Brine Shrimp Larvae Research Articles

In vitro anthelmintic, antibacterial and cytotoxic effects of extracts from plants used in South African ethnoveterinary medicine

Many plants are used for ethnoveterinary purposes in South Africa, particularly in rural areas. Extracts of 17 plant species employed to treat infectious diseases were prepared using three solvents and the antibacterial activity of the extracts was determined against two Gram-positive and two Gram-negative bacteria. Anthelmintic activity was evaluated against the free-living nematode Caenorhabditis elegans and toxicity was determined using the brine shrimp larval mortality test. Most of the plant extracts demonstrated antibacterial activity, with the best minimum inhibitory concentration (MIC) being 0.1 mg mL −1. More than a third of the extracts displayed anthelmintic activity. Toxic effects against brine shrimp larvae were shown by 30% of extracts, with the lowest LC 50 recorded as 0.6 mg mL −1. The promising biological activity displayed by a number of plant extracts supports the ethnoveterinary use of these plants but in vivo tests are required to ascertain fully their medicinal properties and potential toxicity.

Is the use of <i>Gunnera perpensa</i> extracts in endometritis related to antibacterial activ

Rhizome extracts of Gunnera perpensa are used in traditional remedies in South Africa to treat endometritis both in humans and animals. An investigation was undertaken to determine whether this plant possesses antibacterial activity, which may explain its efficacy. Gunnera perpensa rhizome extracts were prepared serially with solvents of increasing polarity and tested for antibacterial activity. Test bacteria included the Gram-positive Enterococcus faecalis and Staphylococcus aureus and the Gram-negative Escherichia coli and Pseudomonas aeruginosa. A moderate to weak level of antibacterial activity in most of the extracts resulted, with the best minimal inhibitory concentration (MIC) value of 2.61 mg ml(-1) shown by the acetone extract against S. aureus. The extracts were also submitted to the brine shrimp assay to detect possible toxic or pharmacological effects. All the extracts were lethal to the brine shrimp larvae at a concentration of 5 mg ml(-1). The acetone extract was extremely toxic at 1 mg ml(-1), with some toxicity evident at 0.1 mg ml(-1). The remainder of the extracts generally displayed little activity at concentrations lower than 5 mg ml(-1). In summary, the results indicate that although the extracts demonstrated a level of pharmacological activity, the relatively weak antibacterial activity is unlikely to justify the use of G. perpensa rhizomes in the traditional treatment of endometritis. Rather, the slightly antibacterial nature of the rhizomes may contribute to an additive effect, along with their known uterotonic activity, to the overall efficacy of the preparation.

Bioactive metabolites in improved cowpea seeds

The International Institute of Tropical Agriculture (IITA) has developed some pest and disease resistant cowpeas. From these the seeds of 8 cowpea cultivars were extracted with ethanol, and partitioned into chloroform and water-soluble fractions, the water-soluble fraction was further extracted with ethyl acetate. Residues from ethanol, chloroform and ethyl acetate soluble fractions for each of the 8 cowpea cultivars were screened against brine shrimp larvae. The seed extracts of cowpea cultivars IT93K ‐ 596 ‐ 9 ‐ 12, IT90K ‐ 277 ‐ 2 and IT93K ‐ 452 ‐ 1 were found to be most active, indicating that they contain cytotoxic compound(s).

Phytotoxin Production and Phytoalexin Elicitation by the Phytopathogenic Fungus Sclerotinia sclerotiorum

