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1.
After intramammary infection, polymorphonuclear neutrophil leukocytes (PMN) are the first cells recruited into the mammary gland. Rapid recruitment of and bacterial phagocytosis and killing by PMN are the most effective defenses against establishment of bacterial infection. In addition to their phagocytic and bactericidal properties, PMN may play a key supportive role through secretion of cytokines during the innate immune response. We sought to determine whether bovine PMN produce cytokines in response to stimulation by lipopolysaccharide (LPS). To investigate the effects of LPS on the expression of cytokines secreted by bovine PMN, we measured the expression of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-12, and interferon (IFN)-gamma by ELISA after stimulation with different concentrations of LPS, and secretion of IL-8 after co-stimulation with LPS and either TNF-alpha or IL-1beta. Bovine PMN were shown to secrete TNF-alpha , IL-1beta, IL-12, IL-8 and IFN-gamma in response to LPS. Co-incubation of PMN with LPS and TNF-alpha increased secretion of IL-8 when compared to LPS alone. It was concluded that LPS stimulation up-regulates the secretion of cytokines by bovine PMN, and that co-incubation of LPS with TNF-alpha had an additive effect on the secretion of IL-8. These data show that bovine PMN, in addition to their phagocytic and bactericidal properties, may play a supportive role in the innate immune response to infection by Gram-negative bacteria through their ability to produce immuno-regulating cytokines.  相似文献   

2.
The local and systemic immune response to a formolized E. coli oral vaccine was investigated in 13 gnotobiotic piglets. Beginning at ten days of age animals received a daily dose of 1010 or 1011 bacteria, on ten consecutive days. Intestinal loop tests with one animal of each group on day 26 showed protection which was more pronounced in the animal dosed 1010 bacteria compared with the other immunized piglet. Immunoglobulin class-specific antibodies to O and K antigens were determined by ELISA technique. In serum no IgG or IgA antibodies were found, whereas IgM-anti O149 antibodies in both immunized groups reached their highest level at day 4 of dosing and decreased thereafter. IgM-anti K88 antibodies were first detected at day 10 of dosing. Both immunized groups had comparable serum levels at days 20 and 30. Also in gut secretion the IgM antibody response was predominant, and higher levels were found in the 1010 group than in the 1011 group. IgG and IgA antibody response were also detected in secretion.  相似文献   

3.
The host defense competence of uterine-derived polymorphonuclear leukocytes (PMN) from mares considered resistant (grade I uteri) and susceptible (grade III uteri) to chronic endometritis was evaluated for phagocytic and killing (bactericidal) capacities, using a fluorochrome assay. Peripheral blood PMN from noncategorized mares and from grade I and grade III mares were used as controls. Uterine-derived PMN from mares with grade I uteri were functionally competent for phagocytosis and killing of Candida albicans, whereas uterine-derived PMN from mares with grade III uteri had significantly less phagocytic and killing capacities (P greater than or equal to 0.0001). Results of the present study, together with data obtained from chemotactic responsiveness and deformability assays of a previous study, indicated an overall deficiency in the host defense mechanism of uterine-derived PMN from mares with grade III uteri obtained 12 hours after induced Streptococcus zooepidemicus infection. This deficiency may account for the susceptibility of mares with grade III uteri to chronic endometritis.  相似文献   

4.
To examine the effect of parity on polymorphonuclear neutrophils (PMN) function, phagocytic and bactericidal activity of the PMN isolated from blood and milk against Staphylococcus aureus was compared between groups of 6 primiparous and 6 multiparous healthy dairy cows during early lactation using bacteriological and PMN-pathogen interaction assays. Latex-stimulated luminol-amplified chemiluminescence (CL) and viability of these PMN were also investigated. The phagocytosis and killing of S. aureus by blood were remarkably higher than those of milk PMN. Similarly, the CL and viability in blood PMN were markedly higher than in milk PMN. Both in blood and in milk the phagocytosis of S. aureus by PMN in primiparous cows was substantially higher than in multiparous cows. The killing activity of blood PMN against S. aureus was 42.3+/-3.4% and 23.2+/-1.7% in primiparous and multiparous, respectively. Milk PMN killed only 20.7+/-2% S. aureus in primiparous and 10.2+/-1.3% in multiparous cows. Blood and milk PMN CL and milk PMN viability were significantly higher in primiparous cows. The pronounced reduction in phagocytic and bactericidal activity in blood and milk-resident PMN from multiparous cows, in part, resulted from the pronounced decrease of PMN viability and free radicals production capacity; this suggests that heifers' udders could be more protected against S. aureus, which remains to be tested in the field.  相似文献   

