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Until the latter part of the nineteenth century, there were no domestic animals other than pigs, dogs and poultry in the island of New Guinea. From 1889 onwards, occupying authorities, missionaries and settlers from Germany, the UK, Japan and Australia imported ruminants, pigs and horses. Some of these importations were from Asia. This paper describes some outcomes of those importations and the potential hazards for Australia entailed in them., (© 2019 Australian Veterinary Association.)

Objectives: To establish multiple strains of Dichelobacter nodosus in two flocks of sheep and to assess the virulence of five of these strains., Methods: In experiment 1, sheep were challenged with five D. nodosus strains, which varied in both virulence and serotype. In experiment 2, consisting of four replicates (paddock groups), sheep were challenged with seven different strains of D. nodosus. In experiment 3, sheep were challenged with one of five D. nodosus strains., Results: In experiment 1, at 28 days post challenge, four of the five challenge strains were present. Multiple-strain infections were present in 27 feet, with isolates from three serogroups being recovered from 5 feet, and four serogroups from 1 foot. Challenged hind feet were more frequently affected (P < 0.001). In experiment 2, four of the seven strains were recovered from one replicate and three strains from the remaining three replicates. Significantly more hind feet were affected (262/471, 55.6%) than front feet (198/481, 42%) (P < 0.001). Clinically, in both experiments 1 and 2 the footrot resembled an intermediate form, despite the inclusion of a virulent strain of D. nodosus. In experiment 3, this virulent strain caused a higher prevalence of more severe footrot, a greater mean total foot score and, in Merino sheep, resulted in significantly lower weight gains (P < 0.05). Interaction between D. nodosus strain and breed occurred, with Polwarth sheep being significantly more severely affected by one strain than Merino sheep., Discussion: The clinical expression of multiple-strain infections has implications for both research and control of footrot. A novel method of control is proposed., (© 2017 Australian Veterinary Association.)

Objective: To identify and compare programs for eradicating virulent footrot (VFR) chosen by owners of quarantined sheep flocks in southern New South Wales., Method: Data from 196 sheep flocks in the Wagga Wagga and Young Rural Lands Protection Boards were used to determine the program chosen, the influence of flock size on the program chosen and the effects of the program chosen and the use of contractors on the time in quarantine., Results: The most popular programs in flocks using a single program were: total destocking (61/173; 35.3%) and inspection and culling of affected animals (71/173; 41.0%). Treatment of known infected animals was chosen in 41 flocks and of those, 10 (5.8%) used antibiotics for treatment and 31 (17.9%) used foot-bathing. Combined programs were used in 23 flocks and in 10 flocks a change of program occurred before eradication was achieved. The choice of program was, to some extent, affected by flock size, with owners of small flocks (<500 sheep) more likely to destock. The chosen program strongly influenced the time in quarantine, the shortest time being for destocking (mean 284 days), followed by culling of infected sheep (395 days), treatment with antibiotics (433 days) and finally foot-bathing (502 days). Time in quarantine was significantly shorter when contractors were used., Conclusion: All the options chosen led to the eradication of VFR. However, in this sample both the choice of program and the use of contractors influenced the time taken to achieve eradication and therefore the time in quarantine. Based on time in quarantine, foot-bathing was the least desirable option for the eradication of VFR because of the significantly greater time involved, perpetuation of risk to neighbours and increased cost of inspections. These findings were derived from flocks that were quarantined, but they are relevant to all flock owners considering eradication of VFR., (© 2012 The Authors. Australian Veterinary Journal © 2012 Australian Veterinary Association.)

Objective: To determine if the clinical classification of intermediate footrot (IFR) is changed to virulent footrot (VFR) by a transfer of the infected flock to a region where climatic conditions are more favourable for the transmission of the disease., Design: Clinical examination of two groups of Merino wethers infected with IFR; one group of 309 in a region considered less favourable for footrot and another group of 343 at a second site considered more favourable., Procedures: After characterising the form of footrot at the first site, infection was established at the second site by mixing 142 wethers from the first site with 201 unrelated wethers considered to be free of IFR and VFR. Observations of clinical characteristics were made over a 16 month period during which an outbreak of footrot occurred. Clinical assessments were made by inspecting every foot of every sheep at regular intervals and allocating a footscore. Evidence that the same clonal lines of D. nodosus were responsible for the footrot at both sites was provided by serotyping of isolates and using omp gene RFLP as a molecular epidemiological tool., Results: The disease at the first site was classified as IFR because 7% of the sheep developed a maximum footscore (MFS) of 4, the most severe category, despite relatively low rates of transmission. When the outbreak occurred at the second site, which was more suitable for footrot transmission, the maximum proportion of the flock that developed a MFS of 4 was 3.6%, confirming the initial classification of IFR., Conclusions: When a flock infected with IFR was moved to a region where climatic conditions were more favourable for footrot transmission, the clinical classification of the disease remained the same in both the original flock and in sheep exposed to the infection for the first time.

