Effect of a peri-parturient eprinomectin treatment of dairy cows on milk production.

AIM: To quantify and economically evaluate the effect on milk production of peri-parturient treatment of dairy cows with eprinomectin. METHODS: On 3 farms in separate geographic areas of New Zealand, 849 first-calf heifers and multiparous cows were ranked and paired within parity, date of calving and expected milk production. Within pairs, cows were randomly allocated to treatment with either a commercial formulation of eprinomectin, applied at a dose rate of 500 mug/kg liveweight, or an equivalent volume of vehicle containing no antiparasitic agent and administered at the same dose volume, generally within the first week post-calving. On each farm, trial cows shared the same pasture. Over a single lactation, records were maintained of milk quantity and content. RESULTS: Trichostrongylid eggs were identified in pre-treatment faecal samples from all farms, verifying the presence of gastrointestinal parasites. Overall 25.5% of the cows sampled were positive for nematode eggs, but only 8% had counts 50 eggs per gram of faeces (epg). Daily milk volume, milk protein and milksolids (yield of milk fat + milk protein) were higher for eprinomectin-treated multiparous cows than for controls (milk volume: 20.36 l/day vs 19.76 l/day, p=0.005; milk protein: 0.700 kg/day vs 0.685 kg/day, p=0.012; milksolids: 1.613 kg/day vs 1.583 kg/day, p=0.031, respectively). The daily value of the increased production from eprinomectin-treated multiparous cows was estimated to be NZ0.034 dollar for milk fat (p=0.095) and NZ0.078 dollar for milk protein (p=0.012), equating to NZ0.104 dollars for milksolids (p=0.031), averaged over the whole lactation. No significant difference in milk production was detected between treated and control first-calf heifers. Averaged over the whole herd, the peri-parturient treatment of multiparous cows and first-calf heifers with eprinomectin increased daily milk volume and milk protein production of treated vs control cows (19.28 l/day vs 18.86 l/day, p=0.020, and 0.661 kg/day vs 0.650 kg/day, p=0.047, respectively). CONCLUSION: These data provide evidence that the use of a peri-parturient treatment of eprinomectin on multiparous cows can increase their production of fluid milk and milksolids.

Persistent efficacy of topical eprinomectin against nematode parasites in cattle.

Six studies were conducted to evaluate the persistent efficacy of eprinomectin pour-on against experimental challenges with infective nematode larvae in calves. In each study, calves were randomly assigned to one untreated group and up to four test groups, which were treated with eprinomectin at 500 microg/kg body weight at weekly intervals before single bolus challenge. The calves were necropsied approximately 4 weeks after challenge infection for nematode recovery. Eprinomectin pour-on provided > or =90% efficacy against challenge with Haemonchus placei, Trichostrongylus axei and T. colubriformis at 21 days after treatment and against Cooperia oncophora, C. punctata, C. surnabada, Dictyocaulus viviparus, Nematodirus helvetianus, Oesophagostomum radiatum and Ostertagia ostertagi at 28 days after treatment.

Efficacy of ivermectin in a controlled-release capsule for the control of breech strike in sheep.

OBJECTIVE: To investigate the efficacy of ivermectin in an intraruminal controlled-release capsule (CRC) against blowfly strike. DESIGN: Pen and field trials with controls. ANIMALS: Pen studies: Two breech strike trials involving 60 Romney and 60 Merino sheep. One body strike trial using 100 Merino sheep. Field trials: Eight trials in New Zealand used 1000 Romney and Romney-cross sheep. Fifty Merino lambs in one trial in Australia. PROCEDURE: Pen studies: Sheep were allocated to two equal groups. One was not treated, the other sheep received a CRC that delivered ivermectin at > or = 20 micrograms/kg/day for 100 days. In the breech strike trials, each animal was given an oral laxative 2 days before exposure to adult Lucilia cuprina. In the body-strike trial, the sheep sheep were kept wet to increase susceptibility prior to the release of blowflies. Field trials: Fifty or 200 sheep allocated to equal groups of nontreated or treated with the CRC and grazed at pasture exposed to natural blowfly challenge. RESULTS: Pen studies: Breech strikes developed in 24 of 60 controls but in none of 60 CRC-treated sheep. There was a 35% reduction in the number of CRC-treated sheep struck on the body. Field trials: The average number of breech strikes in CRC-treated sheep was reduced by 86% (P < 0.001). The number of body strikes in the treated groups was a reduced by 27% (P < 0.05). CONCLUSION: The ivermectin CRC is a useful aid in controlling breech strike, but provides only moderate reduction in the incidence of body strike.

