A rapid method for the determination of microbial biomass by dry weight using a moisture analyser with an infrared heating source and an analytical balance

Aims: Microbial biomass is an important biotechnological parameter. The traditional method for its determination involves an oven‐drying step and equilibration to room temperature before weighing, and it is tedious and time consuming. This work studied the utilisation of a moisture analyser consisting of an efficient infrared‐heating module and an analytical balance for the determination of microbial biomass by dry weight. Methods and Results: The method duration depended on the sample volume and was between 7 and 40 min for sample volumes of 1–10 ml. The method precision depended on the total dry weight analysed – 10 mg of total dry weight being sufficient to achieve coefficients of variation of 5% or less. Comparison with the conventional oven method provided a correlation coefficient r² of 0·99. The recovery of an internal standard ranged between 94·2 and 106·4% with a precision of 1·39–4·53%CV. Conclusions: Validation revealed sufficient method accuracy, precision and robustness and was successfully applied to the study of yeast and bacterial growth kinetics. Techniques are discussed that allow for increased method precision at low biomass concentrations, and equations are provided to estimate required drying time and method precision based on sample volume and total sample dry weight, respectively. Significance and Impact of the Study: This work presents a rapid method for the determination of microbial biomass, allowing for the timely implementation of biomass‐based information in biotechnological and laboratory protocols.     

Warming alters the size spectrum and shifts the distribution of biomass in freshwater ecosystems

Organism size is one of the key determinants of community structure, and its relationship with abundance can describe how biomass is partitioned among the biota within an ecosystem. An outdoor freshwater mesocosm experiment was used to determine how warming of～4 °C would affect the size, biomass and taxonomic structure of planktonic communities. Warming increased the steepness of the community size spectrum by increasing the prevalence of small organisms, primarily within the phytoplankton assemblage and it also reduced the mean and maximum size of phytoplankton by approximately one order of magnitude. The observed shifts in phytoplankton size structure were reflected in changes in phytoplankton community composition, though zooplankton taxonomic composition was unaffected by warming. Furthermore, warming reduced community biomass and total phytoplankton biomass, although zooplankton biomass was unaffected. This resulted in an increase in the zooplankton to phytoplankton biomass ratio in the warmed mesocosms, which could be explained by faster turnover within the phytoplankton assemblages. Overall, warming shifted the distribution of phytoplankton size towards smaller individuals with rapid turnover and low standing biomass, resulting in a reorganization of the biomass structure of the food webs. These results indicate future environmental warming may have profound effects on the structure and functioning of aquatic communities and ecosystems.     

An image analysis technique to estimate the cell density and biomass concentration of Trichoderma reesei

Aim:  The objective is to develop an automated image analysis protocol to quantify the cell volume fraction of filamentous fungi ( Trichoderma reesei ) and estimate the biomass concentration.
Methods and Results:  Both dry weight and image analyses were performed on samples collected periodically from 7-l stirred tank fermentations. Using the projected area of lactophenol blue-stained hyphae, the fraction occupied by the cells in a given volume was estimated. Combined with the biomass dry weight obtained by filtration, the method was used to estimate the density of filamentous fungi. Knowing the density of fungi, the algorithm was employed to quantitatively assess the biomass evolution during the course of fermentation even in the presence of solid particles.
Conclusions:  A density of 0.334 g dry weight cm⁻³ was found for T. reesei RUT C-30. The image analysis protocol allowed successful estimation of biomass concentration in the presence or absence of solid particles.
Significance and Impact of the Study:  Methods to quantify biomass during the industrial production of cellulase with T. reesei are often limited due to the presence of solid substrates. The image analysis protocol presented here offers a quick and easy way to estimate biomass concentration of filamentous micro-organisms in insoluble medium.     

