Morphological variability,molecular phylogeny,and biological characteristics of the nematophagous fungus Duddingtonia flagrans

This study aims to investigate the molecular phylogenetic analysis, morphological variability, nematode‐capturing ability, and other biological properties of Chinese Duddingtonia flagrans isolates. We isolated 13 isolates of D. flagrans and found features that have never been reported before, such as two to three septa incluing club‐shaped conidia. Meanwhile, we conducted molecular phylogenetic analysis of the seven isolates and tested the radical growth of the isolates under different pH values, temperatures, and media. The capturing ability against infective larvae (L3) of Cooperia spp. in yak was detected in vitro. Finally, one isolate was selected for scanning electron microscopy (SEM) to investigate the trap formation process. The fungal sequence was obtained and submitted to GenBank (Accession no. KY288614.1, KU881774.1, KP257593.1, KY419119.1, MF488979.1, MF488980.1, and MF488981.1), and the tested isolates were identified as D. flagrans. Except for three isolates, the radial growth of the other isolates on 2% corn meal agar and 2% water agar exhibited faster growth than on other media. The fungus could not grow at 10 and 40°C but grew within 11 to 30°C. Moreover, it did not grow at pH 1–3 and 13–14, but instead at pH 4–12. In the in vitro experimental, L3s were reduced by 94.36%, 88.15%, and 91.04% for SDH035, DH055, and F088, respectively. SEM results showed that at 8 hr post addition of nematodes, some of the latter were captured. In the later stages of the interaction of the fungus with nematodes, a large number of chlamydospores were produced, especially on the predation trap. Results of the present study provided information about the molecular phylogenetic analysis, morphological variability, nematode‐capturing ability, and other biological properties of Chinese Arthrobotrys flagrans isolates before administering them for biocontrol.     

Growth rate and trapping efficacy of nematode-trapping fungi under constant and fluctuating temperatures

The effect of temperature on radial growth and predatory activity of different isolates of nematode-trapping fungi was assessed. Four isolates of Duddingtonia flagrans and one isolate of Arthrobotrys oligospora were inoculated on petri dishes containing either corn-meal agar (CMA) or faecal agar and then incubated for 14 days under three different constant and fluctuating temperature regimes. The radial growth was similar on the two substrates at each temperature regime. All fungal isolates showed a higher growth rate at a constant 20 °C. At 10° and 15 °C, all D. flagrans isolates showed very similar patterns of radial growth at both constant and fluctuating temperatures. At 20 °C, they grew significantly faster at constant than at fluctuating temperatures. A. oligospora grew significantly faster than all D. flagrans isolates except when incubated at a fluctuating 20 °C. Spores of each fungal isolate were added to faecal cultures containing eggs of Cooperia oncophora at a concentration of 6250 spores/g faeces. The cultures were incubated for 14 days at the same temperature regimes described above. Control faeces (without fungal material) were also cultured. More larvae were recovered from the fungus-treated cultures incubated at a constant 10° or 15 °C than from those incubated at the respective fluctuating temperatures, except for one D. flagrans isolate. Incubation at 20 °C showed the opposite effect. The general reduction observed in the number of nematode larvae due to fungal trapping was 18–25% and 48–80% for a constant and fluctuating 10 °C, 70–96% and 93–95% for a constant and fluctuating 15 °C, and 63–98% and 0–25% for a constant and fluctuating 20 °C, respectively. Received: 15 December 1998 / Accepted: 16 February 1999     

Biological control of sheep gastrointestinal nematodiasis in a tropical region of the southeast of Brazil with the nematode predatory fungi Duddingtonia flagrans and Monacrosporium thaumasium

Formulations in matrix of sodium alginate (pellets) of the nematode predatory fungi Duddingtonia flagrans and Monacrosporium thaumasium were evaluated in the biological control of sheep gastrointestinal nematodiasis. Three groups (1, 2, and 3), each one with eight sheep of the Santa Inês breed, at the ages of 15–48 months, were placed in paddocks of Brachiaria decumbens for 5 months. In group 1, each animal received 1 g/10 kg of live weight (l.w.) of pellets of D. flagrans (0.2 g of fungus/10 kg l.w.). In group 2, each animal received 1 g/10 kg of l.w. of pellets of the fungus M. thaumasium (0.2 g of fungus/10 kg l.w.), twice a week, for 5 months. In group 3 (control), the animals received 1 g/10 kg of live weight of pellets without fungus. The monthly averages of the egg countings per gram of feces of the animals of groups 1 and 2 treated were 71.6% and 61.1% smaller, respectively, in comparison to the animals of group 3 (control). The treatment of sheep with pellets containing the nematophagous fungi D. flagrans and M. thaumasium may be used as an alternative for the control of sheep gastrointestinal nematodiasis.     