Field trials evaluating ivermectin controlled-release capsules for weaner sheep and for breeding ewes.

OBJECTIVE: To confirm the efficacy of ivermectin released from a controlled-release capsule administered to young sheep and to breeding ewes under field conditions. DESIGN: Randomised field trials. PROCEDURE: In each of ten field trials 25 weaned lambs were treated with ivermectin controlled-release capsules and 25 remained untreated. Eight similar field trials were conducted using adult ewes. Efficacy against infections of gastrointestinal nematodes was assessed by faecal egg counts and faecal larval culture. Body weights were recorded and faecal soiling of the breech wool (dags) was assessed. RESULTS: Nematode faecal egg counts in the two groups were not different (P = 0.13) before treatment in the weaner trials or before treatment in the ewe trials (P = 0.49), but thereafter eggs in the untreated sheep persisted, whereas counts in sheep given capsules were negligible (P < or = 0.01). In the weaner trials, dag scores for the two groups were not different at the start of the trials (P = 0.18) but at the end, untreated sheep had significantly more dags (P = 0.04) than treated sheep. In the ewe trials, dag scores remained low in both groups. Weaners treated with the capsule gained 1.4 kg (95% CL: 0.7, 3.1) more weight over the 16 week trial period compared to untreated weaners (P = 0.01). Both groups of ewes lost weight as a result of parturition but the mean loss by week 16 was greater for untreated (3.7 kg) (95% CL: -5.1, -2.2) than for treated ewes (1.8 kg) (95% CL: -3.3, -0.4). The mean change in ewe body weight for the two groups was however not significant (P = 0.07). Differentiation of nematode larvae recovered from cultures of faeces from untreated animals indicated that the capsules were effective against the common parasites of sheep. CONCLUSION: The capsule was efficacious against gastrointestinal nematodes judging from faecal egg counts. It has the potential to significantly reduce contamination of pasture with nematode eggs. Treated weaners had less dags for 16 weeks and gained more weight than untreated weaners.

Demonstration of the sustained anthelmintic efficacy of a controlled-release capsule formulation of ivermectin in weaner lambs under field conditions in New Zealand.

Ten field trials were conducted in the North and South Islands of New Zealand to evaluate the anthelmintic efficacy and production responses attributable to treatment of weaner lambs with an intra-ruminal controlled-release capsule formulation of ivermectin. A total of 800 Coopworth, Perendale and Romney lambs weighing on average 20.8-34.8 kg were used. Lambs were either untreated or treated shortly after weaning with an ivermectin controlled-release capsule which delivers ivermectin at 0.8 mg per day for 100 days (minimum dose rate 20 microg/kg/day). Bodyweights, faecal nematode egg counts and dag scores (assessment of faecal soiling in the breech area) were determined before treatment and at about 4,8, 12, 14 and 16 weeks after treatment. Sheep treated with the Ivermectin capsule gained significantly more weight (11.6 kg) over the 16 weeks of the trials compared to untreated sheep (7.3 kg) (p < 0.01). Before treatment, faecal strongylid and Nematodirus spp. egg counts were equivalent (p > 0.10) but, at each time point thereafter, egg counts in ivermectin capsule-treated sheep were significantly lower (p < 0.01 or p < 0.05). Dag scores were not different at the start of the trial (p > 0.10), but at the end of the trial control sheep had significantly greater dags (p < 0.05) than sheep treated with the ivermectin capsule. These findings indicate that treated animals contributed significantly fewer nematode eggs to the contamination of pasture and therefore pasture contamination should be significantly reduced for at least 112 days. The productivity of the ivermectin capsule-treated sheep over the I6 weeks of the trials was also significantly increased compared to salvage-treated controls. Furthermore, the presence of dags, which predispose sheep to blowfly strike in the breech area and result in production losses due to the costs of dagging and downgrading of breech wool, were also significantly (p < 0.05) reduced in the ivermectin capsule-treated sheep.

