Zebrafish Larvae as a Model to Evaluate Potential Radiosensitizers or Protectors.

Zebrafish are extensively used in several kinds of research because they are one of the easily maintained vertebrate models and exhibit several features of a unique and convenient model system. As highly proliferative cells are more susceptible to radiation-induced DNA damage, zebrafish embryos are a front-line in vivo model in radiation research. In addition, this model projects the effect of radiation and different drugs within a short time, along with major biological events and associated responses. Several cancer studies have used zebrafish, and this protocol is based on the use of radiation modifiers in the context of radiotherapy and cancer. This method can be readily used to validate the effects of different drugs on irradiated and control (non-irradiated) embryos, thus identifying drugs as radio sensitizing or protective drugs. Although this methodology is used in most drug screening experiments, the details of the experiment and the toxicity assessment with the background of X-ray radiation exposure are limited or only briefly addressed, making it difficult to perform. This protocol addresses this issue and discusses the procedure and toxicity evaluation with a detailed illustration. The procedure describes a simple approach for using zebrafish embryos for radiation studies and radiation-based drug screening with much reliability and reproducibility.

Effects of Dissolved Organic Carbon, Ultraviolet Light and their Co-Exposure on Deepwater Horizon crude oil acute toxicity to larval red drum (Sciaenops ocellatus).

In the aquatic environment, ubiquitous natural factors such as ultraviolet light (UV) and dissolved organic carbon (DOC) are likely to influence crude oil toxicity. The present study examined the interactive effects of DOC, UV, and DOC-UV co-exposure on the acute toxicity of Deepwater Horizon crude oil in larval red drum (Sciaenops ocellatus). Although DOC alone did not influence crude oil toxicity, it mildly reduced UV photo-enhanced toxicity. Environ Toxicol Chem 2020;39:2509-2515. © 2020 SETAC.

Effect of gamma radiation as a post-harvest disinfestation treatment against life stages of the coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae).

Purpose: Gamma radiation is mainly used for disinfesting insect pests as an alternative for harmful fumigants. The specific dose of radiation is known to affect different developmental stages of insect pests. The study was conducted to determine the effective irradiation doses for inhibition of developmental stages and adult longevity of the coffee berry borer, Hypothenemus hampei (Ferrari). Materials and methods: Irradiation was carried with the following doses: five levels between 0.01 and 0.16 kGy for eggs, seven levels between 0.10 and 2.00 kGy for larva and prepupa, six levels between 0.10 and 1.60 kGy for pupa and ten levels between 0.10 and 3.20 kGy for adults. Results: Egg development was completely arrested at 0.160 kGy. A dose of 2.00 kGy caused 100% mortality in the first and second instar larva and 98.99% mortality in prepupa. The dose of 1.60 kGy prevented adult eclosion from the irradiated pupa. The adult mortality was 100% at 3.20 kGy. Conclusion: A dose of 3.20 kGy could successfully provide complete security from all developmental stages of H. hampei and prevent yield loss in green coffee as well as the spread of the pest.

The study of ameliorative effect of dietary supplementation of vitamin C, vitamin E, and tryptophan on Labeo rohita (Cyprinidae) fry exposed to intense light.