The fungus Sclerotinia sclerotiorum (Lib.) de Bary causes rot disease in a vast range of plant families, including Cruciferae (Brassicaceae). We investigated the production of phytotoxins by S. sclerotiorum by using a bioassay-guided isolation, as well as the phytoalexins produced by the resistant wild crucifer Erucastrum gallicum under elicitation by S. sclerotiorum and other agents. We established for the first time that S. sclerotiorum produces a somewhat selective phytotoxin, sclerin, which is phytotoxic to three cruciferous species (Brassica napus, B. juncea, and Sinapis alba) susceptible to Sclerotinia stem rot disease, causing severe necrosis and chlorosis, but not to a resistant species (Erucastrum gallicum). In addition, we have shown that oleic acid, the major fatty acid isolated from sclerotia of S. sclerotiorum is responsible for the toxic activity of extracts of sclerotia to brine shrimp larvae (Artemia salina). Phytoalexin elicitation in leaves of E. gallicum led to the isolation of three known phytoalexins: indole-3-acetonitrile, arvelexin, and 1-methoxyspirobrassinin. Considering that resistance of E. gallicum to S. sclerotiorum is potentially transferable to B. rapa, a susceptible canola species, and that arvelexin, and 1-methoxyspirobrassinin are not produced by B. rapa, these phytoalexins may become useful markers for resistance against S. sclerotiorum.

New Sesquiterpenes from Pluchea arabica

Two new sesquiterpenes, godotol A (1) and godotol B (2), were isolated from Pluchea arabica. Their structures were determined by analysis of NMR data. The absolute configurations of 1 and 2 were established by Mosher ester methodology. The godotols displayed weak activity against bacteria and the brine shrimp larvae. They were also inactive in the DPPH antioxidant assay.

Antibacterial and brine shrimp lethality tests of biflavonoids and derivatives of Rheedia gardneriana

The hydroalcoholic extract of the leaves from Rheedia gardneriana yielded volkensiflavone ( 1), fukugetin ( 2), fukugiside ( 3), GB2a-I-7- O-glucoside ( 4) and epicatechin ( 5). Compounds 1–5, and some derivatives of 1 and 2 were evaluated for lethality to brine shrimp larvae and for antibacterial activity.

Scale-bearing nanoflagellates from southern Tasmanian coastal waters, Australia. I. Species of the genus Chrysochromulina (Haptophyta)

A comprehensive taxonomic survey of Chrysochromulina was undertaken in 52 samples from southern Tasmanian coastal waters. Scale morphology of both field samples and enrichment cultures was examined using transmission electron microscopy. Twenty-eight known species of Chrysochromulina were recorded. These included C. adriatica, C. alifera, C. apheles, C. brevifilum, C. chiton, C. cyathophora, C. ephippium, C. ericina, C. hirta, C. leadbeateri, C. mantoniae, C. minor, C. novae-zelandiae, C. pachycylindra, C. parkeae, C. polylepis, C. pringsheimii, C. pyramidosa, C. simplex, C. spinifera, and Chrysochromulina sp. (eyelash). There were seven new records for Australian waters: C. acantha, C. aff. brachycylindra, C. aff. camella, C. fragaria, C. mactra, C. scutellum and C. aff vexillifera. All species, except C. novae-zelandiae, had been previously reported from the northern hemisphere. A further nine undescribed species are documented. However, lack of complete cells prevented new species descriptions. Two known toxic species, Chysochromulina polylepis and C. leadbeateri, responsible for massive fish kills in Scandinavia, were found in this survey. Toxicity testing of six Chrysochromulina species isolated from Tasmanian waters (C. acantha, C. apheles, C. hirta, C. scutellum, C. simplex, Chrysochromulina sp. [eyelash]) was undertaken, using larval brine shrimp (Artemia) bioassays. None of the strains tested was found to be toxic.

Cyclitol Derivatives from the Sponge Sarcotragus Species.

AbstractFor Abstract see ChemInform Abstract in Full Text.

