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Biomedical subjects

M R Williams

Publications and source records attributed to M R Williams.

At least 91 records · Page 5Linked to original sources

The opsonic activity of bovine milk whey for the phagocytosis and killing by neutrophils of encapsulated and non-encapsulated Escherichia coli.

Encapsulated strains of Escherichia coli were found to be more resistant to phagocytosis and killing by bovine neutrophils; requiring in the order of 100 times more serum opsonins than non-encapsulated strains. Mid-lactation pooled whey from cows with no history of mastitis was opsonic for non-encapsulated strains, but had no effect on encapsulated organisms. In contrast, early lactation pooled whey (5-10 days post-partum) was opsonic for all strains of E. coli. It is concluded that since early lactation milk contains sufficient opsonins, severe E. coli mastitis at this stage of lactation is not due to opsonic deficiency.

Animals↗

Increase in specific opsonic activity in bovine milk following experimental Escherichia coli mastitis.

An infection in a single bovine mammary quarter with a capsulated Escherichia coli stimulated a long lasting opsonic activity for the phagocytosis and killing of the homologous strain by polymorphonuclear leucocytes. The induced activity was associated with increases in the IgM fraction and an appearance of activity in the IgG2 fraction. The capsular polysaccharide was the major inducing antigen, with the immune whey showing opsonic activity for other strains of E coli with antigenically similar capsules.

Animals↗

Enhanced killing of penicillin-treated S. aureus by host defences: effects of amoxycillin, cloxacillin and nafcillin in vitro and in experimental mastitis.

The effects of pretreatment of Staphylococcus aureus with subMIC or 10 x MIC amoxycillin, cloxacillin and nafcillin on subsequent killing by bovine neutrophils or sensitivity to lysostaphin were examined in vitro, using penicillinase positive and negative strains. All penicillins increased the susceptibility of S. aureus to bovine neutrophils; with amoxycillin this was most marked with the penicillinase negative strain. Cloxacillin pretreatment was significantly more effective than nafcillin or amoxycillin in some tests and equally effective in others. The sensitivity of S. aureus to lysostaphin was increased following cloxacillin or nafcillin exposure. In a mouse model of mastitis, more cloxacillin and nafcillin-treated S. aureus (at 1/4 MIC) were killed than untreated bacteria 1 hr after infection but by 4 hr no differences were found. No differences were seen after intramammary infection with 10 x MIC penicillin-treated S. aureus. In glands with established chronic mastitis both cloxacillin and nafcillin at high doses failed to kill staphylococci. These findings indicate that prior exposure to penicillins increases staphylococcal susceptibility to killing by neutrophils but following phagocytosis, intracellular bacteria are protected from the lethal action of penicillins.

Amoxicillin↗

A role for IgM in the in vitro opsonisation of Staphylococcus aureus and Escherichia coli by bovine polymorphonuclear leucocytes.

The phagocytosis and killing of Escherichia coli (strain P4) and Staphylococcus aureus (strain M60) by bovine polymorphonuclear lymphocytes (PMN) suspended in phosphate buffered saline, requires the presence of calcium ions and opsonins. The highest dilution of normal decomplemented adult sera in which the opsonins are still active is approximately 1/1000 for S aureus and 1/200 for E coli. In fetal and precolostral sera heat labile factors are also required for opsonising E coli but these are not required for S aureus. IgG1 and IgG2 from adult sera did not opsonise the bacteria even though receptors for IgG2 anti-erythrocyte antibodies have been reported on bovine and ovine PMN. A systematic separation of adult serum proteins was carried out by salt fractionation, anion exchange and gel filtration chromatography. The results suggest strongly that the opsonin in adult bovine sera is IgM.

Animals↗

Variation among cows in the ability of their blood polymorphonuclear leucocytes to kill Escherichia coli and Staphylococcus aureus.

Suspensions of polymorphonuclear leucocytes (PMN) from fresh bovine blood contained about 90 per cent neutrophils and 10 per cent eosinophils. Such suspensions had no bactericidal activity against Escherichia coli or Staphylococcus aureus, in the absence of an opsonising agent, in Hanks' balanced salt solution. Suitable opsonins were present in dilute (0.5 per cent), heat-inactivated serum or in 5 per cent whey. After incubation with PMN for two hours in the presence of the dilute serum the activity of PMN from several cows against the two organisms was highly correlated (r = 0.98) but varied significantly (P less than 0.001) among the animals from less than 2 to 30 per cent survival of the bacteria. This bactericidal activity did not alter if PMN from a single cow were incubated with sera from other animals, indicating that the critical factor was the cells and not the serum. The ranking of the animals by the bactericidal activity of their PMN was unaltered if 5 per cent whey was used as the opsonin and the leucocytes were sonicated after incubation.

Animals↗

The relatioship between serum immunoglobulin levels and specific antibody production in cows.

