A modelling approach to the development of an active management strategy for the Queen Elizabeth II reservoir

This article outlines the outcome of work that set out to provide one of the specified integral contributions to the overarching objectives of the EU- sponsored LIFE98 project described in this volume. Among others, these included a requirement to marry automatic monitoring and dynamic modelling approaches in the interests of securing better management of water quality in lakes and reservoirs. The particular task given to us was to devise the elements of an active management strategy for the Queen Elizabeth II Reservoir. This is one of the larger reservoirs supplying the population of the London area: after purification and disinfection, its water goes directly to the distribution network and to the consumers. The quality of the water in the reservoir is of primary concern, for the greater is the content of biogenic materials, including phytoplankton, then the more prolonged is the purification and the more expensive is the treatment. Whatever good that phytoplankton may do by way of oxygenation and oxidative purification, it is eventually relegated to an impurity that has to be removed from the final product. Indeed, it has been estimated that the cost of removing algae and microorganisms from water represents about one quarter of its price at the tap. In chemically fertile waters, such as those typifying the resources of the Thames Valley, there is thus a powerful and ongoing incentive to be able to minimise plankton growth in storage reservoirs. Indeed, the Thames Water company and its predecessor undertakings, have a long and impressive history of confronting and quantifying the fundamentals of phytoplankton growth in their reservoirs and of developing strategies for operation and design to combat them. The work to be described here follows in this tradition. However, the use of the model PROTECH-D to investigate present phytoplankton growth patterns in the Queen Elizabeth II Reservoir questioned the interpretation of some of the recent observations. On the other hand, it has reinforced the theories underpinning the original design of this and those Thames-Valley storage reservoirs constructed subsequently. The authors recount these experiences as an example of how simulation models can hone the theoretical base and its application to the practical problems of supplying water of good quality at economic cost, before the engineering is initiated.

Mechanism of red colouration in Tovel-See (”Conclusions” only) [Translation from: Archiv fur Hydrobiologie 38 299-302, 1941]

Cases of red colouration in small lake basins, due to the abundant appearance of microorganisms have long been known. Usually it is caused by a fast, sudden, intensive propagation (so called ”bloom”) of Cyanophycae and bacteria. (e.g. Oscillatoracae, thiobacteria etc.). An exception to this is the red colouration of Tovel-See, an alpine lake basin in the Dolomites of the Brenta group (Trentino), lying at a height of 1178 m and hidden in the woodland of a valley. Here the red bloom has a double rhythm: a daily and a yearly rhythm. The colouration of one part of the lake takes place in the warmest months of the year (i.e. July, August, September) and in the middle hours of the day. The immediate origin of the bloom has been known for a long time: it is caused by the Peridinacae Glenodinium sanguineum. This paper describes the phenomenon of red colouration of the lake and discusses its conditions.

Radioautographic method of determination of number of bacteria in water-bodies [Translation from: Informatsionnyi Byulleten Biologiya Vnutrennikh Vod No.26, 65-67, 1975]

The radioautographic method of determination of the number of autotrophic microorganisms was initially suggested for counting methane-oxidizing bacteria. With the help of this method colonies of hydrogen-oxidizing bacteria are differentiated even more clearly from heterotrophic. Under laboratory conditions it was shown that colonies grown on membrane filters from a pure culture of thionic bacteria on a nutrient medium with radio- active carbonate, give better prints on film. This method was tested by the authors for determining the number of these bacteria in the meromictic Lake Vae de San Juan during the expedition to Cuba in the summer of 1973. The study showed that that the thionic bacteria are found throughout the pelagial. It proved that the thionic bacteria can be well considered in water-bodies by the radioautographic method.