The stress ameliorating effect of dietary supplementation of vitamin C, vitamin E, and tryptophan on rohu Labeo rohita fry was evaluated. Rohu fry (1.1 ± 0.03 g) were cultured under five different feeding regimes: enriched with 0.08% vitamin C (D1), 0.02% vitamin E (D2), 1.42% tryptophan (D3), a combination of these three ingredients at similar doses (D4), and control diet (D5). Rohu fry of D5 were divided into two groups-exposed to experimental light (D5FL) and ambient light (114 ± 4 lx, D5AL). All fry (except D5AL) were exposed at light intensity of 3442 ± 648 lx. Feeding of rohu with enriched diets significantly (P < 0.05) enhanced the survival rate and average weight. A 15-25% higher survival and 1.3-1.8-fold higher average weight were recorded in rohu fed with enriched diet compared to D5FL treatment. Supplementation of vitamin C in diet (D1) of rohu resulted in 4.1-fold and 6.9-fold higher nitric oxide synthase and reduced glutathione (GSH) levels, respectively compared to the D5FL treatment. The tryptophan-enriched diet (D3) showed 5.8-fold higher melatonin and 4.4-fold lower cortisol levels in rohu compared to the D5FL treatment. Significantly (P < 0.05) higher nitric oxide synthase, GSH and melatonin, and lower cortisol, glucose, thiobarbituric acid reactive substances, carbonyl protein, glutathione S-transferase, and glutathione peroxidase levels were found in D4 diet fed rohu compared to the other treatments. Reduced level of stress in D4 treatment resulted in best performance of rohu in terms of less swimming activity and higher survival and growth compared to the other treatments.

Dietary supplementation of vitamin C: an effective measure for protection against UV-B irradiation using fish as a model organism.

The development of UV-B protective mechanisms in aquacultural species is essential for the sustainable production of healthy aqua crop. Freshwater carp Catla catla larvae (13.5 ± 1.12 mg) were fed with a diet containing 0.5% vitamin C (D1) and a control diet (D2) for 40 days. Each group was exposed to two doses of UV-B irradiation: 360 (5 min, D15 min and D25 min) and 720 mJ cm-2 (10 min, D110 min and D210 min) for 15 days. Significantly (p < 0.05) higher survival and average weight were recorded in D1 compared to D2 exposed to the same dose. Also, significantly (p < 0.001) higher nitric oxide synthase and lower thiobarbituric acid reactive substances and heat shock protein 70 levels were recorded in D15 min compared to the other groups. A direct relationship was found between the dose of UV-B and DNA fragmentation in muscles. DNA damage indices such as tail DNA, tail extent moment and olive tail moment were significantly (p < 0.01) lower in D15 min. Thus, supplementation of vitamin C in the diet provides UV-B protection to larvae.

Phototoxic effects of two common marine fuels on the settlement success of the coral Acropora tenuis.

Coral reefs are at risk of exposure to petroleum hydrocarbons from shipping spills and uncontrolled discharges during extraction. The toxicity of petroleum hydrocarbons can substantially increase in the presence of ultraviolet radiation (UVR), therefore spills in shallow coral reef environments may be particularly hazardous to reef species. Here we investigated the sensitivity of coral larvae (Acropora tenuis) to dissolved hydrocarbons from heavy fuel oil (HFO) and diesel in the absence and presence of UVR. Larval settlement success decreased with increasing concentrations of dissolved HFO, and co-exposure to UVR doubled the toxicity: 50% effect concentrations (EC50) decreased from 96 (-UVR) to 51 (+UVR) total petroleum aromatic hydrocarbons (TPAH). Toxic thresholds for HFO were similar to concentrations reported during marine spills: EC10s of 24 (-UVR) and 15 (+UVR) µg l-1. While less toxic, diesel also reduced settlement and exhibited phototoxicity: EC10s of 122 (+UVR) and 302 (-UVR) µg l-1. This study demonstrates that the presence of UVR increases the hazard posed by oil pollution to tropical, shallow-water coral reefs. Further research on the effects of oils in the presence of UVR is needed to improve the environmental relevance of risk assessments and ensure appropriate protection for shallow reef environments against oil pollution.

Photo-induced toxicity in early life stage fiddler crab (Uca longisignalis) following exposure to Deepwater Horizon oil.

