Fecundity and longevity of Argentine ant (Hymenoptera: Formicidae) queens in response to irradiation

Irradiation is a post‐harvest quarantine treatment option to control ants and other hitchhiker pests on fresh horticultural products traded between countries. As little is known about irradiation effects on ants, radiotolerance of the Argentine ant, Linepithema humile (Mayr) (Hymenoptera: Formicidae: Dolichoderinae), was studied to determine a dose sufficient for its control. Queens collected from Buenos Aires, Argentina, were irradiated with 30, 60, 90 Gy or left untreated as controls, and then followed for 8 weeks to evaluate their survival and fecundity. Overall queen survival and brood viability decreased with increasing irradiation dose. The number of eggs was reduced by 50%, 69% and 56% in the 30, 60 and 90 Gy doses, respectively, compared with untreated control queens. The percentage of eggs that developed into larvae decreased from 41.1% in the control to 22.5%, 1.4%, and 0% in the 30, 60, and 90 Gy treatments, respectively. Thus, the number of larvae was reduced by 69% in the 30 Gy treatment compared with the control, only one larva was observed in the 60 Gy treatment, and none in the 90 Gy treatment. Only one pupa was observed in the 30 Gy treatment and none in the 60 and 90 Gy treatments during the 8‐week experiment. Queens irradiated with 60 and 90 Gy had significantly reduced longevity compared with queens treated with lower doses or untreated queens. Radiation dose ≥90 Gy stopped brood development in Argentine ant queens and should be sufficient as a phytosanitary treatment. The radiotolerance of Argentine ant appears to be similar to that of two other important invasive ants.     

Ionizing radiation for quarantine control of Opogona sacchari (Lepidoptera: Tineidae)

A discriminating irradiation dose of 150 gray (Gy) was used to determine the most tolerant immature stages of Opogona sacchari (Bojer) (Lepidoptera: Tineidae). Based on adult emergence, early and late pupae were determined to be the most tolerant stages, and they were significantly more tolerant than eggs, neonate larvae, and larvae that were 1, 2, or 3 wk old. Irradiation treatment of eggs, neonates, 1-wk-old larvae, 2-wk-old larvae, 3-wk-old larvae, early pupae, and late pupae at 150 Gy resulted in a 96, 96, 95, 73, 61, 8, and 9% reduction in adult emergence, respectively. Pupae were treated with irradiation doses between 60 and 400 Gy. Emergence to the adult stage was significantly reduced by irradiation, averaging 90% in experimental controls and 29% in the 400-Gy treatment. Egg production was also reduced by irradiation, although the average age of pupae at the time of irradiation had a larger effect on fecundity. In total, 2,527 pupae treated with 120 Gy eclosed and produced 47,221 eggs and three F1 larvae. In the 150-Gy treatment, 2,927 adults in total emerged from the 4,626 insects treated as pupae. These adults laid 62,878 eggs, none of which hatched. We conclude that a minimum dose of 150 Gy should be sufficient for sterilization of immature O. sacchari infesting export commodities.     

Influence of larvae and workers on egg production of queens of the pharaoh's ant,Monomorium pharaonis (L.)

Summary Social regulation of egg production and weight in queens was studied in relation to presence and absence of larvae and workers in the pharaoh's ant,Monomorium pharaonis (L.).Results were obtained by counting eggs and weighing queens under various conditions.The results confirm the existence of a positive feed-back loop between mated queens and their larvae as evident from a correlation (Y = 4.575 * X + 6.452) between the number of large worker larvae (X) and the egg yield (Y). This correlation seems to relate to the queens preferential feeding on larval secretions. Queens without larvae maintained a low level of egg production of about 6 eggs/day. Queens deprived of larvae as well as workers stopped producing eggs within 24 hours.Repletes, special workers, with greatly distended gasters functioned as a buffer retarding decline of egg production by feeding the queens during short periods without larvae.     