Antibody responses were measured, by the Farr ammonium sulphate technique, to two unrelated antigens-human serum albumin and sperm whale myoglobin. Differences between cows in their mean responses to both antigens were highly significant (P less than 0.001) but there were no differences between Jerseys and Friesians. Preinoculation IgG1, IgG2 and IgM levels were correlated with subsequent specific antibody production. The most significant relationship was between total immunoglobulin (IgG1 plus IgG2 plus IgM) and specific antibody production (r=0.76, P less than 0.01). But there was a negative correlation between the total Ig levels and the relative avidities of the antibodies produced (r=-0.65). These data suggest that in antibody production there is a balance between quality and quantity; cows with normally high serum immunoglobulin concentrations generally produce more antibody in response to challenge but this increase only compensates for the relatively lower avidity of their antibody for the antigen.

Animals↗

Changes in serum immunoglobulin levels in Jerseys and Friesians near calving.

IgG1 and IgM levels were significantly lower in bovine serum from four weeks before, until at least two months after calving, whereas IgG2 levels were elevated at this time. The shapes of the changes in Ig levels were different for first and second calving and it appeared that IgG1 levels were secreted earlier, from serum to colostrum, in heifers. Changes in management caused marked changes in Ig levels particularly after third calving when the cows were transferred from a balanced complete diet to grass or hay. IgG1 levels increased by 130 per cent and stayed high whereas the increase in IgM was transitory, and IgG2 levels largely unaffected. In Friesians the IgG2 levels increased with age as anticipated from previous studies but in Jerseys the levels appeared to decline with age. Mean IgG2 levels in Jerseys were also significantly lower than in Friesians.

Animal Feed↗

The absorption of immunoglobulin from colostrum by bottle-fed lambs.

Factors affecting the absorption of immunoglobulin from colostrum by new born lambs were studied. Merino, Scottish Blackface, Cheviot, Southdown, Border Leicester, Finnish Landrace and Finnish Landrace x Dorset Horn lambs were removed from their mothers before sucking and bottle-fed with measured amounts of colostrum, either from individual ewes, or from pools of colostrum, some of which had been frozen for up to two years. Afterwards the lambs were fostered onto other ewes or reared on milk substitute. Their serum IgG1 concentrations before and after colostrum intake, were compared with IgG1 concentrations in conventionally reared lambs. The concentrations in lambs given a single feed of 150 to 200 g 1 hour after birth, or 30 g/kg at 1 hour and 7 hours compared favourably with those in control lambs. Lambs fed 50 to 100 g at 1 hour had a high proportion of very low concentrations and lambs fed 30 g/kg at 1 h, or at 1 and 24 h had low mean concentrations but no very low individual concentrations. The concentrations after a feed at 1 hour were positively correlated with the total amounts of IgG1 in the colostrum and negatively correlated with the birth weights. Only very small amounts were absorbed at 24 hours. Absorption was more efficient in males than in females and in Cheviots than in other breeds. It was unaffected by the age of the colostrum. Similar proportions of bottle-fed and control lambs died before 6 months of age, but relatively more bottle-fed lambs died from respiratory infections. The IgG1 concentrations and growth rates of the bottle-fed lambs were not significantly correlated.

Absorption↗

The effects of passive antibodies to egg albumin on active immunity in lambs to Brucella abortus and egg albumin.

The long-term effects of colostrum on active immunity to two unrelated antigens are described. Lambs were fed with pooled colostrum--to equalise passive immunity--with or without added antibodies to egg albumin (Ea). There were significant breed differences in the response both to Brucella abortus measured at one month of age, and to Ea, measured at three months of age, although there was no significant correlation between the responses to the two antigens, either within or between breeds. Surprisingly, whereas antibodies to Ea caused a four-fold reduction in antibody production to B abortus, they did not affect the overall mean response to Ea. But the timing of the response to Ea was significantly affected, suggesting that the low persisting concentrations of antibody had caused qualitative changes in the response.

Animals↗

Effects of energy intake during late pregnancy and of genotype on immunoglobulin transfer to calves in suckler herds.

Blue-Grey (Shorthorn x Galloway) and Hereford x Friesian cows were given levels of feeding, during the last 12 weeks of pregnancy, varying between 75.2 per cent and 171.5 per cent of the maintenance requirements in one experiment and between 65 per cent and 125 per cent of the maintenance requirements in a second experiment. The concentrations of the immunoglobulin classes IgG1, IgG2 and IgM, in their colostrum and in their calves' serum were measured. The IG1 and IgM, but not the IgG2, concentrations were significantly higher in the Blue-Grey than in the Hereford x Friesian colostrum and serum. The relative concentrations of the three classes in the serum were almost identical to those in the colostrum, suggesting that absorption by the calves was non-selective. The amounts of immunoglobulin transferred to the calves were not affected by the cows' levels of feeding in the first year, but there was a small increase in transfer with increasing levels of feeding during the second year. Cows that transferred large amounts to their calves in the first year tended to do so in the second year. The milk intake and growth of the calves during the first 42 days were significantly correlated with their immunoglobulin concentrations.

Animals↗

Changes in IgG2 levels with age in British cattle.