5.
The interactions of 2 capsular serotype A and 4 serotype D strains of Pasteurella multocida with rabbit polymorphonuclear neutrophils (PMN) were compared in vitro, using a PMN phagocytic and bactericidal assay. Bacteria and rabbit PMN were incubated for 15 minutes. The suspensions were subjected to differential centrifugation and the percentage of phagocytosis (cell association) was determined from the number of viable noncell-associated bacteria. The cell pellets and the associated bacteria were resuspended and PMN bactericidal activity was calculated from the number of remaining viable cell-associated bacteria at 45 and 75 minutes after the start of the assay. Test bacteria were not opsonized or were opsonized with immune serum containing active complement. One type A strain was ingested and killed by PMN in the presence and absence of opsonins. The 5 remaining strains were resistant to PMN killing, but only the type A strain resisted phagocytosis. Resistance of the type A strain was attributed to the hyaluronic acid capsule, since pretreatment of the bacteria with hyaluronidase rendered opsonized bacteria susceptible to ingestion and killing. The pattern of resistance of the 4 type D strains was different from that of the resistant type A strain. Both opsonized and nonopsonized type D bacteria became cell associated, but none were killed by PMN. The mechanism of resistance of these 4 strains to PMN bactericidal activity is currently unknown.  相似文献   

6.
The influence of the pH of suspending medium on bovine neutrophil (PMN) function was assessed in tests of phagocytosis and killing of Staphylococcus aureus. Intracellular killing was markedly inhibited by moderate extracellular acidification whereas phagocytosis was little affected, except at the lowest pH level (pH 5.0). The killing of S. aureus by extracts of isolated PMN lysosomal granules showed a similar pH dependence and was optimal at pH levels above neutrality. Survival of S. aureus within PMN from different cows varied significantly and the relative differences in PMN bactericidal efficiency were maintained at all pH levels. The acidification of extracellular medium during incubation which resulted from metabolic activity of the PMN themselves, increased with increasing ratios of bacteria:PMN and varied significantly among cows. Addition of methylamine (10 mM) to elevate phagolysosomal pH inhibited phagocytosis and had no effect on intracellular survival of S. aureus. However, a lower concentration (1.5 mM) did not affect phagocytosis, but reduced bacterial survival without altering the relative differences in efficiency of PMN from different cows. It is suggested that the acidity of the extracellular medium may both reflect and influence the pH changes occurring within PMN phagosomes and, thereby, modulate the efficiency of intracellular destruction of S. aureus.  相似文献   

7.
Cattle submitted to the University of Minnesota for surgical correction of left displaced abomasum (LDA) were examined for the in vitro phagocytic and bactericidal activities of their polymorphonuclear leukocytes (PMN). The PMN from cattle with LDA with or without concurrent infection had depressed phagocytic function when compared with PMN from healthy animals (controls). Those with concurrent infection had phagocytic activities lower than those in the group of cattle with LDA without any concurrent infection, and the former group was also observed to have depressed intracellular killing. Cattle with LDA complicated by infection were the only group in which phagocytic function was altered during surgical correction of LDA (and recovery). Treatment of PMN from both groups of affected cattle with levamisole in vitro enhanced intracellular killing, but had no effect on phagocytosis.  相似文献   