The 2010 explosion of the Deepwater Horizon (DWH) oil rig led to the release of millions of barrels of oil in the Gulf of Mexico. Oil in aquatic ecosystems exerts toxicity through multiple mechanisms, including photo-induced toxicity following co-exposure with UV radiation. The timing and location of the spill coincided with both fiddler crab reproduction and peak yearly UV intensities, putting early life stage fiddler crabs at risk of injury due to photo-induced toxicity. The present study assessed sensitivity of fiddler crab larvae to photo-induced toxicity during co-exposure to a range of environmentally relevant dilutions of high-energy water accommodated fractions of DWH oil, and either <10, 50, or 100% ambient sunlight, achieved with filters that allowed for variable UV penetration. Solar exposures (duration: 7-h per day) were conducted for two consecutive days, with a dark recovery period (duration: 17-h) in between. Survival was significantly decreased in treatments the presence of >10% UV and relatively low concentrations of oil. Results of the present study indicate fiddler crab larvae are sensitive to photo-induced toxicity in the presence of DWH oil. These results are of concern, as fiddler crabs play an important role as ecosystem engineers, modulating sediment biogeochemical processes via burrowing action. Furthermore, they occupy an important place in the food web in the Gulf of Mexico.

Co-exposure to sunlight enhances the toxicity of naturally weathered Deepwater Horizon oil to early lifestage red drum (Sciaenops ocellatus) and speckled seatrout (Cynoscion nebulosus).

The 2010 Deepwater Horizon oil spill resulted in the accidental release of millions of barrels of crude oil into the Gulf of Mexico. Photo-induced toxicity following co-exposure to ultraviolet (UV) radiation is 1 mechanism by which polycyclic aromatic hydrocarbons (PAHs) from oil spills may exert toxicity. Red drum and speckled seatrout are both important fishery resources in the Gulf of Mexico. They spawn near-shore and produce positively buoyant embryos that hatch into larvae in approximately 24 h. The goal of the present study was to determine whether exposure to UV as natural sunlight enhances the toxicity of crude oil to early lifestage red drum and speckled seatrout. Larval fish were exposed to several dilutions of high-energy water-accommodated fractions (HEWAFs) from 2 different oils collected in the field under chain of custody during the 2010 spill and 3 gradations of natural sunlight in a factorial design. Co-exposure to natural sunlight and oil significantly reduced larval survival compared with exposure to oil alone. Although both species were sensitive at PAH concentrations reported during the Deepwater Horizon spill, speckled seatrout demonstrated a greater sensitivity to photo-induced toxicity than red drum. These data demonstrate that even advanced weathering of slicks does not ameliorate the potential for photo-induced toxicity of oil to these species. Environ Toxicol Chem 2017;36:780-785. © 2016 SETAC.

Bioenergetics growth model for the effect of gamma irradiation and plant extract (Barnoof) on the progeny of Black cutworm, Agrotis ipsilon (Hufnagel).

This study investigated the effects of two substerilizing doses of gamma radiation, 100 and 150 Gray (Gy), and/or the plant extract Conyza dioscorides (Barnoof) in two solvents on certain biological aspects and the energy budget of the Black cutworm, Agrotis ipsilon (Hufnagel). Data revealed that the treatment combination of gamma radiation with the plant extract significantly increased reproduction compared with the control at all treatment levels (doses and concentrations). Most of the treatments increased the values of developmental/day and larval duration in the F(1) progeny compared with the progeny from the control treatment, although the percentage survival was decreased in all treatments. These values were more obvious in the combination treatments compared with either gamma radiation or plant extract treatments alone. The coefficient of metabolizable energy (C.M.E.) was not affected by any treatments. No consistent effect on the efficiency of storage of ingested energy [E.S.I.(E.)] and the efficiency of storage of metabolizable energy [E.S.M.(E.)] when the F(1) progeny were treated with plant extract alone or with the plant extract combined with the 100 Gy dose of radiation was noted, while they were both significantly increased at all treatment levels when the 150 Gy dose of radiation was combined with the plant extract. The results obtained are discussed in terms of their implications for the best substerilizing dose of radiation on parental male pupae of A. ipsilon.

Influence of gamma irradiation on productivity indices of the edible Emperor moth caterpillar, Cirina forda (Lepidoptera: Saturniidae).

