Prediction of metabolisable energy value of broiler diets and water excretion from dietary chemical analyses

Thirty various pelleted diets were given to broilers (8/diet) for in vivo measurements of dietary metabolisable energy (ME) value and digestibilities of proteins, lipids, starch and sugars from day 27 to day 31, with ad libitum feeding and total collection of excreta. Water excretion was also measured. Amino acid formulation of diets was done on the basis of ratios to crude proteins. Mean in vivo apparent ME values corrected to zero nitrogen retention (AMEn) were always lower than the AMEn values calculated for adult cockerels using predicting equations from literature based on the chemical analyses of diets. The difference between mean in vivo AMEn values and these calculated AMEn values increased linearly with increasing amount of wheat in diets (P = 0.0001). Mean digestibilities of proteins, lipids and starch were negatively related to wheat introduction (P = 0.0001). The correlations between mean in vivo AMEn values and diet analytical parameters were the highest with fibre-related parameters, such as water-insoluble cell-walls (WICW) (r = −0.91) or Real Applied Viscosity (RAV) (r = −0.77). Thirteen multiple regression equations relating mean in vivo AMEn values to dietary analytical data were calculated, with R² values ranging from 0.859 to 0.966 (P = 0.0001). The highest R² values were obtained when the RAV parameter was included in independent variables. The direct regression equations obtained with available components (proteins, lipids, starch, sucrose and oligosaccharides) and the indirect regression equations obtained with WICW and ash parameters showed similar R² values. Direct or indirect theoretical equations predicting AMEn values were established using the overall mean in vivo digestibility values. The principle of indirect equations was based on the assumption that WICW and ashes act as diluters. Addition of RAV or wheat content in variables improved the accuracy of theoretical equations. Efficiencies of theoretical equations for predicting AMEn values were almost the same as those of multiple regression equations. Water excretion was expressed either as the water content of excreta (EWC), the ratio of water excretion to feed intake (WIR) or the residual value from the regression equation relating water excretion to feed intake (RWE). The best regression predicting EWC was based on sucrose, fermentable sugars (lactose + oligosaccharides) and chloride variables, with positive coefficients. The best equations predicting WIR or RWE contained the sugar and chloride variables, with positive coefficients. Other variables appearing in these equations were AMEn or starch with negative coefficients, WICW, ‘cell-wall-retained water’, RAV or potassium with positive coefficients.     

Molecular detection of native and invasive marine invertebrate larvae present in ballast and open water environmental samples collected in Puget Sound

Non-native marine species have been and continue to be introduced into Puget Sound via several vectors including ship's ballast water. Some non-native species become invasive and negatively impact native species or near shore habitats. We present a new methodology for the development and testing of taxon specific PCR primers designed to assess environmental samples of ocean water for the presence of native and non-native bivalves, crustaceans and algae. The intergenic spacer regions (IGS; ITS1, ITS2 and 5.8S) of the ribosomal DNA were sequenced for adult samples of each taxon studied. We used these data along with those available in Genbank to design taxon and group specific primers and tested their stringency against artificial populations of plasmid constructs containing the entire IGS region for each of the 25 taxa in our study, respectively. Taxon and group specific primer sets were then used to detect the presence or absence of native and non-native planktonic life-history stages (propagules) from environmental samples of ballast water and plankton tow net samples collected in Puget Sound. This methodology provides an inexpensive and efficient way to test the discriminatory ability of taxon specific oligonucleotides (PCR primers) before creating molecular probes or beacons for use in molecular ecological applications such as probe hybridizations or microarray analyses. This work addresses the current need to develop molecular tools capable of diagnosing the presence of planktonic life-history stages from non-native marine species (potential invaders) in ballast water and other environmental samples.     

Comparison between procedures using SDS for shotgun proteomic analyses of complex samples

Filter-aided sample preparation (FASP) and a new sample preparation method using a modified commercial SDS removal spin column are quantitatively compared in terms of their performance for shotgun proteomic experiments in three complex proteomic samples: a Saccharomyces cerevisiae lysate (insoluble fraction), a Caenorhabditis elegans lysate (soluble fraction), and a human embryonic kidney cell line (HEK293T). The characteristics and total number of peptides and proteins identified are compared between the two procedures. The SDS spin column procedure affords a conservative fourfold improvement in throughput, is more reproducible, less expensive (i.e. requires less materials), and identifies between 30 and 107% more peptides at q≤0.01, than the FASP procedure. The peptides identified by SDS spin column are more hydrophobic than species identified by the FASP procedure as indicated by the distribution of GRAVY scores. Ultimately, these improvements correlate to as great as a 50% increase in protein identifications with two or more peptides.     