8.
9.
Antibacterial activity of cefquinome against equine bacterial pathogens   总被引:8,自引:0,他引:8  
Cefquinome is known for its use as an antibacterial drug in cattle and pigs. The objective of this study was to evaluate the antibacterial activity of cefquinome against equine pathogenic bacteria. The minimum inhibitory concentration (MIC) of cefquinome was determined for a total of 205 strains, which had recently been isolated in Europe from diseased horses (respiratory infection, foal septicaemia). The bactericidal activity was tested against 19 strains using the time killing method. The post-antibiotic effect (PAE) and post-antibiotic sub-MIC effect (PA SME) were determined against 12 strains. Cefquinome showed high activity against Actinobacillus equuli and streptococci (MIC(90) of 0.016 and 0.032microg/mL), Enterobacteriaceae (MIC(90)=0.125microg/mL) and staphylococci (MIC(90)=0.5microg/mL). The activity was limited against Rhodococcus spp. and Pseudomonas spp. Cefquinome was shown to be a time dependent bactericidal antibiotic against the target pathogens, killing occurring at a concentration close to the MIC. A PAE of 0.5-10h was calculated against streptococci whereas no PAE was observed for Escherichia coli. A longer PA SME was determined for streptococci (3.3 to >24h with a killing effect) and E. coli (0.5-13.9h). Cefquinome was shown to have a broad spectrum of activity which covers many equine pathogens.  相似文献   

10.
The mechanism of resistance of Capsular Type D strains of Pasteurella multocida to killing by rabbit polymorphonuclear neutrophils (PMN) was studied using an in vitro assay that differentiates intra- from extracellular bacteria. Two Capsular Type D strains (3761 and 3766), resistant to killing by rabbit PMN, and one Type A strain (R1), susceptible to PMN destruction, were compared. After combining opsonized bacteria and PMN, the Capsular Type D Strains 3761 and 3766 remained extracellular while the Capsular Type A Strain R1 was internalized by PMN. Thus, both Type D strains were resistant to phagocytosis by rabbit PMN.  相似文献   

11.
Lactoferrin (LF) is a glycoprotein found in milk, neutrophil granules, secretions and selected organs of mammals. Lactoferrin exhibits antibacterial, antiviral, fungicidal, immunoregulatory and other functions. Although fish are devoid of this protein and its cell receptors, LF effect on the immune mechanisms of fish has been demonstrated. The objective of this study was to investigate the effect of bovine lactoferrin, applied in vitro, on the activity of head kidney and spleen leukocytes in three freshwater fish species: rainbow trout (Oncorhynchus mykiss), European eel (Anguilla anguilla) and wels catfish (Silurus glanis). The obtained results validate LF beneficial effect on the respiratory burst of phagocytes in rainbow trout and wels catfish despite the fact that the potential killing activity against Aeromonas hydrophila was not stimulated in any of the studied species. Bovine lactoferrin enhanced the proliferation of T-lymphocytes in rainbow trout and European eel, as well as of B-lymphocytes in rainbow trout.  相似文献   

12.
Detergent-extracted recombinant Omp31 (rOmp31 extract) from Brucella melitensis produced in Escherichia coli was previously identified as a protective immunogen against B. ovis in mice. In this study, we evaluated the immunogenicity of rOmp31extract in rams. This immunogen was emulsified in an oil adjuvant and administered three times with 4 and 8 weeks intervals. Antibody response was measured in serum by whole B. ovis ELISA. Specific antibodies to purified rOmp31 (pET-Omp31) were detected by Western blotting and indirect ELISA. In addition, isotype specific antibodies were measured in tears. Serum bactericidal activity against B. ovis in the presence of complement was measured in vitro. Cellular immune response was explored by intradermal testing with purified rOmp31. Immunization with rOmp31 extract induced IgG specific antibodies in serum able to bind to whole B. ovis cells. Furthermore, strong inhibition in a competitive ELISA (with an Omp31-specific monoclonal antibody) suggested that a proportion of Omp31-specific antibodies were directed against a loop containing a protective epitope. Serum antibodies killed efficiently B. ovis in vitro in the presence of either guinea pig or ovine serum. Tears had both IgG and IgA antibodies to equivalent titers. Finally, immunized rams showed skin reactivity to Omp31. These data demonstrate that B. melitensis Omp31, a protective antigen identified in the mouse model, induces antibody and cellular immune mechanisms in sheep.  相似文献   

