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.     

Evaluation of Nitric Acid Extraction of Elements from Soils and Sediments in Two watersheds in East Tennessee

Determining the total elemental concentrations of soils requires a total dissolution method, in which hydrofluoric acid (HF) is commonly used. However, this method is tedious and risky due to the dangerous reagent HF. This study compared a single acid extraction technique using a nitric acid (HNO₃) extraction method to total dissolution (HF + microwave assisted aqua regia) for evaluating the total elemental concentrations in soils and sediments. The two methods were used to analyze the elemental content of soils and sediments from the Oostanaula Creek and Pond Creek watersheds in East Tennessee. Twenty-one elements (Al, Ba, Ca, Co, Cr, Cu, Fe, K, La, Li, Mg, Mn, Nd, P, S, Si, Sr, Ti, V, Zn, and Zr) were subsequently measured using inductively coupled plasma optical emission spectrometry (ICP-OES). The extraction efficiency for HNO₃ relative to the total was compared and discussed. In general, HNO₃ can extract the elements in absorbed phases and those residing in non-silicate minerals. Six elements (Ba, Co, Fe, Ca, Mn, and P) generated significant correlations between total and HNO₃ in both OC and PC watersheds. Finally, the elements were grouped according to the interrelationships of their total elemental concentrations based on the dendrogram plots, suggesting their geochemical association in soils and sediment forming minerals.     

Earthworm populations in septic system filter fields and potential effects on wastewater renovation

Wastewater renovation in septic-system filter fields can be affected by preferential flow through soil macropores. Anecic earthworm species make deep vertical burrows that may reduce renovation by acting as preferential flow paths that decrease effluent contact with the soil matrix. On the other hand, endogeic earthworms make largely horizontal burrows that may enhance wastewater renovation by distributing the effluent over a larger area. Additionally, the moist, nutrient-rich environment in filter fields may increase earthworm populations by enhancing their survival. Therefore, our objectives were to determine earthworm numbers and biomass with distance from soil treatment trenches, and identify species present to estimate potential effects on wastewater renovation. Five septic systems were investigated. At each site, earthworm populations were measured using formalin extraction at 10 locations along each of three 7-m long transects perpendicular to the trenches. There were an average of 6.4 times more earthworms and 5.4 times more earthworm biomass within 1 m of the trench than in the background (3.5–7.0 m from the trenches) in 13 of the 15 transects. This suggests that earthworms may have a significant effect on the movement of effluent. Because only epigeic and endogeic species were observed, the potential for reduced renovation and groundwater contamination at these sites is likely low. This may not be the case in areas with large numbers of anecic earthworms.     

What is your diagnosis? Peritoneal fluid from an Arabian horse after colic surgery

A 16-year-old castrated male Arabian horse was presented to the Purdue University Veterinary Teaching Hospital with a 4-hour history of colic. Initial examinations provided strong evidence for small intestinal obstruction. Abdominal surgery revealed a strangulating lipoma, and 25 feet of small intestine were resected. Postoperatively, the horse developed obstructive ileus due to adhesion formation, which required a second laparotomy. During and after surgery, the abdomen was lavaged with sodium carboxymethylcellulose (CMC). One week after the second surgery, evaluation of peritoneal fluid revealed an inflammatory exudate, with many macrophages containing amorphous to granular, pink to magenta phagocytosed material. Extracellular aggregates of the material were also observed. The material was consistent with CMC. To our knowledge, this report is the first to demonstrate the phagocytosis of CMC by peritoneal fluid macrophages.     