Comparison of two filtration-elution procedures to improve the standard methods ISO 10705-1 & 2 for bacteriophage detection in groundwater, surface water and finished water samples

Aim: To select a reliable method for bacteriophage concentration prior detection by culture from surface water, groundwater and drinking water to enhance the sensitivity of the standard methods ISO 10705‐1 & 2. Methods and Results: Artificially contaminated (groundwater and drinking water) and naturally contaminated (surface water) 1‐litre samples were processed for bacteriophages detection. The spiked samples were inoculated with about 150 PFU of F‐specific RNA bacteriophages and somatic coliphages using wastewater. Bacteriophage detection in the water samples was achieved using the standard method without and with a concentration step (electropositive Anodisc membrane or a pretreated electronegative Micro Filtration membrane, MF). For artificially contaminated matrices (drinking and ground waters), recovery rates using the concentration step were superior to 70% whilst analyses without concentration step mainly led to false negative results. Besides, the MF membrane presented higher performances compared with the Anodisc membrane. Conclusion: The concentration of a large volume of water (up to one litre) on a filter membrane avoids false negative results obtained by direct analysis as it allows detecting low number of bacteriophages in water samples. Significance and Impact of the Study: The addition of concentration step before applying the standard method could be useful to enhance the reliability of bacteriophages monitoring in water samples as bio‐indicators to highlight faecal pollution.     

Assessing reliability of microsatellite genotypes from kit fox faecal samples using genetic and GIS analyses

Noninvasive faecal DNA sampling has the potential to provide a wealth of information necessary for monitoring and managing endangered species while eliminating the need to capture, handle or observe rare individuals. However, scoring problems, and subsequent genotyping errors, associated with this monitoring method remain a great concern as they can lead to misidentification of individuals and biased estimates. We examined a kit fox scat data set (353 scats; 80 genotypes) for genotyping errors using both genetic and GIS analyses, and evaluated the feasibility of combining both approaches to assess reliability of the faecal DNA results. We further checked the appropriateness of using faecal genotypes to study kit fox populations by describing information about foxes that we could deduce from the 'acceptable' scat genotypes, and comparing it to information gathered with traditional field techniques. Overall, genetic tests indicated that our data set had a low rate of genotyping error. Furthermore, examination of distributions of scat locations confirmed our data set was relatively error free. We found that analysing information on sex primer consistency and scat locations provided a useful assessment of scat genotype error, and greatly limited the amount of additional laboratory work that was needed to identify potentially 'false' scores. 'Acceptable' scat genotypes revealed information on sex ratio, relatedness, fox movement patterns, latrine use, and size of home range. Results from genetic and field data were consistent, supporting the conclusion that our data set had a very low rate of genotyping error and that this noninvasive method is a reliable approach for monitoring kit foxes.     

Testing the reliability of noninvasive genetic sampling by comparing analyses of blood and fecal samples in Barbary macaques (Macaca sylvanus).

Genetic studies of wild animal populations are often hindered by difficulties in obtaining blood samples. Recent advances in molecular biology have allowed the use of noninvasive samples as sources of DNA (e.g., hair or feces), but such samples may provide low-quality DNA and prevent the determination of true genotypes in subsequent DNA analysis. We present a preliminary study aimed at assessing the reliability of using fecal samples for genotyping in Barbary macaques (Macaca sylvanus). The test was performed on samples of blood and feces from 11 captive animals, using three dinucleotide microsatellites. The CTAB DNA extraction method was found to be the most relevant for Barbary macaque feces, yielding successful amplification at all loci for 70% of PCRs. All the fecal samples tested gave correct genotypes at least once for each locus when referenced against blood-derived genotypes. An average of 18.3% of PCRs displayed spurious genotypes (false homozygous or false allele). The minimum theoretical probability required to obtain a 100% accurate genotype is 0.74, based on the criterion that a correct genotype is assessed only if it was observed at least twice. The observed probability of obtaining a correct genotype from three PCRs, based on our genotyping results, was greater (0.81 on average) than the minimum threshold. In conclusion, our comparison of blood and fecal samples showed that fecal sampling is a reliable tool for the further study of wild Barbary macaque populations.     