Demonstration of the sustained anthelmintic activity of a controlled-release capsule formulation of ivermectin in ewes under field conditions in New Zealand.

Ten field trials were conducted in the North and South Islands of New Zealand to evaluate the anthelmintic efficacy of an intraruminal controlled-release capsule formulation of ivermectin. A total of 810 Coopworth, Perendale, Romney or Coopworth ' Romney ewes, weighing on average 42-70 kg, were used. Ewes were either untreated or treated shortly before lambing in late winter-early spring (eight trials) or in late spring (two trials) with an ivermectin controlled-release capsule which delivers ivermectin at 1.6 mg per day for 100 days (minimum dose rate 20microg/kg/day). Bodyweights, faecal nematode egg counts and dag scores were determined before treatment and at about 2 and/or 4, 6 or 8, 10 or 12, 14 and 16 weeks after treatment. Ewes treated with the ivermectin controlled-release capsule gained on average 1.1kg more than untreated sheep over the 16 weeks of the trials, but this difference was not significant (p > 0.10). Before treatment, faecal strongylid egg counts were equivalent (p > 0.10), but at each time point thereafter, egg counts in ivermectin controlled-release capsule treated sheep were significantly lower (p < 0.01; p < 0.05 at Week 2). Dag scores were not different at the start of the trial (p > 0.10), but at the end of the trial ivermectin controlled-release capsule treated ewes had significantly lower scores (p < 0.01) than untreated ewes. These findings indicated that treated animals shed significantly fewer nematode eggs and therefore pasture contamination with nematode eggs should be significantly reduced for at least 112 days. The control of dags should result in reduced direct losses due to the decreased value of dag wool, and indirect losses due to the cost of dagging sheep and the cost associated with the treatment and control of flystrike initiated by dags in the breech area.

Bovine IgG2a antibodies to Haemophilus somnus and allotype expression.

Bovine IgG2a has been implicated in protection against pyogenic infections, including those caused by Haemophilus somnus. To further investigate the role of IgG2a in defense against H. somnus, IgG1 and IgG2a antibodies were purified from antiserum against an immunodominant 40 kDa outer membrane protein (p40) of H. somnus, which was previously shown to passively protect calves against H. somnus pneumonia. The passive protective capacity of anti-p40 IgG1 or IgG2a was evaluated in vivo in calves. Purified anti-p40 IgG1 or IgG2a was incubated with H. somnus for 15 min before intrabronchial inoculation of calves. Bacteria incubated with anti-p40 IgG1 or IgG2a were inoculated into one caudal lung lobe and bacteria incubated with IgG1 or IgG2a from the respective preimmunization serum were inoculated into the contralateral lobe. The volumes of pneumonia in the right and left lungs were determined 24 h later. The difference in volume of pneumonia with H. somnus preincubated in IgG1 pre- and postimmunization anti p40 was less (16 cm3, P = 0.298) than the difference in volume of pneumonia with H. somnus preincubated in IgG2a pre- and postimmunization anti p40 (30 cm3, P = 0.146). Although the differences in lesion size between pre- and postimmunization serum were not statistically significant, the trend suggests IgG2a may be more protective than IgG1. To examine this further, the peptide specificity of these IgG1 and IgG2a antibodies to p40 was examined. After limited proteolysis of p40, IgG2a antibodies reacted with 2 peptides not recognized by IgG1 antibodies. Other peptides were recognized by both isotypes. Since these studies suggested that IgG2a may be important in protection against infection, we then investigated some aspects of the role of the 2 IgG2a allotypes, A1 and A2. In retrospective studies of age differences in expression of IgG2a allotypes, no heterozygotes were detected in calves of 60 d old or less, and fewer heterozygotes were detected in calves 61-120 d old than in cattle older than 270 d (P < 0.01). In a subsequent prospective study of the time course of allotype expression, Holstein calves shown to be heterozygotes expressed the IgG2aA1 allotype early but the IgG2aA2 allotype was not usually detected until 3 to 4 mo of age. Thus, both the retrospective and the prospective studies showed age related differences in expression of the IgG2aA1 and A2 allotypes. This could have implication in protection.