岷江上游亚高山林区老龄林地上生物量动态变化

中国川西亚高山森林中的天然林大部分为成过熟的老龄林,对其生物量动态研究有助于了解其碳储量的动态变化规律.利用全国森林资源连续清查的27个固定样地数据,基于地上各器官生物量与树干胸径(D)和树高(h)的异速生长方程,估算了岷江上游亚高林山老龄林地上生物量密度的动态变化特征及其时空变化规律.结果表明,(1)从1988～2002年期间,老龄林地上生物量密度净增量为(27.311±15.580)Mg·hm-2,平均每年增长率为(1.930±1.091 )Mg·hm-2·a-1,平均每年枯损率为(2 271±1.424)Mg·hm-2·a-1;(2)地上生物量变化受各径级保留木生长量、枯损量及进界生长量影响,其中20～40cm径级保留木生长量与生物量净增量最大,>80cm径级生物量增量最小,40～60cm和60～80cm径级生物量在调查期间净增量出现负增长.(3)岷江上游老龄林地上生物量动态变化具有时空异质性,同一样地在不同调查间隔期或同一调查期间不同样地间生物量变化不同,不仅有增量数值大小差异,还表现为生物量增量的正负差异.     

Diatoms can be an important exception to temperature–size rules at species and community levels of organization

Climate warming has been linked to an apparent general decrease in body sizes of ectotherms, both across and within taxa, especially in aquatic systems. Smaller body size in warmer geographical regions has also been widely observed. Since body size is a fundamental determinant of many biological attributes, climate‐warming‐related changes in size could ripple across multiple levels of ecological organization. Some recent studies have questioned the ubiquity of temperature–size rules, however, and certain widespread and abundant taxa, such as diatoms, may be important exceptions. We tested the hypothesis that diatoms are smaller at warmer temperatures using a system of geothermally heated streams. There was no consistent relationship between size and temperature at either the population or community level. These field data provide important counterexamples to both James’ and Bergmann's temperature–size rules, respectively, undermining the widely held assumption that warming favours the small. This study provides compelling new evidence that diatoms are an important exception to temperature–size rules for three reasons: (i) we use many more species than prior work; (ii) we examine both community and species levels of organization simultaneously; (iii) we work in a natural system with a wide temperature gradient but minimal variation in other factors, to achieve robust tests of hypotheses without relying on laboratory setups, which have limited realism. In addition, we show that interspecific effects were a bigger contributor to whole‐community size differences, and are probably more ecologically important than more commonly studied intraspecific effects. These findings highlight the need for multispecies approaches in future studies of climate warming and body size.     

Application of an aqueous two‐phase micellar system to extract bromelain from pineapple (Ananas comosus) peel waste and analysis of bromelain stability in cosmetic formulations

Bromelain is a set of proteolytic enzymes found in pineapple (Ananas comosus) tissues such as stem, fruit and leaves. Because of its proteolytic activity, bromelain has potential applications in the cosmetic, pharmaceutical, and food industries. The present study focused on the recovery of bromelain from pineapple peel by liquid–liquid extraction in aqueous two‐phase micellar systems (ATPMS), using Triton X‐114 (TX‐114) and McIlvaine buffer, in the absence and presence of electrolytes CaCl₂ and KI; the cloud points of the generated extraction systems were studied by plotting binodal curves. Based on the cloud points, three temperatures were selected for extraction: 30, 33, and 36°C for systems in the absence of salts; 40, 43, and 46°C in the presence of KI; 24, 27, and 30°C in the presence of CaCl₂. Total protein and enzymatic activities were analyzed to monitor bromelain. Employing the ATPMS chosen for extraction (0.5 M KI with 3% TX‐114, at pH 6.0, at 40°C), the bromelain extract stability was assessed after incorporation into three cosmetic bases: an anhydrous gel, a cream, and a cream‐gel formulation. The cream‐gel formulation presented as the most appropriate base to convey bromelain, and its optimal storage conditions were found to be 4.0 ± 0.5°C. The selected ATPMS enabled the extraction of a biomolecule with high added value from waste lined‐up in a cosmetic formulation, allowing for exploration of further cosmetic potential. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:937–945, 2015     

