Prophylactic efficacy of an ivermectin sustained-release bolus against challenge exposure with gastrointestinal and pulmonary nematode infective larvae in calves

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).     

Studies on the ultrastructural morphology of Bacteroides nodosus.

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.     

Distribution and prevalence of footrot in Bhutan

The first cases of footrot in Bhutan were reported in sheep in 1990 at the National Sheep Breeding Centre (NSBC), which supplies breeding animals to village sheep flocks throughout Bhutan. Despite the presence of footrot at the Centre the distribution of apparently disease-free sheep continued. Cases of footrot were reported in village flocks soon after the disease was diagnosed at NSBC. A national survey was designed to establish the distribution and prevalence of footrot in Bhutan. This detected footrot in 19/94 village sheep flocks surveyed. The 19 affected flocks were distributed among nine different administrative districts whereas the villages selected were in 13 of a total of 16 sheep growing districts. The highest within-flock prevalences were among the seven flocks sampled in Bumthang district (mean 20.4%). The prevalence of the disease within flocks was generally much lower in other affected districts and in three districts a single affected animal was identified in the sample of 14 sheep examined in each village. Nationally, footrot prevalence was estimated to be 3.1% (95% CI 2.16-4.04%). There was a positive association between the receipt of animals from NSBC and the presence of footrot. The prevalence of the disease was higher in flocks with a migratory system of management than in those using a sedentary system. The relative risk of there being footrot in a migratory flock was nine-times higher than in a non-migratory flock. Only one strain of Dichelobacter nodosus (serogroup B) was identified among the 234 isolates obtained from the 19 affected flocks. Sheep with footrot healed quickly when treated with a vaccine made from this strain.     

Anthelmintic activities of ivermectin against immature and adult Dictyocaulus viviparus

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).     

Disease resistance in Merino sheep. III. Genetic variation in resistance to footrot following challenge and subsequent vaccination with an homologous rDNA pilus vaccine under both induced and natural conditions