Field suppression of the invasive ant Wasmannia auropunctata (Hymenoptera: Formicidae) in a tropical fruit orchard in Hawaii

The little fire ant, Wasmannia auropunctata (Roger) (Hymenoptera: Formicidae), is an invasive ant that forms supercolonies when it successfully invades new areas. W. auropunctata was first reported in Hawaii in 1999, and it has since invaded a variety of agricultural sites, including nurseries, orchards, and pastures. Amdro (hydramethylnon; in bait stations), Esteem (pyriproxyfen; broadcast bait), and Conserve (spinosad; ground spray) were tested for their efficacy against W. auropunctata in a rambutan, Nephelium lappaceum L. and mangosteen, Garcinia mangostana L., orchard by making treatments every 2 wk for 16 wk. Relative estimates of ant numbers in plots was determined by transect sampling using peanut butter-baited sticks. Significant treatment effects were observed on weeks 13-17, with reductions in ant counts occurring in the Amdro and Esteem treatments. During this period, the reduction in ant numbers from pretreatment counts averaged 47.1 and 92.5% in the Amdro and Esteem plots, respectively, whereas ant numbers in the untreated control plots increased by 185.9% compared with pretreatment counts. Conserve did not cause a reduction in ant counts as applied in our experiment. No plots for any of the treatments achieved 100% reduction. Pseudococcidae were counted on branch terminals at 4-wk intervals. The two predominant species, Nipaecoccus nipae (Maskell) and Nipaecoccus viridis (Newstead) were significantly lower in the Amdro and Esteem treatments on week 16 compared with controls. Many W. auropunctata were found nesting in protected sites in the orchard trees, which may have compromised the ground-based control methods. Absolute density estimates from shallow core samples taken from the orchard floor indicated the W. auropunctata supercolony exceeded 244 million ants and 22.7 kg wet weight per ha.     

Effects of gamma radiation on different developmental stages of the oriental tobacco budworm,Helicoverpa assulta (Lepidoptera: Noctuidae)

Ionizing radiation is increasingly used as an alternative to post‐harvest crop fumigation by methyl bromide. We studied the effects of gamma irradiation on Helicoverpa assulta (Lepidoptera: Noctuidae) at different stages of development to determine the minimal dose for the prevention of normal emergence of adults. We selected five doses of gamma rays (100, 200, 300, 400 and 500 Gy) based on preliminary experiments and irradiated eggs, larvae, pupae and adults. A dose of 100 Gy to eggs allowed 21.83% of larvae to pupate, but these all died during the pupal stage. A dose of 100 Gy to last‐instar larvae caused larval or pupal death, or the emergence of abnormal adults; no normal adults developed. Irradiation of pupae with doses of 300 Gy and above resulted either in their death or emergence of abnormal adults; however, after 100 or 200 Gy, normal adults emerged and F₁ eggs were produced, but no eggs hatched. Following irradiation of adults, eggs were produced at all doses, although the numbers were significantly decreased compared to untreated controls (P < 0.05; 69.45–125.50 vs. 475.05 eggs per female); however, none of the eggs hatched. As prevention of normal emergence is a key outcome for measuring the effectiveness of radiation, then the 100 Gy dose was effective for irradiation of eggs and larvae, and 300 Gy for pupae.     

Effect of electron beam irradiation on developmental stages of Plutella xylostella (Lepidoptera: Plutellidae)

The effect of electron beam irradiation on each developmental stage of diamondback moth, Plutella xylostella L. (Lepidoptera: Plutellidae), was examined. Eggs, larvae, pupae, and adults were irradiated at target doses of 30, 50, or 100 Gy or they were left untreated as controls in replicated experiments. When eggs and pupae were irradiated with 100 Gy, emergence rates greatly decreased and, although some adults laid eggs, they did not hatch. Egg hatching of irradiated larvae and adult decreased with increasing irradiation doses. However, electron beam irradiation did not kill P. xylostella directly. Adult longevity was not affected. Reciprocal crosses between irradiated and unirradiated moths demonstrated that females were more radiosensitive than males in hatchability; however, the difference was not significant. In addition, electron beam-irradiated larvae showed typical DNA fragmentation in a dose-dependent manner compared with cells from unirradiated larvae. Our findings suggest that electron beam irradiation induces abnormal development and reproduction of P. xylostella; therefore, it may contribute to effective disinfestation and quarantine treatments of P. xylostella.     

