Phytosanitary irradiation using X-rays prevents reproduction in the semi-slug Parmarion martensi (Stylommatophora: Ariophantidae), a host of the human pathogenic nematode Angiostrongylus cantonensis (Rhabditida: Angiostrongylidae).

BACKGROUND: Phytosanitary irradiation is used to control insect pests of quarantine concern on exported fresh horticultural products. Generic irradiation doses of 150 and 400 Gy are approved for tephritid fruit flies and all other insects, respectively. Other invertebrates such as gastropods (snails and slugs) may be classified as quarantine pests and require a disinfestation treatment. Parmarion martensi Simroth (Stylommatophora: Ariophantidae) is a semi-slug quarantine pest sometimes found on fresh sweet potatoes and other fruits and vegetables exported from Hawai'i to the continental USA. Also, P. martensi is a host of the parasitic nematode Angiostrongylus cantonensis (Rhabditida: Angiostrongylidae), the causative agent of neuroangiostrongyliasis or rat lungworm disease in humans. We conducted a study to determine if phytosanitary irradiation could control P. martensi and thereby reduce the risk of transmitting A. cantonensis in the USA. RESULTS: Two-, 12-, and 21-week-old P. martensi were treated with X-ray radiation at a dose of 150 or 400 Gy or left untreated as controls then held in the laboratory for up to 250 days. Survivorship and reproduction were recorded every 2-3 days and individual weights were measured biweekly. Irradiation at 150 and 400 Gy reduced growth and increased the mortality rate compared to untreated controls and prevented reproduction. CONCLUSION: Phytosanitary irradiation treatment at doses ≥150 Gy will prevent the establishment of viable populations of P. martensi. The literature on radiation tolerance in gastropods suggests that the internationally approved generic dose for tephritid fruit flies of 150 Gy may be effective against many slug and snail pest species.

The growth effects of gamma-ray irradiation on third-stage larvae of Angiostrongylus cantonensis in snail.

In order to clarify growth effects of gamma-ray irradiation on the third-stage larvae of Angiostrongylus cantonensis, snails naturally infected with larvae of A. cantonensis were irradiated with a cobalt-60 source, and the total dosage of exposure was 0.1 kGy. Third-stage larvae were collected from each group of irradiated and non-irradiated snails and then administered to rats. A significant reduction was observed in the number of recovery worms from rats sacrificed at 45 days after infection compared with the control group. Moreover, there was an increase in the percentage of female worms, and the body length of the irradiated group's worms was significantly shorter than that of the control group. In ultrastructural observations by transmission electron microscope, the cuticle was thickened and the subcuticle showed partial vacuolization and separation from muscle in the irradiated group. In addition, the muscles myofibril became shorter, the microvilli of intestinal mucosa were shorter and irregular, the terminal web disappeared, and the phagocytic vesicles, mitochondria, lysosomes and secretion granules decreased in number, respectively. Focusing on the mitochondria of the epithelial cells of the intestine, there were degenerative changes in the irradiated group showing vacuolization. The gonads of female worms recovered from the irradiated group revealed degenerative vacuolization, and the gonads of male worms could not be found.

Effect of irradiation on the viability of Angiostrongylus cantonensis and A. costaricensis infective larvae.

Infective larvae (third-stage larvae) of both Angiostrongylus cantonensis and A. costaricensis from their snail intermediate host were subjected to either X-ray or gamma-ray irradiation. The viability of the irradiated larvae was assayed by oral inoculation of the larvae into rodents (A. cantonensis in mice and rats, A. costaricensis in mice only). From the results of worm recovery, the minimal dose of irradiation that inhibited the infectivity of the third-stage larvae of A. cantonensis and A. costaricensis was 2 and 4 kGy, respectively.

Killing effect of gamma irradiation on Angiostrongylus cantonensis in snails.

Cobalt-60 gamma irradiation was used to kill the larvae of Angiostrongylus cantonensis in four groups of snails that were irradiated at a rate of 15.4 Gy/min for 16.23 min (0.25 kGy), 32.47 min (0.50 kGy), 48.70 min (0.75 kGy), and 64.94 min (1.00 kGy), respectively. Third-stage (infected stage) larvae were isolated from these snails and 50 were administered to each experimental animal via an esophageal tube. At a dosage of 0.25 kGy, the mean +/- SD number of worms recovered from 30 rats and 30 mice was 15.6 +/- 6.5 and 11.3 +/- 6.2, respectively. These were significantly different from the number of worms recovered from the nonirradiated (control) groups, which were 28.9 +/- 9.6 (P < 0.05) and 16.6 +/- 10.3 (P < 0.05) for rats and mice respectively. Worms and cellular infiltrations in lung tissue of rats were histopathologically identified in the 0.25-kGy-irradiated group as well as in the control group. Worms and eosinophilic meningitis were also noted in the brain tissue of mice in control groups. No worms or pathologic findings were observed in the heart, lung, or brain tissue of rats or mice infected with larvae irradiated at or exceeding 0.50 kGy. It is therefore concluded that a dosage of 0.50 kGy is sufficient to prevent successful infection in mice and rats.