Objective: To determine if intermediate footrot (IFR) can be eradicated from a flock of sheep by inspection and culling of cases during a non-transmission period and if prior antibiotic treatment or vaccination increases the likelihood of eradication., Procedure: A replicated field experiment that compared the three eradication strategies was followed by an observational study of the best of these applied in a commercial flock of 3000 sheep., Results: In the replicated experiment, IFR was eradicated either by inspection and culling alone, or when combined with vaccination. Eradication failed when the sheep were treated with parenteral antibiotics before inspection and culling during the non-transmission period. In the whole-flock program, eradication by repeated inspection and culling of footrot cases during the non-transmission period was successful and the flock remained free of infection 3 years later., Conclusions: IFR can be eradicated by inspection and culling but latent infections, which may persist undetected for at least 34 weeks, require surveillance inspections to be repeated during the non-transmission phase of the program. The use of parenteral antibiotics as an aid to the eradication of IFR is contraindicated.

Goats are an important natural host for footrot and are infected with Dichelobacter nodosus that have virulence characteristics similar to those of sheep strains. However, the humoral response of goats to D. nodosus antigens and the possibility of a serological diagnosis of footrot in goats have not been studied. With the aim of evaluating a diagnostic ELISA test, we investigated the primary immune response of goats to experimental and natural infection, the memory response in recovered animals, and the transfer and persistence of colostral antibodies in kids. Footrot stimulated the goat's immune system and, as in sheep, under-running lesions were the primary stimulus for production of anti-D. nodosus antibodies. The immune response could be detected in ELISA using either fimbrial or outer membrane protein (KSCN) antigens of D. nodosus. Antibody titres resulting from infection declined quickly after recovery and reached pre-infection levels within 3-4 months. Previously affected animals, however, mounted a memory response when injected with purified D. nodosus antigens. Antibody levels attained after anamnestic challenge were correlated with the maximum levels attained during infection, and were therefore indicative of the infection status. Anti-D. nodosus antibodies were also transferred to kids via colostrum, but these antibodies did not persist and therefore were unlikely to interfere with the diagnostic ELISA after 3 months of age. Though these ELISA tests were highly specific, their sensitivity was rather low. Therefore, they are only suitable for a herd diagnosis of footrot in goats and are dependent on the development of advanced under-running infections in a proportion of affected goats.

Dichelobacter nodosus is the essential causative agent of footrot in sheep. The major D. nodosus-encoded virulence factors that have been implicated in the disease are type IV fimbriae and extracellular proteases. To examine the role of the fimbriae in virulence, allelic exchange was used to insertionally inactivate the fimA gene, which encodes the fimbrial subunit protein, from the virulent type G D. nodosus strain VCS1703A. Detailed analysis of two independently derived fimA mutants revealed that they no longer produced the fimbrial subunit protein or intact fimbriae and did not exhibit twitching motility. In addition, these mutants were no longer capable of undergoing natural transformation and did not secrete wild-type levels of extracellular proteases. These effects were not due to polar effects on the downstream fimB gene because insertionally inactivated fimB mutants were not defective in any of these phenotypic tests. Virulence testing of the mutants in a sheep pen trial conducted under controlled environmental conditions showed that the fimA mutants were avirulent, providing evidence that the fimA gene is an essential D. nodosus virulence gene. These studies represent the first time that molecular genetics has been used to determine the role of virulence genes in this slow growing anaerobic bacterium.

Objective: To determine the infectivity of ovine and caprine strains of Dichelobacter nodosus for both sheep and goats., Design: Pen experiments in which 20 sheep and 19 goats were challenged directly with the two strains, and transmission experiments on pasture, using donors infected by experimental challenge., Results: Sheep and goat strains of D nodosus infected both animal species in experimental challenges. Animals so infected transmitted footrot to both sheep and goats on pasture plots. A significantly smaller proportion of goats than sheep was infected when challenged with either strain. The interval between exposure and development of footrot in goats was longer than in sheep when recipient animals were exposed to infected donors on pasture. The disease was less invasive in goats than in sheep., Conclusions: With the strains of D nodosus used there was no evidence of host specificity. Direct transmission of footrot can occur between sheep and goats in the same environment. There is a need to include goats in ovine footrot eradication programs and vice versa.

