Oral vaccination of brushtail possums with BCG: Investigation into factors that may influence vaccine efficacy and determination of duration of protection

AIMS: To determine factors that may influence the efficacy of an oral pelleted vaccine containing Mycobacterium bovis bacille Calmette-Guérin (BCG) to induce protection of brushtail possums against tuberculosis. To determine the duration of protective immunity following oral administration of BCG.

METHODS: In Study 1, a group of possums (n=7) was immunised by feeding 10 pellets containing dead Pasteur BCG, followed 15 weeks later with a single pellet of live Pasteur BCG. At that time, four other groups of possums (n=7 per group) were given a single pellet of live Pasteur BCG orally, a single pellet of live Danish BCG orally, 10 pellets of live Pasteur BCG orally, or a subcutaneous injection of live Pasteur BCG. For the oral pelleted vaccines, BCG was formulated into a lipid matrix, and each pellet contained approximately 10⁷ colony forming units (cfu) of BCG, while the vaccine injected subcutaneously contained 10⁶ cfu of BCG. A sixth, non-vaccinated, group (n=7) served as a control. All possums were challenged by the aerosol route with a low dose of virulent M. bovis 7 weeks after vaccination, and killed 7–8 weeks after challenge. Protection against challenge with M. bovis was assessed from pathological and bacteriological findings.

In Study 2, lipid-formulated live Danish BCG was administered orally to three groups of possums (10–11 per group), and these possums were challenged with virulent M. bovis 8, 29 or 54 weeks later. The possums were killed 7 weeks after challenge, to assess protection in comparison to a non-vaccinated group.

RESULTS: The results from Study 1 showed that vaccine efficacy was not adversely affected by feeding dead BCG prior to live BCG. Feeding 10 vaccine pellets induced a level of protection similar to feeding a single pellet. Protection was similar when feeding possums a single pellet containing the Pasteur or Danish strains of BCG. All vaccinated groups had significantly reduced pathological changes or bacterial counts when compared to the non-vaccinated group. In Study 2, oral administration of Danish BCG induced protection against challenge with M. bovis, which persisted for at least 54 weeks after vaccination. Some protection was observed in possums challenged 54 weeks after vaccination, but this protection was significantly less than that observed in groups vaccinated 29 or 8 weeks prior to challenge. There was a strong relationship between the proportion of animals producing positive lymphocyte proliferation responses to M. bovis antigens and protection against challenge with M. bovis.

CONCLUSIONS: Factors considered potentially capable of interfering with vaccination, including feeding dead BCG to possums prior to feeding live BCG, feeding multiple doses of BCG at one time, and changing strains of BCG, were shown not to interfere with the acquisition of protective immune responses in possums. Protection against tuberculosis was undiminished up to 29 weeks after vaccination with BCG administered orally. It is concluded that vaccination of possums by feeding pellets containing BCG is a robust and efficient approach to enhance the resistance of these animals to tuberculosis.     

Bartonella species detection in captive, stranded and free-ranging cetaceans

We present prevalence of Bartonella spp. for multiple cohorts of wild and captive cetaceans. One hundred and six cetaceans including 86 bottlenose dolphins (71 free-ranging, 14 captive in a facility with a dolphin experiencing debility of unknown origin, 1 stranded), 11 striped dolphins, 4 harbor porpoises, 3 Risso's dolphins, 1 dwarf sperm whale and 1 pygmy sperm whale (all stranded) were sampled. Whole blood (n = 95 live animals) and tissues (n = 15 freshly dead animals) were screened by PCR (n = 106 animals), PCR of enrichment cultures (n = 50 animals), and subcultures (n = 50 animals). Bartonella spp. were detected from 17 cetaceans, including 12 by direct extraction PCR of blood or tissues, 6 by PCR of enrichment cultures, and 4 by subculture isolation. Bartonella spp. were more commonly detected from the captive (6/14, 43%) than from free-ranging (2/71, 2.8%) bottlenose dolphins, and were commonly detected from the stranded animals (9/21, 43%; 3/11 striped dolphins, 3/4 harbor porpoises, 2/3 Risso's dolphins, 1/1 pygmy sperm whale, 0/1 dwarf sperm whale, 0/1 bottlenose dolphin). Sequencing identified a Bartonella spp. most similar to B. henselae San Antonio 2 in eight cases (4 bottlenose dolphins, 2 striped dolphins, 2 harbor porpoises), B. henselae Houston 1 in three cases (2 Risso's dolphins, 1 harbor porpoise), and untyped in six cases (4 bottlenose dolphins, 1 striped dolphin, 1 pygmy sperm whale). Although disease causation has not been established, Bartonella species were detected more commonly from cetaceans that were overtly debilitated or were cohabiting in captivity with a debilitated animal than from free-ranging animals. The detection of Bartonella spp. from cetaceans may be of pathophysiological concern.     

