镰刀菌Fusarium solani菌株对卤虫Artemia salina的毒性

寻找能杀伤肿瘤细胞而对正常细胞无毒的抗癌药物极具挑战性。具有细胞毒性的植物或者真菌可能含有抗肿瘤的化合物。卤虫无节幼体的致死性可作为筛选抗肿瘤化合物的试验。本研究运用从不同农作物种子分离的8株镰刀菌（Fusarium solani）培养滤液来测试卤虫的细胞毒性效果。结果表明,5株菌株（TS、S-29、B-17、C-10和W-5）对卤虫显示高毒性;3株菌株（SR、T-9和L-25）显示低毒性,且毒性随着培养滤液的稀释而减弱。5株菌株（TS、B-17、SR、T-9和L-25）按照1∶10稀释能导致30％以上的死亡率。NaOH中和后的滤液毒性略微降低,表明培养滤液的pH值可能影响毒性。这些菌株冻干的滤液相对于未冻干的滤液毒性较低。只在3株温和毒性的菌株中得到正己烷可溶萃取物;氯仿可溶萃取物的量极微而不能作进一步处理。各菌株的毒性效果各不相同。从镰刀菌（F. solani）分离的化合物可开发为毒性化合物。     

A new black fly isolate of Bacillus thuringiensis autoagglutinating strain highly toxic to Simulium pertinax (Kollar) (Diptera, Simuliidae) larvae

Formulations containing the entomopathogenic Bacillus thuringiensis serovar israelensis strain IPS-82 has been widely applied for mosquito control around the world. Strain IPS-82 is highly active against Aedes aegypti but less active against other well-known vectors such as Culex quinquefasciatus and Simulium spp. larvae. Eighteen strains of B. thuringiensis were isolated from Simulium pertinax larvae naturally occurring in rivers of Southeast Brazil with one demonstrating special toxic effects. Simulated field tests against S. pertinax larvae showed that the native Brazilian autoagglutinating B. thuringiensis (LFB-FIOCRUZ 1035) has an LC50 at least 25 times lower than the standard IPS-82 strain. The same bacterial preparation was also tested against Ae. aegypti larvae in laboratory trials and the LC50 values obtained with LFB-FIOCRUZ 1035 were at least three times lower than the one for the IPS 82 strain. The results indicate that this strain is more toxic than the standard B. thuringiensis serovar israelensis (H14) in the two Dipteran species tested. It is noteworthy that differences between LC50 values were more pronounced in S. pertinax larvae, the source of the original isolation.     

Influence of photoperiod and temperature on reproductive mode in the Brine shrimp, Artemia franciscana

Brine shrimp, Artemia, exhibit two modes of reproduction: oviparity (diapause cyst production) and ovoviviparity (live larvae release). Environmental conditions determining these developmental routes are poorly understood, so we investigated the effects of photoperiod and temperature on reproductive mode. Nauplii of A. franciscana were hatched from cysts produced in the Great Salt Lake, Utah, and raised in 2% natural sea salt water under photoperiods of 24, 14, 12, or 10 h at 28 degrees or 20 degrees C. Mating pairs of mature shrimp were isolated and reared continuously under those conditions. The mode of reproduction shown by each pair was determined daily throughout their life span, and found to be greatly affected by photoperiod, and less influenced by temperature. The relative degree of oviparity increased as the photoperiod became shorter at both temperatures. In contrast, the degree of ovoviviparity was higher as the photoperiod became longer at both temperatures. The critical photoperiod appears to be between 12 and 14 h. For all photoperiods examined, the degree of oviparity was higher at 28 degrees C than at 20 degrees C, whereas the degree of ovoviviparity was greater at 20 degrees C than at 28 degrees C.     

Screening fusarium strains isolated from overwintered Canadian grains for trichothecenes

A survey of 38 samples of Canadian overwintered grains showed that 14 (37 %) contained viableFusarium. Of a total of 38Fusarium isolates, cultured on autoclaved corn, 20 (from 7 grain samples) showed toxicity to brine shrimp larvae and 12 (from 5 samples) produced levels of trichothecenes detectable by thin layer chromatography. The principal trichothecene found was T-2 toxin, produced by 10 strains and accompanied in half of these by neosolaniol; some of these strains were identified asF. sporotrichioides Sherbakoff. Two strains ofF. poae (Peck) Wollenw. formed small amounts of diacetoxyscirpenol. T-2 toxin was the most toxic of 8 trichothecenes tested on brine shrimp larvae; the wide range of toxicities limits the usefulness of this bioassay as a general screening method for trichothecenes.     