Calves were vaccinated with cloned Moraxella bovis pili of serogroup C (experiment 1) or B (experiment 2) either as a monovalent formulation or as part of a multivalent preparation with pili of six other serogroups. Within 4 weeks of the second vaccine dose vaccinated calves and non-vaccinated controls were challenged via the ocular route with either virulent M. bovis strain Dal2d (serogroup C) or M. bovis strain 3WO7 (serogroup B) in experiments 1 and 2, respectively. Calves vaccinated with multivalent vaccines had significantly lower antibody titres than those vaccinated with monovalent preparations. Nevertheless, the levels of protection against infectious bovine keratoconjunctivitis (IBK) achieved with multivalent vaccines were 72% and 83% for the groups challenged with M. bovis strains of serogroups B and C, respectively. The serogroup C monovalent vaccine gave 100% protection against experimentally induced IBK and M. bovis isolates cultured from the eyes 6 days post-challenge were identified as belonging solely to serogroup C. Unexpectedly, only 25% protection was achieved against homologous strain challenge of calves that received the monovalent serogroup B vaccine. Furthermore, the majority of M. bovis isolates recovered from calves in this group belonged to serogroup C, as did half of those isolates cultured from the multivalent vaccinates. The remaining bacterial isolates from the latter group, together with all isolates from the non-vaccinated controls, belonged to serogroup B. Results are consistent with the hypothesis that derivatives of the serogroup B challenge inoculum had expressed serogroup C pilus antigen within 6 days of the challenge, possibly as a result of pilus gene inversion occurring in response to the presence of specific antibody in eye tissues and tears.

Protection conferred by a cell-free preparation from a haemolytic Moraxella bovis isolate, UQV 148NF, was compared to an equivalent fraction from a non-haemolytic M. bovis isolate, Gordon 26L3, and to a recombinant DNA-derived pili vaccine. Three groups of ten calves were vaccinated twice with one of the three preparations and, together with ten non-vaccinated calves, challenged with virulent M. bovis isolate Dal 2d. Compared to the control group, significant protection was observed in the group receiving the pili vaccine and the group receiving the preparation from haemolytic isolate, UQV 148NF.

Pili (fimbriae) were prepared from Moraxella bovis strain Dalton 2d (Dal2d) and from a derivative of Pseudomonas aeruginosa K/2PfS that contained a plasmid-borne Dal2d pilin gene and produced pili having serogroup-specific identity to Dal2d. Nine calves were vaccinated with two doses each of 30 micrograms authentic M. bovis Dal2d pili in oil adjuvant and 10 calves were vaccinated with a similar dose of P. aeruginosa-derived Dal2d pili in the same formulation. All 19 calves and 10 non-vaccinated controls were challenged by instillation of 1 x 10(9) virulent M. bovis Dal2d cells into both conjunctival sacs 19 days after the second vaccine dose. The serological response to vaccination and the degree of protection against experimentally induced infectious bovine keratoconjunctivitis (IBK) were assessed. None of the nine calves vaccinated with authentic M. bovis Dal2d pili developed IBK while two of those vaccinated with P. aeruginosa-derived Dal2d pili developed lesions which accounted for a mean group lesion score of 0.3. In contrast, 9 of the 10 non-vaccinated calves developed IBK lesions, the majority of which were progressive, required early treatment and accounted for a mean group lesion score of 1.5. These results demonstrate the potential of a relatively low dose of pili produced by recombinant DNA technology for development of an effective vaccine against IBK.

Dichelobacter nodosus is an essential causative agent of ovine footrot, a disease of major economic significance. Four oligonucleotides complementary to variable regions of the 16S rRNA of D. nodosus were identified, synthesized and tested for their specificity and sensitivity as probes for the detection of D. nodosus. In hybridization reactions using total RNA as the target nucleic acid, three probes were found to be both sensitive and species-specific. When these probes were used as primers in PCR reactions, on both purified D. nodosus DNA and whole cells, the sensitivity of detection was increased by several orders of magnitude. Using PCR, it was possible to detect the presence of D. nodosus by direct examination of lesion material from footrot infected sheep.