Efficacy of Duddingtonia flagrans and Arthrobotrys robusta in controlling sheep parasitic gastroenteritis

The aim of this study was to evaluate the efficacy of formulations of sodium alginate matrix (pellets) of the nematode predatory fungi, Duddingtonia flagrans (AC001 isolate) and Arthrobotrys robusta (I-31 isolate), in the biological control of sheep gastrointestinal nematode infections. Thirty young Bergamacia ewes were allocated into three groups: In group 1 (control), the animals received 2 g/10 kg of live weight (l.w.) of pellets without fungus; in group 2, each animal received 2 g/10 kg of l.w. of pellets of D. flagrans (0.2 g of fungus/10 kg l.w.); and in group 3, each animal received 2 g/10 kg of l.w. of pellets of A. robusta (0.2 g of fungus/10 kg l.w.). The animals of each group were kept separately under rotational grazing. Pellets, with or without fungi, were mixed with 1 kg animal food and administered twice a week for 6 months. There was no significant difference in mean live weight and packed cell volume among groups (P > 0.05). Mean nematode fecal egg counts (FEC) did not significantly differ between the control and the remaining groups, except in one or two collections, when FEC was higher in the control group than in group 2 and group 3, respectively. The group that received A. robusta pellets needed less salvage anthelmintic treatments. Haemonchus contortus was the predominant species recovered from tracer lambs. The nematophagous fungi, D. flagrans and A. robusta, did not provide satisfactory results in the prophylaxis of parasitic gastroenteritis in sheep, under the conditions of the present study.     

Comparative efficacy of the nematode-trapping fungus Duddingtonia flagrans against Haemonchus contortus, Teladorsagia circumcincta and Trichostrongylus colubriformis in goat faeces: influence of the duration and of the temperature of coproculture

Amongst the alternative strategies to the use of anthelmintics, the administration of Duddingtonia flagrans spores has already proved its efficacy in reducing the number of developing larvae of several nematode species in goat faeces. In this trial, the efficacy of this fungus against the larvae of the three major nematode species of goats was compared in various conditions of coproculture. Twelve strongyle free goats were experimentally infected with either Trichostrongylus colubriformis, Teladorsagia circumcincta or Haemonchus contortus larvae. Half of the animals received an oral dose of 5×10⁵ Duddingtonia chlamydospores/kg BW daily for 27 days, whereas the remaining was kept as control goats. From the 7th day of administration onwards, individual coproscopical examinations as well as coprocultures, which were incubated 4, 7, 10 or 14 days at 21 or 28°C, were performed. The reduction in developing larvae due to the activity of Duddingtonia ranged from 62.8 to 99.5% compared to control. The trapping efficacy depended on temperature (better activity of the fungus at 21 than at 28°C) and on duration (larval reductions lower after 4 days than after 7, 10 and 14 days of coproculture). Teladorsagia larvae were the least trapped, and Haemonchus larvae were the most trapped.     

Isolation and characterization of the nematophagous fungus Arthrobotrys conoides

The spread of organic farming and the development of resistance to anthelmintics by parasites, especially in small ruminants, have necessitated the search for alternative methods of nematode control. Biological control using nematophagous fungi is one option; however, few studies have been conducted with native strains. The present study was divided into two phases. In the first phase, we aimed to isolate, identify, and assess the in vitro predatory activity of nematophagous fungi that had been isolated on Trichostrongylidae third-instar larvae. In the second phase, the isolate with superior predatory activity in vitro was molecularly characterized, and its morphological plasticity was observed using scanning electron microscopy (SEM) on Haemonchus third-instar larvae. Of the 56 soil samples from different regions of Paraná State, Brazil, 57 fungal strains were recovered, of which four exhibited predatory activity. Two pure isolates were obtained: the CED and LIN strains. After demonstrating 96.35 % predatory activity for the CED strain, this strain was selected and characterized using molecular criteria by sequencing the rDNA internal transcribed spacer and was identified as Arthrobotrys conoides (GenBank ID: JN191309). Morphological patterns in this strain during the interaction between the fungus and the nematode were revealed by SEM, in which two extensions of the infection bulb that was used to pierce the nematode's cuticle were clearly visible.     

