Oil production by six microalgae: impact of flocculants and drying on oil recovery from the biomass

Oil production in batch photoautotrophic cultures of the following microalgae is reported: the freshwater microalgae Chlorella vulgaris, Choricystis minor, and Neochloris sp.; the marine microalgae Nannochloropsis salina and Cylindrotheca fusiformis; and C. vulgaris grown in a full-strength seawater medium. In all cases, the solvent extraction of lipids from freeze-dried biomass is compared with extraction from the fresh biomass paste. For all algae, the oils could be extracted equally effectively from freeze-dried samples and the paste samples (67–88 % moisture by weight). Moisture content determinations of the biomass using the freeze-drying method and the high-temperature oven drying were found to be equivalent for all algae. The biomass recovered by flocculation with metal salts (aluminum sulfate, ferric chloride) followed by centrifugation had a certain amount of the flocculant irreversibly bound to it. Washing failed to remove the adsorbed flocculants. For all algae, the adsorbed flocculants did not interfere with oil recovery by solvent extraction. The solvent system of chloroform–methanol–water proved highly effective for quantitative extraction of the lipids from all algae.     

Changes in defense allomone compositions of <Emphasis Type="Italic">Eutrichodesmus elegans</Emphasis> and <Emphasis Type="Italic">Eutrichodesmus</Emphasis><Emphasis Type="Italic">armatus</Emphasis> (Polydesmida: Haplodesmidae) during different stages of their life cycles

A mixture of E- and Z-(2-nitrovinyl)benzenes is a known allomone of two adult haplodesmid millipedes, Eutrichodesmus elegans (Miyosi) (Polydesmida: Haplodesmidae) and Eutrichodesmus armatus (Miyosi), as is (2-nitroethyl)benzene in E. armatus. However, the proportions of these compounds have not yet been studied in detail at the nymph stage. In the present study, the ratios of these three nitro compounds were shown to change during ontogenetic development. (2-Nitroethyl)benzene was newly detected as the second major component of the mixture in both species at stage I, just after eggs hatched (mean 43.0% in E. armatus and 7.8% in E. elegans), decreasing rapidly to less than 0.1% during growth. These changes occurred in a species-specific manner; field-collected E. armatus maintained a characteristic mixture of E- and Z-(2-nitrovinyl)benzenes (59.9–98.2 and 40.0–1.4%, respectively) during all stages including the adult stage. On the other hand, E. elegans contained E-(2-nitrovinyl)benzene as the major component (98.7–99.7%) with Z-(2-nitrovinyl)benzene as a trace component (less than 1.2%), while a minute amount of (2-nitroethyl)benzene was always retained during all nymph and adult stages. No volatiles were detected in eggs before hatching, and sequential changes of composition were observed among the three compounds after emergence in both species.     

Metatrancriptomic analysis from the Hepatopancreas of adult white leg shrimp (<Emphasis Type="Italic">Litopenaeus vannamei)</Emphasis>

The amoeba, Mayorella viridis contains several hundred symbiotic green algae in its cytoplasm. Transmission electron microscopy revealed strong resemblance between symbiotic algae from M. viridis the symbiotic Chlorella sp. in the perialgal vacuoles of Paramecium bursaria and other ciliates. Although it is thought that the M. viridis and symbiotic algae could be model organisms for studying endosymbiosis between protists and green algae, few cell biological observations of the endosymbiosis between M. viridis and their symbiotic algae have been published. In this study, we characterized the specificity of endosymbiotic relationships between green algae and their hosts. Initially, we established stable cultures of M. viridis in KCM medium by feeding with Chlorogonium capillatum. Microscopic analyses showed that chloroplasts of symbiotic algae in M. viridis occupy approximately half of the algal cells, whereas those in P. bursaria occupy entire algal cells. The symbiotic algae in P. bursaria contain several small spherical vacuoles. The labeling of actin filaments using Acti-stain? 488 Fluorescent Phalloidin revealed no relationship between host actin filaments and symbiotic algal localization, although the host mitochondria were localized around symbiotic algae. Symbiotic algae from M. viridis could infect algae-free P. bursaria but could not support P. bursaria growth without feeding, whereas the original symbiotic algae of P. bursaria supported its growth without feeding. These data indicated the specificity of endosymbiotic algae relationships in M. viridis and P. bursaria.     