Phthalate ester hydrolases and phthalate ester toxicity in synchronously developing larvae of the brine shrimp (Artemia)

Di-n-butyl phthalate (DNBP) is toxic to synchronously developing larvae of the brine shrimp, $\text{Artemia}$. The LD50 for 24 h exposure is approximately 30 μM (8 ppm). DNBP is concentrated by larvae, and maximal uptake of DNBP precedes the onset of mortality. At the time of maximal uptake, most of the DNBP remains in the form of the diester. By the time of maximal mortality nearly all of the DNBP has been converted to the monoester, n-butanol and possibly other polar metabolities. n-Butanol and mono-n-butyl phthalate directly incubated with larvae were nontoxic when tested in concentrations at which DNBP was toxic. Soluble enzyme(s) extracted from the hatched larvae, but not from the dormant embryos, can convert DNBP to its monoester and n-butanol. The amount of enzyme activity increased with larval development in parallel with the kinetics of acute toxicity. The enzyme may be significant in the developmental program as well as in the mediation or moderation of the toxic effects of DNBP in $\text{Artemia}$ larvae.     

Macrocyclic trichothecenes produced by Stachybotrys isolated from Egypt and Eastern Europe

Twenty seven isolates of Stachybotrys chartarum, S. albipes, S. kampalensis and S. microspora from Egypt and Eastern Europe were tested for production of macrocyclic trichothecenes. Twenty of the 27 isolates, grown on rice seeds, were toxic to brine shrimp larvae. Based on TLC and HPLC analyses, 5 macrocyclic trichothecenes (verrucarin J, roridin E, satratoxins F, G & H) as well as trichoverrols were identified. When grown in liquid culture on rice extract medium, only 3 isolates were toxic and produced verrucarin J, roridin E and satratoxins G & H. Extracts from mycelial mats were more toxic than culture filterates of two isolates grown on rice extract and both contained the same macrocyclic trichothecenes (285.5 mg/4 L), in addition to trichoverrols A & B (31 mg/4 L) found in mycelial mats only. When grown on 3% sucrose Czapek's medium supplemented with peptone and yeast extract (still cultures), all isolates were non-toxic to brine shrimp and no trichothecenes could be detected in the extracts.     

The accumulation,distribution, and toxicological effects of dietary arsenic exposure in lake whitefish (Coregonus clupeaformis) and lake trout (Salvelinus namaycush)

A 20-day experiment was conducted to compare the accumulation, distribution, and toxicological effects of dietary As, as arsenate, in lake whitefish (LWF, Coregonusclupeaformis) and lake trout (LT, Salvelinus namaycush). Results of this experiment were used to design an experiment of longer duration in which one of the fish species was selected and exposed three times per week to lower dietary As doses. In the present study each treatment group was exposed to a combination of one of three doses of As (0, 100, or 1000 μg As/g) and one of two types of diet, no brine shrimp (NS) or with brine shrimp (WS) for a total of eight dosing events. Brine shrimp were added to determine whether their presence enhanced consumption of As-contaminated food. Modified feeding behavior occurred in both fish species fed As contaminated diets, with the exception of the 100 μg As/g NS food. Brine shrimp addition did not affect feed consumption of the As contaminated diets. Significant As accumulation occurred in stomach, pyloric caeca, intestine, liver, kidney, and gallbladder, but not in bile or muscle. As exposure did not have a significant effect on hepatic and renal metallothionein concentrations. Concentrations of lipid peroxides were only significantly elevated in the plasma of LT fed the 1000 μg As/g WS food. Liver somatic indices decreased significantly in both species, whereas hematological parameters were not affected in either species. Histological lesions occurred in gallbladder, liver, kidney, pyloric caeca and intestine from LWF. These lesions were not observed in LT; however, gallbladders were not examined in this species. Weight gain was lower in both species fed As contaminated diets, however, condition factors were not affected.     