The impact of molecular biology on assessment of water quality: advantages and limitations of current techniques

The advent of molecular biology has had a dramatic impact on all aspects of biology, not least applied microbial ecology. Microbiological testing of water has traditionally depended largely on culture techniques. Growing understanding that only a small proportion of microbial species are culturable, and that many microorganisms may attain a viable but non-culturable state, has promoted the development of novel approaches to monitoring pathogens in the environment. This has been paralleled by an increased awareness of the surprising genetic diversity of natural microbial populations. By targeting gene sequences that are specific for particular microorganisms, for example genes that encode diagnostic enzymes, or species-specific domains of conserved genes such as 16S ribosomal RNA coding sequences (rrn genes), the problems of culture can be avoided. Technical developments, notably in the area of in vitro amplification of DNA using the polymerase chain reaction (PCR), now permit routine detection and identification of specific microorganisms, even when present in very low numbers. Although the techniques of molecular biology have provided some very powerful tools for environmental microbiology, it should not be forgotten that these have their own drawbacks and biases in sampling. For example, molecular techniques are dependent on efficient lysis and recovery of nucleic acids from both vegetative forms and spores of microbial species that may differ radically when growing in the laboratory compared with the natural environment. Furthermore, PCR amplification can introduce its own bias depending on the nature of the oligonucleotide primers utilised. However, despite these potential caveats, it seems likely that a molecular biological approach, particularly with its potential for automation, will provide the mainstay of diagnostic technology for the foreseeable future.

Immunodetection of planktonic algae

Interest in the identification and characterisation of cyanobacteria and dinoflagellates in aquatic environments is increasing rapidly due to the perceived roles of these organisms in primary production and nuisance aspects in terms of water treatment and public health. Techniques for the identification and quantification of these organisms currently are limited, and the application of molecular approaches provides fundamental taxonomic information and techniques of practical value. Antigenic properties of algal cells may be useful taxonomic markers. Immunodetection techniques utilise the specificity of the antibody/antigen association as a probe for recognising and distinguishing between microorganisms according to their cell- surface chemistry. Immunofluorescent detection of unicellular cyanobacteria and dinoflagellates has been studied with success in marine and freshwater ecosystems and a range of techniques and results are presented and discussed. The most recent advances in the study of planktonic algae have come with the application of continuous flow cytometric methods (CFC). Flow cytometry makes use of the autofluorescence properties of the algal cells, which alone can be used to demonstrate their presence and permit their quantification in natural water samples. When used in conjunction with immunolabelling techniques, the potential of CFC analysis is broadened to study the serological/strain composition of plankters in natural populations. Changes in algal strains represented within and between waters over periods of time are reported and discussed, along with the ecological issues thus raised.

The diversity and ecological role of protozoa in fresh waters

Protozoa feed on and regulate the abundance of most types of aquatic microorganisms, and they are an integral part of all aquatic microbial food webs. Being so small, aerobic protozoa thrive at low oxygen tensions, where they feed (largely unaffected by metazoan grazing) on the abundance of other microorganisms. In anaerobic environments, they are the only phagotrophic organisms, and they live in unique symbiotic consortia with methanogens, sulphate reducers and non-sulphur purple bacteria. The number of extant species of protozoa may be quite modest (the global number of ciliate species is estimated at 3000), and most of them probably have cosmopolitan distributions. This will undoubtedly make it easier to carry out further tasks, e.g. understanding the role of protozoan species diversity in the natural environment.

The role of probiotics in aquaculture in Nigeria - a review

Aquaculture is beset by many problems especially diseases caused by bacteria as the major deteriorating factors. The use of vaccines and antimicrobial agents have been centered on disease control, but are associated with problems The development of antibiotic resistance among the microorganisms have become a global concern as a result of indiscriminate use of antibiotics. Several alternative suggestions for disease prevention have been on probiotics for its efficacy, low cost, less side effects and accessible to farmers. Probiotics is gaining a high priority in the developed countries with the aim of replacing conventional drugs. The principal bacterial groups tested as probiotic bacteria in culture of shrimps, crabs, oysters, fish and humans are Vibrio, Pseudomonas, Bacillus, Bifidobacteria and several Lactobacilli. Experiments have mainly been conducted with fish larvae, adult fish, crustaceans and animals where significant reduction in mortalities has been obtained. The purpose of this review is to create awareness of the role of probiotics in disease control in aquaculture as alternative to antibiotics.