Species Richness and Invasion Vectors: Sampling Techniques and Biases

During a European Union Concerted Action study on species introductions, an intercalibration workshop on ship ballast water sampling techniques considered various phytoplankton and zooplankton sampling methods. For the first time, all the techniques presently in use worldwide were compared using a plankton tower as a model ballast tank spiked with the brine shrimp and oyster larvae while phytoplankton samples were taken simultaneously in the field (Helgoland Harbour, Germany). Three cone-shaped and 11 non-cone shaped plankton nets of different sizes and designs were employed. Net lengths varied from 50 to 300 cm, diameters 9.7–50 cm, and mesh sizes 10–100 μm. Three pumps, a Ruttner sampler, and a bucket previously used in ballast water sampling studies were also compared. This first assessment indicates that for sampling ballast water a wide range of techniques may be needed. Each method showed different results in efficiency and it is unlikely that any of the methods will sample all taxa. Although several methods proved to be valid elements of a hypothetical `tool box' of effective ship sampling techniques. The Ruttner water sampler and the pump P30 provide suitable means for the quantitative phytoplankton sampling, whereas other pumps prevailed during the qualitative trial. Pump P15 and cone-shaped nets were the best methods used for quantitative zooplankton sampling. It is recommended that a further exercise involving a wider range of taxa be examined in a larger series of mesocosms in conjunction with promising treatment measures for managing ballast water.

A butyrylcholinesterase in the early development of the brine shrimp ( Artemia salina) larvae: a target for phthalate ester embryotoxicity?

The phthalate ester insensitive blue-green algae ( Synechococcus lividus) were used as a food source to extend the survival of synchronously hatched brine shrimp ( Artemia salina) larvae allowing measurement of a reduced toxic response to phthalate esters at late post-hatching stages of development. The maximum acute toxicity due to di- n-butyl phthalate (DNBP) correlated with the expression of a phthalate ester-hydrolyzing enzyme. The purified enzyme was identified as a butyrylcholinesterase due to its rapid inactivation by low concentrations (10 −7 M) of diisopropyl fluorophosphate and inhibition by physostigmine (IC 50=6×10 −7 M) and tetraisopropylpyrophosphoramide (I-OMPA, IC 50=5×10 −6 M) but not by BW284c5. Apparently competition of the phthalates with the endogenous substrates of the enzyme led to development-dependent toxicity.

Cyclitol Derivatives from the Sponge Sarcotragus Species

In ourcontinuing study on the cytotoxic compounds of Sarcotragussp., the cyclitol derivative sarcotride A (1) has been isolatedalong with two new congeners (2 and 3). The grossstructures of the compounds were elucidated by the aid ofCOSY, HMQC, and HMBC experiments. The isolation,structure elucidation, and biological activity of thecompounds are described herein.The methanol extract of the sponge displayed cytotoxi-cities against five human tumor cell lines (see ExperimentalSection) and showed toxicity to brine shrimp larvae (LD

Feeding and wounding responses in Hydra suggest functional and structural polarization of the tentacle nervous system

The nervous system of Hydra, a freshwater cnidaria, occurs as dispersed, or diffuse, nerve net throughout the animal. It is widely accepted that in a diffuse nervous system an external stimulus is conducted in all directions over the net. Here I report observations that hydra tentacles respond to feeding and wounding stimuli in a unidirectional manner. Upon contact of a tentacle with a brine shrimp larva during feeding, tissue on the proximal side of the point of contact contracted strongly, whereas tissue on the distal side contracted only very weakly. Feeding a tentacle to which a second tentacle was grafted to the proximal end in the reversed orientation showed that unidirectional conduction, once initiated, was blocked by the reversal of polarity, demonstrating that the distal to proximal polarity of tissue is crucial for unidirectional conduction. Unidirectional conduction was obtained also by mechanically pinching the tissue. The response of tentacles devoid of neurons examined was bidirectional, demonstrating that the nervous system is responsible for the unidirectional responses. These observations suggest that polarized property of the nerve net in hydra tentacles is responsible for the unidirectional tentacle contraction.

