Development of a continuous cell line from larval Atlantic cod (Gadus morhua) and its use in the study of the microsporidian,Loma morhua

In vitro cell culture methods are crucial for the isolation, purification and mass propagation of intracellular pathogens of aquatic organisms. Cell culture infection models can yield insights into infection mechanisms, aid in developing methods for disease mitigation and prevention, and inform commercial‐scale cultivation approaches. This study details the establishment of a larval cell line (GML‐5) from the Atlantic cod (Gadus morhua) and its use in the study of microsporidia. GML‐5 has survived over 100 passages in 8 years of culture. The line remains active and viable between 8 and 21°C in Leibovitz‐15 (L‐15) media with 10% foetal bovine serum and exhibits a myofibroblast phenotype as indicated by immuno‐positive results for vimentin, α‐smooth muscle actin, collagen I and S‐100 proteins, while being desmin‐negative. GML‐5 supports the infection and development of two microsporidian parasites, an opportunistic generalist (Anncaliia algerae) and cod‐specific Loma morhua. Using GML‐5, spore germination and proliferation of L. morhua was found to require exposure to basic pH and cool incubation temperatures (8°C), in contrast to A. algerae, which required no cultural modifications. Loma morhua‐associated xenoma‐like structures were observed 2 weeks postexposure. This in vitro infection model may serve as a valuable tool for cod parasitology and aquaculture research.     

Starving the soil of plant inputs for 50 years reduces abundance but not diversity of soil bacterial communities

If soil communities rely on plant-derived carbon, is biodiversity lost when this primary source is removed? Soil microbial and mesofaunal communities at the Rothamsted Highfield site were compared under a mixed grass sward, arable rotation and a section maintained as a bare-fallow for the past 50 years by regular tillage. Organic matter reserves have been degraded and microbial and mesofaunal numbers and mite diversity have declined in this unique bare-fallow site, where fresh carbon inputs have been drastically reduced. However, it supports a species-rich metabolically active bacterial community of similar diversity to that in soil maintained as grass sward. Thus in contrast to soil mesofauna, bacterial diversity (but not abundance) is apparently independent of plant inputs.     

Impact of Macrotermes termitaria as a source of heterogeneity on tree diversity and structure in a Sudanian savannah under controlled grazing and annual prescribed fire (Burkina Faso)

Macrotermes termitaria are conspicuous features of savannah ecosystems in the Sudanian and Sahelian zones of West Africa. The mounds, alive or abandoned, are a major source of heterogeneity in the landscape. The purpose of the present study was to assess the impact of termitaria on tree community in a state forest of the Sudanian regional centre (Tiogo forest, Burkina Faso), under controlled burning and grazing experiments. A comparative inventory was carried out in a split-plot experiment (16 subplots of 2500 m²): 8 subplots where fire regime and grazing were controlled and 8 subplots exposed to grazing and with annual prescribed fire since 1992. All tree individuals (≥1.5 m) were recorded, both on termitaria and outside and their basal area at stump level was measured. A total of 61 observed (or 65.7 ± 2.4 estimated) tree species were recorded on 28 Macrotermes subhyalinus mounds (54 observed species or 60.8 ± 3.3 estimated), the immediate surroundings (44 observed and 59.0 ± 0.0 estimated species) and the rest of subplots (56 observed and 63.6 ± 0.0 estimated). Specific density was higher on mounds in comparison with the surroundings (P < 0.05). Results showed that termitaria played a key role in maintaining higher species diversity as compared to their surroundings (P < 0.05). Differences in species diversity between termitaria and immediate surroundings appeared more pronounced in disturbed plots (submitted to both fire and grazing). Some species, such as Tamarindus indica, Boscia senegalensis, Cadaba farinosa, Capparis sepiaria and Maerua angolensis are found solely on termitaria. Besides, the density of trees was significantly higher on termitaria compared to the surrounding (P < 0.05), as well as total basal area per unit of 100 m² area (P < 0.05). We concluded that Macrotermes termitaria play an important role as a source of heterogeneity in this Sudanian savannah woodland ecosystem. This role is particularly important in ecosystems under stresses. Termitaria acted as refuge for tree vegetation. The density and dynamics of M. subhyalinus termitaria should, therefore, be taken into account in the global strategy of the forest resources management and conservation.     