Cytotoxic 16-β-[(d-xylopyranosyl)oxy]oxohexadecanyl triterpene glycosides from a Malagasy plant, Physena sessiliflora

Brine shrimp lethality assay-guided separation of the MeOH extract of leaves of Physena sessiliflora, which is endemic to Madagascar, afforded eight triterpene glycosides, Physenoside S1–4 and 16-β-[(d-xylopyranosyl)oxy]oxohexadecanyl homologues, Physenoside S5–8. Structural elucidation of these compounds was based on both spectroscopic analyses and chemical properties. Physenoside S7 and S8 have significant cytotoxic activities in the brine shrimp lethality assay.     

A quorum sensing-disrupting brominated thiophenone with a promising therapeutic potential to treat luminescent vibriosis

Vibrio harveyi is amongst the most important bacterial pathogens in aquaculture. Novel methods to control this pathogen are needed since many strains have acquired resistance to antibiotics. We previously showed that quorum sensing-disrupting furanones are able to protect brine shrimp larvae against vibriosis. However, a major problem of these compounds is that they are toxic toward higher organisms and therefore, they are not safe to be used in aquaculture. The synthesis of brominated thiophenones, sulphur analogues of the quorum sensing-disrupting furanones, has recently been reported. In the present study, we report that these compounds block quorum sensing in V. harveyi at concentrations in the low micromolar range. Bioluminescence experiments with V. harveyi quorum sensing mutants and a fluorescence anisotropy assay indicated that the compounds disrupt quorum sensing in this bacterium by decreasing the ability of the quorum sensing master regulator LuxR to bind to its target promoter DNA. In vivo challenge tests with gnotobiotic brine shrimp larvae showed that thiophenone compound TF310, (Z)-4-((5-(bromomethylene)-2-oxo-2,5-dihydrothiophen-3-yl)methoxy)-4-oxobutanoic acid, completely protected the larvae from V. harveyi BB120 when dosed to the culture water at 2.5 μM or more, whereas severe toxicity was only observed at 250 μM. This makes TF310 showing the highest therapeutic index of all quorum sensing-disrupting compounds tested thus far in our brine shrimp model system.     

Differences in the susceptibility of American white shrimp larval substages (Litopenaeus vannamei) to four vibrio species

The rapid expansion of commercial culture of penaeid shrimp is threatened by Vibrio diseases affecting survival and growth. These opportunistic microorganisms are considered part of the normal ecosystem of penaeid shrimp and cause diseases only under conditions that favor them over the host. Shrimp larvae show different susceptibility to these pathogenic agents. In the present work, we report on a comparative study of the susceptibility of all American white shrimp (Litopenaeus vannamei) larval substages to four potentially pathogenic Vibrio species (V. harveyi, V. parahaemolyticus, V. alginolyticus, and V. penaeicida). Strains of these bacterial species were used to infect nauplii, protozoea I-III, mysis I-III, and postlarvae 1 by immersion challenge at 10(3), 10(5), or 10(7) cfu mL(-1) for 30 min. V. alginolyticus infection had no significant effect on survival rate, compared to control, in all shrimp larvae and at all doses tested. Shrimp larvae infected with V. alginolyticus showed a high survival rate compared to other Vibrio species at the three dose levels. V. penaeicida produced a significant mortality effect (P < 0.01) in all shrimp substages and only in postlarvae 1 at low infection dose (10(3) cfu mL(-1)). V. harveyi and V. parahaemolyticus induced significant mortality rates (P < 0.01) only at high doses in shrimp larvae. In summary, shrimp larvae demonstrated an age susceptibility that depends on the Vibrio species and dose level.     

Optical motility test for the detection of trichothecenes using brine shrimps

A new optical bio-assay using brine shrimp larvae (Anemia salina L.) has been successfully applied for monitoring the presence of highly toxic trichothecenes. The motility of the swimming animals was determined instead of their lethality. This permits more objective and precise results. The inhibition of the motility can be seen earlier, than the death of animals in toxicity tests. The relationship between time and dose is indicated for two different concentrations. This test can also be applied in the control of other harmful compounds.     