Loss of phenotypic plasticity generates genotype-caste association in harvester ants

Caste differentiation and reproductive division of labor are the hallmarks of insect societies. In ants and other social Hymenoptera, development of female larvae into queens or workers generally results from environmentally induced differences in gene expression. However, several cases in which certain gene combinations may determine reproductive status have been described in bees and ants. We investigated experimentally whether genotype directly influences caste determination in two populations of Pogonomyrmex harvester ants in which genotype-caste associations have been observed. Each population contains two genetic lineages. Queens are polyandrous and mate with males of both lineages , but in mature colonies, over 95% of daughter queens have a pure-lineage genome, whereas all workers are of F1 interlineage ancestry. We found that this pattern is maintained throughout the colony life cycle, even when only a single caste is being produced. Through controlled crosses, we demonstrate that pure-lineage eggs fail to develop into workers even when interlineage brood are not present. Thus, environmental caste determination in these individuals appears to have been lost in favor of a hardwired genetic mechanism. Our results reveal that genetic control of reproductive fate can persist without loss of the eusocial caste structure.     

The unusual social organization of the antPachycondyla tridentata (Formicidae,ponerinae)

Colonies of the ponerine antPachycondyla tridentata from Malaysia occur with and without queens. In a total of 7 colonies we found more than 80% of the workers to be mated, irrespective of the presence or absence of queens. This is a hitherto unknown social organisation in ants. Queens and workers competed equally for reproduction. In the colonies investigated several ants were laying eggs. Behavioral observations revealed persistent dominance interactions between colony members. A few ants, but not necessarily a queen, occupied top positions. Removal of the most dominant ants led to a new hierarchy in which subordinate ants with developed ovaries were attacked significantly more frequently than non-reproductive ants. On the average, callows were more aggressive than older subordinate ants, displacing most of the older laying workers in one colony. Nestmate recognition tests revealed that non-reproductive ants were much more aggressive towards foreign ants than were ants with developed ovaries.     

Effects of gamma irradiation on the life stages of yellow flower thrips, Frankliniella schultzei (Trybom) (Thysanoptera: Thripidae)

Irradiation at a minimum absorbed dose of 250 Grays (Gy) has been approved by the USDA as a quarantine treatment for certain fruits in Hawaii to control four species of tephritid fruit flies. Subsequent research must determine whether this dose is sufficient to control other quarantine pests, such as mealybugs, thrips, mites, beetles, moths, and scale insects, on other commodities with export potential that are approved for irradiation treatment for fruit flies. This study demonstrated that irradiation at 250 Gy caused non‐emergence of eggs and pupae, failure of larval development, and sterility of adults of yellow flower thrips, Frankliniella schultzei (Trybom). Adults were the most resistant stage tested, with 100% mortality at 57, 36 and 30 days post‐treatment for the 250, 350 and 400 Gy treatments, respectively. Untreated adults survived up to 66 days. After receiving an irradiation dose of 250 Gy, no one‐ to two‐day old eggs hatched successfully, while 3–4‐day old eggs hatched but did not develop beyond the larval stage. Of the controls, 96.0% of 1–2‐day old eggs and 75.9% of the 3–4‐day old eggs hatched and survived through pupation. No first or second instar larvae treated with a target dose of 250 Gy were able to pupate. When pupae were irradiated at 250 Gy, 37% emerged as adults and all were sterile compared to 88.3% emergence of controls.     

Genetic caste determination in Pogonomyrmex harvester ants imposes costs during colony founding

Some populations of Pogonomyrmex harvester ants comprise genetically differentiated pairs of interbreeding lineages. Queens mate with males of their own and of the alternate lineage and produce pure-lineage offspring which develop into queens and inter-lineage offspring which develop into workers. Here we tested whether such genetic caste determination is associated with costs in terms of the ability to optimally allocate resources to the production of queens and workers. During the stage of colony founding, when only workers are produced, queens laid a high proportion of pure-lineage eggs but the large majority of these eggs failed to develop. As a consequence, the number of offspring produced by incipient colonies decreased linearly with the proportion of pure-lineage eggs laid by queens. Moreover, queens of the lineage most commonly represented in a given mating flight produced more pure-lineage eggs, in line with the view that they mate randomly with the two types of males and indiscriminately use their sperm. Altogether these results predict frequency-dependent selection on pairs of lineages because queens of the more common lineage will produce more pure-lineage eggs and their colonies be less successful during the stage of colony founding, which may be an important force maintaining the coexistence of pairs of lineages within populations.     

