Aerosol delivery of synthetic DNA containing CpG motifs in broiler chicks at hatch under field conditions using a commercial-scale prototype nebulizer provided protection against lethal Escherichia coli septicemia.

Synthetic DNA containing CpG motifs (CpG-ODN) are potent innate immune stimulators in neonatal and adult broiler chickens against bacterial septicemia. We have recently demonstrated that intrapulmonary (IPL) delivery of CpG-ODN as microdroplets under laboratory conditions can protect neonatal chickens against lethal Escherichia coli septicemia. The objectives of this study were to develop a commercial-scale poultry nebulizer (CSPN) that can deliver CpG-ODN as microdroplets in neonatal broiler chicks in the hatcheries and study the efficacy of CSPN in inducing immune-protective effects under different environmental conditions in 2 geographical locations in Canada. Three field experiments were conducted in commercial poultry hatcheries during different seasons of the year in Saskatchewan and British Columbia, Canada. Neonatal broiler chicks (n = 8,000/experiment) received CpG-ODN by the IPL route in the CSPN chamber for 30 min, and control chicks received distilled water (DW) for 30 min. Broiler chicks (CpG-ODN-240 chicks/experiment and DW-40 chicks/experiment) were randomly sampled from all locations of the CSPN after nebulization and challenged with a lethal dose of E. coli to examine the CpG-ODN nebulization induced protection. We found a significant level (P < 0.05) of protection in broiler chicks against E. coli challenge, suggesting that the newly built CSPN successfully delivered CpG-ODN via the IPL route. We found that when the CSPN was maintained at humidex 28°C or below and relative humidity (RH) between 40 and 60%, neonatal birds were significantly (P < 0.05) protected against E. coli septicemia after IPL delivery of CpG-ODN. By contrast, protection in chicks was adversely affected when the CSPN was maintained at the humidex of 29°C or higher and RH of 70%. Overall, the present study successfully built a CSPN for CpG-ODN delivery in chicks at the hatchery and revealed that the temperature, humidity, and humidex were critical parameters in CSPN for efficient delivery of CpG-ODN.

In vitro demonstration of in utero larval development in an oviparous parasitic nematode: Haemonchus contortus.

The life-cycle of Haemonchus contortus, a pathogenic stomach nematode of sheep, is typical of those of the other members of the superfamily Trichostrongyloides, all of which require a period of development outside the definitive host. Classically, gravid H. contortus, known as strictly oviparous, releases her eggs into the abomasal lumen. The eggs are passed out in the faeces in which they hatch into the 1st-stage larvae and then develop into the infective 3rd-stage larvae. We have developed a method to study the fate of eggs within gravid worms. Using this procedure, we have shown that H. contortus may also exhibit viviparity with apparently normal development from the egg to the 4th larval stage taking place within the gravid female maintained in vitro. On the basis of these observations we speculate that viviparity might occur in vivo with consequent autoinfections; if so, this might explain some puzzling clinical and epidemiological features of haemonchosis, as well as the incomplete efficacy of current control measures.

Seasonal fluctuation and inhibited development of populations of Dictyocaulus filaria in ewes and lambs.

The fluctuation of populations of Dictyocaulus filaria in sheep was studied under field conditions in which animals are housed during the winter and grazed from late spring to autumn. A comparison was made between residual pasture contamination with overwintered larvae, the fecal larval deposition by ewes from June and both of these factors combined as sources of infection for spring born lambs. Ewes and lambs were killed serially over a year and worms were recovered from the lungs and counted. It was found that during the stabling period most of the ewes were carrying moderate numbers of D. filaria. However, while the vast majority of lungworm populations in the winter was inhibited in development at the early fifth larval stage, virtually all worms in the spring were adults. Any one source of infection studied contributed to the acquisition of important burdens of D. filaria by lambs as well as ewes. Worm counts reached peak in all lambs by November and this pointed to only one important Dictyocaulus generation per grazing season. It would also appear that larvae picked up by ewes and lambs as the grazing season advanced had become inhibited in development with the inhibition rate being most marked in autumn.

Gastrointestinal nematode populations in stabled ewes of Rimouski region.

Nematode populations is stabled ewes of the Rimouski region were studied by means of fecal worm egg counts, fecal culture of larvae, and worm counts at necropsy. It was found that during the winter strongyle egg counts were low, Trichostrongylus eggs being most numerous, The stronglye egg counts increased following lambing and reached peak in June. Ostertagia spp was the principal contributor to this "spring-rise", with substantial contribution from Trichostrongylus and Haemonchus contortus. The bulk of adult worm populations in winter, however, was made up of Trichostrongylus, whereas the great majority of the populations of Ostertagia spp, H. contortus and Nematodirus spp were inhibited in development at the fourth larval stage. All the worms recovered at necropsy in spring were adults, coinciding with the "spring-rise".

Seasonal fluctuations of nematode populations in breeding ewes and lambs.

The seasonal changes in nematode populations of a flock of sheep in the Montreal area were determined using serial fecal egg counts, fecal culture of larvae and necropsy worm counts. It was found that Ostertagia spp.,Nematodirus spp., Trichostrongylus agei, Trichostrongylus spp. and Chabertia ovina over-wintered on pasture and could initiate patent infections the following spring. The development of populations of H. contortus was typical of that seen with most of the other species and was characterized by the following series of events. In early winter when the study was started with stabled pregnant ewes, most of the populations were immature and the egg counts were low and remained so throughout the entire winter. However, in the spring, following lambing, large numbers of adult worms were seen with a consequent decrease in immatures and a sudden increase in egg counts. When the ewes and lambs were pastured together, the egg counts in ewes dropped consequent to "self-cure", the "spring-rise" providing the major source of overwhelming infections for lambs with deaths by the end of July. As the season progressed larvae taken in by both ewes and lambs did not mature, and by early fall, most of the worm population consisted of immature forms. It appeared that H. contortus could not have more than two generations in ewes or lambs in a single grazing season.