Kinetics of capture and infection of infective larvae of trichostrongylides and free-living nematodes Panagrellus sp. by Duddingtonia flagrans

Duddingtonia flagrans, a nematode-trapping fungus, has been investigated as an agent for biological control against infective larvae of gastrointestinal nematode parasites of production animals. The initial process of nematode-trapping fungi infection is based on an interaction between the trap structure of the fungus and the surface of the nematode cuticle. This report investigates by light and scanning electron microscopy the kinetics of capture and infection during the interaction of D. flagrans with the infective larvae (L₃) of trichostrongylides and the free-living nematode Panagrellus sp. D. flagrans was cultivated for 7 days in a Petri dish containing agar–water. L₃ and Panagrellus sp. were inoculated in the Petri dishes and the samples consisting of agar–L₃–fungi and agar–Panagrellus sp.–fungi were collected after 10, 20, 30, 40, 50, 60, and 70 min and 3, 4, 5, 10, 15, 20, and 25 h of interaction. All samples were observed by light microscopy. The samples with 1, 5, 15, and 25 h of interaction were also analyzed by scanning electron microscopy. The interaction was monitored up to 25 h. An initial differentiation of predation structures was observed after 30 min of interaction. The presence of traps and of captured L₃ or Panagrellus sp. occurred after 70 min. The live captured nematodes were observed up to 3 h of interaction. However, after 4 h, all Panagrellus sp. were dead. It took 15 h of interaction for the fungus to invade the L₃, and the presence of hyphae inside the nematode near the region of penetration was evident. At this time, the hyphae had filled the whole body of Panagrellus sp. The complete occupation of the body of L₃ occurred at 20 h of interaction and with 25 h the nematode was completely damaged except for the cuticle. Although the double cuticle of L₃ slows the penetration of D. flagrans, it was possible to verify that the process of trap formation and capture occurs quickly when both nematodes were tested, suggesting that the organisms would eventually be killed once in contact with the fungi encouraging the use of the fungus as a biological control agent.     

In vitro influence of temperature on the biological control activity of the fungus Duddingtonia flagrans against Haemonchus contortus in sheep

Recently, research for alternative methods to combat gastrointestinal parasites has increased, and the biological control activity of the fungus Duddingtonia flagrans stands out. In this study, the possible influence of temperature on the nematophagous activity of D. flagrans, after gastrointestinal passage, against Haemonchus contortus in sheep was analysed. Four female sheep, between 2 and 3 years of age and weighing between 40 and 50 kg, were used. Two sheep were parasitised with H. contortus, while two other sheep were dewormed. Before the collection of faeces, one of the dewormed animals received a dosage of 1?×?10⁶ chlamydospores of D. flagrans, lyophilised in gelatin capsules, for three consecutive days. The faeces were collected with collector bags, mixed, and then separated as samples with (fungus; 800 eggs per gram (EPG) of faeces) or without fungus (control; 900 EPG). Each sample (five replicates) was maintained in a biochemical oxygen demand incubator under different temperatures (5, 10, 15, 20, 25, 30, or 35 °C) for 21 days, followed by determination of the larval recovery. Compared to the control group, the best temperature for fungal action was 30 °C, while no larvae were recovered at 5 °C. At 10 °C, fungal action was detected, yet there was no significant difference in the percent larval reduction between all temperatures, demonstrating that larval presence seems to be the main factor affecting the nematophagous action of D. flagrans. Temperature does not appear to be a limiting factor in the biological control activity of D. flagrans against H. contortus, but larval presence, which was not observed at 5 °C, is mandatory. At low temperatures, which are typically suboptimal conditions for fungal and larval development, the lyophilised D. flagrans reduced the number of H. contortus larvae, which demonstrates the biological control potential and the potential use of D. flagrans in the subtropics.     

