Measurement of immunoglobulins in reproductive tract fluids of bulls

Seminal, seminal vesicular, urethral and preputial fluids from bulls of two different age groups were assessed for quantitative differences in immunoglobulins. Selected markers were measured in individual samples to differentiate locally derived immunoglobulins from those present as a result of trauma or secretions from other accessory glands.Immunoglobulin levels in reproductive tract fluids from older bulls (5–6 years) were higher than those of younger bulls (3–4 years) and preputial fluids contained the highest concentration of immunoglobulins of all fluids examined. Similarities existed, however, among all fluids in the relative concentrations of immunoglobulins. IgG was generally in highest concentration, though the predominant subclass varied. A marked predominance of IgG₂ over IgG₁ occurred in preputial fluid samples of older bulls. IgA was in second highest concentration, and levels were often equal to or greater than those in serum. IgM was in low concentration and occasionally undetectable. IgG/IgA ratios did not exceed 5 in most of the reproductive fluids, whereas serum ratios were usually over 100. Proportional contents of albumin and immunoglobulin in reproductive tract fluids by comparison with those in serum indicated that substantial quantities of IgG as well as IgA were synthesized locally or derived by selective transport. Increased numbers of plasma cells in the lamina propria of the preputial and penile mucosa of older bulls were correlated with higher immunoglobulin concentrations in preputial fluid from older bulls, suggesting that differences in local synthesis were responsible.     

The differential localization of IgA,IgM and IgG in the gut of suckled neonatal piglets

The localization of immunoglobulins IgG, IgM and IgA in tissue sections prepared from the ileum of neonatal and adult swine were compared. Eighty percent of the immunoglobulin-containing lymphoid cells in the lamina propria of conventional adult German Landrasse swine were IgA-positive with lower numbers of IgM cells and occasionally an IgG cell. Anti-μ and α-chain reagents also stained the cytoplasm of the crypt epithelial cells. By comparison to these adult control tissues, the ileum of unsuckled neonates contained no immunoglobulins although after the ingestion of colostrum, the entire cytoplasm of the villus epithelial cells stained intensely when tested for IgG with only faint staining for IgM and IgA. On the other hand, IgA and IgM were readily localized on what appears to be only the apical border of the crypt epithelial cells but in contrast to the adult, the cytoplasm of these cells was unlabelled. IgG was absent from the crypt region. We interprete these findings to indicate an important, selective role for the villus epithelium in the absorption into the neonatal circulation of colostral IgG and probably IgA and IgM, and a specialized role for the crypt epithelium in adsorbing colostral IgA and IgM; possibly by complexing with mucin-bound secretory component.     

Comparison of inoculation regimes for the experimental production of swine erysipelas arthritis: I Clinical, pathological and bacteriological findings

The arthritic form of swine erysipelas was induced in pigs by multiple intravenous inoculation of 2 strains of Erysipelothrix rhusiopathiae. The strains differed significantly in their arthritogenicity but not in the number of cases of lameness induced. The use of 3 intravenous inoculations instead of 5 did not significantly affect the outcome. In a second trial, the more arthritogenic strain was injected in ten-fold dilutions from 5 x 10(9) to 5 x 10(4) organisms. Pigs receiving the lower doses showed high variability in their arthritic responses that precluded sensitive analysis of the dose effects on the number of arthritic and infected joints. However, slaughter weights showed a significant negative correlation with dose. Mean slaughter weights in treatment groups varied by 14.6 kg per pig, an average weight loss of 3 kg per pig for each ten-fold rise in dose of the highly virulent strain, and significantly correlated with the number of arthritic and infected joints. Culture of homogenised synovial membrane through selective horse meat-serum broth containing kanamycin, neomycin and vancomycin identified 66% and 59% more infected joints than primary blood agar culture of synovial fluid or synovial membrane homogenate, respectively.     

Quantitation of bovine immunoglobulins in culture fluids by use of sandwich radioimmunoassay with monoclonal antibodies

Bovine immunoglobulin isotype-specific murine monoclonal antibodies were used in sandwich radioimmunoassays to detect and quantitate bovine IgG1, IgG2, IgM, and IgA in culture fluids. The concentrations of bovine immunoglobulins in unknown samples were extrapolated from standard curves generated with bovine monoclonal immunoglobulins. The lowest detection limits for the bovine immunoglobulin isotypes ranged from 65 to 270 ng/ml.     