SUMMARY: Eight traits representing clinical indicators of resistance to footrot were examined in 1562 Merino sheep, representing the progeny from 162 sires in four major bloodlines. Over a 4-year period, sheep were exposed to virulent isolates of Dicbelobacter nodosus under both an experimental challenge in which footrot was induced, and a separate natural challenge involving a different isolate of D. nodosus. Five footrot traits and three healing traits were each recorded on seven occasions following induced challenge, and on five occasions following natural challenge. All sheep were vaccinated with a primary and booster injection of an homologous rDNA pilus vaccine, 9 and 6 weeks after initiation of the induced and natural challenge respectively. The major fixed effects which influenced variation in resistance were (in order of importance) time of inspection after challenge, year and group in which sheep were challenged, and sex of the animal. Date of birth, birth-rearing type and age or dam were unimportant in the expression of footrot. Half-sib heritability estimates of resistance to footrot were low to moderate for single observations recorded pre-vaccination (0.07-0.22), and slightly lower for inspections made after vaccination (0.07-0.15). Repeatability estimates for footrot traits during a challenge ranged from 0.31 to 0.70 for inspections pre-vaccination, and 0.19 to 0.35 for inspections post-vaccination. Genetic correlations among footrot traits recorded at repeat inspections were high for observations pre-vaccination (range 0.87-1.00) and slightly lower for observations made after vaccination (0.52-1.00). Heritability estimates derived from repeat measurements approached 0.30 for most traits, except for traits describing healing, which had a heritability of almost zero. Heritability estimates of liability to footrot ranged between 0.09 and 0.41 depending on the time after challenge when the inspections were made. The genetic correlation between induced and natural footrot ranged from 0.14 to 0.95, depending on the period over which inspections were made, with an average of 0.67. In addition to within-flock genetic variation in resistance to footrot, significant differences were observed between different bloodlines within the experimental flock. It was concluded that there is substantial genetic variation in resistance to challenge with virulent isolates of D. nodosus. However, practical restrictions of exploiting available genetic variation may limit the widespread adoption of direct selection. ZUSAMMENFASSUNG: Krankheitsresistenz in Merinos III. Genetische Variabilit?t in Moderhinke Resistenz nach Infektion und folgender Impfung mit homologer rDNA pilus Vakzine unter induzierten und natürlichen Bedingungen Acht Merkmale, die als klinische Hinweise auf Moderhinkeresistenz betrachtet werden, wurden in 1562 Merino Schafen aus 162 Vatertieren von vier wichtigen Linien untersucht. über eine 4-Jahresperiode wurden die Schafe virulenten Isolaten von Dichelobacter nodosus unter Versuchsbedingungen ausgesetzt und eine getrennte natürliche Infektion mit verschiedenen Isolaten von D. nodosus durchgeführt. Fünf Moderhinkemerkmale und drei Gesundungsmerkmale wurden nach Infektion bei sieben Gelegenheiten festgehalten und an fünf nach natürlicher Infektion. Alle Schafe wurden mit einer prim?ren und einer booster Injektion homologer rDNA pilus Vakzine geimpft, 9 und 6 Wochen nach der induzierten und natiirlichen Infektion. Die wichtigsten fixen Effekte, welche die Variabilit?t der Resistenz beeinflussen, waren, nach Wichtigkeit gereint, Zeit der Prüfung nach Impfung, Jahr und Gruppe in welcher Schafe geimpft wurden und Geschlecht. Geburtsdatum, Aufzuchttyp und Mutterschaf-alter waren im Hinblick auf Moderhinke unwichtig. Halbgeschwister-Heritabilit?tssch?tzungen ihrer Resistenz waren niedrig bis mittel für Einzelbeobachtungen vor der Impfung (0,07-0,22) und geringfügig geringer für Beurteilung nach Impfung (0,07-0,15). Wiederholbarkeitssch?tzungen für Moderhinkemerkmale bewegten sich von 0,31 bis 0,70 für Inspektionen vor und 0,19-0,35 für Inspektionen nach Impfung. Genetische Korrelationen zwischen Moderhinkemerkmalen bei verschiedenen Untersuchungen waren fur Beobachtungen vor der Impfung hoch (0,87-1) und geringfügig niedriger nachher (0,52-1). Heritabilit?tssch?tzungen von wiederholten Messungen erreichten 0,30 für die meisten Merkmale au?er für jene, welche Heilung beschreiben, die nahezu keine Heritabilit?t zeigen. Heritabilit?tssch?tzungen für Moderhinkeempfindlichkeit variierten zwischen 0,09 und 0,41 in Abh?ngigkeit von der Untersuchungszeit nach den Impfungen. Die genetische Korrelation zwischen induzierter und natürlicher Moderhinke schwankte von 0,14 bis 0,95 in Abh?ngikeit von der Dauer der Beobachtungsperiode, durschnittlich 0,67. Zus?tzlich zur genetischen Variabilit?t innerhalb der Herde wurden signifikante Unterschiede zwischen verschiedenen Linien innerhalb der Versuchsherde gefunden. Darauf wird es geschlossen, da? substantielle genetische Variabilit?t für Resistenz gegenüber virulenten Isolaten von D. nodosus existiert. Allerdings k?nnen praktische Hindernisse die Ausnutzung der vorhandenen genetischen Variabilit?t durch direkte Selektion einschr?nken.     

Anthelmintic activity of the macrocyclic lactone F28249-alpha in sheep

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.     

Eradication of virulent footrot from sheep and goats in an endemic area of Nepal and an evaluation of specific vaccination

Programmes based on the identification and treatment of cases and the culling of animals refractory to treatment had failed to eradicate virulent footrot from two districts in the western region of Nepal. From 1993 to 1996 vaccination against two endemic virulent strains of Dichelobacter nodosus was tested for its potential to contribute to the eradication of footrot from the region. Only sheep and goats which had been free of signs of footrot at three inspections at monthly intervals before their annual migration to alpine pastures were eligible for inclusion. From November 1992, the treatment of cases identified during inspections included the injection of specific vaccine. Successfully treated cases migrated with their flocks but were excluded from the vaccine trial. Non-responding cases were culled. Forty combined flocks of sheep and goats (approximately 9500 animals) were used initially to compare three vaccination regimens. Eleven flocks (sheep and goats) were treated with two doses of specific vaccine (group A), nine (sheep and goats) were treated with commercial vaccine followed by specific vaccine (group B) and 10 (sheep and goats) were treated with two doses of commercial vaccine (group C) in March to April 1993 before the annual migration; 10 flocks (sheep and goats) remained unvaccinated (group D). Only sheep and goats free of signs of footrot were allowed to migrate. Nevertheless, virulent footrot recurred in many flocks three months later. However, its prevalence was significantly lower in group A than in the other three groups combined. Groups A, B and C then received the specific vaccine before their migrations in 1994 to 1996; group D remained unvaccinated. The annual programme of inspection and identification and treatment of cases continued for seven years, but the vaccinations ceased after four years. There was no recurrence of virulent footrot after November 1993. After the first season the virulent strains of D nodosus used in the specific vaccine could no longer be isolated, although antigenically distinct, benign strains of the organism persisted in cases of benign footrot.     