Efficacy of an energy block containing Duddingtonia flagrans in the control of gastrointestinal nematodes of sheep

The efficacy of the nematode-trapping fungus Duddingtonia flagrans incorporated into an energy block was evaluated for the control of gastrointestinal nematodes in sheep. Four naturally parasitised sheep with average nematode egg counts of 2,470 eggs per gram grazed by pairs on two similar parasite-free paddocks for 30 days. During that period, one pair of sheep (treated animals, T1) received an energy block containing chlamydospores of D. flagrans at a dose of 200,000 chlamydopores/kg bw/day, while the second pair (control animals, C1) received a fungus-free energy block. The animals in both groups were taken off the paddocks after contaminating the pastures for a month with either nematode eggs plus fungal chlamydospores (T1) or nematode eggs alone (C1). Twelve parasite-free sheep were divided into two groups of six animals each, the treated group (T2) was placed on the paddock previously contaminated with parasites and fungus, while the control group (C2) was placed on the parasite-only paddock. These two groups grazed on their respective paddocks during 30 days and were then housed for 15 days, after which period they were slaughtered in order to determine the parasite burden present in each animal. Results showed that animals in group T2 harboured significantly less nematodes than their counterpart in group C2. The efficacy of D. flagrans was 92% against the total parasite burden, 100% against Haemonchus contortus and Teladorsagia circumcincta, 89.9% against Trichostrongylus colubriformis, 87.5% against Cooperia onchopora, and 90% against Trichostrongylus axei. No efficacy was detected against Nematodirus spathiger, Trichuris ovis and T. skrjabini.     

Resistance of different fungal structures of Duddingtonia flagrans to the digestive process and predatory ability on larvae of Haemonchus contortus and Strongyloides papillosus in goat feces

The dynamics of the passage of conidia, chlamydospores, and mycelia of the fungus Duddingtonia flagrans through the digestive tracts of goats was evaluated. Four groups with five goats each were formed. In the group conidia, each animal received 1 × 10⁶ D. flagrans conidia per kilogram of live weight. In the group chlamydospore, each animal received 1 × 10⁶ chlamydospores per kilogram of live weight. In the group mycelia, each animal received 1 g of mycelium mass per kilogram of live weight. In the control group, the animals received no fungal structure. Feces were obtained 3 h before and 12, 24, 30, 36, 42, 48, 60, 72, 84, and 96 h after the inoculation. The feces were placed in Petri dishes containing water-agar. The Petri dishes were examined to detect the fungus and trapped nematodes. A second trial evaluated the effect of the fungal structures on the number of gastrointestinal larvae of Haemonchus contortus and Strongyloides papillosus harvested from the fecal cultures of the goats. The feces were obtained from the goats in the 12–24, 24–30, 30–36, 42–48, 60–72, 72–84, and 84–96 intervals after the inoculation. D. flagrans survived the digestive process of the goats and maintained its predatory activity, being observed from 12 to 96 h before inoculation in the animals that received chlamydospores and conidia.     

Effect of artificial gastrointestinal fluids on the excystation and metacystic development of Entamoeba invadens

The effect of artificial gastric fluid (AGF), containing 0.5% pepsin and 0.6% hydrochloric acid, pH 1.8, in distilled water, on the excystation and metacystic development of Entamoeba invadens was examined. Excystation, which was assessed by counting the number of metacystic amoebae after inducing excystation, was enhanced by pretreatment of cysts with AGF for 30 to 60 min at 37°C but not 26°C. Longer exposure of cysts to AGF significantly reduced their viability. Significant enhancement of excystation was observed by pretreatment of cysts with distilled water only at 37°C. In addition, 0.6% hydrochloric acid had a comparable enhancing effect on excystation to AGF. Metacystic development, when determined by the number of nuclei in amoeba, was slightly enhanced by pretreatment with AGF. An artificial intestinal fluid (AIF), containing 1% pancreatin, 1% sodium bicarbonate, and 5% ox bile, pH 8.0, in distilled water, had a significant toxic effect on cysts, where 1% pancreatin had neither an enhancing effect on excystation nor a toxic effect on cysts, whereas 5% ox bile had a toxic effect on cysts. Pretreatment of cysts with AGF followed by AIF had a similar toxic effect on cysts to that by AIF only. These results suggest that gastric fluid but not intestinal fluid at 37°C contributes to enhancing excystation for Entamoeba infection.     

