The antibody response to bovine lymphocyte alloantigens

Antibodies were raised against lymphocyte cell-surface antigens by multiple immunisations with purified lymphocytes or by the exchange of skin allografts. Eighteen of 21 cattle immunised with lymphocytes raised a detectable cytotoxic antibody response. The serum antibody from 10 responders recognized only common lymphocyte antigens, those antigens which are present on all peripheral blood lymphocytes. One animal responded only to B lymphocyte antigens while 7 others responded to both classes of antigens. The amount of antibody produced varied greatly between individuals; antibody titres ranged from 1 to 1028. Antibody raised early in the response was sensitive to treatment with 2-mercaptoethanol (2-ME) suggesting that IgM was the predominant class of immunoglobulin. Subsequently antibody became resistant to this treatment suggesting the appearance of IgG. The antibody responses following the exchange of skin grafts were very similar in all 12 cattle studied. High titred antibody to common lymphocyte antigens was detected in the serum 14 days after grafting. The early antibody activity was sensitive to 2-ME treatment but became totally resistant within 14 days. Total peak antibody titres ranged from 128-2048. Antibody to B lymphocyte antigens was identified in 8 of the 12 cattle. The responses to B lymphocyte antigens were similar to those against the more widely distributed common lymphocyte antigens with respect to time of antibody appearance, time of peak titre and sensitivity to 2-ME. Peak titres ranged from 2 to 32. The change in antibody specificity with time was also studied. Sera from 11 of the 18 cattle which had responded against lymphocytes showed an increase or broadening in reaction frequency as immunisations increased, suggesting the production of antibody to secondary specificities. In the cattle which had been skin grafted, the broadest reaction patterns were seen 14 to 21 days after grafting. The broadest reaction patterns were seen when the antibody responses were at their highest titre levels and narrowed as titres decreased.     

Failure to detect antibody to bovine respiratory syncytial virus in bovine fetal serum.

Sera obtained from 147 bovine fetuses estimated to be between 120 and 270 days of gestation at an abattoir were tested for antibody to bovine respiratory syncytial virus. Antibody to bovine respiratory syncytial virus was not detected in any of the sera examined. Based on the results of this study and a review of the literature, it appears that transplacental infection by bovine respiratory syncytial virus does not occur, or is uncommon.     

Influence of orally administered CpG-ODNs on the humoral response to bovine serum albumin (BSA) in chickens

Synthetic oligodeoxynucleotides containing CpG motifs (CpG-ODNs) have been reported to be effective mucosal adjuvants in mice when given orally. Studies on their effectiveness in chickens are currently very limited. This study investigated whether CpG-ODNs could adjuvant the immune response to BSA when given orally to a commercial line of SCWL chickens. In two experiments, performed over time, chickens were given selected concentrations of CpG-ODNs with BSA followed by 6 consecutive days of ad libitum access to drinking water containing 1.4 mg/ml BSA. Serum responses, and in some cases intestinal specific antibodies, were measured out to 33 days post-immunization. Birds receiving a single dose of CpG-ODN had consistently higher IgG, IgM, and IgA titers in the serum, dependent upon dose, and in specific areas of the intestine when compared to the non-immunized and BSA only groups. These findings suggest that a single oral CpG-ODN administration can accelerate the kinetics of antigen specific antibodies of all three isotypes in commercial-strain chickens immunized via the drinking water using common protein antigen.     

Dietary L-carnitine supplementation enhances the lipopolysaccharide-induced acute phase protein response in broiler chickens

The objective of the present study was to evaluate the potential effects of dietary L-carnitine supplementation on acute phase protein response upon a lipopolysaccharide (LPS) challenge of male broiler chickens receiving a commercial broiler diet supplemented with 15 or 100 mg L-carnitine/kg or an unsupplemented (control) diet from 14 days of age onwards. At 28 days of age, eight chickens per dietary treatment were weighed and subcutaneously injected with 300 microg LPS from E. coli (100 microg LPS/ml saline) or 3 ml saline (unsupplemented group only). During the next 10 days, blood samples were taken repeatedly and analysed for their hemopexin (HX) and alpha-1 acid glycoprotein (AGP) levels. Extra dietary L-carnitine did not affect broiler performance. At day 1 postinjection, plasma HX and AGP levels were significantly increased in all treatment groups. However, the elevations in circulating HX and AGP levels were more pronounced in the L-carnitine supplemented chickens, especially in the 100mg L-carnitine group. It is concluded that extra L-carnitine in the diet of broiler chickens enhances or advances the acute phase protein response. The exact mode of action needs to be elucidated but seems to be consistent with a glucocorticoid mimicking effect.     