13.
The phenotypic characteristics are described for the activity of polymorphonuclear leucocytes NMN) obtained by either lavage of the cavity system of juvenile mammary glands stimulated with a synthetic muramyl dipeptide analogue or isolation from the peripheral blood. Attention was paid to the variability of characteristics and its sources, and to correlations among them. The following characteristics were investigated in 27 clinically healthy, unbred Bohemian Red Pied x Holstein heifers: migration activity in situ, number of phagocytosing PMN, phagocytotic index, bactericidal activity of PMN and unstimulated and zymosan-stimulated luminol-dependent chemiluminescence. Considerable individual variation was found in the characteristics. Significant differences between blood PMN and PMN from lavages after influx induction were found for bactericidal activity (P < 0.05) and chemiluminescence (P < 0.01). A significant correlation between blood PMN and mammary gland PMN was found only for the number of phagocytosing cells (r = 0.329; P < 0.01). Highly significant positive correlations (P < 0.01) were demonstrated between the number of phagocytosing PMN [a], phagocytotic index [b], and bactericidal activity [c] in both blood PMN (r(ab) = 0.602; r(ac) = 0.565; r(bc) = 0.529) and mammary gland PMN (r(ab) = 0.730, r(ac) = 0.618, r(bc) = 0.589). No significant correlation was demonstrated for non-stimulated (NS), zymosan-stimulated (ZS), or opsonized zymosan-stimulated (OZS) chemiluminescence with any of the other characteristics of phagocytotic activity, in either blood PMN or mammary gland PMN (P > 0.05). The animal was a highly significant source of variability for all the phagocytotic activity characteristics (P < 0.01). Udder quarter was a non-significant source of variability for all the characteristics of phagocytotic activity except for NS chemiluminescence (P < 0.05) and ZS or OZS chemiluminescence (P < 0.01). However, udder quarter was a non-significant source of variability of chemiluminescence indices ZS/NS and OZS/NS (P > 0.05). It has been demonstrated that in situ migration activity, the number of phagocytosing PMN, phagocytotic index, bactericidal activity of PMN and chemiluminescence indices of PMN collected from juvenile mammary glands of unbred heifers after influx induction can be regarded as candidate early markers of resistance to mammary infections.  相似文献   

14.
Enterotoxigenic Escherichia coli (ETEC) strains that produce K88 (F4)+ fimbria are important causes of diarrhea and post-diarrheal septicemia in swine. ETEC O8:K87, a serotype represented by a number of these strains, is typically serum resistant. Strain-specific antibodies are known to activate alternative C pathway-mediated killing of other serum-resistant E. coli [Hill, A.W., Shears, A.L., Hibbitt, K.G., 1978. The requirement of specific antibody for the killing of E. coli by the alternate complement pathway in bovine serum. Immunology 34, 131-136], but their antigenic targets have not been determined. We tested the hypothesis that anti-K87 antibodies activate alternative pathway-mediated killing of ETEC O8:K87. Pigs were immunized with ETEC O8:K87 strain 2534-86 cells or purified K87 polysaccharide. Post-, but not pre-immunization sera killed 2534-86 cells, and absorption with 2534-86 cells or by K87 affinity chromatography eliminated bactericidal activity. Complementation of absorbed serum with anti-K87 antibodies restored bactericidal activity, confirming the ability of these antibodies to activate C-mediated serum killing. Serum from age-matched, non-vaccinated control pigs also killed 2534-86. This activity was eliminated by absorption with 2534-86 cells, but not K87 affinity chromatography, indicating that specific non-capsular antibodies are also able to activate C-mediated killing. In all cases, Mg-EGTA-treated serum was as effective as non-treated serum in killing, suggesting that bactericidal activity was mediated predominantly if not exclusively via the alternative C pathway.  相似文献   

