inhibitor but not a JNK inhibitor. The mislocalization of tricellulin at tricellular contact areas was confirmed in MECs treated with IL-1ß or anisomycin. These results indicate that IL-1ß is a key cytokine that adversely affects the TJs between MECs by activating p38.