Longevity in vitro of Ditylenchus dipsaci (Kiihn) Filipjev from narcissus, JE Bosher

Tags: room temperature, King Alfred, C. Refrigerator, storage temperature, B. C. Previous, narcissus, wool, temperature, nematodes, lyophilization, Ditylenchus dipsaci, Copyright, glass vials, narcissus bulbs, Canada Agriculture, BOSHER Plant Pathology Laboratory, Helminthological Society
Content: JULY, 1960]
Helminthological Society
127
Longevity in vitro of Ditylenchus dipsaci (Kiihn) Filipjev from Narcissus J. E. BOSHER Plant Pathology Laboratory, Beseardi Branch, Canada Agriculture, Saanichton, B. C. Previous studies (Hasting, 1942) of survival of the pre-adult stage of Diti/leuclius (lipsaci in the form of nematode "wool" as found in the basal region of narcissus bulbs have shown 100% mortality after storage in glass vials at room temperature for a period of four years. Fielding (1951) records survival of /). (lipsaci in dried plant tissues for a period of 23 years, indicating that maintenance of life in these nematodes is affected by obscure factors of environment. Further studies (Bosher and McKeen, 1954) showed that D. dipsaci in the dry state from narcissus "wool" and in certain media survived freezing to -80° C. followed by vacuum dehydration (lyophilization) and storage hi raciio in sealed tubes for a period of up to 28 days. It was postulated that lyophilization may prove to be of value as a method for the maintenance of stock cultures for laboratory investigations. Survival of these nematodes in relation to environment has been further investigated as follows. MATERIALS AND METHODS Clusters of nematode "wool" from narcissus collected in 1951 were divided into two portions and placed in separate screw-cap glass vials. One portion was placed on a laboratory shelf at room temperature and one lot was stored in a household type refrigerator at 2°-4° C. Portions of the clusters were removed at 2-year intervals until 1958 and the percentage of motile nematodes recorded after 48 hours immersion in shallow tap water as shown in Table 1. Sealed tubes of nematodes i-n- raciio prepared in 1953 by lyophilization at -SO0 C., held in storage at room temperature for 5 years, were opened and viability of the nematodes was determined by immersion in shallow tap water. Table 2 shows the percentage of nematodes that regained motility as compared with similar samples examined in 1953 shortly after lyophilization.
Table 1. Eevival of J)i~tylencltnt< diptiaei from narcissus "wool" in vitro in relation to time and storage temperature.
Storage
% revival after storage for:
1 yr.
3 yrs. 5 yrs. 7 yrs.
Boom temperature, approx. 21° C. Refrigerator at 2° -4° C.
8(5
38
3
0
89
86
81
78
RESULTS Xematodes that revived from the material stored at low temperature regained active motility within 24 hours after being placed in water. The small percentage that revived after five years at room temperature exhibited comparatively feeble movement between 24 and 48 hours after immersion. Xematodes from tubes of the lyophilized series were poured into pots containing bulbs of narcissus var. King Alfred from nematode-free stock. Examination of the bulbs after one year's growth showed populations of T). dipsaci of all stages from eggs to adults in bulbs inoculated with nematodes from lyophilized dry wool and dry wool in beef serum. Xo nematodes were found in bulbs inoculated with I), dipsaci treated as dry wool in sucrose or in water.
Copyright © 2011, The Helminthological Society of Washington
128
PROCEEDINGS OF THE
[VOL. 27, No. 2
Table 2. Bevival of Ditylenchus dipsaci from narcissus "wool" subsequent to lyophilizatioii at -80° C. in relation to time of storage in vacuo at room temperature.
Nematode state
% revival after storage for:
28 days
5 years
Dry wool
80
20.4
Dry wool in beef serum
80
11.8
Dry wool in 50% sucrose
90 #
0
Dry wool in water
30
0
#12 days storage.
DISCUSSION The results presented herein are a further indication of the remarkable resistance to unfavorable environment of the pre-adult stage of D. dipsaci in the dry state. Storage at low temperature is indicated as a more effective method for maintenance of visability of these nematodes than lyophilizatioii at extreme cold followed by storage in vacua.
LITERATURE CITED BOSHER, J. E. and W. E. McKEEN. 1954. Lyophiliaztion and low TEMPERATURE STUDIES with the bulb and stem nematode Ditylencltus dipsaci (Kiihn 1858) Filipjev. Proc. Helminth. Soc. Wash. 21:113-117. FIELDING, M. J. 1951. Observations on the length of dormancy in certain plant infecting nematodes. Proc. Helminth. Soc. Wash. 18:110-112. HASTINGS, R. J. 1942. Longevity of congelations of bulb nematode, dipsaci. (Kiihn) Filipjev, from narcissus. Sci. Agr. 23:1-3.
Isolation of Trichojnonas gallinae from the White-winged Dove, Zenaida a. asiatica Louis N. LOCKE and William H. KIEL, JR. Bureau of Sport Fisheries and Wildlife, Laurel, Maryland. On September 13, 1959, a series of 17 throat swabbings were obtained from white-winged doves shot by hunters near Edinburg-, Texas. The swabs were placed in tubes of Diamond's trichomonad medium (Jour. Parasit., 43: 488-490, 1957) and mailed to the Patuxent Research Refuge for examination. Upon arrival at the refuge, two and three days later, the tubes were placed in an incubator at 37.5° C. The following day the cultures were examined for trichomonads. Six of the 17 were positive for Tricliomonas gallinae; all six were swabbings from the 10 adult doves that on External Examination were normal and fat. No Tricliomonas gallinae was isolated from the 7 immature birds. Although T. gallinae frequently has been isolated from mourning doves, this is, to our knowledge, the first report of its isolation from the whitewinged dove. Thanks are extended to Mr. David Blankinship, Texas Game and Fish Commission, for his aid in collecting several of the samples.
Copyright © 2011, The Helminthological Society of Washington

JE Bosher

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