scholarly journals Cold tolerance mechanisms of the free-living stages of Trichostrongylus colubriformis (Nematoda)

1989 ◽  
Vol 145 (1) ◽  
pp. 353-369
Author(s):  
D. A. Wharton ◽  
G. S. Allan

1. All free-living stages of the nematode parasite of sheep, Trichostrongylus colubriformis Giles, survived exposure to freezing temperatures in contact with water, with the exception of the first-stage juvenile (J1). The third-stage juvenile (J3) was the most resistant stage. The order of relative survival of the different stages was different from that of the lowest F50 (the temperature at which 50% froze), suggesting that an ability to supercool was not the only determinant of survival. 2. The F50 was shown to be a good measure of the degree of supercooling and to extend greatly the lower size limit of organisms that could be measured. 3. The J3 uses a freeze-avoiding strategy by supercooling when in air or covered by liquid paraffin. In water it uses a mixture of freeze-avoiding and freeze-tolerant strategies, with a proportion of the population surviving freezing caused by exogenous ice nucleation. 4. Removal of the J3 sheath results in a shift from freeze avoidance to freeze tolerance, with an overall reduction in survival. A major function of the sheath may be to reduce the probability of exogenous ice nucleation.

1996 ◽  
Vol 199 (2) ◽  
pp. 465-471 ◽  
Author(s):  
J Mugnano ◽  
R Lee ◽  
R Taylor

During the autumn, the third-instar larvae of the gall fly Eurosta solidaginis acquire freeze tolerance and their crystallization temperatures increase into the -8 to -10 °C range. Despite conflicting reports, efficient endogenous ice nucleators have not been identified in this freeze-tolerant insect. We found large crystalloid spheres within the Malpighian tubules of overwintering larvae. Energy-dispersive X-ray microanalysis and infrared spectroscopy indicated that the spherules were a hydrate of tribasic calcium phosphate. To test for ice-nucleating activity, we placed the calcium phosphate spherules in 10 µl of Schneider's insect medium and cooled them in a refrigerated bath. The addition of spherules increased the crystallization temperature of Schneider's medium by approximately 8 C, from -18.4±0.8 °C to -10.1±0.9 °C (mean ± s.e.m., N=20). Ice-nucleating activity (-10.9±0.9 °C) was also demonstrated in fat body cells suspended in 10 µl of Schneider's medium. Both calcium phosphate spherules and fat body cells have ice-nucleating activity sufficiently high to explain whole-body crystallization temperatures. Furthermore, other crystalloid deposits, commonly found in diapausing or overwintering insects, also exhibited significant ice-nucleating activity. These endogenous crystalloid deposits represent a new class of heterogeneous ice nucleators that potentially regulate supercooling and promote freeze tolerance in E. solidaginis and possibly in other overwintering insects.


2005 ◽  
Vol 77 (2) ◽  
pp. 83-92 ◽  
Author(s):  
C. Richard ◽  
J.-G. Martin ◽  
S. Pouleur

In order to know which species of Fusarium are ice nucleating and to determine the factors affecting their pathogenicity, ice nucleation activity (INA) was examined in Fusarium oxysporum, F. sporotrichioides, and F. tricinctum. Positive controls (lna+) used were F. acuminatum and F. avenaceum. The test for fungal INA was done with a simple and rapid tube nucleation assay. Twelve out of the 42 F. oxysporum isolates, and 8 out of 14 F. tricinctum isolates were lna+. No INA was detected in F sporotrichioides. In this test the threshold freezing temperature tended to increase with culture age, reaching a peak of -1°C in a few samples, which is as high as the warmest INA reported for bacteria, and higher than the INA detected in pure cultures of free-living fungi, lichen fungi, lichen algae and cyanobacteria. This is the first report of INA for F oxysporum.


