Molecular characterization of the uncultivatable hemotropic bacterium Mycoplasma haemofelis

ABSTRACT: Mycoplasma haemofelis is a pathogenic feline hemoplasma. Despite its importance, little is known about its metabolic pathways or mechanism of pathogenicity due to it being uncultivatable. The recently sequenced M. haemofelis str. Langford 1 genome was analysed and compared to those of other available hemoplasma genomes.Analysis showed that in hemoplasmas genes involved in carbohydrate metabolism are limited to enzymes of the glycolytic pathway, with glucose appearing to be the sole energy source. The majority of the pentose phosphate pathway enzymes that catalyze the de novo synthesis of ribonucleotides were absent, as were cell division protein FtsZ and chaperonins GroEL/ES. Uncharacterized protein paralogs containing putative surface expression motifs, comprised 62% of M. haemofelis and 19% of Mycoplasma suis genome coverage respectively, the majority of which were present in a small number of unstructured islands. Limited mass spectrometry and immunoblot data matched a number of characterized proteins and uncharacterized paralogs, confirming their expression and immunogenicity in vivo.These data have allowed further characterization of these important pathogens, including their limited metabolic capabilities, which may contribute to their uncultivatable status. A number of immunogenic proteins, and a potential mechanism for host immune system evasion, have been identified.     

Experimental infection of hamsters with avian paramyxovirus serotypes 1 to 9

ABSTRACT: Avian paramyxoviruses (APMVs) are frequently isolated from domestic and wild birds throughout the world and are separated into nine serotypes (APMV-1 to -9). Only in the case of APMV-1, the infection of non-avian species has been investigated. The APMVs presently are being considered as human vaccine vectors. In this study, we evaluated the replication and pathogenicity of all nine APMV serotypes in hamsters. The hamsters were inoculated intranasally with each virus and monitored for clinical disease, pathology, histopathology, virus replication, and seroconversion. On the basis of one or more of these criteria, each of the APMV serotypes was found to replicate in hamsters. The APMVs produced mild or inapparent clinical signs in hamsters except for APMV-9, which produced moderate disease. Gross lesions were observed over the pulmonary surface of hamsters infected with APMV-2 & -3, which showed petechial and ecchymotic hemorrhages, respectively. Replication of all of the APMVs except APMV-5 was confirmed in the nasal turbinates and lungs, indicating a tropism for the respiratory tract. Histologically, the infection resulted in lung lesions consistent with bronchointerstitial pneumonia of varying severity and nasal turbinates with blunting or loss of cilia of the epithelium lining the nasal septa. The majority of APMV-infected hamsters exhibited transient histological lesions that self resolved by 14 days post infection (dpi). All of the hamsters infected with the APMVs produced serotype-specific HI or neutralizing antibodies, confirming virus replication. Taken together, these results demonstrate that all nine known APMV serotypes are capable of replicating in hamsters with minimal disease and pathology.     

Biosecurity and vaccination strategies to minimise the effect of an equine influenza outbreak on racing and breeding

Three biosecurity and relief-and-recovery initiatives adopted by the NSW horse racing industries reduced the economic and social disruption caused by the disease and subsequent movement controls during the 2007 Australian equine influenza (EI) incursion. The first was the creation of biosecure horse training and racing precincts around the Sydney area to permit racing to continue with healthy horses. Infection was excluded for 3-5 weeks and race meetings were conducted safely during this period. The second was a vaccination program of racehorses at these and other precincts to maintain an ongoing healthy pool of racehorses. Vaccination commenced too late to enable viable racing to continue in Sydney in the short term, but assisted in managing an early return to racing throughout the state before EI-free status had been regained. The third was the establishment of approved quarantine stations to facilitate the movement of racing and breeding horses out of high-risk regions. The difficulties in establishing and managing these initiatives in the face of the EI incursion are discussed.     