Peripheral blood lymphocyte subsets in fleece rot-resistant and -susceptible sheep.

OBJECTIVE: To compare haematological values and lymphocyte phenotypes in the peripheral blood of fleece rot-resistant and -susceptible sheep. PROCEDURE: Experiments were conducted on 2- and 3-year-old Merino rams, flock 1 (17 rams) and flock 2 (32 rams), respectively. Within each flock, individual rams were classified as fleece rot-resistant or -susceptible, based on established criteria. Total and differential white cell counts, and indirect fluorescent antibody tests specific for B cells and T cells were performed on all sheep. The concentration of various subsets of circulating lymphocytes was then determined in each sheep. RESULTS: There were no significant differences between fleece rot-resistant and -susceptible sheep from either flock in the mean total or differential white cell counts. However, fleece rot-resistant rams in flock 1 did have a significantly higher concentration of circulating SBU-T1+ cells than fleece rot-susceptible rams from the same flock. No such difference was noted in the rams from flock 2. While all rams in flock 1 were free of clinical fleece rot, 24 rams in flock 2 (comprising all 17 fleece rot-susceptible and 7 of 15 fleece rot-resistant animals) had clinical signs of the disease. Fleece rot-free rams in this flock (irrespective of their classification as fleece rot-resistant or -susceptible) had significantly higher concentrations of circulating SBU-T1+ cells compared with fleece rot-affected animals. They also had significantly higher concentrations of circulating B cells, and total lymphocytes. CONCLUSIONS: An examination of peripheral blood lymphocyte subsets in fleece rot-resistant and -susceptible sheep revealed a possible association between resistance to fleece rot and the concentration of circulating SBU-T1+ cells.

Effect of simulated rain, coat length and exposure to natural climatic conditions on the efficacy of a topical formulation of eprinomectin against endoparasites of cattle.

A series of five controlled studies involving 114 cattle were conducted in Australia, North America and the United Kingdom to examine the effect of simulated rain, coat length and exposure to natural climatic conditions, on the efficacy of a topical formulation of eprinomectin against nematode parasites of cattle. In all trials infections were induced with a range of bovine nematode species and treatment was applied when the majority of nematodes were mature. In one study, simulated rain was applied to cattle ending one hour before treatment or beginning one, three or six hours after treatment. In a second study cattle had short (1 cm) or long (3-6 cm) haircoats at the time of treatment. Three other studies were conducted using cattle housed indoors or exposed to various natural climatic conditions. Nematode counts were determined using standard techniques and the efficacy of treatment was assessed relative to vehicle-treated controls. Regardless of the timing of simulated rain relative to treatment, eprinomectin was at least 99.9% effective (P < 0.01) against Haemonchus placei, Ostertagia ostertagi. Trichostrongylus axei and Cooperia spp. There were also no differences (p > 0.10) in efficacy between treatment administered to dry or wet cattle, or treatment administered before or after simulated rainfall. Efficacies against O. ostertagi, T. axei, Cooperia ancophora and Dictyocaulus viviparus were > 99.5% (p < 0.01) regardless of the length of the haircoat at the application site. Exposure of treated cattle to sunshine and precipitation had no effect on anthelmintic efficacy (p > 0.10) with efficacies of greater than 99.5% being maintained against H. placei, O. ostertagi (adult and fourth-stage larvae), T. axei, Cooperia spp., Nematodirus helvetianus (adult and inhibited fourth-stage larvae) and Oesophagostomum radiatum. These findings indicate that eprinomectin (500 micrograms/kg) in a topical formulation is a safe and highly effective nematocide for cattle regardless of their coat length and this high level of efficacy is maintained in cattle exposed to a wide variety of climatic conditions.

Control of gastro-intestinal parasitism in sheep with ivermectin delivered via an intraruminal controlled-release capsule.