1991 ◽  
Vol 261 (6) ◽  
pp. R1346-R1350 ◽  
Author(s):  
J. P. Costanzo ◽  
R. E. Lee

Erythrocytes from the freeze-tolerant wood frog (Rana sylvatica) were subjected to in vitro tests of freeze tolerance, cryoprotection, and osmotic fragility. The responses of cells from frogs acclimated to 4 or 15 degrees C were similar. Erythrocytes that were frozen in saline hemolyzed at -4 degrees C or lower. The addition of high concentrations (150 and 1,500 mM) of glucose or glycerol, cryoprotectants produced naturally by freeze-tolerant frogs, significantly reduced cell injury at -8 degrees C, but concentrations of 1.5 or 15 mM were ineffective. Hemolysis was reduced by 94% with 1,500 mM glycerol and by 84% with 1,500 mM glucose; thus glycerol was the more effective cryoprotectant. Mean fragility values for frog erythrocytes incubated in hypertonic and hypotonic saline were 1,938 and 49 mosM, respectively. Survival in freeze tolerance and cryoprotection experiments was comparable for erythrocytes from frogs and humans, suggesting that these cells may respond similarly to freezing-related stresses. However, the breadth of osmotic tolerance, standardized for differences in isotonicity, was greater for frog erythrocytes than for human erythrocytes. Our data suggest that erythrocytes from R. sylvatica are adequately protected by glucose under natural conditions of freezing and thawing.


2012 ◽  
Vol 21 (4) ◽  
pp. 372-378 ◽  
Author(s):  
Raquel Abdallah da Rocha ◽  
Patrizia Ana Bricarello ◽  
Gilberto Pedroso da Rocha ◽  
Alessandro Francisco Talamini do Amarante

This experiment aimed to assess the recovery of infective larvae (L3) of Trichostrongylus colubriformis from Brachiaria decumbens cv. Australiana, Cynodon dactylon cv. Coast-cross and Panicum maximum cv. Aruana. The experimental module comprised six plots, with two plots per herbage species. Larval survival was assessed from autumn to winter, under the effect of two herbage-paring heights (5 and 30 cm). TThe paring was carried out immediately before contamination with faces containing T. colubriformis eggs. The feces and herbage were collected at one, two, four, eight, 12 and 16 weeks after feces had been deposited in the experimental plots. In general, larvae were recovered from both herbage and feces until the 16th week. The longer persistence of these larvae in the environment was probably due to warmer temperatures. The number of L3 recovered from the pasture was not influenced by the height of plants, except for Brachiaria and Aruana herbage in the fourth week. Regarding the concentrations of larvae per kg of dry matter (L3/kg DM), recovery was higher from low pasture in all three herbage species. During the autumn, the development and survival of the T. colubriformis free-living stages were not affected by the different herbage species.


2020 ◽  
Vol 98 (2) ◽  
pp. 145-153 ◽  
Author(s):  
Jing Zhang ◽  
Liam J. Hawkins ◽  
Kenneth B. Storey

The wood frog is one of the few freeze-tolerance vertebrates. This is accomplished in part by the accumulation of cryoprotectant glucose, metabolic rate depression, and stress response activation. These may be achieved by mechanisms such as DNA methylation, which is typically associated with transcriptional repression. Hyperglycemia is also associated with modifications to epigenetic profiles, indicating an additional role that the high levels of glucose play in freeze tolerance. We sought to determine whether DNA methylation is affected during freezing exposure, and whether this is due to the wood frog’s response to hyperglycemia. We examined global DNA methylation and DNA methyltransferases (DNMTs) in the liver and muscle of frozen and glucose-loaded wood frogs. The results showed that levels of 5-methylcytosine (5mC) increased in the muscle, suggesting elevated DNA methylation during freezing. DNMT activities also decreased in muscle during thawing, glucose loading, and in vitro glucose experiments. Liver DNMT activities were similar to muscle; however, a varied response to DNMT levels and a decrease in 5mC highlight the metabolic role the liver plays during freezing. Glucose was also shown to decrease DNMT activity levels in the wood frog, in vitro, elucidating a potentially novel regulatory mechanism. Together these results suggest an interplay between freeze tolerance and hyperglycemic regulation of DNA methylation.


1978 ◽  
Vol 64 (6) ◽  
pp. 1024 ◽  
Author(s):  
Z. I. Khan ◽  
W. Dorsman

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