A multivalent footrot vaccine has been developed, containing pilus antigens produced in recombinant Pseudomonas aeruginosa and representing all nine serogroups of Dichelobacter (Bacteroides) nodosus commonly recognised in the field. The responses of sheep to the multivalent vaccine have been compared with those to monovalent vaccines representing only a single serogroup. Antigenic competition between serogroups occurred in sheep immunised with the multivalent formation, but high levels of protection were still achieved. The study showed that in multivalent footrot vaccines, antigenic competition is predominantly due to the presence of a family of immunologically-related pilus antigens rather than to interference by extraneous proteins.

K-agglutination, pilus-enzyme-linked immunosorbent assay (ELISA) and outer membrane protein-ELISAs were used to assess humoral responses after vaccination with a commercial, multivalent, ovine foot rot vaccine (Dichelobacter nodosus whole cells) in three groups of nine-month-old lambs of markedly different bodyweight, nutritional history and dietary protein supply. Mean bodyweights of lambs in low (L), medium (M) and high (H) bodyweight/nutrition groups were 22, 32 and 48 kg, respectively, at the time of vaccination. Few significant differences in humoral responses to vaccine antigens were found between groups. However, lambs in group H tended to have lower levels of antibody to a greater number of component antigens than did lambs in the other groups. These results suggest that low bodyweight due to poor nutrition is unlikely to affect the response of sheep to multivalent foot rot vaccines.

The macrolytic lactone F28249-alpha was titrated in experimentally infected sheep and found to be highly effective against most of the common gastrointestinal nematodes as a single oral dose, given at a rate of 0.025, 0.05, or 0.1 mg/kg. Specifically, maximal activity was evident at even the lowest dosage against adult Haemonchus contortus, Ostertagia circumcinta, Trichostrongylus axei, and T colubriformis and L4 O circumcinta. Activity against Oesophagostomum columbianum was also high at all dosages, with a calculated ED95 of 0.029 mg/kg. Cooperia curticei was eliminated at 0.1 mg/kg, but control was erratic at the lower dosages. The greatest weakness of this compound was its activity against C oncophora. The activity against this parasite was weak (less than or equal to 85%) at all dosages, and the dosage-response curve was flat, suggesting dosages substantially higher than those given would be necessary for high-order control of this species.

A mixture of lincomycin and spectinomycin was investigated as a treatment for footrot in sheep. In a controlled clinical trial 92.5% of acute and chronic cases of virulent footrot were cured following a single intramuscular injection of a mixture containing 50 mg lincomycin and 100 mg spectinomycin/ml at a dose rate of 1 ml/10 kg bodyweight. No improvement in clinical response was observed in groups of sheep treated on 3 successive days with this dose rate nor in another group treated once at a dose rate 1 ml/3.3 kg bodyweight. Cure effectiveness of each of the 3 treatment groups relative to untreated controls was 89%, 95% and 95%. Efficacy of lincomycin/spectinomycin was compared with that of penicillin/streptomycin in the treatment of footrot on 2 farms in south western New South Wales. Assessments made 14 to 17 d after treatment showed that on one farm all 122 ewes treated with lincomycin/spectinomycin had recovered while 170 of 175 ewes treated with penicillin/streptomycin recovered in the same period. On the second farm 87 of 90 ewes treated with lincomycin/spectinomycin recovered, compared with 184 of 190 sheep in the same flock treated with penicillin/streptomycin. Supportive footbathing did not seem to improve the clinical response in either treatment group and the paring done was sufficient only to establish diagnosis and to remove grossly overgrown horn.

Paraherquamide, an oxindole alkaloid metabolite of Penicillium paraherquei, was tested against the common gastrointestinal nematode species of sheep at 0.25, 0.5, 1.0, and 2.0 mg/kg, per os. It was highly efficacious (greater than or equal to 98% reduction) as a single oral treatment dosages greater than or equal to 0.5 mg/kg against adult Haemonchus contortus, Ostertagia circumcincta, Trichostrongylus axei, Trichostrongylus colubriformis and Cooperia curticei, and the L4 stage of Cooperia spp. Noteworthy is the fact the isolate of H. contortus used was ivermectin-resistant and the isolate of T. colubriformis used was ivermectin- and benzimidazole-resistant. This suggests a different mode of action for paraherquamide relative to ivermectin and the benzimidazoles. The adult stage of Oesophagostomum columbianum was the dosage-limiting parasite with 79% efficacy recorded at the highest treatment level (2.0 mg/kg). Extrapolation from the O. columbianum response curve suggests a dosage in excess of 4.0 mg/kg would be required to attain 95% efficacy.