Microalgal Biomass and Lipid Production on Dairy Effluent Using a Novel Microalga, <Emphasis Type="Italic">Chlorella</Emphasis> sp. Isolated from Dairy Wastewater

The present study focused on cost-effective production of microalgal biomass and lipid production on dairy effluent. The novel microalga, Chlorella sp. isolated from the dairy effluent showed high growth and lipid production on the undiluted and two-fold diluted dairy effluent which were four to five times higher than those of Chlorella vulgaris (control). The high growth of Chlorella sp. was thought to be possibly due to its heterotrophic growth capacity, high turbidity, COD, nutrients and trace elements. In contrast, C. vulgaris showed poor heterotrophic and photoautotrophic growth under the highly turbid conditions of dairy effluent. Both Chlorella sp. and C. vulgaris showed similar total FAME (mg FAME/g algal cells). The fatty acid composition analysis revealed that both Chlorella sp. and C. vulgaris possessed major C18 and C20 fatty acids which will be used for biodiesel production. Overall, the novel microalga, Chlorella sp. isolated from the dairy effluent showed high potential for cost-effective algal cultivation and lipid production on dairy effluent without any modification of process.     

Diversity and enzyme activity of <Emphasis Type="Italic">Penicillium</Emphasis> species associated with macroalgae in Jeju Island

A total of 28 strains of 19 Penicillium species were isolated in a survey of extracellular enzyme-producing fungi from macroalgae along the coast of Jeju Island of Korea. Penicillium species were identified based on morphological and β-tubulin sequence analyses. In addition, the halo-tolerance and enzyme activity of all strains were evaluated. The diversity of Penicillium strains isolated from brown algae was higher than the diversity of strains isolated from green and red algae. The commonly isolated species were Penicillium antarcticum, P. bialowiezense, P. brevicompactum, P. crustosum, P. oxalicum, P. rubens, P. sumatrense, and P. terrigenum. While many strains showed endoglucanase, β-glucosidase, and protease activity, no alginase activity was detected. There was a positive correlation between halo-tolerance and endoglucanase activity within Penicillium species. Among 19 Penicillium species, three species–P. kongii, P. olsonii, and P. viticola–have not been previously recorded in Korea.     

Effects of using cyanobacteria and fertilizer on growth and yield of rice,Pathum Thani I: a pot experiment

The effects of cyanobacteria and chemical fertilizer on growths and yields of rice (Oryza sativa L.) cv. Pathum Thani 1 were studied in pot trials. Nine treatments were set up without cyanobacteria and fertilizer (control), inoculated with cyanobacteria Nostoc carneum TUBT04 (T1), inoculated with Nostoc commune TUBT05 (T2), and inoculated with combination of N. carneum TUBT04 and N. commune TUBT05 (T3), full dose of recommended use of fertilizer (T4), half dose of recommended use of the fertilizer (T5), half dose of fertilizer combined with N. carneum TUBT04 (T6), half dose of fertilizer combined with N. commune TUBT05 (T7), and half dose of fertilizer combined with N. carneum TUBT04 plus N. commune TUBT05 (T8). Each treatment divides into five replications. Growth parameters including length and fresh and dry weights of shoot and root were measured in the seedlings. The number of spikes per plant and the amount and weight of rice grains per spike were measured after harvesting. The supplement with cyanobacteria promoted rice seedling growth and yield compared to the control treatment. Inoculation with cyanobacteria showed significant increase in root length (p = 0.41). In addition, the combined biofertilizer with a half of the recommended dose of chemical fertilizer significantly enhanced rice production including total number and weight of grain per spike and a total weight of 100 grains (p < 0.001). Therefore, using a half dose of chemical fertilizer with cyanobacteria is suggested to decrease rice production cost of farmers without any effects on rice quantity and quality.     

Influential environmental gradients and patterns of fish assemblages in a West African basin

Twenty-two sites were monitored in the Lower Sassandra River Basin (Côte d’Ivoire, West Africa) to examine patterns in fish species assemblage structure along environmental gradients. Hierarchical clustering of the species presence–absence data identified four types of assemblages corresponding to the man-made Lake Buyo, the main channel, and the two major tributaries. Canonical Correspondence Analysis (CCA) indicated that canopy closure, leaves-wood, aquatic plants, temperature, width, total dissolved solids and depth were the environmental factors most strongly correlated with variation in the fish assemblages. Five species: Papyrocranus afer, Micralestes elongatus, Parailia pellucida, Tilapia busumana and T. mariae were reported from the basin for the first time. The investigation of the variation in species richness led to the assessment of a major anthropogenic disturbance induced by a palm oil factory. In conjunction with the fact that the Sassandra River hosts an endemic species, Synodontis koensis, it was concluded that this basin is of high conservation priority.     