Analysis of the diet of the tadpole shrimp, Triops sinensis, in paddy fields of Shouchang River watershed

The tadpole shrimp, Triops sinensis, was discovered in the paddy fields of the Shouchang River watershed. The gut contents of the shrimp from these field populations were analyzed in order to quantify their dietary patterns. Some species of tadpole shrimps were found to prey on mosquito larvae as well as uproot weeds. Mosquito larvae and weeds were seldom found in the paddy fields which the tadpole shrimp inhabited in the Shouchang River watershed. The results further showed that the larvae of these tadpole shrimp mainly feed on algae and rice lamina. The tadpole shrimp could feed on animal food including protozoa, nematodes, rotifers, crustaceans, and aquatic insect larvae, though the animal food was subsidiary.     

Mycoflora and mycotoxins of peanut (Arachis hypogaea L.) seeds in Egypt. 1-sugar fungi and natural occurrence of mycotoxins

Sixty-four species and 2 varieties which belong to nineteen genera of fungi were identified from 40 peanut seed samples collected from different places in Egypt by using a dilution-plate method on glucose-Czapek's medium. The most frequent genera were Aspergillus (21 species & 2 varieties), Penicillium (16 species) and Fusarium (6 species). A. flavus, A. fumigatus, A. niger, P. chrysogenum and F. oxysporum were the most common fungal species. Forty seven percent of the samples proved to be toxic to brine shrimp (Artemia salina) larvae. Thin-layer Chromatographic analysis revealed that peanuts were contaminated by aflatoxins (11 samples), thrichothecene-toxins (10 samples), zearalenone (one sample) and citrinin (one sample). We believe that this is the first report of the natural occurrence of zearalenone and trichothecene-toxins in peanuts.     

Cell surface NADH oxidase activity of brine shrimp oscillates with a period of 25 min and is entrained by light

Plants have a surface NADH oxidase that measures time by oscillating with a 24-min period. The period is synchronized by light. With plants, a new maximum is observed exactly 12 min after the beginning of the light exposure. These experiments were to determine if animals exhibited a cell surface NADH oxidase having a similar periodicity and to answer the question, does the periodicity in animals respond to light? Using brine shrimp as a model, the findings show that plants and animals exhibit similar oscillating NADH oxidase activity and that the periodicity in this invertebrate animal does respond to light. Brine shrimp were grown for two to three days and transferred to darkness for 45 min. After return to light for one min, NADH was added and measurements of NADH oxidation were recorded over 50 min. The brine shrimp exhibited a cell surface NADH oxidase that oscillated with a period of 25 min. After being subjected to light, the brine shrimp showed a new maximum in NADH oxidation between 12 to 13 min after the beginning of the light exposure and again at 37 min and at 25 min intervals thereafter. The findings demonstrate that the periodic oscillations in NADH oxidation of brine shrimp are light entrainable.     

Analysis of the diet of the tadpole shrimp, Triops sinensis, in paddy fields of Shouchang River watershed

The tadpole shrimp, Triops sinensis, was discovered in the paddy fields of the Shouchang River watershed. The gut contents of the shrimp from these field populations were analyzed in order to quantify their dietary patterns. Some species of tadpole shrimps were found to prey on mosquito larvae as well as uproot weeds. Mosquito larvae and weeds were seldom found in the paddy fields which the tadpole shrimp inhabited in the Shouchang River watershed. The results further showed that the larvae of these tadpole shrimp mainly feed on algae and rice lamina. The tadpole shrimp could feed on animal food including protozoa, nematodes, rotifers, crustaceans, and aquatic insect larvae, though the animal food was subsidiary.     