Successful vaccination of sheep against footrot and attempts to eradicate the disease depend on there being a limit to the antigenic diversity of the causative bacterium, Bacteroides nodosus. Fimbrial antigenic variation was therefore investigated in vivo, both under conditions of chronic infection and under the pressure of a vaccine-induced immune response, to ascertain whether this represented an obstacle to such goals. Material was available from 5 experiments and although B. nodosus appeared to have undergone changes in its fimbrial antigens in one of these, the possibility that superinfection was responsible for the variation detected could not be ruled out because all sheep in this case were maintained at pasture. Overall, the results provided no evidence of fimbrial antigenic shift in B. nodosus in vivo and in conclusion, the survival of the organism in the sheep's foot, both in long-term natural infection and following vaccination, must therefore be related to factors other than the ability to undergo antigenic variation in order to evade the host's immune response.

In vivo ivermectin resistance was selected in an isolate of Trichostrongylus colubriformis (TcR) already known to be benzimidazole resistant. This was accomplished in sheep by using levels of ivermectin calculated to reduce the fecal egg output from each generation of T. colubriformis by congruent to 95%. The first indication of ivermectin resistance was observed with the F10. A dosage-titration trial comparing the parent TcR with the ivermectin-selected F21 demonstrated that the latter was congruent to 20 times more resistant to oral ivermectin therapy in experimentally infected sheep than was the parent isolate. Treatment of the F16 generation with 50 mg/kg of thiabendazole resulted in only 54% egg reduction and confirmed that benzimidazole resistance was stable.

22,23-Dihydroavermectin B1, ivermectin, derived from avermectin B1 by selective hydrogenation using Wilkinson's homogenous catalyst [Ph3P)3RhCl], was shown to be a highly effective drug for the treatment of a wide variety of metazoan parasitic diseases in animals.

The feet of sheep which had been given two doses of oil adjuvant foot rot vaccine were completely resistant to direct challenge with a virulent homologous culture of Bacteroides nodosus four weeks after vaccination, partially resistant after eight weeks but completely susceptible after 12 weeks. Sheep given two doses of alum precipitated vaccine, or a dose of oil emulsion vaccine followed by a dose of alum precipitated vaccine, were resistant four weeks but not eight weeks later. Foot rot in affected, vaccinated sheep was more severe in those which had received the alum precipitated vaccine than in those given the oil emulsion vaccine, but vaccinated sheep were always less severely affected than controls. Where oil emulsion vaccine provided the primary stimulation, agglutinin titres were 18,000 four weeks after revaccination with either vaccine, falling to 4000 after a further four weeks. Significantly poorer titres were observed when two doses of alum precipitated vaccine only were given.

Recombinant Pseudomonas aeruginosa cells containing the Bacteroides nodosus fimbrial subunit gene under the transcriptional control of a strong promoter produce large amounts of B. nodosus-type fimbriae. We have carried out vaccination trials which show that these fimbriae are just as effective as either natural fimbriae or whole cell preparations of B. nodosus in inducing protective immunity against homologous footrot challenge. The recombinant-produced fimbriae are also effective therapeutically in accelerating the rate of healing of pre-existing footrot lesions. These results confirm that the structural subunit of the fimbrial strand is a primary protective antigen against footrot, and demonstrate the practicality and potential of recombinant DNA approaches to the development of new vaccines against B. nodosus and other Type 4 fimbriate pathogens.

A series of methyl imidazo-[11,2-a]pyridine-2-carbamates was synthesized for anthelmintic testing. The preparation of this class of compounds was simplified by utilization of a novel one-step condensation of the appropriately substituted 2-aminopyridine and methyl chloroacetylcarbamate. The most potent compound, methyl 6-(phenylsulfinyl)-imidazo[1,2-a]pyridine-2-carbamate, was orally effective against a broad range of helminths in sheep and cattle, at a dosage of 2.5 mg/kg. Limited trials in swine and dogs demonstrated anthelmintic activity at higher dosages. Limited observations in sheep and cattle indicated that, in both species, a single oral dose of 200 mg/kg was well tolerated.