Dark adaptation and conformations of carotenoids in the cells of <Emphasis Type="Italic">Cladophora aegagropila</Emphasis> (L). Rabenh.

Intact cells of freshwater algae Cladophora aegagropila (L). Rabenh. (synonymous to Aegagropila linnaei Kutz.) were investigated by resonance Raman spectroscopy. It was found that incubation in the dark (up to 24 h) leads to changes in the Raman spectroscopy spectrum of this species, namely to changes in the ratio of amplitudes of the I₁₅₂₃/I₁₁₅₅ and I₉₆₀/I₁₀₀₄ bands and in the half width of band in the region of 1523 cm–1. We suggested that the adaptation of algae to the dark alters the conformation of the molecule of the carotenoid by delocalization of π-electrons in the polyene chain of the molecule and changes the orientation of the ring. Moreover, the composition of carotenoids, as well as their location in the cell and microenvironment in the pigment–protein complexes can change: in the absence of illumination, the distribution of carotenoids in algal cells is more uniform. These changes are probably caused either by changes in the location of cell organelles or by carotenoid redistribution between photosynthetic membranes, plastoglobules, and lipophilic formations in the cytoplasm.     

The biogeochemical background and trace metal accumulation by brown algae of the genus <Emphasis Type="Italic">Fucus</Emphasis> in coastal waters of the Sea of Japan,the Sea of Okhotsk,and the White Sea

The ranges of background concentrations of Fe, Mn, Zn, Cu, and Cd in Fucus algae from the Sea of Japan, the Sea of Okhotsk, and the White Sea were determined. The lower background threshold of element concentration was calculated as Me₁₅–2MAD₁₅, the upper background threshold was determined as Me+2MAD. The upper background threshold of the metal contents in Fucus algae can be used as the maximum permissible regional concentration in assessing pollution levels of marine habitats. Comparison of the Me₁₅–2MAD₁₅ values showed that Fucus algae of the White Sea had increased Mn and decreased Cd concentrations compared to those in Pacific algae. The concentrations of Zn were higher in Fucus algae of the Sea of Japan than in macrophytes from the White Sea and the Sea of Okhotsk. The background concentration range of Fe in Fucus algae of the Sea of Japan was much narrower compared to that in their counterparts from the White Sea and the Sea of Okhotsk. The background metal concentrations in Fucus algae are specific to a sea region; their variations are connected not only with the trace element contents in abiotic components of the environment, but also with the functioning of coastal ecosystems.     

Plastid Genome of <Emphasis Type="Italic">Dictyopteris divaricata</Emphasis> (Dictyotales,Phaeophyceae): Understanding the Evolution of Plastid Genomes in Brown Algae

Dictyotophycidae is a subclass of brown algae containing 395 species that are distributed worldwide. A complete plastid (chloroplast) genome (ptDNA or cpDNA) had not previously been sequenced from this group. In this study, the complete plastid genome of Dictyopteris divaricata (Okamura) Okamura (Dictyotales, Phaeophyceae) was characterized and compared to other brown algal ptDNAs. This plastid genome was 126,099 bp in size with two inverted repeats (IRs) of 6026 bp. The D. divaricata IRs contained rpl21, making its IRs larger than representatives from the orders Fucales and Laminariales, but was smaller than that from Ectocarpales. The G + C content of D. divaricata (31.19%) was the highest of the known ptDNAs of brown algae (28.94–31.05%). Two protein-coding genes, rbcR and rpl32, were present in ptDNAs of Laminariales, Ectocarpales (Ectocarpus siliculosus), and Fucales (LEF) but were absent in D. divaricata. Reduced intergenic space (13.11%) and eight pairs of overlapping genes in D. divaricata ptDNA made it the most compact plastid genome in brown algae so far. The architecture of D. divaricata ptDNA showed higher similarity to that of Laminariales compared with Fucales and Ectocarpales. The difference in general features, gene content, and architecture among the ptDNAs of D. divaricata and LEF clade revealed the diversity and evolutionary trends of plastid genomes in brown algae.     