Factors affecting the vulnerability of cane toads (Bufo marinus) to predation by ants

Experimental evidence on the determinants of prey vulnerability is scarce, especially for vertebrates in the field. Invasive species offer robust opportunities to explore prey vulnerability, because the intensity of predation on or by such animals has not been eroded by coevolution. Around waterbodies in tropical Australia, native meat ants (Iridomyrmex reburrus) consume many metamorph cane toads (Bufo marinus, an invasive anuran). We document the determinants of toad vulnerability, especially the roles of toad body size and ant density. Larger metamorphs were attacked sooner (because they attracted more ants), but escaped more often. Overall, smaller toads were more likely to be killed. Ant densities influenced toad responses, as well as attack rate and success. Data on the immediate outcomes of attacks underestimate mortality: more than 73% of apparent ‘escapees’ died within 24 h. Because mortality during this period was independent of toad size, predation was less size selective than suggested by immediate outcomes. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 99 , 738–751.     

Chemiluminescence determination of chlorpheniramine using tris(1,10‐phenanthroline)–ruthenium(II) peroxydisulphate system and sequential injection analysis

A sequential injection (SI) method was developed for the determination of chlorpheniramine (CPA), based on the reaction of this drug with tris(1,10‐phenanthroline)–ruthenium(II) [Ru(phen)₃²⁺] and peroxydisulphate (S₂O₈^2–) in the presence of light. The instrumental set‐up utilized a syringe pump and a multiposition valve to aspirate the reagents [Ru(phen)₃²⁺ and S₂O₈^2–] and a peristaltic pump to propel the sample. The experimental conditions affecting the chemiluminescence reaction were systematically optimized, using the univariate approach. Under the optimum conditions linear calibration curves of 0.1–10 µg/ml were obtained. The detection limit was 0.04 µg/ml and the relative standard deviation (RSD) was always < 5%. The procedure was applied to the analysis of CPA in pharmaceutical products and was found to be free from interferences from concomitants usually present in these preparations. Copyright © 2008 John Wiley & Sons, Ltd.     

Age, diet, growth and reproduction of a Population of Cobitis paludica (de Buen, 1930) in the Palancar Stream (southwest of Europe, Spain) (Pisces: Cobitidae)

A total sample of 99 specimens of the vulnerable fish species Cobitis paludica(de Buen, 1930), was collected on a monthly basis from a tributary of the River Guadalquivir between September 1990 and September 1991. Female longevity was 1 year longer than that of males, the former reaching age-class 5+. The diet of C. paludicaconsisted mainly of Ostracoda and chironomid larvae, benthic prey of small size and occasionally other resources, such as Macrobiotid eggs spawned during winter. Detritus in gastrointestinal content was scarce, which differs from other populations. Back-calculations of length were performed from scale annuli measurements. The relationship between standard length and scale radius was different for males and females. Females were larger than males of the same age. Both back-calculated length and theoretical length, calculated by applying von Bertalanffy's model, were larger for females. The relationship between female standard length (SL) and fecundity (F) was log F= –7.465 + 5.5753 log SL.     

Different patterns of biomass allocation of mature and sapling host tree in response to liana competition in the southern temperate rainforest