This study was aimed at generating baseline information for sustainable genetic improvement of Cirana forda larvae for entomophagy, through the use of gamma irradiation. Eggs of C. forda were irradiated with increasing doses of gamma rays from 0 to 200 Gy and raised through larval instal stages under laboratory conditions. The Body Weight (BW) and Head Capsule Width (HCW) of the larval instar stages were monitored as indices of productivity. Successful larval emergence was recorded for all irradiation doses tested and BW of the 1st and 2nd instar larvae were not significantly (p > 0.05) different between the control and treated groups (range = 0.021 +/- 0.003 g/larva in the 200 Gy treatment to 0.028 +/- 0.003 g/larva in the control group and 0.105 +/- 0.003 g/larva in 20 Gy treatment to 0.172 +/- 0.009 g/larva in the control group, respectively). On the other hand, BW during the 3rd and 4th larval instars were significantly (p < 0.05) lower among the irradiated treatments than control. Pattern of distribution of HCW was different from that of BW; as HCW increased with irradiation dose from 10-50 Gy during the 3rd and 4th larval instars. Also, HCW during the 5th instar larvae among the irradiated treatments (range = 5.256 +/- 0.012 to 5.662 +/- 0.026 mm) were not higher than that of the 6th instar in the control group (6.065 +/- 0.010 mm). These results suggest promising potentials of the use of gamma irradiation in sustainably improving the productivity of C. forda larvae for entomophagy.

Monoaminergic modulation of photoreception in ascidian: evidence for a proto-hypothalamo-retinal territory.

BACKGROUND: The retina of craniates/vertebrates has been proposed to derive from a photoreceptor prosencephalic territory in ancestral chordates, but the evolutionary origin of the different cell types making the retina is disputed. Except for photoreceptors, the existence of homologs of retinal cells remains uncertain outside vertebrates. METHODS: The expression of genes expressed in the sensory vesicle of the ascidian Ciona intestinalis including those encoding components of the monoaminergic neurotransmission systems, was analyzed by in situ hybridization or in vivo transfection of the corresponding regulatory elements driving fluorescent reporters. Modulation of photic responses by monoamines was studied by electrophysiology combined with pharmacological treatments. RESULTS: We show that many molecular characteristics of dopamine-synthesizing cells located in the vicinity of photoreceptors in the sensory vesicle of the ascidian Ciona intestinalis are similar to those of amacrine dopamine cells of the vertebrate retina. The ascidian dopamine cells share with vertebrate amacrine cells the expression of the key-transcription factor Ptf1a, as well as that of dopamine-synthesizing enzymes. Surprisingly, the ascidian dopamine cells accumulate serotonin via a functional serotonin transporter, as some amacrine cells also do. Moreover, dopamine cells located in the vicinity of the photoreceptors modulate the light-off induced swimming behavior of ascidian larvae by acting on alpha2-like receptors, instead of dopamine receptors, supporting a role in the modulation of the photic response. These cells are located in a territory of the ascidian sensory vesicle expressing genes found both in the retina and the hypothalamus of vertebrates (six3/6, Rx, meis, pax6, visual cycle proteins). CONCLUSION: We propose that the dopamine cells of the ascidian larva derive from an ancestral multifunctional cell population located in the periventricular, photoreceptive field of the anterior neural tube of chordates, which also gives rise to both anterior hypothalamus and the retina in craniates/vertebrates. It also shows that the existence of multiple cell types associated with photic responses predates the formation of the vertebrate retina.

Effect of sub-lethal exposure to ultraviolet radiation on the escape performance of Atlantic cod larvae (Gadus morhua).

