Study of nebulization delivery of aerosolized fluorescent microspheres to the avian respiratory tract

This study investigated the delivery of an aerosol of monodisperse microspheres to the respiratory tract of birds following aerosol exposure. Adult domestic pigeons (Columbia livia domestica, n = 5 birds per timed treatment) were exposed to an aerosol of fluorescent 1.0 microm diameter carboxylate microspheres for 0.5, 1, 2, or 4 hr. During the aerosolization period, the birds were free-standing in a plexiglass treatment chamber and the aerosol was delivered using a commercial nebulizer. Immediately following aerosol exposure, the birds were euthanatized and the carcasses were intravenously infused with a modified paraformaldehyde/gluteraldehyde fixative. Evaluation of microsphere distribution was performed using a stereoscopic microscope with an epifluorescent module. The results from this study revealed that the amount of aerosolized particles delivered using a commercial nebulizer was proportional to exposure periods. Aerosol exposure periods of 0.5 hr or 1 hr did not result in a readily observable distribution of 1.0 microm fluorescent microspheres to the cranial thoracic, caudal thoracic, or abdominal air sac membranes. This was partly attributed to the relatively low concentration of the individual monodisperse microspheres in the aerosolized suspension. The 2- and 4-hr exposure periods resulted in readily observable deposition of the 1.0 mirom fluorescent microspheres in the cranial thoracic, caudal thoracic, or abdominal air sac membranes, with the 4-hr exposure period resulting in the greatest number of particles on the membrane surfaces. For each of the exposure periods, there was individual animal variation regarding the distribution and relative number of spheres deposited. This study demonstrates the widespread deposition of particles that had an aerodynamic equivalent diameter of approximately 1 microm and provides a better understanding of particle deposition efficiency within the respiratory system following aerosol exposure in birds.     

The use of multilocus enzyme electrophoresis to characterise intestinal spirochaetes (Brachyspira spp.) colonising hens in commercial flocks

Multilocus enzyme electrophoresis (MLEE) was used to identify, examine genetic relationships and look at disease associations of a collection of 53 intestinal spirochaete isolates previously recovered from the faeces of adult hens on 14 farms in Qld, Australia. The MLEE results were compared with those previously obtained using species-specific PCR amplifications. The isolates were divided into five Brachyspira species groups by MLEE: Brachyspira murdochii (n=17), B. intermedia (n=15), B. pilosicoli (n=14), B. innocens (n=2) and "B. pulli" (n=1). Three new MLEE groups each containing single isolates also were identified. The results of the PCR assay for B. pilosicoli were concordant with the MLEE results, but the 23S rDNA-based PCR for B. intermedia had failed to detect 8 of the 15 isolates. The B. innocens/B. murdochii nox-based PCR had correctly identified all the isolates of B. murdochii, but did not identify either of the two B. innocens isolates. Using MLEE, isolates from two farms (14%) were identified as B. murdochii, whilst the pathogenic species B. intermedia and B. pilosicoli were present in hens from eight (57%) and five (36%) farms, respectively, and were identified together in four (29%) farms. All seven of the farms with production problems or wet litter were colonised with B. intermedia and/or B. pilosicoli. Six farms had multiple spirochaete isolates available for examination. Two broiler breeder farms both had five isolates of B. pilosicoli that shared the same MLEE electrophoretic type (ET), whilst one laying hen farm had three isolates of B. intermedia that all belonged to the same ET. Hence on each of these farms a predominant strain of a pathogenic species was present. On the other farms isolates of the same species were more diverse and belonged to different ETs. These results show that the epidemiology of intestinal spirochaetal infections in broiler breeder and laying hen flocks can vary considerably between farms, although the reasons for these differences were not established.     

Phenotypic characterisation of Australian sheep and cattle isolates of Mannheimia haemolytica,Mannheimia granulomatis and Mannheimia varigena

OBJECTIVE: To perform a comprehensive phenotypic characterisation of 35 isolates of bacteria previously identified as haemolytic Pasteurella-Actinobacillus and obtained from cattle and sheep. DESIGN: The 35 isolates that had been obtained from Australian animals, 30 from cattle and five from sheep, were compared with reference strains of the five recognised species of the genus Mannheimia--M. haemolytica, M. glucosida, M. granulomatis, M. ruminalis and M. varigena. RESULTS: Thirty-four of the isolates could be confidently assigned to three species of the genus Mannheimia. Twenty-nine were M. haemolytica, with 25 being isolated from cattle and four from sheep. All but three of the bovine M. haemolytica were isolated from pneumonic lungs. Of the three remaining bovine M. haemolytica isolates, one was obtained in pure culture from a bovine milk sample and the other two as part of a mixed flora associated with a middle ear infection of a calf suffering mucosal disease. Of the four ovine M. haemolytica isolates, two were isolated in pure culture from milk and two, also in pure culture, from pneumonic lungs. Three bovine isolates were identified as M. granulomatis--one from a tongue abscess, one from a jaw abscess and one from a lung showing suppurative bronchopneumonia. Two bovine isolates were identified as M. varigena--one coming from an udder and the other from a spleen. The available diagnostic records provided no information on whether these isolates were associated with a disease process. The remaining isolate was obtained from an ovine tongue abscess and could not be assigned to a recognised species within the genus Mannheimia. CONCLUSION: The study represents the first time that M. haemolytica, M. granulomatis and M. varigena have been recognised as being present in cattle and sheep in Australia. Veterinary laboratories that encounter Pasteurella-Actinobacillus-like organisms from cattle and sheep should attempt as complete a characterisation as possible to help improve our knowledge of the disease potential of these organsims.     