Influence of temperature on seed germination in papaya under subtropical conditions of India

The effect of temperature on germination and days taken for germination was evaluated in Carica papaya cvs., ‘Pune selection-3’ and ‘Pusa Dwarf’. The highest average seed germination per cent was observed in the month of July (96.8 and 93.0?%), followed by August (94 and 88.5?%), whereas the lowest average seed germination was observed in the month of January (44.2 and 36.8?%) in ‘Pune selection-3’ and ‘Pusa Dwarf’, respectively. Minimum number of days (5.17 and 6.33) was observed for germination in the month of July, whereas the maximum number of days (29.5 and 31) was taken in the month of January in both the cultivars, respectively. The average highest germination and seedling height was observed at temperature 29.8 °C and 29.5 °C during July and August, respectively, whereas the lowest was observed at temperature 14 °C and 15.9 °C during January and December, respectively. The optimum time for nursery sowing was July to August for autumn season crop under subtropical conditions of India.     

Experimental Corynebacterium pseudotuberculosis mastitis in dairy cows

The inoculation of 2000 colony-forming units of Corynebacterium pseudotuberculosis into one teat canal of each of three cows resulted in severe, chronic, pyogranulomatous mastitis. Within three days the cows had a reduced haematocrit, haemoglobin concentration and red cell count. The anaemia was initially normocytic, normochromic and non-regenerative, and was associated with a brief peak of neutrophilia; a regenerative response became evident two to three weeks later. Clinical signs of mastitis appeared seven to 14 days after the inoculation, with a peak of high fever, more severe anaemia, a second peak of neutrophilia and the complete cessation of milk production from all quarters; extensive and severe pyogranulomatous mastitis developed in the inoculated quarters. No other lesions were detected postmortem, and C pseudotuberculosis was cultured from the affected quarters but not from the supramammary lymph nodes and viscera.     

Staphylococcus aureus exosecretions and bovine mastitis

The role of Staphylococcus aureus and coagulase-negative staphylococcal exosecretions in bovine udder infection was tested by monitoring the cows' response to in vivo inoculation of bacterial exosecretions into udder quarters. Twenty Israeli-Holstein dairy cows were included in the study; two or three of the udder quarters of each cow were intracisternally inoculated with 0.04-0.05 mg/quarter (total proteins) of the various sterile bacterial exosecretions in a sterile pyrogen-free saline. Each udder was inoculated with two or three different bacterial exosecretions or placebo (Columbia Broth). Cows were monitored for 96 h post-inoculation for rectal temperature, heart and respiratory rates, alimentary tract activity (rumen contraction), udder temperature, pain, oedema and udder size. Milk samples were examined bacteriologically and for somatic cell count, N-acetyl-D-glucosaminidase (NAGase) activity and somatic cell differentiation. No enterotoxins (beta-G) or toxic shock syndrome toxin-1 were detected in response to any of the bacteria tested. Control quarters or those inoculated with Columbia Broth, showed similar NAGase and somatic cell count values throughout the experiment. Twelve of the 18 strains tested, induced inflammation in the inoculated quarters while six did not. Of the 12 strains causing local inflammation, only six were found significantly different from the control and were considered as high response (group 1). The other six that caused a local inflammation did not differ significantly from the control, and were considered to be moderate response (group 2). The six S. aureus isolates that did not cause an inflammatory response were considered to have low response (group 3). In all quarters inoculated with S. aureus bacterial exosecretions belonging to groups 1 and 2, the polymorphonuclear cells and macrophages were proportionally increased while CD4+ and CD8+ T-lymphocyte populations decreased. One-dimensional NuPAGE (7%) Tris-acetate gel electrophoresisof the bacterial exosecretions revealed four different bands appearing between 36 and 31 kDa, marked from top to bottom as A, B, C and D. An association was found between the combinations of expressed bands and the cow responses: the majority of the cases could be linked to the expression of bands B and C.     