水生态服务功能分析及其间接价值评价

水生态系统服务功能是指水生态系统及其生态过程所形成及所维持的人类赖以生存的自然环境条件与效用。它不仅是人类社会经济的基础资源 ,还维持了人类赖以生存与发展的生态环境条件。根据水生态系统提供服务的机制、类型和效用 ,把水生态系统的服务功能划分为提供产品、调节功能、文化功能和生命支持功能四大类。水生态系统提供的产品主要包括人类生活及生产用水、水力发电、内陆航运、水产品生产、基因资源等 ;调节功能主要包括水文调节、河流输送、侵蚀控制、水质净化、空气净化、区域气候调节等 ;文化功能主要包括文化多样性、教育价值、灵感启发、美学价值、文化遗产价值、娱乐和生态旅游价值等 ;支持功能是上述服务功能产生的基础 ,其对人类的影响是间接的并且需要经过很长时间才能显现出来。评价中 ,将我国陆地水生态系统分为河流、水库、湖泊、沼泽四个类型 ,结合基础数据的可获得性 ,建立了由调蓄洪水、疏通河道、水资源蓄积、土壤持留、净化环境、固定碳、提供生境、休闲娱乐 8项功能构成的水生态系统间接价值评价指标体系并进行了初步的评价与估算 ,结果表明其总的价值为 6 0 38.78× 10 8元 ,相当于供水、发电、航运、水产品生产等水生态系统提供的直接使用价值的 1.6倍     

Comparison of Campylobacter jejuni PFGE and Penner subtypes in human infections and in water samples from the Taieri River catchment of New Zealand

AIMS: To determine the degree of overlap in strain types of Campylobacter jejuni isolated from clinical cases and water samples from the Taieri catchment in the South Island of New Zealand. METHODS AND RESULTS: Thermophilic Campylobacter were collected from human cases of infection, the main stem of the Taieri River and streams within distinct land-use types over a 1-year period. Campylobacter jejuni (187 isolates) and Campylobacter lari (four isolates) were identified using a multiplex polymerase chain reaction protocol. Isolates were typed by the Penner method and pulsed-field gel electrophoresis (PFGE) utilizing two restriction endonucleases. Several serotypes and PFGE types occurred in both water samples and clinical cases when the restriction profiles for each enzyme were considered separately. However, when PFGE profiles and serotyping were combined, there was no overlap between Camp. jejuni types from water and clinical cases. CONCLUSIONS: The results of this study indicate that recreational water in the Taieri catchment is not a major source of campylobacteriosis in the Dunedin area. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests the risk of acquiring campylobacteriosis from surface waters in the Taieri catchment is considerably lower than previously predicted and highlights the necessity of using two endonucleases in PFGE typing.     

In situ sampling of interstitial water from lake sediments

A sampler with a relatively high resolution has been developed, which allows interstitial water to be obtained from lake sediments at well defined depths, without serious disturbance of sediment structure. Oxidation effects are excluded. Sampling time is in the order of a day. Installation requires little additional equipment. The instrument has been developed for use in shallow lakes.     

Comparison of automatic and conventional physical and chemical analyses in Lake Saimaa,Eastern Finland

Hydrobiologia - The Saimaa lake complex (4460&;nbsp;km2) is a mosaic of interconnected basins draining into the Gulf of Finland via Lake Ladoga. Limnologically, most of the basins are...     

