Cellular and humoral immune responses of Senegalese sole,Solea senegalensis (Kaup), following challenge with two Photobacterium damselae subsp. piscicida strains from different geographical origins

The present study aimed to investigate leucocyte responses to inflammation as well as some innate immune parameters of Senegalese sole, Solea senegalensis, following challenge with two strains of Photobacterium damselae subsp. piscicida belonging to the European and Japanese clones described for this bacterium. Pathogenicity assays were performed to assess the virulence of each Photobacterium damselae subsp. piscicida strain for sole. Subsequently, fish were intraperitoneally injected with phosphate‐buffered saline (control) or two concentrations (2 × 10² and 2 × 10⁶ CFU mL^?1) of each bacterial strain and sampled after 6 and 24 h. Results showed that the European isolate induces a higher degree of response than the Japanese strain. While blood neutrophilia and monocytosis correlated well with the increase in neutrophil and macrophage numbers in the peritoneal cavity, fish infected with the European isolate presented higher peritoneal cell numbers than fish challenged with the Japanese strain. In addition, alternative complement pathway activity and respiratory burst of head kidney leucocytes increased significantly in fish infected with the European isolate. The enhanced innate immune response displayed by Senegalese sole challenged with the European isolate is probably due to the higher degree of virulence presented by this Photobacterium damselae subsp. piscicida strain.     

Effects of dietary carbohydrate/lipid ratios on growth performance,body composition and glucose metabolism of fingerling blunt snout bream Megalobrama amblycephala

This study aimed to investigate the optimum dietary carbohydrate/lipid (CHO/L) ratio for fingerling blunt snout bream Megalobrama amblycephala (average initial weight: 6.61 ± 0.03 g). Six isonitrogenous and isoenergetic diets were formulated to contain various CHO/L ratios ranging from 1.62 to 24.20. Each diet was tested in four replicates for 10 weeks. Specific growth rate (SGR), feed conversion ratio and nitrogen and energy retention all improved significantly (P < 0.05) as dietary CHO/L ratios decreased from 24.20 to 5.64 but showed little difference (P > 0.05) as dietary CHO/L ratios ranged from 2.45 to 5.64. Hepatosomatic index increased significantly (P < 0.05) as dietary CHO/L ratios increased, whereas intraperitoneal fat ratio showed an opposite trend (P < 0.05). Opposite to moisture content, lipid content of whole body and carcass all increased significantly (P < 0.05) as dietary CHO/L ratios decreased. Liver lipid content showed little difference (P > 0.05) among all the treatments, while liver glycogen content increased significantly (P < 0.05) with increasing CHO/L ratios. High dietary carbohydrate enhanced the activities of liver hexokinase, pyruvate kinase and glucose‐6‐phosphate dehydrogenase but did not induce hyperglycaemia. Based on the second‐order polynomial regression analysis of SGR, the optimal dietary carbohydrate and lipid contents for fingerling blunt snout bream were 291.7 and 81.4 g kg⁻¹, respectively, with a corresponding dietary CHO/L ratio of 3.58.     

Tail Muscle Free Amino Acid Concentration of Pacific White Shrimp,Litopenaeus vannamei,Fed Diets Containing Protein‐bound versus Crystalline Amino Acids

Tail muscle tissue free amino acids (FAA) concentration is reported for subadult (4.3 g) Litopenaeus vannamei fed diets containing only protein‐bound amino acids, crystalline lysine as a supplement to protein‐bound lysine, and only crystalline amino acids (CAA). FAA were determined in shrimp at 0.00, 0.25, 0.5, 1, 2, 4, and 8 h postfeeding. Highest total tissue concentrations of essential amino acids and individual amino acids were typically observed 4 h postfeeding for shrimp fed intact and crystalline lysine‐supplemented feeds. Those shrimp offered diets containing only CAA showed no tissue maxima. Similar uptake patterns for lysine shown by shrimp fed the intact and crystalline lysine‐supplemented diet indicate the potential use of small amounts of crystalline lysine in shrimp feeds.     

Effect of Stocking Density on the Growth,Survival, and Behavior of Postpuerulus Western Rock Lobster,Panulirus cygnus (George) (Decapoda: Palinuridae)

The effect of four stocking densities (30, 60, 100, and 150 lobsters/m²) on the growth and survival of Panulirus cygnus postpueruli was determined over a 112‐d grow‐out trial. Agonistic behavior at each experimental density was recorded using infrared filming. Survival and growth decreased with increasing density, although this trend was only significant between the lowest (30/m²) and the highest densities (150/m²) (P < 0.05) at the conclusion of the grow‐out trial. Density had no significant effect on apparent feed intake but significantly affected food conversion ratios (FCR) across all four densities, with FCR being best at 30/m² and worst at 150/m². The number and frequency of agonistic encounters per tank differed significantly with density, with agonistic encounters being highest at 150/m² and lowest at 30/m². Agonistic encounters coincided with the crepuscular foraging pattern of postpueruli. The reduction in survival and growth of postpueruli at the highest density (150/m²) can be attributed to the significant increase in the number of agonistic encounters. This study supports that P. cygnus postpueruli should be stocked at densities less than 100/m² in order to minimize the negative effect of density on growth and survival.     