Groups of 25 chickens free of maternal antibody to reticuloendotheliosis virus (REV) were inoculated with either third or seventh passage REV at either one or seven days of age. Some of the birds inoculated at day 1 with REV were inoculated with Salmonella typhimurium either concurrently or six or 13 days later while some of those inoculated with REV at day 7 were inoculated concurrently with S typhimurium. At day old, infection with S typhimurium alone caused the death of 12 of 25 chicks whereas in the dual infection, using the third passage REV, 18 of 25 birds died. Similarly no seven or 14 day old chickens died when challenged with S typhimurium alone, but previous day-old infection with REV caused a respective mortality of eight of 25 and five of 25 birds. With the seventh passage REV a similar pattern was seen. At day old S typhimurium infection alone killed seven of 25 birds whereas combined with virus the mortality was 14 of 25 and while S typhimurium alone killed none of 25 chicks infected at seven days old, the mortality in birds also infected with REV was 14 of 25. Combined virus and bacterial infections did not increase the proportion of feathering defects in birds surviving S typhimurium infections. There was a significantly higher proportion of feathering defects in birds infected with third passage virus compared with seventh passage virus. Although a higher proportion of birds had antibody responses to REV in the seventh than in the third passage group, there was no discernible difference in the effect the different viruses had on chickens' susceptibility to S typhimurium.

The morphology of Bacteroides nodosus was examined with the electron microscope. B nodosus stained with solium phosphotungstate and uranyl acetate possessed fimbriae and in addition organisms negatively stained with sodium phosphotungstate often possessed rings on their surface. Phage-like particles were also observed in negatively stained preparations. In thin sections, B nodosus had a multilayered cell envelope and the type of cell division characteristic of Gram-negative bacteria. The cytoplasmic region contained a diffuse nucleoid area, ribosomes and, sometimes, concentrically arranged membranous lamellae. Fimbriae and capsular material were also seen in sections of B nodosus fixed with glutaraldehyde-osmium. Their visualisation appeared to be enhanced when ruthenium red was incorporated n the glutaraldehyde-osmium fixative but only when sections were stained with heavy metal salts, indicating that the fimbriae and capsule were not predominantly polysaccharide in nature.

Broiler chickens free of maternal immunity to reticuloendotheliosis virus (REV) were used in the experiment. Two groups of 25 chickens were inoculated with REV at one day of age. One of these groups and another group of 25 chickens were inoculated with Eimeria tenella sporulated oocysts at 7 days of age. Chickens inoculated with E. tenella showed bloody diarrhoea from 12 to 14 days of age. Six out of 25 chickens died (P less than 0.05) at 13 and 14 days of age in the dual infected group. At 14 days of age, when chickens were killed, the lesion score in the combined infection group, was statistically different from that in the chickens inoculated with E. tenella alone. Also the weights of the bursa of Fabricius and thymus were lower in the two REV infected groups than in the controls. Although REV infection alone adversely affected the weight gain and feed conversion, with combined infection this effect was much greater. Following REV inoculation most of the chickens showed feathering defects and all the examined chickens were viraemic at 21 days of age. At the same age, all but one chicken failed to show precipitating antigenaemia and about one-half of these chickens showed a very low serum neutralisation titre. None of these chickens showed precipitating antibodies.

A miniature battery-powered radio beacon attached to a stainless steel bolus housing an osmotic pump is described. When the bolus, of sufficient size and density to be retained in the reticulum, is administered to ruminants the radio signal can be received, using a standard AM broadcast receiver up to a range of 1.5 m. The unit allows making a simple and inexpensive verification that the drug delivery system is still within the rumino-reticulum during long-term drug delivery studies and making an external estimation of the internal anatomic location of the device.

Efficacy of ivermectin in a topical formulation was evaluated in cattle against adult gastrointestinal and pulmonary nematode infections (experiment 1, n = 24), the 2nd- and 3rd-stage larvae of Hypoderma spp (experiment 2, n = 12), and the biting louse Damalinia bovis (experiment 3, n = 12). Nematode infections were induced and grubs and lice were naturally acquired. Treatments consisted of a single dose of ivermectin in a topical formulation of 200, 500, or 1,000 micrograms/kg of body weight in experiment 1 or 500 micrograms/kg in experiments 2 and 3. At 1,000 micrograms/kg, ivermectin was 100% effective against Ostertagia ostertagi, Trichostrongylus colubriformis, Oesophagostomum radiatum, Nematodirus helvetianus, Haemonchus placei, and Dictyocaulus viviparus and was greater than 99% effective against Cooperia oncophora, C punctata, and T axei. At 500 micrograms/kg, the efficacy was 100% against C oncophora, C punctata, O ostertagi, T axei, Oes radiatum, N helvetianus, Haem placei, and Dict viviparus and greater than 99% against T colubriformis. At 200 micrograms/kg, the efficacy was 100% against Oes radiatum, Haem placei, and Dict viviparus, greater than 99% for O ostertagi, 96% for C oncophora, 86% for C punctata, 90% for T axei, 85% for T colubriformis, and 71% for N helvetianus. At 500 micrograms/kg, ivermectin was highly effective against the grubs Hypoderma bovis and H lineatum and eliminated the louse Damalinia bovis.