Making a queen: an epigenetic analysis of the robustness of the honeybee (Apis mellifera) queen developmental pathway

Specialized castes are considered a key reason for the evolutionary and ecological success of the social insect lifestyle. The most essential caste distinction is between the fertile queen and the sterile workers. Honeybee (Apis mellifera) workers and queens are not genetically distinct, rather these different phenotypes are the result of epigenetically regulated divergent developmental pathways. This is an important phenomenon in understanding the evolution of social insect societies. Here, we studied the genomic regulation of the worker and queen developmental pathways, and the robustness of the pathways by transplanting eggs or young larvae to queen cells. Queens could be successfully reared from worker larvae transplanted up to 3 days age, but queens reared from older worker larvae had decreased queen body size and weight compared with queens from transplanted eggs. Gene expression analysis showed that queens raised from worker larvae differed from queens raised from eggs in the expression of genes involved in the immune system, caste differentiation, body development and longevity. DNA methylation levels were also higher in 3‐day‐old queen larvae raised from worker larvae compared with that raised from transplanted eggs identifying a possible mechanism stabilizing the two developmental paths. We propose that environmental (nutrition and space) changes induced by the commercial rearing practice result in a suboptimal queen phenotype via epigenetic processes, which may potentially contribute to the evolution of queen–worker dimorphism. This also has potentially contributed to the global increase in honeybee colony failure rates.     

Maternal effect on female caste determination in a social insect

Caste differentiation and division of labor are the hallmarks of social insect colonies [1, 2]. The current dogma for female caste differentiation is that female eggs are totipotent, with morphological and physiological differences between queens and workers stemming from a developmental switch during the larval stage controlled by nutritional and other environmental factors (e.g., [3-8]). In this study, we tested whether maternal effects influence caste differentiation in Pogonomyrmex harvester ants. By conducting crossfostering experiments we identified two key factors in the process of caste determination. New queens were produced only from eggs laid by queens exposed to cold. Moreover, there was a strong age effect, with development into queens occurring only in eggs laid by queens that were at least two years old. Biochemical analyses further revealed that the level of ecdysteroids was significantly lower in eggs developing into queens than workers. By contrast, we found no significant effect of colony size or worker exposure to cold, suggesting that the trigger for caste differentiation may be independent of the quantity and quality of resources provided to larvae. Altogether these data demonstrate that the developmental fate of female brood is strongly influenced by maternal effects in ants of the genus Pogonomyrmex.     

Effects of γ radiation on the reproduction and enteroendocrine cells of the spruce bark beetle <Emphasis Type="Italic">Ips typographus</Emphasis> and prospects for its control

The European spruce bark beetle Ips typographus is one of the most important pests of spruce forests in Europe. The present study investigated the feasibility of a sterile insect technique (SIT) for this pest control. Laboratory-reared males were exposed to various doses of γ radiation (0, 10, 30, 50, 70 and 90 Gy) and allowed to mate with laboratory-reared untreated females. The radiation significantly affected the number of viable offspring. Although females oviposited on average 16.0–18.8 eggs per maternal tunnel in all treatments, the number of larval tunnels per maternal tunnel observed after 4 weeks of development decreased from 17.3 in the control (0 Gy) to 4.5 in the 90-Gy group. Numerous oviposition sites without larval tunnels were observed in galleries when males received high doses (70 and 90 Gy) of γ radiation. Comparison of the sperm viability in the control and irradiated males did not reveal any statistically significant differences. Side effects of the irradiation were examined by immunostaining of the enteroendocrine cells. Two distinct types of cells were revealed with antibodies to neuropeptides allatostatin A and tachykinin in the midgut. The alimentary tracts of males receiving a high dose of radiation (90 Gy) showed a significantly decreased number of tachykinin-like immunoreactive enteroendocrine cells. This observation suggests potential radiation-induced damage of the digestive system, which could lead to a reduction in male fitness. The implications of these findings for successful use of SIT in spruce bark beetle control are discussed.     

Irradiation to ensure quarantine security for Cryptophlebia spp. (Lepidoptera: Tortricidae) in sapindaceous fruits from Hawaii