Mineral‐nitrogen and phosphorus leaching from vegetable gardens in Niamey,Niger

Urban and periurban agriculture (UPA) contributes significantly to meeting increasing food demand of rapidly growing urban populations in West African cities. The often intensive high‐input vegetable production within UPA results in large positive nutrient balances, being presumably linked to strong nutrient leaching which needs quantification. This study aimed at estimating leaching losses of mineral N and P in three representative urban gardens of Niamey, Niger, using ion‐exchange‐resin cartridges installed below the crop rooting zone at 0.6 m soil depth. In 2007, a year with below‐average annual rainfall (425 mm as compared to 542 mm), mean leaching of mineral N amounted to 5.9 and 7.3 kg N ha^–1 for two gardens with > 80% sand fraction and only 2.2 kg N ha^–1 for a garden with 40% silt and clay. Apparent annual P leaching was 0.7 kg P ha^–1 in all three gardens. Additional multiannual studies are necessary to assess the effect of inter‐ and intraannual variation in precipitation on nutrient leaching in intensive UPA vegetable production of semiarid West Africa.     

Application of biochar to soil and N2O emissions: potential effects of blending fast‐pyrolysis biochar with anaerobically digested slurry

Soil applications of recalcitrant biochar offer the possibility of mitigating climate change effects through long‐term carbon sequestration and potentially also by reducing emissions of the potent greenhouse gas nitrous oxide (N₂O). This laboratory study examined the effect of combining a fast‐pyrolysis biochar at small (1% by mass) and large (3%) concentrations with anaerobically digested slurry on soil N₂O and carbon dioxide (CO₂) emissions over a period of 55 days. The results showed that fast‐pyrolysis biochar applied on its own increased N₂O emissions from soil. However, when biochar was applied together with slurry, the larger biochar concentration decreased N₂O emissions by 47%, relative to those from the slurry treatment with the smaller biochar concentration. Reduced N₂O emissions coincided with enhanced soil microbial activity and immobilization of nitrogen. A combined application of biochar and anaerobic digested slurry could therefore be beneficial for cropping systems in terms of soil nitrogen retention while concurrently mitigating N₂O fluxes and sequestering carbon in soil.     

The use of T‐DNA tagging to isolate mutants of Colletotrichum gloeosporioides and Colletotrichum acutatum with reduced virulence against Hevea brasiliensis

Agrobacterium tumefaciens‐mediated transformation (ATMT) is being increasingly recognized as an effective insertional mutagenesis tool in studies of filamentous fungi for gene discovery and functional analysis. We developed and optimized ATMT for 2 Colletotrichum species, Colletotrichum gloeosporioides and Colletotrichum acutatum, the causative agents of Colletotrichum leaf disease in rubber trees in Southern China. A. tumefaciens strain AGL‐1 carrying an ILV1 gene and a green fluorescent protein gene were used to transform the conidia of these 2 Colletotrichum species. The transformation efficiency was correlated with the co‐cultivation duration and bacterial cell concentrations, which reached 300–400 transformants per 1 × 10⁶ conidia after optimization. Southern blot analysis indicated that about 60.0% of the C. gloeosporioides transformants and 46.2% of the C. acutatum transformants had a single copy of T‐DNA in their genomes. Fungal genomic DNA segments flanking the T‐DNA were identified in the transformants through thermal asymmetrical interlaced polymerase chain reaction followed by sequencing. The flanking sequences from 4 C. acutatum and 7 C. gloeosporioides transformants showed moderate or weak homology to the NCBI database entries. Some sequences matching those reported virulence‐related genes. The results suggest that the T‐DNA inserted mutants banks constructed are useful for the discovery of new or important genes and to elucidate their function in the 2 Colletotrichum species.     

Brenneria spp. and Rahnella victoriana associated with acute oak decline symptoms on oak and hornbeam in Iran

Decline diseases of forest trees are complex syndromes not attributable to single causal factors. In Iran, symptoms of decline disease have been observed in a number of native forest species including Quercus castaneifolia (chestnut‐leaved oak), Q. brantii (Persian oak) and Carpinus betulus (hornbeam). The symptoms are prevalent in the northern forests and the Zagros mountain forests. There are parallels between the disease in Iran and acute oak decline (AOD) reported in the UK, specifically the presence of weeping cankers, which have been attributed to a polybacterial complex wherein Brenneria goodwinii is considered a key necrogen. Based on the AOD symptomatology, and as a first step towards discovering potential causal agents of the stem weeping symptoms of affected trees in Iran, necrotic tissues were tested primarily for the presence of B. goodwinii. Symptomatic Q. castaneifolia and C. betulus from the Mazandaran Province and symptomatic Q. brantii from Kohgiluyeh and Boyerahmad Province were sampled. Isolation and culture on a selective medium yielded uniform bacterial colonies. Isolates were characterized using phenotypic and genotypic (DNA sequencing) tests. The isolates were phenotypically identical to members of Pectobacteriaceae and Yersiniaceae, specifically Brenneria and Rahnella spp. The nucleotide sequences of the 16S rRNA and housekeeping genes gyrB, infB and atpD (MLSA) amplified via PCR demonstrated that the isolates from the trees in Iran were indeed B. goodwinii, B. roseae subsp. roseae, Rahnella victoriana and an unknown species of Brenneria. Most bacteria isolated from non‐symptomatic trees were Gram‐positive, and Pseudomonas spp. were dominant, but bacterial species isolated from the diseased trees were not detected in healthy trees. Hypersensitivity response tests were positive, but inoculation on saplings was more variable with internal necrosis developing only once in the test period. Therefore, further testing is required. This is the first report of the incidence of B. goodwinii, B. roseae subsp. roseae, R. victoriana and Brenneria sp. associated with acute oak decline‐like symptoms on Q. castaneifolia, Q. brantii and C. betulus across the western forests of Iran and in the world.