Chromium supplementation enhances antibody response to vaccination with tetanus toxoid in cattle

Nineteen multiparous late-pregnant dairy cows were divided into an experimental group (n = 10) and a control group (n = 9). Animals in the experimental group were fed a diet supplemented with chromium chelate admixed into wheat meal at 5 mg per animal per day. The supplementation was started 4 weeks before calving and stopped by the end of week 3 after calving. All the cows were vaccinated with tetanus toxoid in the fifth and sixth week of lactation. Monitoring of the characteristics of non-specific immunity did not show any significant between-group differences in total and differential leucocyte counts, percentages of lymphocyte subpopulations, activities of lectin-stimulated lymphocytes, phagocytic activities, and the contents of total immunoglobulins and lysozyme in blood sera. Tetanus toxoid-specific antibody titres, those of the IgG2 isotype in particular, were higher in the experimental group than in the control animals. These results indicate that chromium supplementation at the level used in this experiment modulated the regulation of functions of the immune system.     

Selection for immune response in goats: the effect of immunization procedure on antibody response to diphtheria toxoid and human serum albumin.

The serum antibody titers to diphtheria toxoid and human serum albumin were determined in 103 goat kids from lines selected for 12 yr for high or low antibody response to diphtheria toxoid. In the 12th yr, six groups of kids were immunized with different preparations of the antigens. In all groups but one, the antigens were emulsified in Freund's incomplete adjuvant with added sonicated Mycobacterium paratuberculosis. The groups received the following treatments: Group 1 was immunized with both antigens mixed in the same syringe, Group 2 got both antigens injected separately, Group 3 got both antigens injected separately, but with a lower concentration of M. paratuberculosis, Group 4 was immunized with diphtheria toxoid only, Group 5 was immunized with human serum albumin only, and Group 6 was immunized with both antigens mixed, but without any M. paratuberculosis. The animals were immunized at 4 wk of age, and the antibody titers were determined 3 wk later by ELISA and passive hemagglutination. The mean antibody titers to both antigens were different between the selected lines (P less than .03). There was no effect of separate vs combined injections of antigens. However, there were indications of antigen suppression or competition between the antigens. Animals receiving only one antigen seemed to mount a higher antibody response to that antigen than did animals immunized with two antigens.     

Studies on adjuvant-induced arthritis, tumor transplantability, and serologic response to bovine serum albumin in Sendai virus-infected rats.

Sendai virus, one of the most prevalent of the murine viruses, was studied in regard to its capability to alter various functions of rats given a second, unrelated antigenic challenge. Rats given a single foot-pad injection of Freund's complete adjuvant (FCA) had an 85% incidence of adjuvant arthritis. The adjuvant disease was significantly less severe (P less than 0.01) in those rats given 0.05 ml of 10(5.5) median egg infective doses of egg-propagated Sendai virus intranasally 7 days before injection of adjuvant. Rats given Sendai virus concurrently with the FCA, or any time after FCA was injected, did not have a lessened severity of the arthritic reaction, as compared with that in control animals. Sendai virus infection had no detectable effect on median tumor dose requirement for Walker carcinosarcoma cells in rats or on the antibody response to bovine serum albumin.     

The use of concentrated bovine serum albumin in canines.

In humans it has been estimated that for each 2.5 g L^–1 decrease in serum albumin, risk of death increases by 24–56%. Clinical impression suggests this may be similar in veterinary patients. Species‐specific albumin (plasma) is often unavailable and concentrated solutions are not. Our experience using 25% human serum albumin (HSA) in critically ill dogs suggests a positive effect (results submitted), however it is expensive. Bovine serum albumin (BSA) may be a more cost effective and readily available alternative. The purpose of this study was to assess the immediate and long‐term safety of an intravenous dose (500 mg kg^–1) of bovine albumin administered to healthy dogs. Ten mature dogs (eight males, two females, 28 ± 6 kg) were to receive BSA (250 mg mL^–1) twice (BSA1 and BSA2) with 14 days between treatments. Temperature, blood pressure, and pulse and respiration rate were continuously monitored to identify a reaction to BSA. All dogs received BSA1. One dog immediately developed mild urticaria and pruritus, otherwise the infusion was well tolerated. No immediate reaction was noted in the other nine dogs. Two dogs received BSA2. One dog developed a mild immediate reaction similar to that occurring with BSA1, and one dog (the dog immediately reacting to BSA1) developed a severe anaphylactic reaction. Due to these reactions, no other dogs received BSA2. During a two‐week observation of the remaining eight dogs given BSA1, five developed a mild or severe generalized type‐III hypersensitivity reaction. The dog experiencing a mild reaction during BSA2 administration also developed a generalized type‐III hypersensitivity reaction. Delayed reactions occurred 15 ± 2.7 days after BSA exposure. Three dogs did not develop a reaction. All reacting dogs recovered fully. The severity of reactions, and the number of dogs affected, suggests prior (natural) exposure and immunological sensitization to bovine albumin. Bovine serum albumin is not suitable for therapeutic use in dogs.     