UNLABELLED: The amount of ultraviolet (UV) radiation reaching the earth's surface has increased due to depletion of the ozone layer. Several studies have reported that UV radiation reduces survival of fish larvae. However, indirect and sub-lethal impacts of UV radiation on fish behavior have been given little consideration. We observed the escape performance of larval cod (24 dph, SL: 7.6±0.2 mm; 29 dph, SL: 8.2±0.3 mm) that had been exposed to sub-lethal levels of UV radiation vs. unexposed controls. Two predators were used (in separate experiments): two-spotted goby (Gobiusculus flavescens; a suction predator) and lion's mane jellyfish (Cyanea capillata; a "passive" ambush predator). Ten cod larvae were observed in the presence of a predator for 20 minutes using a digital video camera. Trials were replicated 4 times for goby and 5 times for jellyfish. Escape rate (total number of escapes/total number of attacks ×100), escape distance and the number of larvae remaining at the end of the experiment were measured. In the experiment with gobies, in the UV-treated larvae, both escape rate and escape distance (36%, 38±7.5 mm respectively) were significantly lower than those of control larvae (75%, 69±4.7 mm respectively). There was a significant difference in survival as well (UV: 35%, CONTROL: 63%). No apparent escape response was observed, and survival rate was not significantly different, between treatments (UV: 66%, CONTROL: 74%) in the experiment with jellyfish. We conclude that the effect and impact of exposure to sub-lethal levels of UV radiation on the escape performance of cod larvae depends on the type of predator. Our results also suggest that prediction of UV impacts on fish larvae based only on direct effects are underestimations.

Individual-based model of Chironomus riparius population dynamics over several generations to explore adaptation following exposure to uranium-spiked sediments.

Natural populations are chronically exposed to various pollutants over many generations. It is thus crucial to understand and quantify adaptive dynamics of stressed populations in order to increase the relevance of ecotoxicological risk assessment. However, long-term consequences to population exposure are not much studied yet. The present study investigated evolutionary responses of Chironomus riparius populations exposed to uranium (heavy metal pollutant) and to assess the underlying mechanisms. To fulfil our objective, we produced data with organisms exposed to four relevant concentrations of uranium through eight successive generations. We built an individual-based (IBM) model of C. riparius population dynamics to analyse these data and to test several assumptions about the mechanisms involved in the phenotypic changes. The IBM was based on a dynamic energy budget (DEB) model for C. riparius by Pery et al. (2002). DEB models account mathematically for the acquisition and use of energy to describe and predict growth, maintenance, development and reproduction of living organisms. The IBM accounted for the influence of the test conditions on the observations over eight generations and highlighted some trait evolution such as time to emergence and adult size in control conditions. The model was then used to analyse the exposed population data. Our results showed that exposure to uranium led to a phenotypic selection via a differential survival characterised by longer time to emergence and smaller larval maximal size. As a general conclusion, IBMs based on DEB-based modelling developed to analyse multi-generation experiments are very promising for understanding and quantifying long term selection and tolerance mechanisms in a population under toxic stress.

Laser microsurgery in Caenorhabditis elegans.

Laser killing of cell nuclei has long been a powerful means of examining the roles of individual cells in C. elegans. Advances in genetics, laser technology, and imaging have further expanded the capabilities and usefulness of laser surgery. Here, we review the implementation and application of currently used methods for target edoptical disruption in C. elegans.

Effects of dietary uranium on reproductive endpoints--fecundity, survival, reproductive success--of the fish Danio rerio.

Exposure to metal-contaminated water has been shown to result in a number of reproductive abnormalities in adult and larvae fish, such as failure of oocyte maturation and teratogenic effects. Recently, dietary uptake of metals by fish has been recognized as a critical route of exposure, however, the mechanisms of metal uptake and toxicity are poorly understood and in need of further investigation. The objectives of the present study are to quantify uranium (U dietary transfers from spiked artificial diets) in Danio rerio tissues and embryos, as well as establish its effect on reproduction and embryonic development. Uranium's environmental prominence is currently increasing because of new mining and milling activities. Uranium concentrations range from 0.02 µg/L in natural waters to 2 mg/L. The focus of this study was to examine the trophic transfer and effects of U following exposure modalities (dose, exposure duration 1 to 20 d). Two different isotopes were used to distinguish between chemical and radioactivity toxicity of U. Results showed that U trophic transfer was low (0.52%). Uranium tissue distributions showed that accumulation occurred in digestive organs (liver, digestive tract) following dietary exposure. High levels of U were measured in the gonads (female in particular, >20% of relative burden). High U accumulation levels in eggs indicated maternal transfer of the contaminant. Moreover, U trophic exposure led to a reduction in reproduction success as a function of U accumulated levels. High U exposure conditions strongly reduced the total number of eggs (50%) and their viability at 10 d (reduction of the clutch number, low quality of eggs).