Lianas can negatively affect their host tree. The evidence comes from studies in tropical forests, where lianas decrease the growth rate and reproduction of their host tree. This is primarily a consequence of water and nutrient competition, two limiting factors in tropical forests. In contrast, for some areas of southern temperate rainforests the competition for these resources could be less severe, because of the high rainfall and fertile soils. But so far, no study has determined the effect of liana competition over their host tree in southern temperate rainforests. The aim of this study was to evaluate, in field conditions, the effect of liana Cissus striata (Ruiz & Pavon) competition over the growth rate of mature Nothofagus obliqua (Bidr Egefam) host tree. In an experimental approach, we determined whether above‐ and/or below‐ground competition is more important in this interaction. We also looked for compensatory strategies that would allow to trees to respond to liana competition. In field conditions, we found that infested trees have a decrease in their relative growth rate of 26% and a reduction of the leaf area index (LAI) of 20% compared with control trees. In the greenhouse experiment, we found that saplings were water stressed and that there was light competition. Neither competition for water nor light had a significant effect on the growth rate of infested saplings. This could be explained because saplings showed compensatory strategies in response to competition. These strategies were based in the biomass distribution between organs (leaf area, slenderness index) and within leaves (LMA). In conclusion, we found that C. striata has a negative effect over the growth of mature and sapling N. obliqua host trees. This was a consequence of above‐ground and below‐ground competition, but we cannot disentangle which type of competition is more important. Trees respond to liana competition, mature host trees change the canopy architecture and saplings allocate resource between and within organs, which allows them to optimize resource capture.     

Responses of biomass allocation across two vegetation types to climate fluctuations in the northern Qinghai–Tibet Plateau

The Qinghai–Tibet Plateau (QTP) is particularly sensitive to global climate change, especially to elevated temperatures, when compared with other ecosystems. However, few studies use long‐term field measurements to explore the interannual variations in plant biomass under climate fluctuations. Here, we examine the interannual variations of plant biomass within two vegetation types (alpine meadow and alpine shrub) during 2008–2017 and their relationships with climate variables. The following results were obtained. The aboveground biomass (AGB) and belowground biomass (BGB) response differently to climate fluctuations, the AGB in KPM was dominated by mean annual precipitation (MAP), whereas the AGB in PFS was controlled by mean annual air temperature (MAT). However, the BGB of both KPM and PFS was only weakly affected by climate variables, suggesting that the BGB in alpine ecosystems may remain as a stable carbon stock even under future global climate change. Furthermore, the AGB in PFS was significantly higher than KPM, while the BGB and R/S in KPM were significantly higher than PFS, reflecting the KPM be more likely to allocate more photosynthates to roots. Interestingly, the proportion of 0–10 cm root biomass increased in KPM and PFS, whereas the other proportions both decreased, reflecting a shift in biomass toward the surface layer. Our results could provide a new sight for the prediction how alpine ecosystem response to future climate change.     

黑龙江省完达山东部林区东北虎猎物生物量

研究一个地区猎物种群生物量能否满足捕食动物种群数量的需求,这对于了解濒危大型食肉动物是否受到来自于食物缺乏的威胁和制定相应的保护措施极其重要。为了掌握黑龙江省完达山东部林区东北虎食物需求与猎物生物量之间的关系,于2008年冬季至2009早春积雪覆盖期采用随机布设样线,通过收集有蹄类动物在雪地上留下的足迹等活动的方法,在东方红林业局和迎春林业局管辖境内3 692.06 km²的区域布设大样方48个,并在大样方里共布设样线240条开展有蹄类动物种群数量调查,确定东北虎猎物生物量。调查结果表明:研究地区野猪(成体502 606只,亚成体209 210只)、马鹿(成体331 357只,亚成体67 72只)和狍子(成体810 815只,亚成体202 203只)的生物量分别为74 767.50 87 825.00 kg、79 744.50 85 984.50 kg 和 31 337.00 31 525.50 kg,3种有蹄类动物生物量共计1 85 849.00 205 335.00 kg。研究地区猎物总生物量为209 619.89 231 598.24 kg。如果按8%的生物提供给东北虎,3种主要猎物生物量可满足5.22 6.92只东北虎个体的食物需求,研究地区猎物总生物量则可满足5.89 7.81只东北虎个体的食物需求。此外,对足迹遇见率与抽样强度、抽样强度与足迹遇见率的均值标准误差之间关系的分析表明,在完达山东部林区布设120条样线(抽样距离600 km)、150条样线(抽样距离750 km)和115条样线(抽样距离675 km)能满足野猪、马鹿、狍子种群数量调查准确性的最低需求。     