Studies were undertaken to determine whether irradiation treatment at 250 Gy, an accepted treatment for disinfestation of fruit flies in spindaceous fruits from Hawaii, would also disinfest fruit of two species of Cryptophlebia. Cryptophlebia illepida (Butler) was determined to be more tolerant of irradiation than Cryptophlebia ombrodelta (Lower); therefore, C. illepida was the focus for detailed tests. Using the criterion of success in developing to the adult stage, the pattern of tolerance to irradiation in C. illepida was generally eggs < early instars < late instars < pupae. The most tolerant stage potentially occurring in harvested fruits was late (fourth and fifth) instars. Development to adult was reduced slightly in late instars receiving an irradiation dose of 62.5 Gy, whereas development to adult was dramatically reduced in late instars receiving irradiation doses > or = 125 Gy. No C. illepida larvae receiving an irradiation dose > or = 125 Gy emerged as adults and produced viable eggs, indicating sterility can be achieved at doses well below 250 Gy. In large scale tests, when 11,256 late instars were irradiated with a target dose of 250 Gy, 951 pupated (8.4%) and none eclosed as adults. Within the pupal stage, tolerance increased with age; 7- to 8-d-old pupae (the oldest pupae tested) treated with an irradiation dose of 125 Gy produced viable offspring, whereas those treated with a dose of 250 Gy produced no viable offspring. Irradiation of adults with a target dose of 250 Gy before pairing and mating resulted in no viable eggs. Irradiation of actively ovipositing adult females resulted in no subsequent viable eggs. Therefore, the irradiation quarantine treatment of a minimum absorbed dose of 250 Gy approved for Hawaii's fruits will effectively disinfest fruits of any Cryptophlebia in addition to fruit flies.     

Hepatic radiation injury in the rat.

The whole livers of rats were exposed intraoperatively to graded doses (0 to 75 Gy) of 137Cs gamma radiation. At various times (0 to 155 days) after liver irradiation, minimally invasive, nondestructive tests (rose bengal retention and plasma alkaline phosphatase, glutamic-oxaloacetic acid transaminase, glutamic-pyruvic transaminase) were performed on at least half the surviving animals in each dose group to assess developing liver injury. Liver histology was done on animals sacrificed 96 to 100 days after irradiation. Radiation damage to the stomach killed approximately 50% of the animals 30 to 60 days after exposure to doses of 25 Gy or higher. These deaths were significantly reduced when care was taken to shield the stomach during irradiation. Stomach injury did not, however, appreciably affect liver function as measured by rose bengal retention. Whole-liver irradiation to 15 Gy resulted in reduced liver size and minimal histological changes, but did not result in increased rose bengal retention or plasma alkaline phosphatase concentration. The next highest dose group studied (25 Gy) showed significant histological abnormalities and liver injury as measured by increased rose bengal retention and liver enzymes. The latent period for development of hepatic injury, as measured by increased rose bengal retention, was 35 to 42 days and was relatively invariant over the 25- to 75-Gy dose range. Hepatic vein lesions and cellular necrosis were the most prominent histological lesions observed in 25-Gy-irradiated liver.     

Sex allocation conflict in ants: when the queen rules

Insect societies are paramount examples of cooperation, yet they also harbor internal conflicts whose resolution depends on the power of the opponents. The male-haploid, female-diploid sex-determining system of ants causes workers to be more related to sisters than to brothers, whereas queens are equally related to daughters and sons. Workers should thus allocate more resources to females than to males, while queens should favor an equal investment in each sex. Female-biased sex allocation and manipulation of the sex ratio during brood development suggest that workers prevail in many ant species. Here, we show that queens of Formica selysi strongly influenced colony sex allocation by biasing the sex ratio of their eggs. Most colonies specialized in the production of a single sex. Queens in female-specialist colonies laid a high proportion of diploid eggs, whereas queens in male-specialist colonies laid almost exclusively haploid eggs, which constrains worker manipulation. However, the change in sex ratio between the egg and pupae stages suggests that workers eliminated some male brood, and the population sex-investment ratio was between the queens' and workers' equilibria. Altogether, these data provide evidence for an ongoing conflict between queens and workers, with a prominent influence of queens as a result of their control of egg sex ratio.     

Effects of gamma irradiation used to inhibit potato sprouting on potato tuber moth eggs Phthorimaea operculella Zeller (Lep., Gelechiidae)

Abstract: Different age groups of potato tuber moth, Phthorimaea operculella Zeller, eggs were exposed to gamma irradiation at incremental doses up to 150 Gy (the upper dose limit allowed for potato sprout inhibition). Young eggs were more likely to be sensitive to gamma irradiation than older eggs and the sensitivity level declined with age of the eggs. The exposure of 3–3.5-day-old eggs to 150 Gy resulted in a 26% increase in the egg incubation period compared with unirradiated eggs. The minimum dose required to prevent 4–4.5-day-old eggs from hatching was 1 kGy. When eggs were exposed to 75 Gy, 9.7% of larvae survived to the adult stage but emerged as deformed moths. At 100 and 125 Gy, 10.2 and 9.6% of larvae pupated, respectively, although no adult eclosion was recorded, whereas at 150 Gy, the larvae remained in mid-instar stage and eventually died. Gamma irradiation doses applied to inhibit potato sprouting could be also considered as a valuable control tool against potato tuber moth infestations.     