The efficacy of ivermectin delivered by an intraruminal controlled-release capsule against gastro-intestinal nematodes of sheep was evaluated under controlled conditions. In seven Australian studies involving 170 Merino or Merino x Border Leicester sheep, intraruminal capsules developed for 20-40 kg or 40-80 kg sheep, and delivering 0.8 or 1.6 mg of ivermectin/day respectively for 100 days (minimum dose 20 microg/kg/day), were evaluated. Studies were designed to test the therapeutic efficacy against naturally acquired and induced infections treated at the adult and fourth larval stage, and the prophylactic efficacy against naturally acquired and induced infections with third stage infective larvae. The predominant pathogenic nematodes of sheep were represented. Two studies included known benzimidazole- and levamisole-resistant nematode strains. Sheep were necropsied for total nematode counts 21-8.5 days after treatment. The efficacy of the ivermectin controlled-release capsule was generally >99% against all nematode species tested, including those confirmed to be benzimidazole- and levamisole-resistant. High therapeutic activity was demonstrated against existing adult and fourth larval stage nematode infections at the time of treatment, and high prophylactic efficacy was shown against incoming third stage larvae of all species and strains tested.

Field trials in New Zealand confirming the efficacy of eprinomectin applied topically to cattle.

The efficacy and acceptability of eprinomectin in a topical formulation against gastro-intestinal nematodes in cattle was assessed under field conditions. Seven similar commercial dairy farms in the North and South Islands of New Zealand were included in the studies, involving 247 Holstein-Friesian, Jersey or Jersey-cross cattle. Cattle were confirmed by positive faecal nematode egg counts to have natural infections of gastro-intestinal nematodes and were held in separate treatment groups. In each replicate, four animals received eprinomectin (500 microg/kg body weight) and one animal received vehicle solution, all applied topically at 1 ml/10 kg body weight. Faecal samples were collected before, and 14 days after treatment, for faecal trongylid egg counts. Animals treated with eprinomectin had significantly lower (p < 0.05) Day 14 faecal strongylid egg counts than the controls. There were no significant differences (p > 0.10) between treated and control groups for pretreatment strongylid egg counts. No formulation runoff or adverse reactions were observed. These studies showed eprinomectin to be effective against gastro-intestinal nematode infections and safe for use in dairy cattle under natural field conditions.

Efficacy of a topical formulation of eprinomectin against endoparasites of cattle in New Zealand.

Two controlled studies involving 24 cattle were conducted in New Zealand to determine the efficacy of a topical, non-flammable formulation of eprinomectin against induced and naturally acquired nematode infections. In Trial 1, nematode infections were induced on Day -5 with third-stage larvae of Cooperia spp., Haemonchus contortus, Ostertagia ostertagi and Trichostrongvlus colubriformis so that the nematodes would be at the fourth larval stage when the cattle were treated. In Trial 2, cattle had naturally acquired nematode infections as determined by faecal nematode egg counts and larval cultures. The cattle were allocated on Day 0 (Trial 1) or Day 6 (Trial 2) on a stratified random basis according to bodyweight to one of two treatments: untreated control or eprinomectin (0.5% w/v) applied topically at 1 ml/10 kg bodyweight. Necropsies were undertaken on Days 14 and 15 and total nematode counts were done. In Trial 1, cattle treated with eprinomectin had significantly (p < 0.05) fewer Cooperia spp. and O. ostertagi than the controls. Larvae of H. contortus and T. colubriformis did not establish. In Trial 2, cattle treated with eprinomectin had significantly (p < 0.05) fewer of the following parasites than the controls: Haemonchus spp. (adult), Cooperia surnabada (adult), C. oncophora (adult), Cooperia spp. (L,), Ostertagia lyrata (adult), O. ostertagi (adult), Oesophagostomum spp. (adult), T. avei (adult and L1) and Trichuris spp. (adult). Reductions of 100% were observed for Capilfaria spp. (adult), D. viviparus (adult and L,), and Nematodirus helvetianus (adult), but these were not statistically significant (p > 0.05) because four or fewer control animals were infected with these parasites. In Trial 2, efficacies of greater than 99% were observed against all species for which moderate to high burdens occurred in the untreated controls. These findings indicate that eprinomectin in a topical formulation is a highly effective nematocide in cattle.

Efficacy of ivermectin controlled-release capsules for the control and prevention of nasal bot infestations in sheep.

