Influence of porcine intestinal pH and gastric digestion on antigenicity of F4 fimbriae for oral immunisation

Newly weaned piglets can be orally immunised against F4+ enterotoxigenic Escherichia coli (ETEC) infection with F4 fimbriae. However, to efficiently develop a vaccine against ETEC induced postweaning diarrhoea, knowledge of the stability of the F4 fimbriae to different pH and gastric digestion is needed. The gastrointestinal pH in suckling and recently weaned piglets was measured and the stability of F4 fimbriae to different pH and to pepsin was assessed in vitro. In the stomach the lowest pH was found in the fundus gland region. Gastric pH values below 2.5 were not found in suckling piglets or at the day of weaning, in contrast to piglets 1 and 2 weeks postweaning. Along the first half of the small intestine and in the caecum, a negative correlation was found between pH and age. The F4 fimbriae were stable to pH 1.5 and 2 for 2 h, whereas longer incubation periods resulted in conversion of the multimeric forms into monomers. The F4 fimbriae were partially degraded by incubation for 15-30 min in simulated gastric fluid at pH 1.5 and 2, and completely digested from 3 h onwards. At pH 3, the fimbriae maintained their antigenicity for at least 4h. The results demonstrate that gastric digestion will only have a limited impact on oral immunisation since liquid passes through the stomach relatively quickly (50% within 2 h). However, we previously demonstrated that the transit times are prolonged shortly after weaning. Shortly after weaning it could be necessary to protect the F4 fimbriae against gastric digestion to obtain efficient oral immunisation of the piglets.     

Assessment of dexmedetomidine/ketamine anesthesia in golden-headed lion tamarins (Leontopithecus chrysomelas)

OBJECTIVE: To compare the anesthetic and cardiovascular effects of dexmedetomidine/ketamine combinations in golden-headed lion tamarins. STUDY DESIGN: Prospective blinded randomized study. ANIMALS: Six healthy golden-headed lion tamarins, three males and three females were studied (mean body weight +/- SD = 0.498 +/- 0.054 kg). METHODS: The animals were given one of the dexmedetomidine/ketamine combinations (0.005/10, 0.01/10, and 0.01/5 mg kg(-1) IM; treatments 5D/10K, 10D/10K, and 10D/5K, respectively) on three successive occasions. Time to sedation and recumbency, as well as anesthesia, standing, and walking times were recorded. Heart and respiratory rate, arterial blood pressure, hemoglobin saturation, and rectal temperature were recorded during anesthesia. Sedation, muscle relaxation, and auditory response were evaluated after drug administration. RESULTS: Heart rate decreased after two combinations (10D/5K and 10D/10K). Respiratory rate and rectal temperature progressively decreased in all the treatments. Arterial blood pressures were maintained in 5D/10K and 10D/5K treatments and decreased after 10D/10K administration. There were no differences in sedation and recumbency time between the treatments, but anesthesia time was significantly longer in treatment 10D/10K (67.80 +/- 13.30 minutes) compared to treatments 10D/5K (44.54 +/- 8.25 minutes) and 5D/10K (30.60 +/- 6.80 minutes). Standing time was shorter in treatment 10D/10K (11.13 +/- 10.01 minutes) than in treatments 5D/10K (18.20 +/- 10.03 minutes) and 10D/5K (19.12 +/- 14.55 minutes), and walking time was longer in treatment 5D/10K (20.00 +/- 12.46 minutes) compared with treatments 10D/10K (16.00 +/- 3.43 minutes) and 10D/5K (12.40 +/- 5.02 minutes). Anesthetic quality was significantly better 5 and 15 minutes after treatments 10D/10K and 10D/5K than after treatment 5D/10K. Analgesia scores were higher after administration of 10D/10K than after the 5D/10K treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Dexmedetomidine/ketamine is useful for chemical immobilization of golden-headed lion tamarins, but bradycardia and hypotension warrant close monitoring.     

Diagnostic procedures and surgical treatment of craniodorsal coxofemoral luxation in calves

OBJECTIVE: To report use of ultrasonographic examination of the coxofemoral joint, a surgical technique for repair of craniodorsal coxofemoral dislocation, and outcome. STUDY DESIGN: Clinical case reports. ANIMALS: Calves (n=4) with coxofemoral luxation. METHODS: Craniodorsal coxofemoral luxation was diagnosed by physical examination, radiographic, and ultrasonographic findings. Open surgical reduction of the femoral head was performed using a modified caudal approach. RESULTS: Craniodorsal luxation of the femoral head and the presence of an intact femoral neck were confirmed by ultrasonography. All luxations were successfully reduced and reluxation did not occur. At follow-up, 1 heifer had calved and 1 was 5 months pregnant. One calf died of bronchopneumonia 6 days after surgery. One calf had severe coxofemoral degenerative joint disease diagnosed (ultrasonography, radiography, and arthrocentesis) 3 months after surgery and confirmed by necropsy. CONCLUSION: Ultrasonography proved to be a simple and effective non-invasive technique for diagnosis of coxofemoral luxation. Immediate surgical intervention in hip dislocation in calves is necessary to avoid unnecessary trauma to subchondral structures. In calves, open instead of closed surgical reduction appears preferable because it allows access to the acetabular cavity for removal of debris. CLINICAL RELEVANCE: Ultrasonography should be considered a supplementary but not an alternative to radiographic examination for diagnosis of coxofemoral luxation and for follow-up examinations after reduction.     