Maternal-neonatal immunoregulation: suppression of de novo synthesis of IgG and IgA, but not IgM, in neonatal pigs by bovine colostrum, is lost upon storage

Fifty-four neonatal pigs were allotted to 4 groups and reared in an electrically controlled automatic feeding device (autosow). Each group was reared on a different pool of bovine colostrum: fresh, stored 1 month, stored 6 months, and stored 8 years. Bovine and porcine immunoglobulins in the sera of these pigs, and in a group of conventionally reared pigs, were measured periodically during the first 42 days after birth. The maximal concentration of absorbed bovine immunoglobulin was reached between 12 and 18 hours and equaled or exceeded the amount of porcine immunoglobulin absorbed by the conventionally reared pigs. Large differences in the concentrations of the bovine immunoglobulin isotypes among the various pools of colostrum were positively correlated with concentration of these isotypes in the sera of the neonatal pigs fed these pools. Relative to their concentrations in colostrum, approximately 41% of the IgG1, 55% of the IgG2, 29% of the IgM, and 67% of the IgA was absorbed. The IgA was absorbed the best and IgM was least absorbed. Significant trends or differences in absorption were not observed among groups. Neonatal pigs given fresh colostrum, which had a higher fat content, had significantly more weight gain (P less than 0.05). This occurred, despite the fact that the fresh colostrum had the lowest concentration of bovine immunoglobulin. Serum half-lives for bovine IgG1 and IgG2 were significantly less than for porcine IgG (P less than 0.05), whereas the half-lives for bovine and porcine IgM and IgA were similar. De novo-synthesized immunoglobulins were detectable in serum after 6 days; IgM concentrations reached a maximum at 15 days in neonatal pigs given stored, but not fresh, colostrum.(ABSTRACT TRUNCATED AT 250 WORDS)     

Age-related variations in serum concentrations of the third component of complement in swine

Single radial immunodiffusion was used to determine the concentration of the third component of complement (C3) in serum from swine in the following age groups; 36 to 60 hours (neonates), 6 to 7 weeks (weanlings), 4 to 5 months (adolescents) and greater than 1 year (adults). Mean serum C3 concentrations, expressed as the percentage of pooled reference sera from 15 adult swine for the 4 groups were 23, 123, 119, and 98%. With the exception of mean values for weanlings and adolescents, all comparisons of group means were significantly different. Regression models were developed to estimate serum C3 concentration in neonates as a function of litter size, birth weight, and total serum IgG concentration. Increases in birth weight and litter size were accompanied by increased serum C3 concentration, possibly reflecting ontogenic variation of the complement system at the end of gestation. Passive transfer of colostral immunoglobulins, as measured by total serum IgG concentration, was inversely related to serum C3 concentration in neonatal swine, suggesting that colostral absorption of C3 has minimal impact on complement activity in neonatal swine.     

Immunoglobulin concentrations in ovine body fluids.

Ovine IgG, IgM and IgA and antisera specific for these immunoglobulins were prepared. The specific antisera were used to estimate the immunoglobulin concentrations in certain sheep body fluids. IgA was shown to be the major immunoglobulin in saliva, lung and lachrymal fluid, tracheobronchial and nasal secretions while IgG was the predominant immunoglobulin in colostrum, milk, bile and serum.     

Passive transfer of colostral immunoglobulins from ewe to lamb and its influence on neonatal lamb mortality

The transfer of immunoglobulin G (IgG) and immunoglobulin M (IgM) from ewe to lamb was quantitated to determine the occurrence of failure in passive transfer. Concentrations of IgG and IgM in ewe serum did not correlate with those in the colostrum. Colostrum from all ewes contained abundant amounts of immunoglobulins when compared with serum values, with IgG being selectively concentrated over IgM. Absorption through the intestinal tract of the lamb appeared to be a nonselective process, lacking predilection for IgG and IgM. All lambs tested 24 hours after birth absorbed colostral immunoglobulins to some extent; however, 13 (14%) of 91 clinically normal lambs demonstrated some failure of passive transfer. In contrast, failure of passive transfer was found in 27 (46%) of 59 lambs dying of natural causes between 24 hours and 5 weeks of age. Evidence presented emphasizes the importance of absorption of adequate amounts of immunoglobulins to enable the newborn lamb to survive the first few weeks of life.     