15.
The effect of bovine mammary secretion during the early nonlactating period and of antibiotic preparations on bovine polymorphonuclear neutrophil (PMN) phagocytic function and morphology were evaluated in a series of in vitro multifactorial experiments. Benzathine cloxacillin (CL), benzathine cephapirin (CE), sodium novobiocin (NO), and a combination of dihydrostreptomycin with procaine penicillin G (DP) were prepared in the presence and absence of a peanut oil aluminum monostearate vehicle. The PMN were isolated from bovine blood, and the effect of each antibiotic preparation on PMN function and morphology was evaluated in a buffer, fat, skin, and a combination of fat with skim from bovine mammary secretion during the nonlactating period. The fat and skim were diluted with buffer to approximate their concentration in mammary secretion. Phagocytic functions of PMN were monitored by fluorescent microscopy, which made it possible to estimate both ingestion and intracellular killing of bacteria by PMN. Changes in PMN morphology were monitored by transmission electron microscopy. The ability of PMN to ingest and kill Staphylococcus aureus ATCC 25923 was significantly decreased by fat, skim, CL, CE, NO, and DP. Effects of some antibiotics on ingestion and killing of bacteria by PMN were influenced by the addition of vehicle and by interactions with mammary secretion. Neutrophil morphology was altered by fat, skim, CL, CE, NO, and DP. The detrimental effects of CL, CE, NO, and DP on PMN morphology were influenced (some significantly) by the presence of vehicle and interactions with mammary secretion. There were significant correlations among secretion- and antibiotic-induced changes in PMN ingestion of bacteria, PMN killing of bacteria, and PMN morphology.  相似文献   

16.
The humoral and cell-mediated immune (CMI) response to 2 commercial killed Salmonella Enteritidis (SE) vaccines (Layermune and MBL SE4C) was evaluated in laying hens. Layers were distributed in 2 experimental groups. The first received a single immunization at 16 wk of age, while the second experimental group was immunized at 12 wk of age and again at 18 wk of age. Serum immunoglobulin (Ig)G antibodies were measured using a commercial SE ELISA kit and showed persistent levels from 3 to 32 and 34 wk post-vaccination. The vaccination protocol using 2 immunizations showed a higher seroconversion level than the single vaccination. However, our results for bacterial intracellular survival indicated that IgG titers were not linked with bacterial killing. Local IgA production was measured in the intestines and oviducts with an in-house SE whole cell antigen ELISA. Only the MBL SE4C vaccine elicited IgA antibody production when tested on intestine and oviduct mucosal secretions, 3-weeks post-vaccination in both immunization protocol groups. To evaluate the CMI response, the splenic T-cells and B-cells populations were analyzed using flow cytometry. The CD3/B-cell ratio decreased 3 wk after the second immunization in the twice vaccinated Layermune group due to an increase in B-cells.  相似文献   

17.
柞蚕抗菌肽对柑桔黄龙病及溃疡病病原菌的杀菌作用   总被引:9,自引:1,他引:8  
首次报道柞蚕(Antheraeapernyi)抗菌肽对柑桔黄龙病病原类细菌(BacteriallikeOrganism,BLO)及溃疡病黄单胞菌(XanthomonascampestrisPVcitriDye)的杀菌作用,分别在平板培养基上用孔穴法注入5—10μl免疫血淋巴,则出现明显的抑菌圈。用纯化的抗菌肽10μg/ml处理BLO及X.citri20─60min,用电镜及激光共焦扫描显微镜观察到病原菌细胞膜的损坏、内容物泄出乃至死亡的过程,对抗菌肽的杀菌机理进行了讨论。  相似文献   