Genetic relationships between native and introduced populations of the little fire ant Wasmannia auropunctata

Native to much of Central and South America, the little fire ant Wasmannia auropunctata has been rapidly spreading throughout the world. In its introduced range, W. auropunctata is frequently linked with drastic reductions of ant diversity; anecdotal reports of damaging attacks on vertebrates are also common. As it poses an ever-increasing threat to biodiversity, W. auropunctata has emerged as a model system for the study of ecological differences between native and invasive ant populations. These studies have been hampered by a lack of information on the genetic relatedness between native and introduced populations. By investigating the genetic structure of W. auropunctata populations, we provide a framework for conducting phylogenetically independent tests of differences between these ants in their native and invasive ranges. Phylogenetic analyses, based on the cox1 / cox2 region of mtDNA, revealed at least three separate source populations of W. auropunctata distributed across two large clades. Much of the Caribbean region, presumably part of the native range, is inhabited by a clade of ants sharing very similar or identical mtDNA haplotypes, suggesting the possibility of multiple introductions or high levels of gene flow across that area. Most invasive populations in the Pacific were closely related to these ants. The invasive populations in Gabon and New Caledonia arise from another, relatively distantly related clade. Phylogenetically independent contrasts confirm McGlynn's (1999) observation that invasive W. auropunctata populations are smaller than native populations. Given the complex phylogeographical structure of W. auropunctata populations, future comparative work should correct for phylogenetic effects.     

Do army ant queens re-mate later in life?

Queens of eusocial Hymenoptera are inseminated only during a brief period before they start to lay eggs. This has probably been kin-selected because repeated insemination of old queens would normally be against the inclusive fitness interest of their daughter workers. Army ants have been considered to be the only possible exception to this rule due to their idiosyncratic life-history. We studied two distantly related species of army ants, the African Dorylus (Anomma) molestus and the Neotropical Eciton burchellii and present data from microsatellite genotyping, behavioural observations and sperm counts.We also describe the copulation behaviour of African army ants for the first time. Our results strongly suggest that, contradictory to earlier contentions, army ant queens do not mate repeatedly throughout their life and thus do not constitute an exception among the eusocial Hymenoptera in this respect. Sperm counts for males and queens of both species show that army ant queens have to mate with several males to become fully inseminated. However, sperm limitation by queens is unlikely to have been the prime reason for the evolution of high queen-mating frequencies in this group. Received 5 July 2006; revised 26 September 2006; accepted 11 October 2006.     

Demyelination Occurred as the Secondary Damage Following Diffuse Axonal Loss in a Rat Model of Radiation Myelopathy

The main purpose of the present study was to examine the time and dose-dependent course of demyelination in the rat radiation myelopathy model in the first 180 days after irradiation of the spinal cord. An irradiated cervical spinal cord rat model (C2-T2 segment) was generated using a ⁶⁰Co irradiator to deliver 50 Gy and 100 Gy, respectively. The behavioral dysfunction was observed by the forelimb paralysis scoring system. The histological damage in the irradiated spinal cord was examined by hematoxylin/eosin staining, luxol fast blue staining, immunohistochemical analysis, methylene blue/Azure II staining, and uranyl/lead salts staining. The gene expression of oligodendrocyte-related markers were also determined by quantitative real-time PCR. The complete loss of forelimb motor function in all animals was observed at 180 days 50 Gy post-irradiation and at 120 days 100 Gy post-irradiation. We demonstrated that a 50 and 100-Gy single-dose irradiation of the C2-T2 spinal cord segment resulted in diffuse axonal loss and elicited secondary demyelination damage in the spinal cord. We further observed that 100-Gy irradiation reduced the gene expression of myelin oligodendrocyte glycoprotein in irradiated spinal cord. Taken together, our data not only define diffuse axonal loss as the main histological damage but also provide the first evidence that demyelination occurred as the secondary damage in irradiated spinal cord.