抗铅锌功能菌生长菌株和干菌体吸附Pb2+、Zn2+性能优化及机理分析

【目的】针对湖南资兴铅锌矿污染问题,筛选本土耐性菌株用作生物修复。【方法】供试菌株J3筛选自湖南资兴铅锌矿区的尾砂矿矿渣,利用正交实验分析其生长菌株及干菌体最佳吸附条件以优化其吸附效果,同时对相关数据进行动力学拟合以探讨其吸附机理,最后结合形态观察和18S rRNA基因序列分析对其进行分类鉴定。【结果】在最佳条件下,J3生长菌株对Pb2+和Zn2+的去除率分别为92.2%和87.7%;干菌体对Pb2+和Zn2+去除率分别为72.6%和23.8%,反应动力学模型研究表明,生长菌株吸附过程中吸附速率受重金属浓度影响,对Zn2+吸附主要为颗粒内扩散作用;干菌体对Pb2+吸附推测为膜扩散和颗粒内扩散作用,而对Zn2+的吸附则由膜扩散控制。根据形态特征和系统发育分析,J3初步鉴定为虫生轮枝菌。【结论】生长菌株吸附效果好于干菌体,二级动力学方程拟合结果可为生物吸附反应器的设计提供理论参考和实践指导。     

Responses of radish (Raphanus sativus) to drought stress

With this study the effects of water deficit on radish growth, dry mass production and partitioning, source–sink relations, physiological responses and accumulation of secondary metabolites in storage roots and leaves have been investigated. Three water stress treatments were applied: (Control) 100% of water holding capacity (WHC), (WS50) 50% of WHC, (WS30) 30% of WHC. Water stress was effectively acquired starting from 287 growing degree days (GDD) as demonstrated by the lower values of relative water content (RWC), higher leaf temperature and NIR‐based water indices values in water stressed conditions. Water stress reduced storage root dry weight by 27% at WS50 and 53% at WS30 as well as leaf dry matter accumulation (by 23% and 31% in WS50 and WS30, respectively), expansion (by 28% and 45% in WS50 and WS30, respectively) and specific leaf area (by 7% and 20% in WS50 and WS30, respectively) at 403 GDD. The increasing of leaf‐to‐root mass (L/R) and leaf area‐to‐root mass (LA/R) ratios indicated less dry matter allocation into storage organs under water stress conditions. Besides, water shortage increased leaf greenness as estimated by the higher soil–plant analysis development (SPAD) values (+14% and +20% on average for WS50 and WS30, respectively); other reflectance indices only partially confirmed SPAD readings. Substantially, water limitation did not significantly influence total anthocyanin content, ABTS‐radical scavenging activity and total free phenolic compounds in storage roots, as well as the total free phenolic compounds in leaves. Radish showed a strong plasticity in its adaptation to drought thanks to avoidance mechanisms such as constrained leaf development, increased leaf thickness and adjusted source–sink relationships.     

Structure of the food web including the endangered lycaenid butterfly Shijimiaeoides divinus asonis (Lepidoptera: Lycaenidae)

The structure of the food web including the endangered lycaenid butterfly Shijimiaeoides divinus asonis (Matsumura) was analyzed to identify species contributing most to maintaining the equilibrium of the food web. Twenty‐seven species belonging to 17 families fed on Sophora flavescens Aiton, the host‐plant of S. divinus asonis: 15 species were leaf and stem feeders, seven (including S. divinus asonis) fed on flower buds, four were flower feeders and one fed on the seeds of So. flavescens. Of these 27 species, four were omnivores. The natural enemies of S. divinus asonis comprised six insect species, 11 spider species and one entomopathogenic fungus species, including six new predator records. The linkage density, total number of trophic links, connectance, average chain length and predator–prey ratio were 1.617, 97, 0.0548, 2.267 and 0.694, respectively. Exclusion of any of the 15 species with four or more trophic links reduced the connectance of the food web. These 15 species included facultative mutualistic attendant ants and predators of S. divinus asonis, herbivores to So. flavescens, an omnivore feeding on S. divinus asonis and So. flavescens, and prey insects. Therefore, future studies should monitor these 15 species.     