18.
Several isolates of logarithmic-phase organisms of Pasteurella haemolytica were shown to be sensitive to an antibody and complement-mediated killing mechanism in adult bovine serum. Data suggested that the classical complement pathway was important in the induction of bactericidal activity of serum. Sera from calves after colostrum feedings (post-colostral sera) killed only 30% of the bacteria in spite of the presence of high levels of antibodies against P. haemolytica. Addition of post-colostral serum to heat-inactivated adult bovine serum decreased the bactericidal capacity of the latter. It was speculated that this inhibition may have been caused by the presence of blocking antibodies (IgA) found in the post-colostral serum. Undiluted nasal secretions collected from adult cattle were not bactericidal to P. haemolytica. The results also suggest that the bronchoalveolar washings (BAW) from vaccinated calves, in spite of having a high antibody titer, were less bactericidal to P. haemolytica than BAW from sham-vaccinated calves (71.12% vs. 83.12%). The bactericidal factor(s) present in BAW from sham-vaccinated calves was heat stable, not complement dependent, and was not related to lysozyme concentration.  相似文献   

19.
The impact of replacing soybean crude protein (CP) with CP from ensiled taro leaves (ET) on growth performance, carcass traits, and organ weights in Landrace × Yorkshire (LY) and Moo Lath (ML) Lao pigs was studied. Twenty-four castrated male pigs, 12 of each breed, were allocated to the treatments according to a completely randomized 3?×?2 factorial (three levels of ET × two breeds) arrangement with four pigs per treatment. The pigs were kept in individual pens and were fed at 4 % dry matter of body weight for 105 days. The control diet (ET0) was formulated with soybean meal as the main CP source, and in the other two diets, soybean CP was replaced to 25 % (ET25) and 50 % (ET50), respectively, with CP from ensiled taro leaves. Calculated metabolizable energy intake decreased with increasing replacement of soybean CP in the diet, while dry matter intake (DMI), CP intake (CPI), average daily gain (ADG), and feed conversion ratio (FCR) were unaffected. Carcass weight, back fat thickness, and dressing percentage were unaffected by soybean CP replacement, while organ weights (except for spleen) increased (P?<?0.001) when soybean CP was replaced by CP from ensiled taro leaves in the diet. LY pigs had higher (P?<?0.001) DMI, CPI, and ADG and better (P?<?0.001) FCR than ML pigs. LY pigs had higher carcass weight (P?<?0.001), lower back fat thickness (P?<?0.001), and higher organ weights (P?<?0.05–0.001) than the ML pigs. In conclusion, taro leaf silage can replace up to 50 % of soybean CP in the diet of growing Lao LY and ML pigs without negative effects on performance and carcass traits.  相似文献   

20.
The lipoidal amine, N,N-dioctadecyl-N',N'-bis (2-hydroxyethyl) propanediamine (avridine or CP 20,961), formulated in liposomes, was evaluated for its effect on leukocyte kinetics, lymphocyte blastogenesis, and polymorphonuclear leukocyte (PMN) function in dexamethasone-treated and nontreated cattle. In the 1st experiment, cattle were given avridine in a single IM injection of 0.1, 1.0, or 10 mg/kg of body weight. All doses induced swelling at the injection site, a febrile response, and a leukocytosis due to a neutrophilia. Mononuclear cell numbers were normal. All 3 groups of avridine-treated animals had a higher mean lymphocyte blastogenic response to mitogens on the 4 days after administration than did the control nontreated animals. Avridine administration was associated with an enhanced ability of PMN to ingest Staphylococcus aureus and to mediate antibody-dependent cell-mediated cytotoxicity (ADCC). The highest dose (10 mg/kg) was associated with a depression of the ability of PMN to iodinate protein. An effect of avridine on PMN random migration under agarose or nitroblue tetrazolium (NBT) reduction was not observed. In a 2nd experiment, cattle were given no treatment, 0.04 mg of dexamethasone/kg IM, or 10 mg of avridine/kg IM followed 24 hours later by 0.04 mg of dexamethasone/kg. Dexamethasone administration caused a leukocytosis due to a neutrophilia with normal mononuclear cell numbers, an enhancement of PMN random migration under agarose, and an inhibition of NBT reduction, iodination, and ADCC activity of PMN. Dexamethasone did not have a detectable effect on lymphocyte blastogenesis or on ingestion of S aureus by PMN.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

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