Disease resistance in Merino sheep. II. RFLPs in Class IIMHC and their association with resistance to footrot

SUMMARY: Restriction fragment length polymorphism (RFLP) within the Major Histocompatibility Complex (MHC) Class II region was examined in 83 sheep. Two restriction enzymes (TaqI and PvuII) and two human cDNA probes (DQα-folke and DRβ-malta) were used to probe Southern blots from all sheep. In addition, a second human DRβ probe (signe) and the restriction enzyme, EcoRI, were used for 15 sheep to match previously published polymorphism in sheep using the same probe/enzyme combination. All sheep were part of an experiment in which footrot was experimentally induced and then controlled through vaccination with an homologous rDNA pilus vaccine. The four main probe/enzyme combinations detected extensive polymorphism; in total 74 bands were detected of which only 2 were present in all sheep. A similar level of polymorphism was also seen with the EcoRI/DRβ-signe probe/enzyme combination, which was greater than previously published for sheep, cattle or goats. After accounting for cross-hybridising bands, the sheep DRβ region appeared to be more polymorphic than the DQα region. Similarly the TaqI restriction enzyme revealed more polymorphism than the PvuI enzyme (40 and 34 bands respectively). For all 74 bands, 7 main clusters were identified both within and across probe/enzyme combinations. After removing bands/clusters which were of extreme frequency (< 0.10 and > 0.90), a total of 53 bands/clusters were analysed for their potential association with footrot and antibody responses. One band was significantly associated with susceptibility to footrot over the 27-week period during which footrot was measured. In addition, two single bands and one cluster were significantly (P > 0.001) associated with residual footrot infection after vaccination. Nine bands/clusters were associated with antibody titre during infection, and one cluster with antibody titre after vaccination. These results suggest a possible involvement for genetic polymorphism within the ovine Class II region in variation in response to footrot. ZUSAMMENFASSUNG: Krankheitsresistenz bei Merinos, II. RFLP's in Klasse II MHC und ihre Verbindung mit der Moderhinkeresistenz Restriktionsfragmentl?ngenpolymorphismus (RFLP) innerhalb des Haupthistokompatibilit?tskomplexes (MHC) Klasse II Region wurde bei 83 Schafen untersucht. Zwei Restriktionsenzyme (TaqI und PvuII) und zwei humane cDNA-Sonden (DQα-folke und DRβ-malta) wurden für Southern blots bei allen Schafen verwendet. Zus?tzlich wurde eine zweite humane DRβ-Sonde (signe) und das Restriktionsenzym EcoRI bei 15 Schafen zum Vergleich mit früher publizierten Polymorphismus mit den selben Enzym/Sondenkombinationen durchgeführt. Alle Schafe waren Teil eines Versuches, in welchem Moderhinke experimentell induziert und dann durch Vaccination mit homologer rDNA-pilus vaccine bek?mpft worden war. Die 4 Enzym/Sondenkombinationen entdeckten extensiven Polymorphismus. Insgesamt 74 Banden wurden entdeckt, wovon nur zwei in allen Schafen vorhanden waren. Ein ?hnliches Ausma? von Polymorphismus wurde auch mit der EcoRI/DRβ-signe Enzym/Sondenkombination entdeckt, das gr??er war als bisher für Schafe, Rinder oder Ziegen publiziert. Nach Berücksichtigung der Kreuz-Hybridisierungsbande scheint die Schaf-DRβ-Region st?rker polymorph als die DQα-Region zu sein. ?hnlicherweise zeigte das TaqI Restriktionsenzym mehr Polymorphismus als das PvuII Enzym (40 bzw. 34 Bande). Bei allen 74 Banden wurden sieben Hauptkluster identifiziert, sowohl innerhalb als auch über Enzym/Sondenkombinationen. Nach Entfernung von Banden/Kluster mit extremen H?ufigkeiten (< 0,10 bzw. > 0.9) wurden insgesamt 53 Bande/Kluster hinsichtlich ihrer potentiellen Verbindung mit Moderhinke und Antik?rperreaktion analysiert. Ein Band war signifikant mit Moderhinkeempfindlichkeit w?hrend der 27-Wochenperiode assoziiert. Zus?tzlich waren zwei einzelne Bande und ein Kluster signifikant (p > 0,001) assoziiert mit Moderhinkeinfektion nach der Impfung. 9 Band/Kluster waren mit Antik?rperspiegel w?hrend der Infektion verbunden und ein Kluster mit Antik?rperspiegel nach Impfung. Die Ergebnisse deuten auf Zusammenhang des genetischen Polymorphismus innerhalb der ovinen Klasse II-Region mit Variabilit?t im Moderhinkeverlauf.