Quantitation of immunoglobulins in ovine sera and secretions by laser nephelometry. Comparison with the radial immunodiffusion (RID) technique

In twenty-five ovine body fluids (serum, lung fluid and cerebrospinal fluid), the concentrations of IgG1I, IgG2, IgM and IgA were determined by laser nephelometry and radial immunodiffusion (RID). When nephelometric assays are carried out, antisera free from any turbidity are essential. Methods ensuring that goat and rabbit anti-sera will satisfy this requirement are described in the present paper. When sheep immunoglobulins were measured by laser nephelometry, adequate and reproducible results were obtained, comparable with those obtained by RID. Advantages of the nephelometric method include the speed of assay and its sensitivity, allowing precise determination of the very low concentrations of immunoglobulins in CSF.     

The association of extrinsic bovine IgG1, IgG2, SIgA and IgM with the major fractions and cells of milk

The association of purified, iodinated bovine immunoglobulins with the various fractions of whole milk and with cell-free skim milk and with two different mammary cell populations was studied. Associations in whole milk were concentration independent over a 6-fold range and revealed that SIgA and IgM were 5-fold more prevalent in milk fat than IgG1 and IgG2; the concentration of IgM and SIgA was 3-fold and 2-fold higher in fat than whey, respectively. A significant proportion of the IgM, ca 20% and to a lesser extent IgG2, ca 10%, were found in association with the casein pellet. Greater than 85% of the IgG2, greater than 90% of the IgG1, approximately equal to 80% of the SIgA and 70% of the IgM were found in milk whey. Isoelectric precipitation of casein significantly reduced the amount of IgM which associated with fat. When labelled Igs were incubated with milk leukocytes alone, only SIgA and IgM became significantly associated with them. However, when 10(6) cells were added, the amount of SIgA and IgM in the casein-cell pellet was not additive, although the increase for IgM was significant. These Igs also associated with the casein pellet of cell-free skim milk. When whole milk was used, the milk fat competed with cells and casein for association with SIgA and IgM. Homogenization of the fat layer from normal milk with or without added cells, caused significant release of the Igs which sedimented in the pellet.     

黄芪多糖佐剂对猪瘟疫苗免疫效果的影响

本试验旨在探讨以黄芪多糖(astragalan,AG)为佐剂对仔猪猪瘟疫苗诱导猪瘟抗体的影响。将20日龄仔猪40头随机分成4组(n=10),即生理盐水(normal sodium,NS)组、猪瘟疫苗(swine plague vaccine,SPV)组、黄芪多糖注射液(astragalan injection,AGI)组以及黄芪多糖注射液和猪瘟疫苗(astragalan injection and swine plague vaccine,AGI-SPV)组。各组于前腔静脉采血5 mL(用于检测)后,分别于仔猪出生第20、85天连续3 d颈部肌肉注射NS 3 mL×3 d,SPV 1 mL×1 d+NS 3 mL×2 d,AGI 10 mg/kg×3 d,(AGI 10 mg/kg+SPV 1 mL)×1 d+AGI 10 mg/kg×2 d。分别采血检测20(给药前)、35、100 d血清IgG、IgM、IgA、γ-球蛋白、猪瘟抗体(SP-Ab)等。100 d血清测定结果显示,AGI组和AGI-SPV组的γ-球蛋白、IgG、IgM和IgA水平显著高于NS组和SPV组(P＜0.05),SPV组和AGI -SPV组猪瘟抗体阻断率分别显著高于35 d和20 d,且AGI-SPV组高于SPV组(P＜0.05)。 AG作为免疫佐剂能提高猪瘟疫苗对仔猪的免疫力,其作用机制与增加免疫球蛋白和促进猪瘟抗体形成有关。     

The influence of age and breed on the concentrations of serum IgG, IgA and IgM in sows throughout the reproductive cycle

