美洲鲥鱼嗜水气单胞菌病原的分离与鉴定

2021年,山东省临沂市一养殖场养殖的美洲鲥鱼(Alosa sapidissima)突发疾病并出现严重死亡,日死亡率高峰期达到2.5%,累积死亡率约为90%。患病鱼主要症状为体表出血、溃疡,解剖可见腹腔腹水、肝脏暗红,并伴有肠炎。组织病理学检测发现,病鱼肝脏出现弥散性坏死,嗜碱性粒细胞增多和细胞肿胀空泡变性;脾脏出现出血性贫血性坏死灶、核破裂和核固缩;肾脏淋巴细胞坏死脱落,肾小体毛细血管球萎缩,近端小管和远端小管内的细胞出现不同程度的细胞结构消失。发病鱼肉眼和显微镜观察未见明显寄生虫,利用PCR方法检测鲤疱疹病毒2型(Cyprinid herpesvirus 2)、鲈鱼蛙病毒(largemouth bass ranavirus)等淡水鱼类常见病毒均为阴性。细菌分离培养结果显示,从发病鱼的肝脏、肾脏和脾脏中分离得到形态一致的优势菌,命名为AS-AH2101。经16S rRNA测序比对和生理生化鉴定,确定AS-AH2101为嗜水气单胞菌(Aeromonas hydrophila)。毒力基因检测结果显示,AS-AH2101携带气溶素(aerA)、溶血素(hlyA)、丝氨酸蛋白酶(ahpA)、...     

嫁接对西瓜根际微生物种群数量及群落结构的影响

分别采用微生物培养的方法、16S rDNA PCR-RFLP方法和phl D基因PCR-RFLP方法研究了嫁接对西瓜根际微生物种群数量、细菌群落结构及拮抗菌（2, 4-DAPG产生菌）群落结构的影响。结果表明：① 嫁接西瓜根际细菌和真菌数量有所提高,而放线菌数量则有所减少。② 嫁接西瓜与自根西瓜具有1个相同的主要根际细菌基因型;不同砧木嫁接的西瓜与自根西瓜也分别具有不同基因型的细菌。③ 嫁接西瓜具有与自根西瓜不同的主要根际拮抗菌基因型。     

不同处理牛羊粪便堆积发酵试验效果比较

利用发酵菌剂对不同处理的试验Ⅰ组、试验Ⅱ组、试验Ⅲ组、试验Ⅳ组、试验Ⅴ组和对照组进行堆积发酵试验,在发酵过程中比较了不同试验组的温度、水分、挥发性总固体、pH、细菌群落的变化,以及对牧草籽发芽率、虫卵杀除率和总养分的影响。     

不同乳酸菌组合对苜蓿青贮细菌群落结构的影响

为探讨不同乳酸菌互作对苜蓿(Medicago sativa)青贮细菌群落结构的影响,以2种植物乳杆菌、乳酸片球菌、戊糖片球菌及凝结芽孢杆菌形成的6种乳酸菌组合按1.5 mL·kg-1的添加量制作苜蓿青贮,以等量蒸馏水替代添加剂作为对照,45 d后运用高通量测序分析细菌群落结构.结果表明,各苜蓿青贮的优势乳酸菌群均为厚壁...     

Contributions of ammonia-oxidizing archaea and bacteria to nitrification in Oregon forest soils

Ammonia oxidation, the first step of nitrification, is mediated by both ammonia-oxidizing archaea (AOA) and bacteria (AOB); however, the relative contributions of AOA and AOB to soil nitrification are not well understood. In this study we used 1-octyne to discriminate between AOA- and AOB-supported nitrification determined both in soil-water slurries and in unsaturated whole soil at field moisture. Soils were collected from stands of red alder (Alnus rubra Bong.) and Douglas-fir (Pseudotsuga menziesii Mirb. Franco) at three sites (Cascade Head, the H.J. Andrews, and McDonald Forest) on acidic soils (pH 3.9–5.7) in Oregon, USA. The abundances of AOA and AOB were measured using quantitative PCR by targeting the amoA gene, which encodes subunit A of ammonia monooxygenase. Total and AOA-specific (octyne-resistant) nitrification activities in soil slurries were significantly higher at Cascade Head (the most acidic soils, pH < 5) than at either the H.J. Andrews or McDonald Forest, and greater in red alder compared with Douglas-fir soils. The fraction of octyne-resistant nitrification varied among sites (21–74%) and was highest at Cascade Head than at the other two locations. Net nitrification rates of whole soil without NH₄⁺ amendment ranged from 0.4 to 3.3 mg N kg⁻¹ soil d⁻¹. Overall, net nitrification rates of whole soil were stimulated 2- to 8-fold by addition of 140 mg NH₄⁺-N kg⁻¹ soil; this was significant for red alder at Cascade Head and the H.J. Andrews. Red alder at Cascade Head was unique in that the majority of NH₄⁺-stimulated nitrifying activity was octyne-resistant (73%). At all other sites, NH₄⁺-stimulated nitrification was octyne-sensitive (68–90%). The octyne-sensitive activity—presumably AOB—was affected more by soil pH whereas the octyne-resistant (AOA) activity was more strongly related to N availability.     