The potential for crop rotation in controlling diseases in shrimp culture

The use of antibiotics and other chemicals in controlling shrimp pathogens become ineffective as the strains grow more resistant to these chemicals. Moreover, the bacterial pathogen (Vibrio harveyi) produced biofilm coating that protects it from dying and disinfection procedures that are followed during pond preparation. Biological control is being considered as an alternative means of preventing shrimp disease outbreak. The main principle behind biological control is to enhance the growth of beneficial microorganisms which serve as antagonists or target pathogens. The paper discusses shrimp and tilapia crop rotation as a form of effective biological control, a technique which is already being practiced in Indonesia and the Philippines.

Saline-saturated DMSO-EDTA as a storage medium for microbial DNA analysis from coral mucus swab samples

The mucus surface layer of corals plays a number of integral roles in their overall health and fitness. This mucopolysaccharide coating serves as vehicle to capture food, a protective barrier against physical invasions and trauma, and serves as a medium to host a community of microorganisms distinct from the surrounding seawater. In healthy corals the associated microbial communities are known to provide antibiotics that contribute to the coral’s innate immunity and function metabolic activities such as biogeochemical cycling. Culture-dependent (Ducklow and Mitchell, 1979; Ritchie, 2006) and culture-independent methods (Rohwer, et al., 2001; Rohwer et al., 2002; Sekar et al., 2006; Hansson et al., 2009; Kellogg et al., 2009) have shown that coral mucus-associated microbial communities can change with changes in the environment and health condition of the coral. These changes may suggest that changes in the microbial associates not only reflect health status but also may assist corals in acclimating to changing environmental conditions. With the increasing availability of molecular biology tools, culture-independent methods are being used more frequently for evaluating the health of the animal host. Although culture-independent methods are able to provide more in-depth insights into the constituents of the coral surface mucus layer’s microbial community, their reliability and reproducibility rely on the initial sample collection maintaining sample integrity. In general, a sample of mucus is collected from a coral colony, either by sterile syringe or swab method (Woodley, et al., 2008), and immediately placed in a cryovial. In the case of a syringe sample, the mucus is decanted into the cryovial and the sealed tube is immediately flash-frozen in a liquid nitrogen vapor shipper (a.k.a., dry shipper). Swabs with mucus are placed in a cryovial, and the end of the swab is broken off before sealing and placing the vial in the dry shipper. The samples are then sent to a laboratory for analysis. After the initial collection and preservation of the sample, the duration of the sample voyage to a recipient laboratory is often another critical part of the sampling process, as unanticipated delays may exceed the length of time a dry shipper can remain cold, or mishandling of the shipper can cause it to exhaust prematurely. In remote areas, service by international shipping companies may be non-existent, which requires the use of an alternative preservation medium. Other methods for preserving environmental samples for microbial DNA analysis include drying on various matrices (DNA cards, swabs), or placing samples in liquid preservatives (e.g., chloroform/phenol/isoamyl alcohol, TRIzol reagent, ethanol). These methodologies eliminate the need for cold storage, however, they add expense and permitting requirements for hazardous liquid components, and the retrieval of intact microbial DNA often can be inconsistent (Dawson, et al., 1998; Rissanen et al., 2010). A method to preserve coral mucus samples without cold storage or use of hazardous solvents, while maintaining microbial DNA integrity, would be an invaluable tool for coral biologists, especially those in remote areas. Saline-saturated dimethylsulfoxide-ethylenediaminetetraacetic acid (20% DMSO-0.25M EDTA, pH 8.0), or SSDE, is a solution that has been reported to be a means of storing tissue of marine invertebrates at ambient temperatures without significant loss of nucleic acid integrity (Dawson et al., 1998, Concepcion et al., 2007). While this methodology would be a facile and inexpensive way to transport coral tissue samples, it is unclear whether the coral microbiota DNA would be adversely affected by this storage medium either by degradation of the DNA, or a bias in the DNA recovered during the extraction process created by variations in extraction efficiencies among the various community members. Tests to determine the efficacy of SSDE as an ambient temperature storage medium for coral mucus samples are presented here.