Characteristics of bovine spermatozoa after migration through a bovine serum albumin gradient

Experiments were conducted to determine the efficiency with which viable, morphologically normal bovine spermatozoa could be isolated using a discontinuous bovine serum albumin (BSA) gradient. In the first experiment, extended semen was layered on top of a BSA gradient (4% BSA over 10% BSA) contained in a 500-ml separatory funnel. When comparing 1, 3, 5, 7, 14 or 21 X 10(9) spermatozoa applied to the gradient, the percentage of spermatozoa recovered from the lower third of the 10% BSA ranged from 2.9 to 18.5%. The greatest recovery was achieved when 1 X 10(9) sperm cells were applied. Increasing the number of spermatozoa applied to the gradient increased the percentage of spermatozoa remaining in the upper portions of the gradient. Motility of spermatozoa immediately after collection from the 10% BSA layer of the gradient was greater than 90%, regardless of the number of spermatozoa applied. In a second experiment with freeze-thawed separated or unseparated spermatozoa, post-thaw motility (greater than 60%) and acrosomal integrity (greater than 85%) of separated spermatozoa (4 or 10% BSA layer) was greater (P less than .05) than that of unseparated spermatozoa (38 and 66%, respectively). The discontinuous gradient excluded decapitated spermatozoa and spermatozoa with mid-piece and principal piece abnormalities from entering the lower layers. Sperm cells with head abnormalities were not separated. These data indicate that a population of spermatozoa with a high frequency of viable, motile, morphologically-normal bovine spermatozoa can be isolated using a discontinuous BSA gradient.     

Enhanced freezability of goat spermatozoa collected into tubes containing extender supplemented with bovine serum albumin (BSA)

The purpose of this study was to investigate the effects of semen collection into tubes containing extender supplemented with BSA on the cryosurvival of goat spermatozoa. Semen was collected from two goats into empty tubes or tubes containing 10 ml extender supplemented with 0, 0.1, 1, or 5% BSA, and the washed spermatozoa were frozen as pellets in egg yolk-trehalose extender with the addition of 0.04% SDS and 4% glycerol. Sperm motion parameters were evaluated after post-thawing and during a thermal resistance test. The acrosome status of frozen-thawed spermatozoa was also observed using FITC-PNA staining. In frozen semen that was collected into tubes containing extender supplemented with 5% BSA, the post-thawed spermatozoa exhibited a significant improvement in motion parameters and maintained high motility throughout incubation and acrosome integrity, as compared with semen collected into tubes containing extender supplemented with lower concentrations of BSA. In conclusion, semen collection into tubes with a large volume of extender containing high concentrations of BSA dramatically improves the motility and acrosome integrity of frozen-thawed spermatozoa. This suggests that the in vitro functional freezability of spermatozoa is abruptly modified by reducing contact with seminal plasma and by flash contact with BSA at ejaculation.     

Experimental immune complex glomerulonephritis in dogs receiving cationized bovine serum albumin

Eight 16-week-old dogs were used to induce immune complex glomerulonephritis by daily intravenous injections of 120 mg highly cationized bovine serum albumin (pI9.5). Of four control dogs, two received unmodified native anionic bovine serum albumin (pI 4.5) while the other two received only phosphate buffered saline. The renal glomeruli were examined by light, electron (transmission and scanning) and immunofluorescence microscopy at intervals from five to 11 weeks after the start of the injections. Animals receiving cationic antigen all developed generalised diffuse granular deposits of IgG and C3 along the capillary walls; these were detected as early as five weeks and continued until the termination of the experiment at 11 weeks. Ultrastructural studies revealed many electron dense deposits along the subepithelial regions of the glomerular basement membrane. The experimental disease resembled in many respects naturally occurring membranous nephropathy, the most common form of immune complex glomerulonephritis in dogs.     

Characterization of the bovine secondary in vitro antibody response

In the present report, the bovine secondary in vitro antibody response to keyhole limpet hemocyanin is described. The induction of anti-KLH antibody was not dependent upon the presence of mitogen but was antigen specific (KLH vs ovalbumin). Furthermore, this response was dependent upon cell density (10(6) per well), antigen dose (1 to 10(-5) ug/culture) and time in culture (5 days). The antibody produced was specific for KLH as measured in several binding assays. An unresponsive state was detected with high concentrations of KLH (more than 10 ug per culture) which was not due to the formation of immune complexes but to the inactivation of B and/or T cells. The induction of the antibody response was dependent on the presence of macrophages (syngeneic or allogeneic) and their presence could not be replaced with 2-mercaptoethanol.     