Fungal and bacterial N2O production regulated by soil amendments of simple and complex substrates

Fungal N₂O production results from a respiratory denitrification that reduces NO₃⁻/NO₂⁻ in response to the oxidation of an electron donor, often organic C. Despite similar heterotrophic nature, fungal denitrifiers may differ from bacterial ones in exploiting diverse resources. We hypothesized that complex C compounds and substances could favor the growth of fungi over bacteria, and thereby leading to fungal dominance for soil N₂O emissions. Effects of substrate quality on fungal and bacterial N₂O production were, therefore, examined in a 44-d incubation after soils were amended with four different substrates, i.e., glucose, cellulose, winter pea, and switchgrass at 2 mg C g⁻¹ soil. During periodic measurements of soil N₂O fluxes at 80% soil water-filled pore space and with the supply of KNO₃, substrate treatments were further subjected to four antibiotic treatments, i.e., no antibiotics or soil addition of streptomycin, cycloheximide or both so that fungal and bacterial N₂O production could be separated. Up to d 8 when antibiotic inhibition on substrate-induced microbial activity and/or growth was still detectable, bacterial N₂O production was generally greater in glucose- than in cellulose-amended soils and also in winter pea- than in switchgrass-amended soils. In contrast, fungal N₂O production was more enhanced in soils amended with cellulose than with glucose. Therefore, fungal-to-bacterial contribution ratios were greater in complex than in simple C substrates. These ratios were positively correlated with fungal-to-bacterial activity ratios, i.e., CO₂ production ratios, suggesting that substrate-associated fungal or bacterial preferential activity and/or growth might be the cause. Considering substrate depletion over time and thereby becoming limited for microbial N₂O production, measurements of soil N₂O fluxes were also carried out with additional supply of glucose, irrespective of different substrate treatments. This measurement condition might lead to potentially high rates of fungal and bacterial N₂O production. As expected, bacterial N₂O production was greater with added glucose than with added cellulose on d 4 and d 8. However, this pattern was broken on d 28, with bacterial N₂O production lower with added glucose than with added cellulose. In contrast, plant residue impacts on soil N₂O fluxes were consistent over 44-d, with greater bacterial contribution, lower fungal contribution, and thus lower fungal-to-bacterial contribution ratios in winter pea- than in switchgrass-amended soils. Real-time PCR analysis also demonstrated that the ratios of 16S rDNA to ITS and the copy numbers of bacterial denitrifying genes were greater in winter pea- than in switchgrass-amended soils. Despite some inconsistency found on the impacts of cellulose versus glucose on fungal and bacterial leading roles for N₂O production, the results generally supported the working hypothesis that complex substrates promoted fungal dominance for soil N₂O emissions.     

Fractionation of the reference inoculum of epizootic rabbit enteropathy in discontinuous sucrose gradient identifies aetiological agents in high density fractions

Epizootic rabbit enteropathy (ERE) is a major cause of economic loss in intensive rabbit production. Since its first recognition in 1997, much work has been done to determine the pathogenic mechanisms of the disease and to identify the aetiological agent(s). Unfortunately, the quest for aetiology has only met with limited success despite the ability to reproduce the syndrome by inoculation of intestinal contents from field cases. These intestinal inocula contain a huge number of microorganisms which could all be involved in the aetiology of ERE. To decrease the number of putative agents, the French reference inoculum TEC3 was fractionated on a discontinuous sucrose gradient so that seven fractions (supernatant, 10%, 20%, 30%, 40%, 50% and pellet) were obtained. Specific-pathogen-free rabbits were inoculated with three out of these seven fractions (supernatant, 30%, and pellet). The objectives were: (1) to characterise the seven fractions by bacteriological examination; (2) to verify whether the aetiological agent was present in the fractions by inoculation of rabbits; (3) to assign the aetiological agent of ERE to a morphological group of pathogens; (4) to identify a fraction which could replace the reference inoculum TEC3 in applications such as cell cultures or egg inoculation. The results strongly suggest that ERE is a bacterial disease and does not have a viral or parasitic aetiology.     