Brine Shrimp (Artemia salina L.) Larvae as a Screening System for Fungal Toxins

Concentrations resulting in 50% mortality, determined with brine shrimp (Artemia salina L.) larvae exposed to known mycotoxins for 16 hr, were (mug/ml): aflatoxin G(1), 1.3; diacetoxyscirpenol, 0.47; gliotoxin, 3.5; ochratoxin A, 10.1; and sterigmatocystin, 0.54. 4-Acetamido-4-hydroxy-2-butenoic acid gamma-lactone gave no mortality at 10 mug/ml. Used as a screening system involving discs saturated with solutions of known mycotoxins, the larvae were relatively sensitive to aflatoxin B(1), diacetoxyscirpenol, gliotoxin, kojic acid, ochratoxin A, rubratoxin B, sterigmatocystin, stemphone, and T-2 toxin. Quantities of 0.2 to 2 mug/disc caused detectable mortality. The larvae were only moderately sensitive to citrinin, patulin, penicillic acid, and zearalenone which were detectable at 10 to 20 mug/disc. They were relatively insensitive to griseofulvin, luteoskyrin, oxalic acid, and beta-nitropropionic acid. The disc screening method indicated that 27 out of 70 fungal isolates from foods and feeds grown in liquid or solid media produced chloroform-extractable toxic material. Examination of toxic extracts by thin-layer chromatography for 17 known mycotoxins showed that the toxicity of eight isolates could be attributed to aflatoxin B(1) and B(2), kojic acid, zearalenone, T-2 toxin, or ochratoxin A. Nine out of 32 of these fungal isolates grown in four liquid media yielded toxic culture filtrates from at least one medium. Chemical tests for kojic, oxalic, and beta-nitropropionic acids showed the presence of one or two of these compounds in filtrates of seven of these nine isolates.     

Toxicity ofFusarium sambucinum Fuckel sensu lato to brine shrimp

In a screening test sevenFusarium strains out of 17 proved to be toxic towards brine shrimp. Among these, fourF. sambucinum strains as well as threeF. venenatum isolates caused toxic effects. The chemical screening by TLC analysis revealed the presence of sambucinol and diacetoxyscirpenol in the extracts of the toxic isolates ofF. venenatum 64537.     

Toxic lanostane triterpenes from the basidiomycete Ganoderma amboinense

A highly oxygenated new lanostane triterpene, designated as ganodermacetal (1), together with 15 related known compounds, has been isolated from the fruiting bodies of the basidiomycetous fungus Ganoderma amboinense. The structure of the new metabolite was elucidated on the basis of extensive spectroscopic analyses and chemical transformations. Ganodermacetal (1) possesses an uncommon 7,15-acetonide unit. Some of the isolated compounds showed significant toxic activity against brine shrimp larvae.     

Assessment of toxic potential of local Jordanian Bacillus thuringiensis strains on Drosophila melanogaster and Culex sp. (Diptera)

AIMS: To assess the toxic potential of different local Jordanian Bacillus thuringiensis isolates on larvae of Drosophila melanogaster and Culex sp. METHODS AND RESULTS: Scanning electron microscopy revealed the presence of spherical, bi-pyramidal, and bi-pyramidal and cuboidal parasporal bodies produced by the toxic isolates. Spherical inclusions dominated. The toxicity of the isolates to the two insects, determined using 24-well plates or vials, indicated that the 50% lethal concentration (LC50) of the bacterial suspension for D. melanogaster and Culex sp. larvae varied from 4.60 to 8.65, and from 5.30 to 6.74, respectively. CONCLUSION: Comparison of the LC50 values of isolate 82 with those of the reference strain B. t. israelensis showed that this isolate has a higher toxicity potential. SIGNIFICANCE AND IMPACT OF THE STUDY: Some local Jordanian B. thuringiensis isolates exhibit toxic potential that could be used to control some important pests, and could replace chemical pesticides.     

Beyond Symbiosis: Cleaner Shrimp Clean Up in Culture

Cleaner organisms exhibit a remarkable natural behaviour where they consume ectoparasites attached to “client” organisms. While this behaviour can be utilized as a natural method of parasitic disease control (or biocontrol), it is not known whether cleaner organisms can also limit reinfection from parasite eggs and larvae within the environment. Here we show that cleaner shrimp, Lysmata amboinensis, consume eggs and larvae of a harmful monogenean parasite, Neobenedenia sp., in aquaculture. Shrimp consumed parasite eggs under diurnal (63%) and nocturnal (14%) conditions as well as infectious larvae (oncomiracidia) diurnally (26%). Furthermore, we trialled the inclusion of cleaner shrimp for preventative parasite management of ornamental fish, Pseudanthias squamipinnis, and found shrimp reduced oncomiracidia infection success of host fish by half compared to controls (held without shrimp). Fish held without cleaner shrimp exhibited pigmentation changes as a result of infection, possibly indicative of a stress response. These results provide the first empirical evidence that cleaner organisms reduce parasite loads in the environment through non-symbiotic cleaning activities. Our research findings have relevance to aquaculture and the marine ornamental trade, where cleaner shrimp could be applied for prophylaxis and control of ectoparasite infections.