Composition of essential oil of <Emphasis Type="Italic">Artemisia scoparia</Emphasis> Waldst. et Kit. from Buryatia and Mongolia

Essential oil from the epigeal portion of Artemisia scoparia Waldst. et Kit. collected in different regions of Buryatia and Mongolia has been isolated using steam distillation. The yield of essential oil from the stems and leaves of A. scoparia collected in different regions of Buryatia and in the Selenga aymag of Mongolia ranged from 0.06 to 0.40%, that is, the variation was rather high. The composition of the oil was analyzed using chromatography with mass spectrometric detection on a gas chromatography device Agilent Packard HP 6890 N with a quadrupole mass spectrometric detector HP MSD 5973. The total number of compounds detected exceeded 100. The major components of Artemisia scoparia oil were the following: p-cymene (0.6–15.2%), limonene (0.1–6.3%), α-pinene (0.2–10.1%), β-pinene (0.4–8.9%), trans-β-ocimene (0.3–5.4%), caryophyllene (4.6–13.8%), germacrene D (11.5–40.3%), spathulenol (4.0–11.7%), and caryophyllene oxide (4.3–15.6%). Several chemotypes of essential oil from A. scoparia were identified according to the nature of the major components: 1) oil that contained acetylene hydrocarbons (from plants collected in Gilan, Mazandaran, Kashan, and Tigran provinces of Iran, Tajikistan, and the European part of the CIS); 2) oil that contained monoterpenes and aromatic compounds (from plants collected in the Khorasan province of Iran, India, and South Korea); and 3) oil that contained monoterpenes or aromatic and sesquiterpene compounds (from plants collected in Kazakhstan and Mongolia). The major components of A. scoparia essential oil obtained from Buryatian plants belonged to the classes of mono- and sesquiterpene compounds, and thus the oil can be assigned to the third chemotype characteristic of A. scoparia from geographically close regions.     

Effectiveness of cyanobacteria and green algae in enhancing the photosynthetic performance and growth of willow (<Emphasis Type="Italic">Salix viminalis</Emphasis> L.) plants under limited synthetic fertilizers application

The physiological response of plants to triple foliar biofertilization with cyanobacteria and green algae under the conditions of limited use of chemical fertilizers was investigated. Triple foliar biofertilization with intact cells of Microcystis aeruginosa MKR 0105, Anabaena sp. PCC 7120, and Chlorella sp. significantly enhanced physiological performance and growth of plants fertilized with a synthetic fertilizer YaraMila Complex (1.0, 0.5, and 0.0 g per plant). This biofertilization increased the stability of cytomembranes, chlorophyll content, intensity of net photosynthesis, transpiration, stomatal conductance, and decreased intercellular CO2 concentration. Applied monocultures augmented the quantity of N, P, K in plants, the activity of enzymes, such as dehydrogenases, RNase, acid or alkaline phosphatase and nitrate reductase. They also improved the growth of willow plants. This study revealed that the applied nontoxic cyanobacteria and green algae monocultures have a very useful potential to increase production of willow, and needed doses of chemical fertilizers can be reduced.     

Effect of salt stress on the antimicrobial activity of <Emphasis Type="Italic">Ruta chalepensis</Emphasis> essential oils

In the current investigation, the biological activities of essential oils obtained from organs of Ruta chalepensis plants grown under salt stress (0, 50 and 100 mM NaCl) were analyzed. Their chemical composition was often investigated by GC/FID and GC–MS and the antimicrobial activities towards eight bacteria (Salmonella All, Salmonella K, Escherichia coli 45AG, Escherichia coli 45AI, Staphylococcus aureus 9402, Staphylococcus aureus 02B145, Listeria 477 and Pseudomonas aeruginosa ATCC 10145) and five fungi strains (Aspergillus, Saccharomycee crvisiale, Streptomyces griseus, Fusarium solani and Penicillium thomii) were studied. Results revealed that salt increased essential oil production in leaves at 50 and 100 mM NaCl. A total of 20 compounds were identified in leaves, undecan-2-one, nonan-2-one and geijerene being the dominant ones. In stems, 21 compounds were found; they were dominated by decan-2-one, geijerene, nonan-2-one and undecan-2-one. In contrast, roots exhibited a large variation with 25 volatile compounds and octyl acetate, methyl decanoate, phytyl acetate were the major ones. Salt stress induced significant antibacterial activity changes, mainly in leaves and stems. In leaves, the minimum inhibitory and bactericidal concentration decreased at 100 mM NaCl against Listeria 477, the two strains of E. coli (45AG and 45AI) and P. aeruginosa but it increased versus other bacteria. In stems, salt increased oil antibacterial activity against all strains except P. aeruginosa ATCC 10145. Root oil showed the least antibacterial activity under saline conditions versus Listeria 477 and P. aeruginosa ATCC 10145. As regards antifungal activity, NaCl reduced the antifungal activity of essential oils against the majority of fungi strains.     