IgG1, IgG2 and IgM levels were measured in serum samples from 282 cattle of various ages and the results analysed by least squares. There were marked increases with age in IgG2 levels in bulls, and in dairy cattle in the absence of culling for disease or low production traits. There was about a 300% increase between one year and eight years of age for IgG2 compared with the small increase of about 20% for IgG1 but there was no apparent effect of age on IgM levels. There were no differences between Hereford and Friesian bulls for any of the immunoglobulins but there were environmental differences, between AI centres, for IgG1 and IgM. No animals were found to be immunodeficient but IgG2 levels ranged widely from 1 mg/ml to 35 mg/ml in different bulls.

Aging↗

The role of colostrum antibodies and the significance of antigenic competition in young lambs.

Young lambs were fed with a small amount of pooled colostrum and injected at either 1, 3, 6 or 12 weeks after birth with the antigen hen egg albumin (Ea). Five days before the Ea injection the experimental group were given a large dose of pooled vaccines (competing antigens). Blood samples were collected at regular intervals after the Ea injection. The results showed marked antigenic competition of Ea at both 6--and 12--weeks of age but the results were partially confounded by the presence of unexpected passive antibodies to Ea in the pooled fed. The experiment was repeated with a separate group of conventionally reared lambs given the same competing antigens but using human serum albumin (HSA) as the suppressed antigen. In this experiment the results differed since antibody concentrations and avidities to HSA were similar in 1, 6 and 12-week-old animals and there was no evidence for antigenic competition. The data presented illustrate the difficulties both of measuring immune responses in young ruminants and also of interpreting such results.

Animals↗

Problems with radiolabelling egg albumin for the Farr ammonium sulphate technique for antibody estimation.

High concentrations of passive haemagglutinating antibodies were raised in sheep immunised with egg albumin (Ea) in Feund's adjuvant. Further attempts to study the antigen binding capacities and avidities of these antisera by the Farr ammonium sulphate technique were hindered by unsuccessful attempts to label Ea with 125Iodine by either the chloramine T or sodium nitrite-HCl methods. This problem was attributed to the unavailability of tyrosine in the native protein. This was solved by labelling Ea with the Bolton-Hunter reagent which does not require the presence of tyrosine. However labelled Ea binds non-specifically to normal serum to give high control values of circa 40% binding. Despite this it was possible to estimate antibody concentrations and avidities of lamb sera.

Ammonium Sulfate↗

Changes in bovine immunoglobulin levels during a response to homologous erythrocyte membrane antigen.

Changes in serum immunoglobulin levels of IgG1, IgG2 and IgM were studied in cows immunised with a butanol extract of bovine erythrocytes. Donor and recipients were matched to differ at only one known red cell antigen, the V antigen, and after the first course of injections a weak antibody response to V antigen was associated with elevated levels of IgG2 immunoglobulins. A second course of injections of V antigens together with Bordetella pertussis led to a stronger and more longlasting antibody response to V and this was associated with elevated levels of IgG1 and correspondingly decreased levels of IgG2. One cow calved during the period of investigation and it was found that even though serum antibody levels disappeared three weeks before calving there was a high antibody titre in the colostrum which was passed on to the calf. The loss of serum antibodies before calving was associated with a fourfold drop in serum IgG1 and an increase in serum IgG2 levels.

Animals↗

Passive immunity in the lamb. Effects of a second feed of colostrum on antibody absorption from the first feed.

Lambs were taken from their mothers before sucking and fed pooled colostrum (30 g/kg body weight), containing added antibody to egg albumin, 1 h after birth. At 4 h after feeding there was a two-fold range in immunoglobulin (IgG) concentration and a sixteen-fold range in anti-egg albumin activity in their sera. The coefficient of correlation between the IgG and antibody concentrations was significant but low (r = 0.445, P less than 0.02). When a second feed of colostrum without added antibody was given 6 h after the first, both the IgG and the antibody concentrations increased significantly, indicating that there had been increased absorption from the first feed, rather than absorption from the second. The increases in IgG had a significant negative correlation with the IgG concentrations before the second feed (r = 0.760, P less than 0.001). Sera from control lambs fed only untreated colostrum gave only trace, probably non-specific, antibody reactions. The results suggest that single feeds of colostrum are generally inefficient, and that the passive immune status of young lambs may be improved and made more uniform by subsequent feeds, even though these may be themselves deficient in IgG.

Animals↗

Quantitative studies on bovine immunoglobulins.

The influence of serum immunoglobulins on the incidence of calf pneumonia, and the relationship between sub-clinical pneumonia, and calf serum Ig levels, is discussed. Monospecific antisera were used to measure levels of IgG1, IgG2, IgM and IgA in the sera of bull calves aged around 2 1/2 weeks. These calves were selected retrospectively according to their disease record up to six months of age. A clear association was found between low levels of IgG1, IgG2 and IgA in the "2 1/2-week" sample and subsequent susceptibility to pneumonia at around 2 1/2 months of age. Calves showing signs of pneumonia had low levels of IgG1 (45-5 per cent had less than 8 mg per ml compared with "now-pneumonic" calves which had relatively high levels (only 9.5 per cent had less than 8 mg per ml). In contrast, older calves bled at monthly intervals and found to have pneumonic lesions at slaughter had significantly evevated levels of both IgGa and IgG2.

Age Factors↗