An immunohistochemical study of the organization of ganglia and nerve fibres in the mucosa of the porcine intestine

In order to elucidate the organization of the enteric nervous system in the mucous plexus, wholemounts from six intestinal regions in six pigs were studied by vasoactive intestinal peptide, substance P, nitric oxide synthase and neurofilament proteins immunohistochemistry. The mucous plexus of both large and small intestine contained ganglia and isolated neurons. They were many and comparably larger in the caecum and colon, few in the ileum, and fewer and smaller in the jejunum. The mucous plexus was subdivided into the lamina muscularis mucosae and lamina proprial subplexuses, and based on location the latter was subdivided further in order to clarify their variations with respect to the amount, sizes and shapes of ganglia and neurons, sizes and orientation of nerve strands and immunoreactivities. Ganglia were situated at different topographical levels in the lamina muscularis mucosae subplexus, outer proprial and interglandular proprial meshworks in the lamina proprial subplexus with the majority of ganglia occurring in the outer proprial meshwork. The mucous plexus in the intestine of the pig is thus a ganglionated plexus showing marked segmental variation in the amount of intramucosal ganglia and isolated nerve cells. These new observations, calls for a re-examination of the mucous plexus to elucidate the regulatory mechanisms of importance in mucosal functions and consideration of the mucous plexus in the intestine of the pig to be one of the major ganglionated plexuses.     

Temporal patterns and quantification of excretion of Mycobacterium avium subsp paratuberculosis in sheep with Johne's disease

OBJECTIVES: To determine the frequency of excretion of Mycobacterium avium subsp paratuberculosis in Merino sheep with Johne's disease and to quantify excretion in a group of Merino sheep. DESIGN: A pen and laboratory experiment. PROCEDURE: Seven sheep selected from an affected flock on the basis of acid-fast bacilli in the sheep's faeces were housed and total daily faecal output was collected, weighed and subjected to culture for M avium subsp paratuberculosis. An end-point titration method was used to enumerate viable M avium subsp paratuberculosis in a 15 day pooled sample from five sheep that had acid-fast bacilli in their faeces while housed. RESULTS: Four sheep with subclinical multibacillary Johne's disease excreted M avium subsp paratuberculosis each day for 11 days of cultural observation. A further three sheep were intermittent excreters but lacked other evidence of infection with M avium subsp paratuberculosis. The average number of viable bacteria excreted was 1.09 x 10(8) per gram of faeces while total daily excretion was 8.36 x 10(10) viable M avium subsp paratuberculosis per sheep. Examination of faecal smears stained with Ziehl Neelsen was an unreliable means of assessing daily excretion in individual animals except in those with severe lesions. CONCLUSION: Excretion of M avium subsp paratuberculosis in Merino sheep with multibacillary Johne's disease occurred daily, proving that environmental contamination can be continuous on farms with endemic ovine Johne's disease. Faecal culture is a useful method for detecting infection as it does not appear to be affected by the timing of collection of a sample from sheep with multibacillary disease however, to maximise the sensitivity of disease surveillance using faecal culture, sampling rates should be adjusted to take account of the proportions of multibacillary and paucibacillary cases.     

Tansy ragwort poisoning in a horse in southern Ontario

Bizarre behavior, apparent lameness, and colic were noticed in 1 of 3 horses on a pasture overgrown by weeds during a drought. Liver failure and hepatoencephalopathy were diagnosed, caused by pyrrolizidine alkaloid toxicosis associated with consumption of tansy ragwort. The horse made a full recovery when removed from the pasture.     

Subolesin/akirin orthologs from Ornithodoros spp. soft ticks: cloning, RNAi gene silencing and protective effect of the recombinant proteins

Subolesin/akirin is a well characterized protective antigen highly conserved across vector species and thus potentially useful for the development of a broad-spectrum vaccine for the control of arthropod infestations including hard ticks, mosquitoes, sand flies and the poultry red mite Dermanyssus gallinae. Soft ticks could be also targeted by this vaccine if proved that the soft tick subolesin orthologs are conserved and induce protective immune responses too. However, to date no soft tick subolesin orthologs have been fully characterized nor tested as recombinant antigens in vaccination trials. The objectives of the present work were to clone and characterize the subolesin orthologs from two important vector species of soft ticks as Ornithodoros erraticus and O. moubata, to evaluate the effect of subolesin gene silencing by RNAi, and to test the protective value of the recombinant antigens in vaccination trials. The obtained results demonstrate that both soft tick subolesins are highly conserved showing more than 69% and 74% identity with those of hard ticks in their nucleotide and amino acid sequences, respectively. Additionally, we demonstrate that both soft ticks possess fully operative RNAi machinery, and that subolesin gene silencing by dsRNA injection inhibits oviposition indicating the involvement of subolesin in tick reproduction. Finally, vaccination with the recombinant soft tick subolesins induced a partial protective effect resulting in the reduction of the oviposition rate. These preliminary results encourage further studies on the use of recombinant subolesins as vaccines for the control of soft tick infestations, either alone or in combination with other specific molecules.