Sex ratio after insemination of bovine spermatozoa isolated using a bovine serum albumin gradient

This experiment was undertaken to determine if a method reported to successfully enrich the proportion of Y-chromosome-bearing spermatozoa in human semen could be adapted for separation of bovine spermatozoa. Semen was collected from four Angus bulls and aliquots were either separated on discontinuous gradients of bovine serum albumin (BSA) or untreated before processing for cryopreservation. Two hundred seventy-one cows or heifers were assigned randomly to be artificially inseminated (20 X 10(6) sperm/insemination) with separated or unseparated spermatozoa. The proportions of male offspring were 45 and 54% after inseminations with separated or unseparated spermatozoa, respectively. In a second phase of the experiment, pooled semen from three Holstein bulls was either extended and frozen without separation or frozen after separation using the discontinuous BSA gradient. Separated and unseparated spermatozoa were analyzed by flow cytometry to determine the ratio of X- and Y-chromosome-bearing spermatozoa based on differences in DNA content. The ratios of X- and Y-bearing spermatozoa in separated or unseparated samples were indistinguishable. We concluded that the separation method did not enrich the proportion of Y-bearing bovine spermatozoa.     

L-carnitine supplementation in the therapy of canine dilated cardiomyopathy

In evaluating any novel therapeutic agent or intervention, the clinician must question the need for as well as the efficacy and safety of the proposed therapy. In an attempt to address these critical questions with regard to the role of L-carnitine supplementation in the therapy of dilated cardiomyopathy, this article reviews the currently available scientific evidence and clinical experience regarding carnitine metabolism and supplementation in the dog.     

猪流行性腹泻病毒血清IgA抗体间接ELISA方法的建立

建立了检测猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)IgA抗体的间接ELISA方法,旨在为PEDV感染检测与免疫效果评价提供技术手段。以重组PEDV结构蛋白(S1蛋白)作为包被抗原,采用方阵滴定法确定抗原包被浓度、封闭液、稀释液以及待测血清和酶标抗体的最佳工作浓度,建立了PEDV IgA抗体检测间接ELISA方法。进而检测方法的特异性、批内与批间重复性,评价建立方法与国外试剂盒的符合率,分析ELISA检测血清特异性IgA抗体水平与中和抗体的相关性。结果显示,ELISA方法的最佳抗原包被浓度为1 mg/L,封闭液和抗体稀释液为含5%犊牛血清的PBS,待检血清和酶标抗体的工作浓度分别为1∶40与1∶1 500倍稀释。结果表明,该ELISA能够特异地检测PEDV抗体,与猪繁殖与呼吸综合征病毒、猪瘟病毒、伪狂犬病毒、猪圆环病毒2型等病毒的抗血清无交叉反应,批内和批间重复性试验的变异系数低于10%,与现有试剂盒的总符合率为94.8%,血清特异性IgA抗体水平与中和抗体呈正相关(r=0.69,P<0.001)。     

Possible association of antibody responses to human serum albumin and (T,G)-A--L with the bovine major histocompatibility complex (BoLA)

Antibody responses to human serum albumin (HSA) and (T,G)-A--L were determined in 130 young bulls in Norway and the BoLA types of the bulls were defined. Significant associations of some BoLA antigens with immune responsiveness were shown, indicating the likely existence of an immune response (Ir) region linked to the BoLA class I antigens. High response to HSA seems to be a dominant trait. BoLA w2 showed an association with low response to HSA. This may reflect the effect of a specific MHC-associated immune suppressor gene.     

Inhibition of bovine lymphocyte response to phytohemagglutinin by a specific 5-lipoxygenase inhibitor

AA861, a specific 5-lipoxygenase inhibitor, inhibited bovine lymphocyte response to phytohemagglutinin (PHA). Mitogen-stimulated cultures of mononuclear cells produced leukotriene B4 (LTB4) in 24 hours. The production of LTB4 was completely inhibited by concentrations of AA861 that inhibited mitogen-induced 3H-thymidine incorporation. The inhibition of lymphocyte proliferation was totally reversed by LTB4 but not by leukotriene C4 or leukotriene D4. The inhibition of interleukin-2 (IL-2) production by AA861 was also completely reversed by addition of exogenous LTB4 to lymphocyte cultures. Thus, endogenous LTB4 production appeared to be necessary for PHA-induced IL-2 production and lymphocyte proliferation.