Anhydrobiosis-associated nuclear DNA damage and repair in the sleeping chironomid: linkage with radioresistance.

Anhydrobiotic chironomid larvae can withstand prolonged complete desiccation as well as other external stresses including ionizing radiation. To understand the cross-tolerance mechanism, we have analyzed the structural changes in the nuclear DNA using transmission electron microscopy and DNA comet assays in relation to anhydrobiosis and radiation. We found that dehydration causes alterations in chromatin structure and a severe fragmentation of nuclear DNA in the cells of the larvae despite successful anhydrobiosis. Furthermore, while the larvae had restored physiological activity within an hour following rehydration, nuclear DNA restoration typically took 72 to 96 h. The DNA fragmentation level and the recovery of DNA integrity in the rehydrated larvae after anhydrobiosis were similar to those of hydrated larvae irradiated with 70 Gy of high-linear energy transfer (LET) ions ((4)He). In contrast, low-LET radiation (gamma-rays) of the same dose caused less initial damage to the larvae, and DNA was completely repaired within within 24 h. The expression of genes encoding the DNA repair enzymes occurred upon entering anhydrobiosis and exposure to high- and low-LET radiations, indicative of DNA damage that includes double-strand breaks and their subsequent repair. The expression of antioxidant enzymes-coding genes was also elevated in the anhydrobiotic and the gamma-ray-irradiated larvae that probably functions to reduce the negative effect of reactive oxygen species upon exposure to these stresses. Indeed the mature antioxidant proteins accumulated in the dry larvae and the total activity of antioxidants increased by a 3-4 fold in association with anhydrobiosis. We conclude that one of the factors explaining the relationship between radioresistance and the ability to undergo anhydrobiosis in the sleeping chironomid could be an adaptation to desiccation-inflicted nuclear DNA damage. There were also similarities in the molecular response of the larvae to damage caused by desiccation and ionizing radiation.

Ionizing radiation as a phytosanitary treatment against fruit flies (Diptera: Tephritidae): efficacy in naturally versus artificially infested fruit.

Some phytosanitary irradiation treatment research against tephritid fruit flies (Diptera: Tephritidae) has used artificially infested fruit with the unstated and untested assumption that the method adequately simulated a natural situation. We compare grapefruit, Citrus paradisi Macfayden, naturally infested by Mexican fruit fly, Anastrepha ludens (Loew), via oviposition until larvae reached the late third instar versus insertion of diet-reared third instars into holes made in grapefruits 24 h before irradiation; the latter technique has been used in other studies. Both infestation techniques resulted in statistically indistinguishable results, indicating that insertion of diet-reared third instar Mexican fruit fly into holes bored into grapefruit and subsequently sealed 24 h before irradiation would adequately represent natural infestation and could be used to develop a radiation phytosanitary treatment of the insect in grapefruit when prevention of adult emergence is used as the measure of efficacy. Nevertheless, it may not be advisable to extend this conclusion to other fruit fly/fruit combinations without doing appropriate comparison studies. Dissection of puparia from nonirradiated control insects that failed to emerge as adults showed a relatively even distribution of mortality among the developmental stages within the puparium. In contrast, dissection of puparia from irradiated third instars that did not emerge as adults revealed a sharp attenuation in development from cryptocephalic to phanerocephalic pupae demonstrating this transition to be the developmental step most affected by radiation.