The eighth generation of adult Haemonchus contortus, selected by subjecting infected pairs of sheep to suboptimal ivermectin treatment once per generation from parent (P; BBH isolate) through F7 (IV-A; selected isolate), required an approximate 4-fold increase in the ivermectin dose to produce 95% efficacy compared with its contemporary parent isolate. In a dose titration experiment the dose-response curve of the drug pressure-derived isolate, IV-A, was significantly (0.02 less than P less than 0.05) less steep than was the response curve of the parent, BBH, isolate. Potency estimates based upon these nonparallel dose-response curves would not remain constant over a range of efficacy levels but would decrease rapidly at efficacies greater than 95%. Passage of a closed population of the F8 generation of IV-A sequentially through pairs of sheep for an additional 11 generations (F8A-F8K) without additional drug pressure being applied produced no reversion to sensitivity to ivermectin relative to the F7 generation, thus suggesting that the selected "resistance" was stable.

The distribution of 12 Class I ovine lymphocyte antigens (OLA) was examined in 4 flocks of sheep vaccinated against and/or challenged with Bacteroides nodosus, the transmitting agent of footrot. In a flock of 47 Corriedales in New Zealand, which had been specially bred for resistance to footrot, a higher frequency (70.2%) of OLA type SY6 was found compared with 42.9% in 49 unselected Corriedale sheep (P = 0.001). The serum antibody response of 12 selected Corriedale ewes was compared with that of 12 unselected ewes of the same age after vaccination with a multivalent footrot vaccine and the selected ewes had significantly (P = 0.01) higher agglutinin titres than the unselected ewes, 7 weeks after vaccination. In 3 trials involving 108, 120 and 135 Australian Merinos in Victoria, SYlb was associated with a reduction in the number of feet affected with severe footrot (P = 0.05, P = 0.01, P = 0.02) and in 2 of the trials there was a relationship between SY6 and high vaccinal agglutinin titres. This SY6 effect was evident in the first trial 31 days after primary vaccination (P = 0.05) and again 20 days later after secondary vaccination (P = 0.01). In the second trial, when the sheep were vaccinated 49 days after challenge, an association was again found between SY6 and high agglutinin titres (P = 0.05) after primary but not after secondary vaccination. Exposure of 157 vaccinated Merino rams to B. nodosus during a footrot outbreak in New South Wales also showed an association between low infection and SY6 and SYlb.(ABSTRACT TRUNCATED AT 250 WORDS)

Twelve Holstein calves were used to determine the prophylactic efficacy of ivermectin against challenge exposure with gastrointestinal and pulmonary nematodes. Two groups of 6 calves (mean body weight, 205 kg) each were formed by restricted randomization according to body weight. Group-1 calves served as nonmedicated controls. Each calf of group 2 was orally given one prototype sustained-release bolus designed to deliver ivermectin at a continuous daily dose of 8 mg. Third-stage nematode infective larvae were given to the calves on posttreatment days 28 and 42. The calves were euthanatized 77 or 78 days after treatment. Ivermectin was 100% effective (P less than 0.05) in preventing the establishment of infection by Haemonchus placei, Ostertagia ostertagi, Cooperia spp (C punctata, C oncophora, C surnabada), Nematodirus helvetianus, Oesophagostomum radiatum, and Dictyocaulus viviparus and was greater than 99% effective against Trichostrongylus axei. Incidental infection by Trichuris spp was reduced by 94% (P = 0.08).