Participation of the Green Algae <Emphasis Type="Italic">Ulvaria obscura</Emphasis> in Bioremediation of Sea Water from Oil Products

Participation of green algae in bioremediation of sea water from oil products (OPs) has been studied. Decrease in the content of OPs in water is accompanied by their accumulation by the alga Ulvaria obscura (Chlorophyta). It has been assumed that neutralization of OPs in Ulvaria occurs, similarly to Fucus vesiculosus (Phaeophyta), all over the thallome: destruction of OPs starts on the alga surface by epiphytic hydrocarbonoxidizing bacteria providing absorption and subsequent degradation of OPs by plant cells. The role of macrophytes in bioremediation of coastal areas from OPs is considerably higher than previously supposed.     

Antarctic red macroalgae: a source of polyunsaturated fatty acids

The n-3 and n-6 series of polyunsaturated fatty acids (PUFAs) are important for numerous metabolic functions that reduce the risk of inflammation, cancer, and heart diseases. These fatty acids (FAs) have been detected in high concentrations in some species of algae. The amount of the n-3 and n-6 PUFAs is closely associated with abiotic factors, such as solar radiation intensity, salinity, and temperature variation. However, abiotic influence on PUFA levels, along with the physiological function of these molecules in algae, remains inconclusive. In the present study, the quantities of the n-3 and n-6 in Antarctic red algae species Iridaea cordata, Palmaria decipiens, Plocamium cartilagineum, and Pyropia endiviifolia were determined. The lipids were extracted from the macroalgae according to the method followed by Bligh and Dyer (1959) and converted to methyl esters for further analysis using gas chromatography. The main n-6 PUFA identified and quantified in this study of Antarctic red algae were linoleic acid (18:2n-6), dihomo-γ-linolenic acid (20:3n-6), and arachidonic acid (20:4n-6). The eicosapentaenoic acid was detected at high concentrations in all species, with approximately 62.8% of total FA in P. endiviifolia, 75.4% in P. decipiens, 50.4% in I. cordata, and 20.1% in P. cartilagineum. The results corroborate those of the literature and show that PUFAs increased in red algae environments, with increased production of n-3 and n-6 PUFAs.     

The potential of azooxanthellate poriferan hosts to assess the fundamental and realized <Emphasis Type="Italic">Symbiodinium</Emphasis> niche: evaluating a novel method to initiate <Emphasis Type="Italic">Symbiodinium</Emphasis> associations

On coral reefs, Symbiodinium spp. are found in most cnidarian species, but reside in only a small number of sponge species. Of the sponges that do harbor Symbiodinium, most are found in the family Clionaidae, which represents a minor fraction of the poriferan diversity on a reef. Our goal was to determine whether Symbiodinium can be taken up by sponge hosts that do not typically harbor these algal symbionts, and then to follow the fate of any Symbiodinium that enter the intracellular space. We used the filter-feeding capacity of sponges to initiate intracellular interactions between sponge-specialist clade G Symbiodinium and six sponge species that do not associate with Symbiodinium. Using a pulse-chase experimental design, we determined that all of the species we examined captured Symbiodinium, and undamaged intracellular algae were found up to 1 h after inoculation. In a longer-term experiment, Symbiodinium populations in Amphimedon erina persisted in sponge cells for at least 5 d post-inoculation. While no evidence of digestion was detected, the population decreased exponentially after inoculation. We contrast these data with the characteristics of symbiont acquisition and establishment in Cliona varians, which normally harbors Symbiodinium. Explants from experimentally derived aposymbiotic sponges were placed in the field where they acquired Symbiodinium from ambient sources (i.e., we did not inoculate them as in the pulse-chase experiments). We began to detect Symbiodinium cells in C. varians after 12 d, and the algal population increased exponentially until densities approached those typically found in this host (after ~128 d). We discuss the implications of this work in light of growing interest in the evolution of specificity between hosts and symbionts, and the fundamental and realized niche of Symbiodinium.     