A novel herpesvirus associated with chronic superficial keratitis and proliferative conjunctivitis in a great horned owl (Bubo virginianus)

An adult great‐horned owl (Bubo virginianus; GHOW) presented with a history of recurrent corneal ulceration of the right eye (OD). Findings included ulcerative superficial keratitis, proliferative conjunctivitis, and iris pigmentary changes. The ulcer was initially nonresponsive to medical therapy, but showed rapid and appropriate healing following diamond burr debridement. Proliferative conjunctivitis markedly improved following topical antiviral therapy with cidofovir 1%, interferon alpha 2B ophthalmic solutions, and oral l ‐lysine. Histopathologic evaluation of a conjunctival biopsy revealed epithelial features suspicious for viral cytopathic changes and intranuclear structures suspicious for viral inclusions, suggestive of a possible viral‐induced papillomatous conjunctivitis. A novel alphaherpesvirus, referred to as Strigid Herpesvirus 1 (StrHV1), was identified using PCR and gene sequencing. This case represents a new clinical manifestation of a previously unreported herpesvirus in the GHOW. Identification of the herpes virus was critical to administration of appropriate therapy and resolution of the conjunctivitis, and corneal epithelial debridement promoted resolution of the chronic corneal epithelial defect.     

Phosphorus toxicity in the Proteaceae: A problem in post-agricultural lands

South African Proteaceae are adapted to the low soil phosphorus (P) concentrations of the Cape Floristic Region. The efficient P uptake by Proteaceae means that these plants experience phosphorus (P) toxicity at lower rhizosphere [P] than crop plants. This is only problematic when cultivating Proteaceae (and many plants from this region) on previously agricultural land with high residual soil [P]. In this study we hypothesize that P toxicity will result in element imbalances in leaves of Proteaceae and information from this study aims to facilitate ameliorative treatments. Phosphorus toxicity was induced on-farm in Leucadendron ‘Safari Sunset’ (Proteaceae) with subsequent mapping of element distribution in non-necrotic leaf tissue using micro particle-induced X-ray emission spectrometry. Phosphate supply up to 0.01 mM in a fertigation solution resulted in increased stem length of Leucadendron ‘Safari Sunset’ while P concentrations in excess of this resulted in decreased stem length, increased leaf [P] up to 0.25% (w/w) and, between 1 mM and 5 mM P supply, typical P toxicity symptoms were observed. High P supply (5 mM P) resulted in increased leaf [P] in most leaf tissues including the epidermis, where calculations from an equilibrium speciation model indicated that there was 30% more dissolved PO₄³⁻ in the epidermis compared to leaves at low P supply (0 mM added P on soil with 34 mg P kg⁻¹). Concomitantly, bundle sheath and epidermal [Ca] were reduced and 10% more Ca was predicted to be adsorbed and precipitated as hydrapatite at high P supply. High P supply resulted in increased leaf [Cl] and [Mn] in all tissues studied; decreased total leaf [Fe], bundle sheath, xylem, phloem and epidermal [Fe] and decreased total leaf [Zn] and xylem and phloem [Zn]. The observed symptoms of P toxicity in Leucadendron ‘Safari Sunset’ (necrosis in some plants, chlorosis and leaf rosetting) co-occurred with (1) excess PO₄³⁻, which may bind Ca in the epidermis (leading eventually to necrosis); (2) reduced [Fe] and increased [Mn] (leading to chlorosis) and (3) reduced total and vascular [Zn] (leading to leaf rosetting).     

Photoemission evidence for a remnant fermi surface and a d-wave-like dispersion in insulating Ca2CuO2Cl2

An angle-resolved photoemission study is reported on Ca2CuO2Cl2, a parent compound of high-Tc superconductors. Analysis of the electron occupation probability, n(k), from the spectra shows a steep drop in spectral intensity across a contour that is close to the Fermi surface predicted by the band calculation. This analysis reveals a Fermi surface remnant, even though Ca2CuO2Cl2 is a Mott insulator. The lowest energy peak exhibits a dispersion with approximately the &cjs3539;coskxa - coskya&cjs3539; form along this remnant Fermi surface. Together with the data from Dy-doped Bi2Sr2CaCu2O8+delta, these results suggest that this d-wave-like dispersion of the insulator is the underlying reason for the pseudo gap in the underdoped regime.     

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.