The Antimalarial and Antimicrobial Activity and Brine Shrimp Toxicity of <i>Clematis Brachiata</i> Extracts

The in vitro antimalarial activity of the root extract in partly supports the ethnobotanical use of the plant to manage malaria. Clematis brachiata Thunberg (Ranunculaceae) is used in Kenya for the management of headaches, malaria and other febrile illnesses, abdominal disorders, yaws and for skin disorders. Old stems and leaves are chewed for the management of toothaches and sore throats. Extracts of the plant were subjected to tests for antimalarial, antibacterial and antifungal activity. The toxicity of the extracts was assessed using the brine shrimp lethality bioassay. The root extract gave the highest in vitro antimalarial activity against the mulitidrug resistant strain, Plasmodium falciparum VI/S (IC50=39.24 mg/ml). The stem and leaf extracts had insignificant antiplasmodial activity. The leaf, stem and root extracts had no bacterial or fungal inhibitory effects even at very high concentrations of 10 mg/ml. The LD50 values of the stem and leaf methanol extracts against the brine shrimp larvae was 365.60 and 66.5 mg/ml, respectively. (E & C Afr Jnl Pharm Sci: 2002 5(1): 15-18)

Larvicidal Activity of Extracts and Triterpenoids from Lantana camara

Extracts of the leaves, twigs, stems and roots of Lantana camara Linn. (Verbenaceae) were solvent-partitioned and screened for activity in the brine shrimp lethality test (BST). The active fractions yielded known oleanonic acid (1), lantadene A (2) and oleanolic acid (3), which were very toxic to brine shrimp larvae. Compounds 1-3 were not lethal to Spodoptera littoralis Biosduval (Lepidoptera: Noctuidae), Clavigralla tomentosicollis Stal. (Hemiptera: Coreidae) and Aphis craccivora Koch (Homoptera: Aphididae) when tested at 5000µg/ml. Compound 2, however, suppressed the fecundity of C. tomentosicollis at this concentration.

Monoacylglycerol from Punica granatum Seed Oil

The seeds of Punica granatum, known as hap roman in the Arabian Peninsula, are commonly eaten as a dessert. As part of an ongoing project to find nonnutritional natural products which have health benefits, or that can be exploited to protect crops, the chloroform-soluble extract of the fermented seeds of P. granatum was found to be rich in 1-O-trans,cis,trans-9,11,13-octadecatrienoyl glycerol (1). The seed oil is not lethal to brine shrimp larvae. 1-O-isopentyl-3-O-octadec-2-enoyl glycerol (2) and the known cis-9-octadecenoic, octadecanoic, and eicosanoic acids were also detected in small amounts in the seed oil by LC and MS. The structure of 1 was determined from NMR and MS spectral data.

Chemical contamination and toxicity of sediment from a coastal area receiving industrial effluents in Kuwait.

The Shuaiba coastal area (12.5 x 1.5 km) was examined for contamination with total organic carbon, volatile organic matter, total petroleum hydrocarbons, polycyclic aromatic hydrocarbons, cadmium, chromium, copper, lead, nickel, vanadium, and zinc in sediment; their desorption by aqueous elution; and toxicity to aquatic biota. The pollutants were mainly accumulated in the upstream area facing Mina Al-Ahmadi refinery to Shuaiba harbour. Solid-phase Microtox assays showed severe toxicity, and the LC(50) was negatively correlated with most of the chemical parameters, suggesting that toxicity was the function of collective effects of the pollutants present in sediment. Sea water elutriation showed poor desorption of pollutants from sediment, and the elutriates were not found toxic to Microtox and brine shrimp larvae. Whole sediment suspension in sea water reduced the survival of fingerlings in fish bioassays. Action from Shuaiba Area Authority is required to reduce pollutant accumulation in identified depositional area on the Shuaiba coast by facilitating unrestricted water flow in the area and restricting pollutant discharge at source.

A new stilbene glycoside from Elephantorrhiza goetzei

A new stilbene glycoside, 5-methoxy-( E)-resveratrol 3- O-rutinoside ( 1) was isolated from the root bark of Elephantorrhiza goetzei, along with known compounds, namely gallic acid, ( E)-resveratrol, (±)-catechin, (±)-gallocatechin and oleanene triterpenoids. The combined ethyl acetate-methanol extract exhibited high lethality against brine shrimp larvae (LC 50 10.8 ppm) compared to the isolates, some of which were not active.