Biodegradation of <Emphasis Type="Italic">n</Emphasis>-alkanes on oil–seawater interfaces at different temperatures and microbial communities associated with the degradation

Oil biodegradation studies have mainly focused on microbial processes in dispersions, not specifically on the interfaces between the oil and the seawater in the dispersions. In this study, a hydrophobic adsorbent system, consisting of Fluortex fabrics, was used to investigate biodegradation of n-alkanes and microbial communities on oil–seawater interfaces in natural non-amended seawater. The study was performed over a temperature range from 0 to 20 °C, to determine how temperature affected biodegradation at the oil–seawater interfaces. Biodegradation of n-alkanes were influenced both by seawater temperature and chain-length. Biotransformation rates of n-alkanes decreased by reduced seawater temperature. Low rate coefficients at a seawater temperature of 0 °C were probably associated with changes in physical–chemical properties of alkanes. The primary bacterial colonization of the interfaces was predominated by the family Oceanospirillaceae at all temperatures, demonstrating the wide temperature range of these hydrocarbonoclastic bacteria. The mesophilic genus Oleibacter was predominant at the seawater temperature of 20 °C, and the psychrophilic genus Oleispira at 5 and 0 °C. Upon completion of n-alkane biotransformation, other oil-degrading and heterotrophic bacteria became abundant, including Piscirickettsiaceae (Cycloclasticus), Colwelliaceae (Colwellia), Altermonadaceae (Altermonas), and Rhodobacteraceae. This is one of a few studies that describe the biodegradation of oil, and the microbial communities associated with the degradation, directly at the oil–seawater interfaces over a large temperature interval.     

The ontogeny of the oribatid mite <Emphasis Type="Italic">Nanhermannia coronata</Emphasis> (Acari,Oribatida, Nanhermanniidae)

The ontogeny of the oribatid mite Nanhermannia coronata (Oribatida, Nanhermanniidae) was studied in the laboratory. At a temperature of 22–23°C and air humidity 100%, the entire life cycle from egg to adult lasted for 105–124 days. N. coronata juveniles and females are described. The diagnostic characters of the chaetom of N. coronata and N. nana larvae and nymphs are given. Morphological differences between N. coronata and N. nana allow distinguishing these species at all active stages.     

Morphological and molecular characterization of cyanobacteria from a Brazilian facultative wastewater stabilization pond and evaluation of microcystin production

The cyanobacterial population in the Cajati waste stabilization pond system (WSP) from São Paulo State, Brazil was assessed by cell isolation and direct microscope counting techniques. Ten strains, belonging to five genera (Synechococcus, Merismopedia, Leptolyngbya, Limnothrix, and Nostoc), were isolated and identified by morphological and molecular analyses. Morphological identification of the isolated strains was congruent with their phylogenetic analyses based on 16S rDNA gene sequences. Six cyanobacterial genera (Synechocystis, Aphanocapsa, Merismopedia, Lyngbya, Phormidium, and Pseudanabaena) were identified by direct microscope inspection. Both techniques were complementary, since, of the six genera identified by direct microscopic inspection, only Merismopedia was isolated, and the four other isolated genera were not detected by direct inspection. Direct microscope counting of preserved cells showed that cyanobacteria were the dominant members (>90%) of the phytoplankton community during both periods evaluated (summer and autumn). ELISA tests specific for hepatotoxic microcystins gave positive results for six strains (Synechococcus CENA108, Merismopedia CENA106, Leptolyngbya CENA103, Leptolyngbya CENA112, Limnothrix CENA109, and Limnothrix CENA110), and for wastewater samples collected from raw influent (3.70 μg microcystins/l) and treated effluent (3.74 μg microcystins/l) in summer. Our findings indicate that toxic cyanobacteria in WSP systems are of concern, since the treated effluent containing cyanotoxins will be discharged into rivers, irrigation channels, estuaries, or reservoirs, and can affect human and animal health.