Milk leucocyte populations in heifers free of udder infection

Improvement of udder health through a process of genetic selection is related to heritability and the role of the specific trait in the probability of an individual cow developing an infection. It was suggested that different patterns of leucocyte population of the healthy gland are a significant factor in mastitis. Thus, in order to analyse the heritability of a trait and its correlation with udder health, the present study examined the leucocyte populations of uninfected mammary glands, their variability among quarters in a particular cow, and the changes that occur during lactation. Each one of the 20 cows examined was tested on average 3.06 times during lactation. The somatic cell count (SCC)/ml ranged from 12,000 to 151,000, the coefficients of determination (R2) were higher than 0.5 for SCC. No significant differences were found in the dependent variables between the sampled times (test) nor any interaction between the slopes calculated for the cows over time. No significant differences were found among quarters within a cow for any of the dependent variables including SCC. The effect of the cow trait was found to be significant for polymorphonuclear (PMN), macrophage (MO), and T-lymphocyte-bearing CD4+. The number of lymphocytes labelled with the anti-B monoclonal antibodies was negligible. In conclusion the patterns of leucocyte populations in milk together with the variance among cows should enable an analysis of the heritability of this trait and its correlation with udder health in a future study.     

Comment on "Fire-derived charcoal causes loss of forest humus"

Wardle et al. (Brevia, 2 May 2008, p. 629) reported that fire-derived charcoal can promote loss of forest humus and belowground carbon (C). However, C loss from charcoal-humus mixtures can be explained not only by accelerated loss of humus but also by loss of charcoal. It is also unclear whether such loss is related to mineralization to carbon dioxide or to physical export.     

The priming potential of biochar products in relation to labile carbon contents and soil organic matter status

Recognition of biochar as a potential tool for long-term carbon sequestration with additional agronomic benefits is growing. However, the functionality of biochar in soil and the response of soils to biochar inputs are poorly understood. It has been suggested, for example, that biochar additions to soils could prime for the loss of native organic carbon, undermining its sequestration potential. This work examines the priming potential of biochar in the context of its own labile fraction and procedures for their assessment. A systematic set of biochar samples produced from C4 plant biomass under a range of pyrolysis process conditions were incubated in a C3 soil at three discrete levels of organic matter status (a result of contrasting long-term land management on a single site). The biochar samples were characterised for labile carbon content ex-situ and then added to each soil. Priming potential was determined by a comparison of CO₂ flux rates and its isotopic analysis for attribution of source. The results conclusively showed that while carbon mineralisation was often higher in biochar amended soil, this was due to rapid utilisation of a small labile component of biochar and that biochar did not prime for the loss of native organic soil organic matter. Furthermore, in some cases negative priming occurred, with lower carbon mineralisation in biochar amended soil, probably as a result of the stabilisation of labile soil carbon.     

Changes in the flavonoid and phenolic acid contents and antioxidant activity of red leaf lettuce (Lollo Rosso) due to cultivation under plastic films varying in ultraviolet transparency

Red leaf lettuce (Lollo Rosso) was grown under three types of plastic films that varied in transparency to UV radiation (designated as UV block, UV low, and UV window). Flavonoid composition was determined by high-performance liquid chromatography (HPLC), total phenolics by the Folin-Ciocalteu assay, and antioxidant capacity by the oxygen radical absorbance capacity (ORAC) assay. Exposure to increased levels of UV radiation during cultivation caused the leaves to redden and increased concentrations of total phenols and the main flavonoids, quercetin and cyanidin glycosides, as well as luteolin conjugates and phenolic acids. The total phenol content increased from 1.6 mg of gallic acid equivalents (GAE)/g of fresh weight (FW) for lettuce grown under UV block film to 2.9 and 3.5 mg of GAE/g of FW for lettuce grown under the UV low and UV window films. The antioxidant activity was also higher in lettuce exposed to higher levels of UV radiation with ORAC values of 25.4 and 55.1 micromol of Trolox equivalents/g of FW for lettuce grown under the UV block and UV window films, respectively. The content of phenolic acids, quantified as caffeic acid, was also different, ranging from 6.2 to 11.1 micromol/g of FW for lettuce cultivated under the lowest and highest UV exposure plastic films, respectively. Higher concentrations of the flavonoid glycosides were observed with increased exposure to UV radiation, as demonstrated by the concentrations of aglycones after hydrolysis, which were cyanidin (ranging from 165 to 793 microg/g), quercetin (ranging from 196 to 880 microg/g), and luteolin (ranging from 19 to 152 microg/g). The results demonstrate the potential of the use of UV-transparent plastic as a means of increasing beneficial flavonoid content of red leaf lettuce when the crop is grown in polytunnels.