Chemical composition of growing pigs and its relationship with body tissue composition assessed by X-ray-computed tomography

Ninety hybrid (mainly Large White × Landrace) pigs from 2 experimental replicates were used to study the potential use of computed tomography (CT) as a nondestructive technology for estimating the chemical body composition of growing pigs. Body tissue components (lean, fat, and bone) of 6 live pigs from each sex (boars, gilts, and barrows) were assessed by CT imaging before slaughter at approximately 30, 60, 90, 120, and 150 kg of BW. After slaughter, the empty body components were ground and frozen until analyzed for protein, lipid, ash, and moisture content. Several growth functions were evaluated and the allometric function (Y = aBW(b)), which was evaluated as log(10)chemical component weight = b(0) + b(1)log(10)BW, provided the best fit to the data. For each sex, the allometric coefficient (b(1)) for protein (0.92 to 0.99) was close to but less than 1; for ash (1.03 to 1.12), it was close to but greater than 1; for moisture (0.82 to 0.86), it was less than 1, and for lipid (1.61 to 1.71), it was greater than 1. Deposition rates (change in component weight per unit change in BW) for each chemical component were predicted using derivatives of the function. The mean deposition rates for protein and lipid were 0.141 and 0.286 kg/kg of BW gain, respectively. The deposition rate for protein was generally stable across different BW, whereas that for lipid increased as BW increased. In addition, linear, quadratic, exponential, and logistic functions were fitted to the data to study the relationship between the CT data and chemical components. The linear function was assessed to be the best equation, based on the Bayesian information criterion. The prediction equation for protein (kg) = -1.64 + 0.28 × CT lean (kg), and for lipid (kg) = -0.69 + 1.09 × CT fat (kg), had R(2) values of 0.924 and 0.987, respectively. Sex had no effect (P > 0.05) on the prediction of protein and lipid. The effect of BW was not significant (P > 0.05) for the prediction of lipid, but it was significant (P > 0 0.05) for the prediction of protein. However, the addition of BW to the base prediction equation for protein resulted in an increase of only 0.013 in the R(2) value. It was concluded from this study that CT scanning has great potential as a nondestructive technology for estimating the physical and chemical body composition of pigs. Additional research is required to validate the utility and accuracy of the prediction equations.     

Response of three cereal crops in continuous arable or ley-arable rotations to fertilizer nitrogen and soil nitrogen at Rothamsted's Woburn Ley-arable experiment

Rothamsted's Woburn Ley-arable experiment, started in 1938 on a sandy loam soil, provides valuable real-world data on the effects of all-arable and ley-arable rotations. In this study, six rotations were compared from 1973 to 2001. Two had 3-year arable “treatment” crops, two had 3-year leys, and two had 8-year leys; the leys being all-grass given fertilizer nitrogen (Ln3 and Ln8), or grass/clover (Lc3 and Lc8). Here, we present the yields of two test crops, winter wheat (1981–2000) followed by spring barley (1982–1991) or winter rye (1997–2001) in each of the six rotations, and their response to four rates of fertilizer N and soil N. From fitted yield/N response curves, we show that maximum wheat yields were least (7.10 t ha⁻¹) in the AB rotation, slightly higher, but not significantly so (7.65 t ha⁻¹) following Ln leys but significantly higher (8.12 t ha⁻¹) following Lc leys. Significantly less fertilizer N (30 kg ha⁻¹) was needed to achieve the higher yields following Lc leys. Yields of the second cereal following the leys were 0.3–0.8 t ha⁻¹ higher than those in the AB rotation; these increases were not statistically significant. However, significantly less fertilizer N, 26–38 kg N ha⁻¹, was required to achieve those yields. There was no difference found between the type of ley. The initial benefit of the Lc leys was short-lived. If leys are to be introduced into mainly arable farming systems, they may need to be subsidized to make them financially viable.     