Histopathologic and bacteriologic evaluations of cellulitis detected in legs and caudal abdominal regions of turkeys

The objective of this study was to identify the causative agent of cellulitis in turkeys. Eighteen flocks from nine producers were sampled at the local processing plant, and 37 birds with cellulitis on legs or caudal thoracic area were obtained. None of the 37 birds with cellulitis had lesions in other organs. On gross examination, lesions were categorized into two groups: cellulitis with unopened skin lesions (type a) and cellulitis with opened skin lesions (type b). Histopathologically, cellulitis with unopened skin lesions had dermal necrosis with underlying fibrin and inflammatory exudate but cellulitis with open skin lesions had chronic granulomatous/granulation tissue-type reaction associated with foreign material. A complete bacteriologic study was conducted on 25 of 37 birds. Bacteria were isolated from 12 of the 25 birds with cellulitis lesions. No aerobic, microaerophilic, or anaerobic bacteria were isolated from the remaining 13 birds with cellulitis lesions. Escherichia coli was isolated in low numbers in mixed cultures with Proteus mirabilis, Lactobacillus spp., Klebsiella spp., and Staphylococcus spp. in 9 of 12 lesions. The remaining few cases yielded P. mirabilis in pure culture or in mixed culture with Pseudomonas aeruginosa. Types a and b cellulitis lesions in turkeys could be associated with primary contact dermatitis and skin abrasions, respectively. Their occurrence is likely associated with different management practices.     

Severity and persistence of footrot in Merino sheep experimentally infected with a protease thermostable strain of Dichelobacter nodosus at five sites

Objective To test the hypothesis that ovine footrot associated with a thermostable protease strain of Dichelobacter nodosus undergoes self cure or is sustained as an annually recurring disease, depending on the environment.
Design and procedure Forty Merino sheep from a single blood line were infected with a protease thermostable strain of D nodosus a t each of five sites in Western Australia. Footrot lesions and microscopic evidence of D nodosus were recorded every fortnight for 2.5 years, supplemented by laboratory culture. Rainfall, soil and air temperature, pasture quantity and composition and soil types were also recorded. Flocks that apparently self cured were relocated to a more favourable site for footrot in the final spring season.
Results The maximum prevalence of feet with clinical footrot lesions was 80.6, 1.3, 14.4, 3.8 and 88.1% at the five sites. Severe footrot occurred for three consecutive spring seasons at one site that had clay loam soil and at least 3500 kg/ha total pasture dry matter annually. However, the infection was asymptomatic for up to 10 weeks between outbreaks. D nodosus was isolated from flocks for 2.5 years at only two sites, although there was microscopic evidence of the organism at other sites in the final year. A thermolabile variant (strain U6) of D nodosus was isolated from the two sites where footrot persisted.
Conclusion Depending on time and location, ovine footrot induced by a protease thermostable strain of D nodosus either self cured or persisted as annual outbreaks interspersed with periods of asymptomatic infection.     

Acute cortisol responses of calves to four methods of dehorning by amputation

Objective To measure the plasma cortisol response in calves dehorned by four different methods (scoop, guillotine shears, saw, embryotomy wire) for 9 h after dehorning.
Design A physiological study with controls.
Procedure Horn amputation was carried out on calves restrained manually in a race.
Results The four methods of dehorning provoked similarly increased cortisol responses which lasted for 6 h. During the first hour after dehorning the plasma cortisol concentrations were similar to those following ACTH injection. The overall cortisol response to control handling was about 30% of the responses to dehorning.
Conclusions The similarity of the cortisol responses produced by the four methods of dehorning suggests that the distress experienced by calves following dehorning by amputation is similar regardless of method used.     