矿泉水和山泉水中铜绿假单胞菌污染调查及分离菌株毒力基因与耐药性分析

【目的】初步掌握全国矿泉水和山泉水生产过程中铜绿假单胞菌(Pseudomonas aeruginosa)的污染情况。分析矿泉水与山泉水中铜绿假单胞菌的致病性与耐药性。【方法】研究通过对广西、湖北、云南等全国9个省36家水厂进行采样,共采集108个样本,并根据《饮用天然矿泉水检测方法》国家标准(GB/T 8538-2008)检测其铜绿假单胞菌的污染率、污染水平。对分离出的铜绿假单胞菌菌株进行毒力基因与药敏实验。【结果】全国矿泉水水源水、活性碳过滤后水、成品水的污染率分别为16.7%、16.7%、0,污染水平分别为3.7、2.0、0 CFU/250 m L。全国山泉水水源水、活性碳过滤后水、成品水的污染率分别为66.7%、83.3%、5.6%,污染水平分别为5.1、7.3、2.0 CFU/250 m L。对所分离出的36株铜绿假单胞菌进行毒力基因检测和药敏试验显示:exo U、exo S、phz M、tox A、las B检出率分别为25.0%、75.0%、100%、88.8%、100%,但对美国国家临床实验室标准化委员会标准中14种抗生素均无耐药性。【结论】山泉水水源水、活性碳过滤后水、成品水污染率明显高于矿泉水,但污染水平均较低,无大于40.0 CFU/250 m L样品检出。山泉水活性碳过滤后污染率最高,表明大部分企业在活性碳过滤环节存在污染问题。毒力基因exo U、exo S、phz M、tox A、las B在分离到的36株铜绿假单胞菌检出率高,但分离到的菌株对所选取的14种抗生素均无耐药性。     

土地利用、溪流级别与溪流河水理化性质的关系

为研究土地利用对溪流水质的影响 ,对黑龙江省尚志市帽儿山境内阿什河流域的 1～ 5级溪流河水进行采样 ,同时记载森林覆盖度、邻近土地利用类型及地貌特征。对溪水样品的的 p H,浑浊度 ,NH 4- N、NO- 3- N和 PO3 3- P浓度进行分析测定。对不同溪流级别和土地利用类型的溪流河水进行分析。结果表明 :随着溪流级别的升高和土地利用类型由森林变为农田和村镇 ,溪流河水的 p H,浑浊度 ,NH 4- N、NO- 3- N和 PO3- 3- P浓度均显著升高。森林植被保存相对较好的 1、2级溪流水质最佳 ,但溪水中溶解无机磷和无机氮的浓度与美国温带地区相似的低级溪流相比 ,分别高 30倍和 1 0 0倍。河岸带破坏 ,土地利用的变化 ,放牧、樵采等频繁干扰造成的土壤侵蚀和养分流失输入是造成该流域溪流富营养化的主要原因     

Detection of Legionella pneumophila in water samples by species-specific real-time and nested PCR assays

AIMS: Legionella pneumophila is a contaminant of man-made water systems, including potable water, cooling towers, water systems of large buildings, etc. It is the most common causative agent of legionellosis, a respiratory infection, which may give rise to restricted outbreaks. To survey environmental water samples from hospitals and private habitations in Bologna, we developed a species-specific nested and a TaqMan real-time PCR for the detection of L. pneumophila. We compared the two assays and both to cultural isolation. METHODS AND RESULTS: The targeted gene was macrophage infectivity potentiator (mip), conserved in L. pneumophila, and divergent in other legionellae. One assay was based on a nested PCR and the other on a TaqMan real-time PCR protocol. Their sensitivities were 14 % or 5% higher than that of cultural isolation respectively. The detection limits were 1-2 genome equivalents per 50-microl reaction. Specificity was assessed using DNA from nine target and 20 nontarget organisms. CONCLUSIONS: When applied to water samples, both assays detected L. pneumophila at 80% or higher frequency. SIGNIFICANCE AND IMPACT OF THE STUDY: The species-specific molecular diagnosis of L. pneumophila by means of nested PCR does not require a specific instrumentation, exhibits a high sensitivity, and is advantageous over the cultural isolation and real-time PCR detection. It allows to quickly monitor water samples for the risk assessment of environmental contaminations.     

Comparison of membrane filters for recovery of legionellae from water samples.

The procedure currently used for isolating legionellae from environmental samples recommend filtration through a 0.2-microns-pore-size polycarbonate filter. In this study we evaluated the performance of 23 other filters composed of various materials and having various pore sizes. We prefer the 0.2-micron-pore-size Gelman Supor filter because of its high level of recovery, faster filtration rate, and ease of handling.     

Hand-operated box sampler for shallow water

The construction and operation of a hand-operated box sampler is described. The sampler covers an area of 25 × 25 cm and penetrates into the sediment to the depth of 15 cm. It was successfully operated on a sandy bottom, and on sand mixed with gravel up to a water depth of 1 meter.IAEC, Yavne 70600, Israel