Eighteen calves about 3 months old were inoculated with 3,000 Dictyocaulus viviparus infective larvae. Three groups of 6 calves each were formed. Thirteen days after inoculations, 3 of the 6 group 1 control calves were given vehicle subcutaneously (SC) and the group 2 calves were given ivermectin at the dose rate of 200 micrograms/kg, SC. Thirty-five days after inoculation, the remaining 3 calves in group 1 were given vehicle SC and the group 3 calves were given ivermectin at the dose rate of 200 micrograms/kg, SC. Necropsies were performed 42 days after inoculations. A total of 474 D viviparus was recovered from the group 1 control calves, whereas none was recovered from the calves treated when the nematodes were in the 4th stage of development (group 2) or adult stage (group 3).

Avermectins A2a, B1a, and B2a (1, 2, and 3) were acetylated to give 4"- and 23-acetates 4 and 5 and 4",23-diacetate 6 from 1, the 4"-and 5-acetates 7 and 8 and 4",5-diacetate 9 from 2, and triacetate 10 from 3. Structure proof by 300-MHz 1H NMR and mass spectral fragmentation is discussed for 10. Forcing acetylation conditions generated from both 1 and 3 the identical aromatic diacetate 11. Good anthelmintic activities in gerbils and sheep for 4"-acetylated derivatives 4 and especially 7 prompted the preparation of additional 4"-acylated derivatives of 2 with pivaloyl, n-octanoyl, succinoyl, carbamoyl, dimethylcarbamoyl and N-acetylglycyl substituents, prepared from the 5-O-tert-butyldimethylsilyl-protected intermediate 12. Other key intermediates were the trichloroethyoxysuccinoyl derivative 18 and 4-nitrophenyl carbonate 21. Anthelmintic activities against Trichostrongylus colubriformis in gerbils comparable in potency to the natural product 2 are shown by the more polar substituted derivatives 20, 23, and 27. Substitution of the 5-hydroxy group or its loss due to aromatization results in drastically reduced anthelmintic potency.

Ivermectin, a potent antiparasitic agent with activity against internal and external parasites, was delivered to cattle at a controlled zero-order rate for 35 d via orally administered, specially weighted, ALZET 2ML4 osmotic pumps. The osmotic pumps delivered the drug consistently over the trial period. Steady-state levels in plasma were achieved in 7-14 d, and plasma concentration depletion curves were observed to start at approximately day 35, the theoretical delivery lifetime of the osmotic pumps. Bioavailability was estimated to be 40%, and dose rate-plasma steady-state interrelationships were shown to be linear.

Some 2-arylhydrazino- and 2-arylazo-2-thiazolines were synthesized for anthelmintic testing. The most potent compound, 2-(o-tolylazo)-2-thiazoline, was orally effective in sheep against a broad range of helminths.

Anthelmintic efficacies of a series of 6-substituted methyl imidazo[1,2-alpha]pyridine-2-carbamates were compared to similarly substituted benzimidazole-2-carbamates. With only one exception, methyl 6-benzoylimidazo[1,2-alpha]pyridine-2-carbamate, both classes of compounds exhibited similar activity vs. Nematospiroides dubius in mice. Preliminary screening indicated methyl 6-(1,2,2-trichloroethenyl)imidazo[1,2-alpha]pyridine-2-carbamate to be the most potent derivative in the series. However, evaluation in sheep indicated that its anthelmintic spectrum was inferior to methyl 6-(phenylsulfinyl)imidazo[1,2-alpha]pyridine-2-carbamate.

Ivermectin delivered continuously from a rumino-reticular sustained release device was prophylactically effective in preventing establishment of nine nematode parasite species in multiply-exposed cattle. Ivermectin dosages which permitted less than or equal to 1% of infected control calf worm populations to establish ranged from less than 2.5 micrograms kg-1 day-1, which totally prevented infection with Dictyocaulus viviparus and Oesophagostomum radiatum, to approximately equal to 30 micrograms kg-1 day-1 required to suppress Nematodirus helvetianus to the same extent. Between these extremes, in decreasing order of sensitivity to enterical sustained release ivermectin, were Ostertagia ostertagi, Trichostrongylus axei, Haemonchus placei, Cooperia punctata, C. oncophora and T. colubriformis which were maximally affected at less than or equal to 10 micrograms kg-1 day-1 of ivermectin.

A series of dihydroquinoxalino[2,3-b]quinoxalines was synthesized and tested for anthelmintic activity in a model assay. The most promising compound, 5,12-diacetyl-5,12-dihydroquinoxalino[2,3-b]quinoxaline, was orally effective in sheep at a dose of 200 mg/kg against a broad range of helminths.