Differentiation in the ability to utilize Pterostyrax hispida (Ebenales: Styracaceae) among four local populations of the phytophagous ladybird beetle Henosepilachna yasutomii (Coleoptera: Coccinellidae)

The oligophagous ladybird beetle Henosepilachna yasutomii Katakura (Coleoptera: Coccinellidae) shows inter-population differences in its host-use. In this beetle, populations using the deciduous tree Pterostyrax hispida Sieb. et Zucc. (Styracaceae) were recently rediscovered in the Kanto districts of central Honshu, Japan. In the present study, the ability to utilize P. hispida and other host plants was compared among four populations of H. yasutomii occurring on P. hispida and Scopolia japonica Maxim. (Solanaceae), Chelidonium japonicum Thunb. (Papaveraceae), and Solanum tuberosum L. (Solanaceae). As regards the feeding habits of adult beetles, only the populations occurring on P. hispida accepted this plants leaves, which differed distinctly from the feeding habits of the other populations, although the differences among the four populations were not readily apparent with respect to the leaf amounts consumed. The larvae from the populations occurring on P. hispida showed significantly higher survivorship on this plant than did the larvae from the other populations. Considering the host use patterns and the life cycles of beetles under natural conditions, the large abundance of P. hispida leaves throughout the season may have played an important role in selection for the ability to utilize P. hispida observed in the H. yasutomii populations occurring on this woody host.     

亚洲玉米螟体重和体型的地理变异

为探明亚洲玉米螟Ostrinia furnacalis体重和体型地理变异,我们详细比较了来自4个不同地理种群（海南乐东18.8°N, 109.2°E),广西阳朔24.8°N, 110.5°E),江西南昌28.8°N, 115.9°E)和河北廊坊39.5°N, 116.7°E)）亚洲玉米螟的体重、体型大小及其与采集地纬度的关系。结果表明：不同地理种群的亚洲玉米螟卵重随纬度的升高而逐渐增大,符合贝格曼法则(Bergmann’s law), 而雌雄蛹重及成虫体长、后足腿节长和前翅长均随纬度的升高而逐渐减小, 符合反贝格曼法则(Converse Bergmann’s law)。雌虫的前翅显著长于雄虫, 其性体型二型性符合任希法则(Rensch’s rule),即在雌虫体型较大的种群中,雄虫前翅比雌虫前翅增长幅度相对较大。本文结果进一步揭示了即使在同一种类昆虫中,其各个虫态体重和体型的地理变异也可能不同。     

Influences of the reproductive cycle on condition of marine catfishes (Siluriformes,Ariidae) in a coastal area at southeastern Brazil

Marine catfishes use estuaries and bays as part of their life cycle for spawning and feeding purposes, respectively. We compared three species of the family Ariidae in two different environmental conditionsa relatively stable bay site and an estuarine site, which differ in their magnitudes of environmental fluctuation to investigate effects of spawning on condition. We found differences in the parameter b (slope) and in they-intercept (a) among species and sexes. The parameter a varied inversely with b, and thus is not a good indicator to express fish condition. Condition factor did not differ between the two sites for species and sexes when we consider the pooled data. Overall condition was associated with reproductive process, and was high before the reproductive season, decreasing during the spawning period in the estuary, then increasing when fish move to the bay (April–August). Temporal segregation in the reproductive period and small-scale movements between the estuary and the bay are the probable mechanisms to optimize the use of the area by the three species.