PCR-based assay for detection of four coral pathogens

Several microorganisms have been identified as pathogenic agents responsible for various outbreaks of coral disease. Little has been learned about the exclusivity of a pathogen to given disease signs. Most pathogens have only been implicated within a subset of corals, leaving gaps in our knowledge of the host range and geographic extent of a given pathogen. PCR-based assays provide a rapid and inexpensive route for detection of pathogens. Pathogen-specific 16S rDNA primer sets were designed to target four identified coral pathogens: Aurantimonas coralicida, Serratia marcescens, Vibrio shilonii, and Vibrio coralliilyticus. Assays detected the presence of targets at concentrations of less than one cell per microliter. The assay was applied to 142 coral samples from the Florida Keys, Puerto Rico, and U.S. Virgin Islands as an in situ specificity test. Assays displayed a high-level of specificity, seemingly limited only by the resolution of the 16S rDNA.

Cloning and expression of zebra fish ferroportin and investigating its influence on anemia

Iron is required for many microbes and pathogens for their survival and proliferation including Leishmania which cause leishmaniasis. Leishmaniasis is an increasingly serious infectious disease with a wide spectrum of clinical manifestations. These range from localized cutaneous leishmaniasis (CL) lesions to a lethal visceral form. Certain strains such as BALB/c mice fail to control L. major infection and develop progressive lesions and systemic disease. These mice are thought to be a model of non-healing forms of the human disease such as kala-azar or diffuse cutaneous leishmaniasis. Progression of disease in BALB/c mice has been associated with the anemia, in last days of their survival, the progressive anemia is considered to be one of the reasons of their death. Ferroportin (Fpn), a key regulator of iron homeostasis is a conserved membrane protein that exports iron across the duodenal enterocytes as well as macrophages and hepatocytes into the blood circulation. Fpn has also critical influence on survival and proliferation of many microorganisms whose growth is dependent upon iron, thus preparation of Fpn is needed to study the role of iron in immune responses and pathogenesis of micoorganisms. To prepare and characterize a recombinant ferroportin, total RNA was extracted from Indian zebrafish duodenum, and used to synthesize cDNA by RT-PCR. PCR product was first cloned in Topo TA vector and then subcloned into the GFP expression vector pEGFP–N1. The final resulted plasmid (pEGFP-ZFpn) was used for expression of FPN-EGFP protein in Hek 293T cells. The expression was confirmed by fluorescence microscopy and flow cytometery. Recombinant Fpn was further characterized by submission of its predicted amino acid sequences to the TMHMM V2.0 prediction server (hidden Markov model), NetOGlyc 3.1 server and NetNGlyc 3.1 server. Data emphasised that obtained Fpn from indian zebrafish contained eight transmembrane domains with N- and C-termini inside the cytoplasm and harboured 78 mucin-type glycosylated amino acid. The results indicate that the prepared and characterized recombinant Fpn protein has no membrane topology difference compared to other Fpn described by other researcher. Our next aim was to deliver recombinant plasmid (pEGFP-ZFpn) to entrocyte cells. However, naked therapeutic genes are rapidly degraded by nucleases, showing poor cellular uptake, nonspecificity to the target cells, and low transfection efficiency. The development of safe and efficient gene carriers is one of the prerequisites for the success of gene therapy. Chitosan and alginate 139 polymers were used for oral gene carrier because of their biodegradability, biocompatibility and their mucoadhesive and permeability-enhancing properties in the gut. Nanoparticles comprising Alginate/Chitosan polymers were prepared by pregel preparation method. The resulting nanoparticles had a loading efficiency of 95% and average size of 188 nm as confirmed by PCS method and SEM images had showed spherical particles. BALB/c mice were divided to three groups. The first and second group were fed with chitosan/alginate nanoparticles containing the pEGFP-ZFpn and pEGFP plasmid, respectively (30 μgr/mice) and the third group (control) didn’t get any nanoparticles. The result showed BALB/c mice infected by L.major, resulted in higher hematocryte and iron level in pEGFP-ZFpn fed mice than that in other groups. Consentration of cytokines determined by ELISA showed lower levels of IL-4 and IL-10 and higher levels of IFN-γ/IL-4 and IFN-γ/IL-10 ratios in pEGFP-ZFpn fed mice than that in other groups. Morover more limited increase of footpad thickness and significant reduction of viable parasites in lymph node was seen in pEGFP-ZFpn fed mice. The results showed the first group exhibited a highr hematocryte and iron compared to the other groups. These data strongly suggests the in vivo administration of chitosan/alginate nanoparticles containing pEGFP-ZFpn suppress Th2 response and may be used to control the leishmaniasis .