OBJECTIVE: To investigate the therapeutic and prophylactic efficacy of an ivermectin controlled-release capsule against nasal bots (Oestrus ovis) in sheep. DESIGN: Trial 1--A pen study with controls. Trial 2--A field study with controls. ANIMALS: Trial 1--Forty Merino wethers with natural infestations of nasal bot were used. Trial 2--One hundred nasal bot-free wethers were used. PROCEDURE: Trial 1--Ten randomly selected animals were slaughtered and the heads split and examined to confirm bot infestation. Fifteen animals were allocated to untreated controls and 15 to treatment with a controlled-release capsule delivering ivermectin at > or = 20 micrograms/kg/day for 100 days. Twenty-nine days after treatment the sheep were killed and examined for nasal bots. Trial 2--Nasal bot-free sheep were allocated to two groups of 45 animals. One group was untreated the other sheep were treated with capsules as above. The sheep were grazed as a single group exposed to natural challenge from O ovis. Ninety days after treatment the animals were slaughtered and examined for nasal bot infestation. RESULTS: Trial 1--Live O ovis larvae were recovered from 60% of control sheep. No live larvae were collected from treated sheep. Trial 2--Forty-one percent of untreated sheep harbored nasal bot infestations. No live larvae were collected from any treated animal. CONCLUSION: Treatment with a single ivermectin controlled release capsule was 100% effective against existing infestations of O ovis and as a prophylactic treatment for this parasite.

An evaluation of the quality and physical properties of calf leather following topical application of eprinomectin.

The gross and histological appearance of skins obtained from twelve 6-10 month-old Friesian X Hereford calves 14-15 days after they were treated with a topical formulation of 0.5% eprinomectin administered at 1 ml per 10 kg bodyweight was compared to that of skins from twelve untreated controls. No significant differences were observed and there were also no significant differences in the quality and physical properties of leather produced from the skins of treated and untreated groups, indicating that topical eprinomectin has no deleterious effects on the leather quality of calf skins.

An ivermectin tablet for sheep: efficacy against gastro-intestinal nematodes and a bioavailability comparison with a liquid ivermectin formulation.

An ivermectin tablet for oral administration to sheep was developed for use in countries where it is customary to treat sheep with anthelmintic tablets. Tablets require no special administration equipment, and offer convenience for storage and transport. The ivermectin tablet, which delivers 10 mg of ivermectin (200 micrograms kg-1 in a 50 kg sheep), had similar bioavailability to a liquid formulation of ivermectin (IVOMEC Liquid for Sheep) as determined by peak plasma ivermectin concentrations and area under the concentration curve in plasma (P > 0.10). In dose confirmation trials in which nematode infections were induced in helminth-naive sheep, animals treated with the ivermectin tablet had significantly fewer adult and fourth-stage larval nematodes than untreated control sheep (P < 0.01) with efficacies > 99% against all nematode species tested. In six field trials evaluating the efficacy of the ivermectin tablet in 240 Merino sheep, the reductions in faecal nematode egg counts ranged between 98 and 100%, as determined by comparison of pre- and post-treatment counts for the ivermectin-treated group.

Immunodominant antigens of zoospores from ovine isolates of Dermatophilus congolensis.

Zoospores of Dermatophilus congolensis were analysed by SDS-PAGE and western blotting. The electrophoretic profiles of zoospores from 13 isolates of D. congolensis were similar but not identical when stained with Coomassie blue or silver. Immunodominant polypeptides with apparent molecular masses of 76 and 31 kDa were identified in western blots of 13 of 13 and 12 of 13 isolates respectively of D. congolensis reacted with hyperimmune, ovine, antizoospore sera. Identical immunodominant polypeptides were observed in western blots reacted with sera obtained from naturally infected sheep. Initial characterisation of the 76 and 31 kDa polypeptides indicated that they were probably surface exposed because (i) antibodies eluted from the surface of live zoospores after adsorption of hyperimmune antizoospore serum, reacted principally against the 76 and 31 kDa subunit polypeptides in western blots, (ii) adsorption of hyperimmune antizoospore serum with live zoospores resulted in significant diminution of reactivity against both the 76 and 31 kDa polypeptides in western blots, (iii) indirect fluorescent immunostaining of zoospores with antiserum prepared against gel-purified 76 kDa polypeptide, resulted in intense staining of the zoospore outer coat. Immuno-gold electron microscopy of negatively stained zoospores with antiserum prepared against gel-purified 31 kDa polypeptide identified this antigen as a flagella subunit.