Effects of oral administration of sodium citrate or acetate to pigs on blood parameters, postmortem glycolysis, muscle pH decline, and quality attributes of pork

The objective of this study was to determine the effects of oral administration of sodium citrate (CIT) or acetate (ACE) to pigs on blood parameters, postmortem glycolysis, pH decline, and quality attributes of pork. Previous studies have shown that CIT has the potential to inhibit phosphofructokinase (PFK), a key enzyme in postmortem muscle glycolysis. In Exp. 1, CIT, ACE, or water was orally administered (0.75 g/kg of BW) to 24 pigs. After a 30-min rest, pigs were exercised, and blood samples were taken at 45 and 75 min after oral treatment. Citrate and ACE tended (P = 0.08) to increase blood pH and increased (P = 0.02) bicarbonate levels immediately after exercise. After a 30-min rest, blood pH of pigs administered ACE tended (P = 0.09) to remain higher, whereas blood pH of CIT-treated pigs was similar to that of control pigs. Bicarbonate levels in ACE- and CIT-treated pigs were still greater (P < 0.05) than those of control pigs at 75 min after oral treatment. In Exp. 2, 30 pigs were administered CIT, ACE, or water 45 min before stunning (electric plus captive bolt). Antemortem treatments had no effect (P > 0.10) on muscle pH or postmortem concentrations of the glycolytic metabolites of glucose-6 phosphate, fructose-6 phosphate, fructose-1,6 bisphosphate, glyceraldehyde-3 phosphate, dihydroxyacetone phosphate, or lactate. Minor, but inconsistent, differences in quality attributes were found in LM chops, and no differences in quality attributes were found between control and CIT- or ACE-treated pigs for inside and outside semimembranosus muscles (P > 0.10). There was no significant inhibition of the PFK enzyme by orally administered CIT or ACE; however, the PFK glycolytic metabolite data analysis indicated that PFK was a main regulatory enzyme in postmortem muscle.     

Spatial patterns of large natural fires in Sierra Nevada wilderness areas

The effects of fire on vegetation vary based on the properties and amount of existing biomass (or fuel) in a forest stand, weather conditions, and topography. Identifying controls over the spatial patterning of fire-induced vegetation change, or fire severity, is critical in understanding fire as a landscape scale process. We use gridded estimates of fire severity, derived from Landsat ETM+ imagery, to identify the biotic and abiotic factors contributing to the observed spatial patterns of fire severity in two large natural fires. Regression tree analysis indicates the importance of weather, topography, and vegetation variables in explaining fire severity patterns between the two fires. Relative humidity explained the highest proportion of total sum of squares throughout the Hoover fire (Yosemite National Park, 2001). The lowest fire severity corresponded with increased relative humidity. For the Williams fire (Sequoia/Kings Canyon National Parks, 2003) dominant vegetation type explains the highest proportion of sum of squares. Dominant vegetation was also important in determining fire severity throughout the Hoover fire. In both fires, forest stands that were dominated by lodgepole pine (Pinus contorta) burned at highest severity, while red fir (Abies magnifica) stands corresponded with the lowest fire severities. There was evidence in both fires that lower wind speed corresponded with higher fire severity, although the highest fire severity in the Williams fire occurred during increased wind speed. Additionally, in the vegetation types that were associated with lower severity, burn severity was lowest when the time since last fire was fewer than 11 and 17 years for the Williams and Hoover fires, respectively. Based on the factors and patterns identified, managers can anticipate the effects of management ignited and naturally ignited fires at the forest stand and the landscape levels.     

Effect of Insemination–Ovulation Interval and Addition of Seminal Plasma on Sow Fertility to Insemination of Cryopreserved Sperm

In swine, the use of frozen-thawed (FT) sperm for artificial insemination (AI) is limited because of poor sow fertility, possibly associated with a post-thaw capacitation-like status resulting in fewer fully viable sperm. Sow fertility to AI with FT sperm may improve with deeper deposition of sperm within the female tract, insemination very close to ovulation, or reversal of cryocapacitation by seminal plasma (SP). We performed two experiments to examine these suggestions. In experiment 1, 122 multiparous Yorkshire sows received 600 IU equine chorionic gonadotrophin at weaning and 5 mg pLH 80 h later to control time of ovulation. The predicted time of ovulation (PTO) was 38 h after pLH injection. Thereafter, sows were assigned on the basis of parity to a single AI of FT sperm at 2 h before PTO, or at 12 h before PTO, or FT sperm supplemented with 10% SP at 12 h before PTO. Control sows received fresh semen at 12 h before PTO. All semen doses were adjusted to 3 x 10(9) live cells and deposited into the cervix. Experiment 2 employed 99 multiparous crossbred sows and repeated the treatments of experiment 1 except that all FT inseminations were intrauterine. In both experiments, farrowing rates were lower (p < 0.01) following FT inseminations with no effect of time of insemination or of supplemental SP. In experiment 1, litter size was smaller following FT insemination (p < 0.05), but no effect on litter size was evident in experiment 2. Supplemental SP had no effect on litter size in either experiment. The lack of effect of either SP or timing of FT insemination on sow fertility suggests that the non-lethal sperm cryoinjury affecting fertility involves more than just cryocapacitation.