Modelling long‐term carbon dynamics in soils reconstituted with large quantities of organic matter

In urban conditions, the widescale availability of organic matter to be recycled and the necessity for soil reconstitution (Technosol) has led to the input of very large quantities of organic matter (up to 50% v/v). The long‐term degradation of these large quantities of organic matter in the soil is not well known. We monitored, over a 60‐month period, the total carbon (C) content and the particulate and biochemical fractions of reconstituted soils placed in 600‐litre boxes under natural conditions. The top layer was a sandy loam amended with 20 or 40% of sphagnum peat or organic compost (sewage sludge, wood chip compost or green waste compost) lying on a layer of sandy loam. We measured C mineralization over time under controlled conditions and built a long‐term model to simulate carbon dynamics where exogenous organic carbon (EOC) was divided into two biodegradable compartments. The model yielded the proportions of EOC that either resisted degradation or contributed to soil organic C storage by mineralization and/or humification. Organic matter degradation was linked to its maturity and to its contents in certain particulate and biochemical fractions but was independent of how much of a given organic matter was introduced. We found a good correlation between the degradable organic compartment and the lignin and cutin‐like, hemicellulose and cellulose‐like fractions larger than 1 mm. The model showed that a large part of initial EOC was still present in the soil after 5 years in a potentially biodegradable but resistant compartment. The degradation of that compartment by mineralization or humification is therefore expected to take longer.     

Apoptotic effects of tamoxifen on leukocytes from horse peripheral blood and bronchoalveolar lavage fluid

A reduction in inflammatory cell apoptosis is an important concept in the maintenance of inflammation and a potential target for the resolution of inflammation in many inflammatory diseases. Dysregulation of apoptosis has been implicated in a range of diseases, including tumors, neurodegenerative disorders and autoimmunity, and may also be implicated in allergic asthma. In horses, recurrent airway obstruction (RAO) is an asthma-like condition that is characterized increased survival neutrophil bronchial. Tamoxifen is a synthetic, non-steroidal, anti-estrogen agent that is widely used for treating all stages of breast cancer and has been approved for the prevention of breast cancer in high-risk women. The observed efficacy of tamoxifen has been attributed to both growth arrest and the induction of apoptosis. Therefore, the aim of our study was to evaluate the ability of tamoxifen to induce apoptosis in vitro in granulocytic cells from peripheral blood and in mononuclear cells from bronchoalveolar lavage fluid (BALF) in horses. Flow cytometry using commercial AnnexinV-FITC and propidium iodide was used to quantify early and late apoptotic leukocytes, respectively. The results showed a significant increase in early apoptosis in peripheral blood and bronchial granulocytic cells treated with tamoxifen. The rate of early apoptosis of mononuclear cells from blood and BALF when incubated with tamoxifen was significantly lower compared with granulocytic cells. We did not observe a direct effect of tamoxifen on late apoptosis in any of the in vitro assays in the cell types used here. These results indicate that the apoptotic mechanisms under these experimental conditions would affect only blood and BALF granulocytic cells, particularly in early apoptosis. Finally, further in vitro and in vivo studies are needed to better understand apoptotic mechanisms because tamoxifen could be used to treat chronic, inflammatory pathologies associated with granulocytes and allergic diseases, such as asthma or equine RAO.     

Conventionalization of the organic sesame network from Burkina Faso: shrinking into mainstream

This research examines the structure and development of the organic sesame network from Burkina Faso to explain the declining trend in organic sesame export. The paper addresses particularly the question whether the organic sesame network is structurally (re)shaped as a conventional mainstream market or whether it still presents a real alternative to conventional sesame production and trade. It is found that over the last decade organic sesame is increasingly incorporated into mainstream market channels. But contrary to the well-known case of conventionalization in California, where organic agriculture grew into mainstream agro-food arrangements, this study illustrates a case where organic sesame agriculture shrank into mainstream agro-food arrangements. The weak coherence between the production and marketing nodes in the organic sesame chain resulted in failures to vertically mediate information, balance power relationships in and across sesame chains, build trust, and limit price volatility and speculation, resulting in a shrinking organic sesame market. For developing a viable alternative to conventional sesame trading, relations between production and trading nodes in the organic networks need to be strengthened through public–private partnerships, combined with other public and legal reinforcement.