The influence of age and breed on the concentrations of IgG, IgA and IgM in the sera of sows throughout the reproductive cycle was investigated in 4137 sows which had had 0 – 20 gestations and representing three breeds: Swedish Landrace, German Landrace and L-12 (=Swedish Landrace × Large White). Data revealed an increase in total immunoglobulins (Ig), IgM and IgG serum levels with increasing gestation number; the latter contributed > 80% to total Ig levels. IgG was significantly increased up to the fourth gestation, whereas IgM showed a significant increase only to the third gestation. IgA showed only minor differences. Age-dependent increases in serum IgM were ascribed to the incraased probability of antigenic exposure during suckling, while failure to observe this change in serum IgA was ascribed to its role as a local Ig. Breed differences were observed to be significant for all three Ig's. It is concluded that establishment of group norms for serum Ig's should consider age and breed difference as well as stage of gestation or lactation.     

The absorption of colostral immunoglobulins in newborn piglets. III. Influence of the duration of colostrum administration

The gastrointestinal tract of newborn piglets is permeable for intact immunoglobulins ingested with the colostrum. The duration of this passage was investigated by administering hourly rations of 25 ml of either porcine or bovine colostrum for 6, 12, 18 or 24 hrs after birth. The plasma concentrations of the subclasses porcine IgG, IgM and IgA or bovine IgG1, IgG2, IgM and IgA were determined at 12, 18 and 24 hrs after birth and on days 3 and 6. Feeding periods of 6 hrs resulted in plasma Ig levels of the same order of magnitude as observed in natural rearing. These levels were not substantially increased after prolonged feeding. The 6% gain from 6 to 12 feedings seen with porcine colostrum as compared with a gain of 24% for bovine colostrum points at an earlier closure of the intestinal wall for the species-specific proteins. There was no further increase of Ig permeation after 12 hourly feedings. Growth performances and losses were identical in all groups.     

Quantitation of the immunoglobulins in reproductive tract secretions of the mare

IgG, IgA, IgM and albumin concentrations were measured in serum, follicular fluid and oviductal, uterine and intestinal secretions of the horse. Follicular protein concentrations were found to be dependent on serum concentration and molecular size. Of the immunoglobulins only IgG was detectable in oviductal secretions, but IgG:albumin ratios did not differ significantly from those in serum. IgG, IgA and IgM were measured in uterine secretions, with IgG predominant. Serum transudation into uterine secretions was minimal. In intestinal secretions, IgA levels were slightly higher than IgG, with albumin and IgM at low levels. In five mares with histories of chronic metritis, IgG, IgA and albumin concentrations were significantly elevated in uterine secretions.     

Humoral and cellular responses in calves experimentally infected with bovine leukemia virus (BLV)

In order to elucidate whether infection by Bovine Leukemia Virus (BLV) might induce an immunodeficient state, we inoculated sixteen calves with BLV. The calves were followed up for two years and were tested for humoral and cellular responses using various parameters, namely the appearance of antibodies to the BLV antigens, the changes in the numbers of lymphocytes involved, and the ratio between the two main populations of lymphocytes. Antibodies to the BLV antigens were of both the IgG and the IgM classes of immunoglobulins. The levels of antibodies of the IgM class were higher than those of IgG. There was a temporary decrease of reactive antibodies to the BLV antigens, to below detectable levels, during the 14-24 weeks post infection. A significant decrease in the level of plasma IgM was found in all BLV infected calves exhibiting lymphocytosis, while the level of IgG in the plasma of all experimental calves did not diverge significantly from the initial values, throughout the experiment. BLV infection was followed by lymphocytosis of B-cells in most infected calves, which persisted for the whole course of the experiment, while a decrease in the population of T-cells in peripheral blood was observed for a period of several months in all infected calves.     