冀北水稻田土壤细菌群落结构和多样性研究

为探讨冀北水稻田土壤细菌群落结构和多样性,以承德地区水稻主产区（河北隆化,河北滦平）稻田土壤为试验材料,采用高通量测序技术,对水稻大田土壤细菌群落结构和多样性进行研究。结果表明：不同地区水稻田土壤微生物多样性存在差异,其中样本ZYY.CKD9中微生物多样性最高且有益菌较多,主要有硫杆菌属（Thiobacillus）、拟衣藻属（Dechloromonas）和地杆菌属（Geobacter）;但也存在共有细菌菌群,如Kaistobacter、溶杆菌属（Lysobacter）、乳酸杆菌属（Lactobacillus）、Candidatus_Solibacter、硫杆菌属（Thiobacillus）、红长命菌属（Rubrivivax）、拟衣藻属（Dechloromonas）、Anaeromyxobacter、硝化螺旋菌属（Nitrospira）等;PCA分析结果显示,土壤细菌群落结构空间相关性不明显。这为冀北水稻田土壤修复、病害防治及功能菌肥开发,促进水稻高产栽培提供依据。     

Fungal biomass in pastures increases with age and reduced N input

Previous studies have shown that soil fungal biomass increases towards more natural, mature systems. Shifts to a fungal-based soil food web have previously been observed with abandonment of agricultural fields and extensification of agriculture. In a previous field experiment we found increased fungal biomass with reduced N fertilisation. Here, we explore relationships between fungi, bacteria, N input and grassland age on real dairy farms in the Netherlands. We hypothesised that also in pastures that are still in production there is a negative relationship between fungal biomass and fertilisation, and that fungal biomass increases with grassland age in pastures that are still in production. We expected the fungal/bacterial biomass ratio to show the same responses, as this ratio has often been used as an indicator for management changes. We sampled 48 pastures from eight organic dairy farms. Sites differed in age and fertilisation rate. We determined fungal and bacterial biomass, as well as ergosterol (a fungal biomarker). Fungal and bacterial biomass and ergosterol, showed a negative relationship with N application rate, and correlated positively with organic matter percentage. In old pastures, fungal biomass and ergosterol were higher than in younger pastures. Because bacterial biomass responded in the same way as fungal biomass, the F/B ratio remained constant, and can therefore—in our data set—not be used as an indicator for changing management. We conclude that the changes in fungal and bacterial biomass were driven by changes in organic matter quality and quantity. The negative relationship we found between N application rate and fungal biomass adds to earlier work and confirms the presence of this relationship in pastures with relatively small differences in management intensities. Earlier studies on shifts in fungal biomass focused on ex-agricultural fields or restoration projects. Here we show that fungal biomass is also higher in older agricultural pastures.     

Effects of substrate sterilization,fungicide treatment,and mycorrhization helper bacteria on ectomycorrhizal formation of pedunculate oak (Quercus robur) inoculated with Laccaria laccata in two peat bare-root nurseries

Summary Pedunculate oak seedlings (Quercus robur) inoculated with the ectomycorrhizal fungus Laccaria lacata were grown for 1 year on fertilized sphagnum peat in two nurseries. Three factors affecting microbial populations in the substrate were studied, fungicide treatment of the seeds, peat disinfection before sowing (methyl bromide or steam pasteurization), and inoculation with mycorrhization helper bacteria. Treatment of acorns with Iprodione had no depressive effect on mycorrhiza formation. Both disinfection techniques were equivalent, stimulating or depressing mycorrhiza formation depending on the initial microflora in the peat. The introduction of two previously selected mycorrhization helper bacteria (one Pseudomonas fluorescens and one unidentified fluorescent pseudomonad), isolated from L. laccata sporocarps associated with Douglas fir—L. laccata ectomycorrhizas in other nurseries, significantly increased the mycorrhizal rate from 30 to 53% of the short roots. The implications of these results for the controlled mycorrhization of planting stocks and the specificity of mycorrhization helper bacteria are discussed.