Phenotypic phase variation in Haemophilus somnus lipooligosaccharide during bovine pneumonia and after in vitro passage.

A high rate of phenotypic variation in the lipooligosaccharide (LOS) electrophoretic profile of Haemophilus somnus occurred in most isolates obtained at approximately weekly intervals from three calves intrabronchially challenged with a cloned isolate of H. somnus 2336. Daily subculturing for 2 weeks resulted in at least one major alteration in the LOS electrophoretic profiles for strain 2336 and both additional disease isolates examined, but no change occurred in the LOS electrophoretic profiles for any of three commensal isolates examined. None of the LOSs from any of the postchallenge intrabronchial isolates reacted with rabbit antiserum to the challenge strain LOS in immunoblotting, but LOSs from two nasopharyngeal isolates did. Antigenic variation in the extracted LOSs of most of the isolates was supported by the results of an enzyme-linked immunosorbent assay. Preimmune serum from each of the calves did not react with any of the isolates or the challenge strain, whereas sera obtained 35 days after challenge reacted with the challenge strain and zero to five additional isolates and sera obtained 74 days after challenge reacted with two to six additional isolates. Recognition of LOSs from isolates obtained near the end of the 70-day experiment by day-74 sera was related to clearance of the bacteria from the lungs. Isolates demonstrating major electrophoretic changes showed variations in the composition of the oligosaccharide, but not lipid A, moiety of their LOSs. The oligosaccharide of the LOS of each isolate was composed predominantly of glucose but varied substantially in the contents of galactose, arabinose, xylose, mannose, and 3-deoxy-D-manno-octulosonic acid. Therefore, the LOS of H. somnus is capable of undergoing compositional and antigenic variations, which may act as an important virulence mechanism for evading host immune defense mechanisms.

Characterization of immunodominant surface antigens of Haemophilus somnus.

An immunodominant Haemophilus somnus outer membrane protein with an apparent molecular mass of 40 kDa on Western blots (immunoblots) of gradient sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels was characterized because a monospecific antibody against this antigen was protective. This monospecific antibody was used for immunoaffinity purification of the antigen. The immunoaffinity-purified antigen reacted with a polyclonal antibody to the 40-kDa antigen but not with a monoclonal antibody (3G9) which reacted with the 40-kDa antigen in gradient gels. On 8 or 10% gels, the approximately 40-kDa antigen was resolved as two bands, a 40-kDa band which reacted with the protective monospecific polyclonal antibody (p40) and a band of lower molecular mass which reacted with monoclonal antibody 3G9. The latter antigen was designated p39. Both antigens were conserved in all H. somnus isolates tested. The specific antibodies were also used to detect cross-reacting antigens in other gram-negative bacteria. Antibody to p40 reacted with proteins of 55 to 28 kDa, with the greatest intensity shown among proteins from other members of the family Pasteurellaceae. Antibody to p40 was reduced by absorption with live H. somnus or other members of the family Pasteurellaceae, so the antigen appears to be surface exposed. Antibody to p39 only cross-reacted with a broad band (38 to 40 kDa) in Haemophilus agni. Since H. agni is not a bovine pathogen and since convalescent-phase serum from H. somnus-infected animals did recognize p39, the latter may be a good immunodiagnostic antigen, if the lack of cross-reactivity with antigens in other gram-negative bacteria is confirmed with a polyclonal antibody to p39. The cross-reactivity of antiserum to p40 with antigens of members of the family Pasteurellaceae and the ability of this antiserum to protect against H. somnus pneumonia indicate that p40 may be a useful vaccine antigen for H. somnus disease and perhaps even diseases caused by other members of the family Pasteurellaceae.

Streptococcus suis serotypes associated with disease in weaned pigs.

Streptococcus suis was recovered from 9 outbreaks of septicaemia and meningitis in weaned pigs between 1979 and 1983. Fifteen isolates from 7 outbreaks were identified as S. suis type 9, and 3 isolates from 2 outbreaks as S. suis type 2. Three further isolates of S. suis type 2 and an isolate of S. suis type 3 were recovered from cases of bronchopneumonia in weaned pigs from 4 other piggeries.