Cardiac troponins

Objective: To review the use of cardiac troponins as biomarkers for myocardial injury in human and veterinary medicine. Data sources: Data sources included scientific reviews and original research publications. Human data synthesis: Cardiac troponins have been extensively studied in human medicine. Finding an elevated cardiac troponin level carries important diagnostic and prognostic information for humans with cardiovascular disease. Troponin assays are used primarily to diagnose acute myocardial infarction in patients with ischemic symptoms such as chest pain. However, elevated blood levels may be found with any cause of myocardial injury. Veterinary data synthesis: Several studies have shown that cardiac troponins are sensitive and specific for myocardial damage in veterinary patients and may have utility in diagnosis and prognosis for certain disease states. Human assays may be used in most animals due to significant homology in the troponin proteins between species. Conclusions: Cardiac troponins are sensitive and specific markers of myocardial injury although they do not give any information regarding the mechanism of injury. They have redefined how acute myocardial infarction is diagnosed in humans. Their use in the clinical management of veterinary patients is limited at this time. Further prospective studies are warranted.     

Sucrose assists selection of high‐quality oocytes in pigs

We investigated whether high‐quality in vitro matured (IVM) oocytes can be distinguished from poor ones based on the morphological changes after treatment with hyperosmotic medium containing 0.2 mol/L sucrose in pigs. We hypothesize that IVM oocytes maintaining round shape have higher quality than mis‐shapened oocytes following dehydration. Oocyte quality was verified by determining embryonic developmental competence using in vitro fertilization, nuclear transfer and parthenogenetic activation. In all cases, the round oocytes had greater (p < .05) developmental competence than that of mis‐shapened oocytes in terms of blastocyst rate and total cell number in blastocysts obtained after 6 days of in vitro culture. We also confirm that round aged oocytes are higher in quality than mis‐shapened aged oocytes. In an attempt to find out why high‐quality oocytes maintain a round shape whereas poorer oocytes become mis‐shapened following sucrose treatment, we examined the arrangement of actin microfilaments and microtubules. Abnormal organization of these cytoskeletal components was higher (p < .05) in mis‐shapened oocytes compared to round oocytes after 52 hr of IVM. In conclusion, sucrose treatment helps selection of high‐quality oocytes, including aged oocytes, in pigs. Abnormal cytoskeleton arrangements partly explain for low developmental competence of mis‐shapened oocytes.     

In search of the first flower: A jurassic angiosperm, archaefructus, from northeast china

Angiosperm fruiting axes were discovered from the Upper Jurassic of China. Angiosperms are defined by carpels enclosing ovules, a character demonstrated in this fossil. This feature is lacking in other fossils reported to be earliest angiosperms. The fruits are small follicles formed from conduplicate carpels helically arranged. Adaxial elongate stigmatic crests are conspicuous on each carpel. The basal one-third of the axes bore deciduous organs of uncertain affinities. No scars of subtending floral organs are present to define the individual fertile parts as floral units, but the leaf-like structures subtending each axis define them as flowers. These fruiting axes have primitive characters and characters not considered primitive.     

Forest fires: An example of self-organized critical behavior

Despite the many complexities concerning their initiation and propagation, forest fires exhibit power-law frequency-area statistics over many orders of magnitude. A simple forest fire model, which is an example of self-organized criticality, exhibits similar behavior. One practical implication of this result is that the frequency-area distribution of small and medium fires can be used to quantify the risk of large fires, as is routinely done for earthquakes.     

SPRINKMOD – pressure and discharge simulation model for pressurized irrigation systems. 3. Sensitivity to lateral hydraulic parameters and leakage

A computer model, SPRINKMOD, was tested with field data collected from a simple sprinkler irrigation system in operation. The original data for laterals and the amount and way leakage is considered were modified to evaluate the model sensitivity. The model predicted the pump station flow rate within 2% and the pump station pressure head within 5% with the original data collected. For this irrigation system, no practical effect was observed in the system operating point by changing the lateral pipe characteristics, lateral leakage amount and distribution, lateral pipe roughness and lateral local loss coefficient for flow past a riser outlet. The amount of leakage had more effect on the model simulation of pressure heads than the way leakage was considered, localized or distributed along the laterals. The use of a variable local loss coefficient, K _r, along the 350-m hand-move laterals had a negligible effect on both system operating point and distribution of pressure heads along the laterals. Received: 14 February 1998     

Compensation of horizontal temperature and salinity gradients in the ocean mixed layer

Establishment of the temperature-salinity relationship in the ocean has concerned oceanographers for decades because of its importance for understanding ocean circulation. High-resolution measurements in the ocean mixed layer are used to show that temperature and salinity gradients on horizontal scales of 20 meters to 10 kilometers tend to compensate in their effect on density. These observations support the notion of a horizontal mixing in the mixed layer that depends on density gradient.