Immunoglobulins and immunoglobulin-containing cells in the reproductive tract of normal rams

The levels of the immunoglobulins IgA, IgG1, IgG2, and IgM were measured in serum and fluid from various locations in the reproductive tract of normal rams. These fluids included semen, preputial washings, and fluid from the accessory sex glands (ASG), vasa deferens, rete testes, and tissue fluid from the seminal vesicles, bulbourethral glands, epididymal tails and efferent ducts. In addition, the prevalence of specific Ig-containing cells (ICC) was measured in sections of formalin fixed tissues stained by an indirect peroxidase-antiperoxidase labelling technique. Mean IgA levels in semen (1.23 mg/ml) and ASG fluid (0.46 mg/ml), were higher than in serum (0.19 mg/ml) and were at levels higher than IgG1 or IgG2 levels in semen, ASG fluid, and preputial washings, thus confirming the existence of a local immune system primarily in the ASG of ram genitalia. Relatively low concentrations of IgA and IgG in other genital fluids and IgG levels in these fluids were consistent with diffusion from serum. The relatively high prevalence of IgA-containing cells in bulbourethral (56% of all ICC) and prostate (49%) glands confirmed these tissues as major sites of local Ig production. ICC were also found in large numbers beneath pelvic urethral and preputial epithelia, but these were predominantly IgG-containing (88 and 72% respectively).     

Quantitation of sheep IgG1, IgG2, IgA, IgM and albumin by radioimmunoassay

A solid phase radioimmunoassay procedure for the measurement of immunoglobulins (IgG1, IgG2, IgA and IgM) and albumin in sheep body fluids (serum, intestinal lymph, caudal mediastinal lymph, bile, mammary secretions and intestinal secretions) is described. This method was found to be easy to perform, rapid, sensitive and reproducible. Results obtained were consistent with those previously reported using radial immunodiffusion and nephelometric techniques.     

The Influence of Sow Colostrum Trypsin Inhibitor on the Immunoglobulin Absorption in Newborn Piglets

The influence of sow colostrum trypsin inhibitor (SCTI) on the immunoglobulin absorption from the gut of 16 newborn colostrum-deprived piglets was investigated in a paired feeding experiment.Three times at 1 h intervals the piglets were fed an experimental diet consisting of sow milk, purified swine serum immunoglobulins containing agglutinins against Bordetella bronchiseptica, and purified SGTI (diet I) or saline (diet II). The serum concentrations of IgG, IgM, IgA, and antibodies for B. bronchiseptica were measured by single radial immunodiffusion and by a tube agglutination procedure and used to evaluate the immunoglobulin absorption. Four and 6 h after the first experimental meal, blood samples from the piglets given SGTI in their diet had a generally higher level of IgG, IgA and aggutinins against B. bronchiseptica than blood samples from the piglets d no SGTI. No real differences were found in the IgM levels. Although the piglets fed no SGTI all showed a considerable immunoglobulin absorption, the SCTI was found to have a statistically significant positive influence on the IgG and IgA absorption.     

Quantification of immunoglobulins in respiratory tract secretions of the horse

Lavage techniques were used to obtain secretions from the nasal cavity, trachea and bronchi of conscious horses. The techniques, which utilised fibreoptic endoscopy for recovery of tracheal and bronchial secretions, were well tolerated by the horses. The recovery rates of the lavaged fluids were acceptable, but were lowest for bronchial secretions, and there was minimal contamination by blood. The fluids were analysed for IgG and IgM by single radial immunodiffusion, and for IgA and albumin by rocket immunoelectrophoresis. Relative to albumin there was significantly more IgA and IgM, and significantly less IgG, in the nasal cavity than the trachea. IgA and IgM levels were greatest in the nasal cavity and decreased progressively to the bronchi, whereas IgG levels showed the reverse trend. The immunoglobulin: albumin ratios of secretions taken from many levels of the tract were significantly higher than those of serum, suggesting local production of immunoglobulin in the respiratory tract.     

Normal profile of immunoglobulins in sera and tracheal washings of chickens

Concentration and distribution of the three immunoglobulins in the sera and tracheal washings of a chicken population was studied. The mean IgM, IgG and IgA concentrations in serum were 1.35, 5.09 and 0.31 mg/ml, respectively. The distribution of IgM and IgG in birds irrespective of age was almost normal whereas that of IgA was skewed. All the three immunoglobulins were present in tracheal washings but the level of IgM was barely detectable. The IgG was predominant in the tracheal washings but higher IgA : IgG ratio compared to that of serum indicated local IgA production in the chicken respiratory tract.