A new screening test for toxicity testing of dental materials

Objectives: The development of a micro plate assay for cytotoxicity testing of dental materials based on a bioassay using brine shrimp larvae (artemia salina) as sensitive organisms. Methods: Brine shrimp larvae are commonly used for cytotoxicity assays in pharmacology. These larvae are sensitive to toxic substances. The ratio between dead larvae (no motility) and living larvae (high motility) in comparison to a control without any toxic substances is used to estimate the toxicity of the test solutions. The test materials (Arabesk ®, Solitaire ®, Pertac ® II, Tetric ®, Herculite ® and the compomer materials Dyract ®, Hytac ®, Compoglass ®) were polymerized and consecutively milled. After incubation of 1 g in 4 ml distilled water at 37°C for 48 h, the solid materials were separated by centrifugation. The solutions were equibrilated with NaCl to a salt content of 25 g/l. Aliquots of 200 μl were distributed in eight micro wells and 50 μl of a artemia salina containing ( n=8–14) solution were added to each well. As controls eight wells with 250 μl salt solution containing a comparable number of brine shrimp were used. At baseline, after 2, 5, 24 and 48 h, the dead shrimp were counted using a stereo microscope. Finally all shrimps were sacrificed using Na-acid (5%) and counted to get the number of shrimps per well. Results: All compomers and Solitaire caused 100% brine shrimp lethality after 24 h and showed significantly ( p<0.01, signed rank test) higher toxicities than the remaining composites. With the exception of Pertac II, all composites showed significantly higher toxic values than the control. Pertac II did not show any differences from the controls used. Conclusions: This new technique has some advantages for toxicity testing of restorative materials, because it can quickly be carried out at low costs. The disadvantage is the high quantity of material used and the low sensitivity.

Abnormal Contamination of Roquefort Cheese in Egypt

Abo-Gharbia, M.M. and El-Sawi, N.M. 1999. Abnormal contamination of Roquefort cheese in Egypt. J. Appl. Anim. Res., 16: 65–74. Roquefort cheese samples of Egyptian and Danish origin were tested for contamination. Chemical analyses revealed that the Roquefort cheese samples were rich in metals. Na was generally high but K, Mn, Ca, Cu, Cd and Ni were detected in low concentrations. Lead was found in Egyptian source only. Fifty-one fungal species belonging to 17 genera were collected and identified from 25 samples of cheese on two isolation media. Three bacterial genera (Streptococcus, Pseudomonas and Staphylococcus) in addition to coliform were identified in variable degree of counts (6.8, 0.6, 219.5 and 2.50 colonies mg g1) and occurrence (17, 3, 19 and 13 samples/25 samples). Four biogenic amines were detected in Roquefort cheese. Histamine was the most dominant in Egyptian and Danish samples. Tryptamine, cadaverine and putrescine appeared in Egyptian source with variable degree of occurrence and concentration. But, tryptamine was only detected in two samples of Danish source. Ten ethyl acetate extracts of Egyptian cheese samples had toxicity to brine shrimp larvae. Based on TLC and UV spectrophotometric analyses, roquefortin, aflatoxin B1 and B1 were detected.

Cytotoxic constituents of Saussurea lappa.

The crude extract of Saussurea lappa displayed significant lethality to brine shrimp larvae. Investigation of the causative components by bioactivity-directed fractionation resulted in the isolation of three C17-polyene alcohols. Based on various nmr spectral data, these compounds were identified as shikokiols which had been previously isolated from Cirsium nipponicum and/or Centaurea aegyptica. These C17-polyene alcohols exhibited moderate cytotoxicities against the human tumor cell lines, A549, SK-OV-3, SK-MEL-2, XF498, and HCT15.