Application of encapsulated nisin and sodium citrate for shelf life increasing of Rainbow trout (Onchorhynchus mykiss) fillet packaged with MAP and their effect on Staphylococcus aureus

Nisin is a widely used naturally occurring antimicrobial effective against many pathogenic and spoilage microorganisms. It has been proposed that reduced efficacy of nisin in foods can be improved by technologies such as encapsulation to protect it from interferences by food matrix components. The aim of this study was using of spray dried encapsulated nisin with zein in concentration of (0.15 and 0.25 g/kg) and sodium citrate (1.5 and 2.5%) and treatments with both of them to extent the shelf life of filleted trouts packaged by Modified Atmosphere Packaging (45% CO2, 50% N2 ,5% O2) and stored at 4±1 °C for 20 days. Furthermore, to evaluate the antimicrobial efficiency of encapsulated nisin and soudium citrate the trouts fillets was inoculated with Staphylococcus aureus as an index pathogenic bacteria. Assessment of chemical spoilage indexes such as (Proxide value, Thiobarbituric acid, total volatile base nitrogen and pH) , microbial parameters (Total Plate Count, Psychrotrophic count, Lactic acid bacteria count), Staphylococcus aureus cont in treatments which were inoculated with 5 logcfu/g of this bacteria and sensory evaluation of fillets including (smell, color, texture and total acceptability) was carried out in days of 0, 4, 8, 12, 16 and 20. The results revealed that treatment with both exposure of nisin and sodium citrate showed significantly lower chemical spoilage indexes in comparison with controls (vaccum packed and MAP) (P<0.05). Furthermore, (nisin 0.25 g/kg sodium citrate 2.5%) treatment which was exposed to the maximal level used of both materials was significantly the lowest treatment with (Proxide value, Thiobarbituric acid, total volatile base nitrogen and pH) of 9.95 (meq O2/kg) , 1.55 (mgMA/kg), 29.65 (mgN/100g) and 6.65 , respectively and according to the maximal recommended level of this indices , shelf life of fillets in this treatment was esstimated 20 days.The control (vaccum packed) treatment was significantly the highest treatment with (Proxide value, Thiobarbituric acid, total volatile base nitrogen and pH) of 15.17 (meq O2/kg), 3.03 (mgMA/kg), 38.4 (mgN/100g) and 6.95 , respectively and according to the maximal recommended level of this indices , shelf life of fillets in this treatment was estimated 11 days. Also, in microbial point of view (nisin 0.25 g/kg- sodium citrate 2.5%) treatment was the lowest treatment with Total Plate Count, Psychrotrophic count, Lactic acid bacteria count and Staphylococcus aureus count of 6.7, 6.83, 5.25 and 6.04 logcfu/g respectively, and conrol (vaccum packed) treatment was the highest treatment with 9.15, 9.41, 7.7 and 9.01 logcfu/g respectively. According to the lower results of chemical and microbial indices and higher sensory evaluated scores assessed in this research for encapsulated nisin in comparison with free nisin , it was concluded that encapsulation of nisin with zein capsules may improve the efficiency of nisin. The measuremented values of Mass yield, Total solids content of capsules, Encapsulation efficiency, In vitro release kinetics in 200 hour for encapsulated nisin in this study was 49.89, 62, 98.31 and 69% respectively and Encapsulated particle size was lower than 674.21 μm for 90% of particles. As a consequence, nisin , in particular encapsulated nisin, and sodium citrate alone or together with and Modified Atmosphere packaging might be considered as effective tools in preventing the quality degradation of the fillets, resulting in an extension of their shelf life.