Mutations of the withered (whd) gene in Drosophila melanogaster confer hypersensitivity to oxidative stress and are lesions of the carnitine palmitoyltransferase I (CPT I) gene.

Since some oxygen defense mutants of Drosophila melanogaster exhibit a crinkled wing phenotype, a screen was performed on strains bearing mutant alleles conferring a visible wing phenotype to determine whether any were hypersensitive to oxidative stress. One mutant, withered (whd), was found to be sensitive to both dietary paraquat and hyperoxia. New alleles of whd were induced on a defined genetic background and strains carrying these alleles were also found to be sensitive to oxidative stress. To identify the product of the whd gene we used a sequence-based positional candidate approach and by this method we determined that whd encodes carnitine palmitoyltransferase I (CPT I), an enzyme of the outer mitochondrial membrane that is required for the import of long-chain fatty acids into the mitochondria for beta-oxidation. Although this function is not vital under laboratory conditions, whd adults were found to be highly sensitive to starvation and to heavy metal toxicity relative to controls. This work uncovers a novel relationship between fatty acid metabolism and reactive oxygen metabolism. Further, these results in conjunction with past research on whd and on mammalian CPT I support the hypothesis that CPT I serves a vital function in the response to thymine supplementation.

The effect of homeopathically prepared thyroxine on highland frogs: influence of electromagnetic fields.

BACKGROUND: Previous experiments show that amphibian larvae are responsive to homeopathically prepared thyroxine. METHODS: We studied the effect of a highly diluted and agitated thyroxine solution exposed to various electromagnetic fields on metamorphosis in highland Rana temporaria. The devices tested were: microwave oven, mobile phone, airport X-ray, and a red light barcode scanner. Animals were treated either with homeopathically prepared thyroxine (10(-30) parts by weight, 10(-35) in the water in which the animals were kept), or analogously prepared blank solution, or analogously prepared thyroxine exposed to the electromagnetic field of one of the devices tested. Solutions were administered at 48h intervals according to a standardized protocol. RESULTS: Animals treated with the standard test solution thyroxine 10(-30) metamorphosed more slowly than the control animals, ie the effect of the homeopathically prepared thyroxine was opposed to the usual physiological effect of molecular thyroxine. The cumulative number of test animals that had reached the four-legged stage at defined points in time was smaller in the group treated with homeopathically prepared thyroxine at most of the points in time. This was found independently by all three research teams involved. In contrast, this effect did not occur when the thyroxine solution had been exposed to the field of the early model microwave oven, or mobile phone. There was no difference between aqueous or alcoholic solutions were used, and there was, if any, only a small protective effect from aluminum foil. Airport X-ray and red light barcode scanning did not diminish the effect of the homeopathic solution.

Irradiation of mangoes as a postharvest quarantine treatment for fruit flies (Diptera: Tephritidae).

Mangoes infested with third instar larvae were irradiated using Co-60 gamma rays and a dose interval of 2-250 Gy to assess the irradiation dose required to prevent adult emergence of the Mexican fruit fly (Anastrepha ludens), the West Indies fruit fly (A. obliqua), the sapote fruit fly (A. serpentina), and the Mediterranean fruit fly (Ceratitis capitata). Doses of 76.9, 87.3, 91.4 and 112.7 Gy, were estimated to inhibit 99.9968% (probit 9) of adult emergence forA. obliqua, A. serpentina, A. ludens, and C. capitata, respectively. Using mangoes infested with a total of 100,000 larvae of each species, the results obtained in the laboratory were confirmed using a dose of 100 Gy for the Anastrepha species and 150 Gy for C. capitata. No adult emergence was observed for any of the four species compared with approximately 80% emergence in the controls. A dose of 150 Gy is recommended as a generic quarantine treatment against potential infestation of these species in exported mangoes. A minor decrease in the ascorbic acid content was the only adverse effects observed in irradiated mangoes.