Predatory activity of the fungus <Emphasis Type="Italic">Duddingtonia flagrans</Emphasis> in equine strongyle infective larvae on natural pasture in the Southern Region of Brazil

Biological control is an alternative method to reduce the population of parasites through natural predators. A promising option of biological control in the reduction of infective larvae on pasture is the use of nematophagous fungi. In this study, the efficacy of the nematophagous fungus Duddingtonia flagrans in controlling gastrointestinal nematode parasites in field-raised horses was tested. Ten foals with an average age of 12 months were divided in two groups: five males constituted the treated group and five females constituted the control group. Each group was introduced in a field of mixed pasture with approximately 5 ha. The treated group received the fungus D. flagrans at a concentration of 10⁶ chlamydospores per kilogramme of animal body weight daily, mixed with horse food for 5 months. The control group did not receive the fungus. Samples were collected to perform eggs per gramme (EPG) counts weekly. Coproculture and collection of pasture were done monthly for larvae counting. No significant difference was observed in the EPG counting and in the number of larvae recovered from coprocultures, where cyathostomines, Strongylus and Trichostrongylus spp. were found after monthly larvae counting. No significant difference was observed in the EPG counts, and Trichostrongylus sp. was identified. The number of recovered larvae on pasture was significantly lower in the treated group in the last month of treatment, showing a reduction of 73.5% (p < 0.05). As such, the fungus was able to reduce the number of infective larvae in the pasture. Nevertheless, this did not reflect in a decrease of parasitic infection during the 5-month study period.     

Efficacy of Chrysosporium tropicum metabolite against mixed population of adult mosquito (Culex quinquefasciatus, Anopheles stephensii, and Aedes aegypti) after purification with flash chromatography

Chrysosporium tropicum is a keratinophilic fungus and an effective mosquito control agent. This fungus was grown in Richards broth at 27 ± 2°C and a relative humidity of 75% ± 5% for 15 ± 2 days. Filtration was done with Whatman number 1 filter paper, column chromatography, and flash chromatography. Adulticidal efficacy was performed against a mixed population of mosquitoes including Culex quinquefasciatus, Anophelese stephensii, Aedes aegypti at five different concentrations 5:5, 6:4, 7:3, 8:2, 9:1 by adding fungal filtrate of flash chromatography to methanol in different ratio (metabolite/methanol). The experiment was conducted in the cage with an area of 2 × 2 × 3 ft. The mortality in mosquito population was recorded after 8 hours of exposure, and adulticidal activity was tested by probit analysis. The LC₅₀ was determined to be 4.9921 ml. Results of present study confirm that metabolites of C. tropicum can be utilized as alternative biological control agents for adult mosquitoes.     

Xylanase production by Aspergillus versicolor

The xylanolytic system of Aspergillus versicolor is controlled by induction and carbon catabolite repression. Carboxymethylcellulose and wheat bran were the best inducers of xylanolytic activity. When the fungus was grown for 5 days on VOGEL 's liquid medium with wheat bran, the optimal pH and temperature for xylanase production were 6.5 and 30 °C, respectively. Optimal conditions for the xylanolytic activity assay were at pH 6.0 and 55 °C. The half-life at 60 °C of the crude enzyme was 6.5 and 21 minutes, in the absence or presence of substrate, respectively.     

Thermal resistance and viability of asexual spores of thermophilic fungi from composts

The growth-temperature relations and thermal tolerance of the asexual spores of thermophilic fungi commonly found in composting plant materials were determined. The optimum temperature range for germination of the spores was 40°–50°C. The spores of Humicola lanuginosa (Griffon and Maublanc) Bunce were able to survive 60 min exposure to 68°C while those of Mucor pusillus Cooney and Emerson survived only 5 min exposure to 68°C. A determination of the thermal death rates of the spores at 65°C showed a rapid loss of viability of those of Torula thermophila Cooney and Emerson. The thermal tolerance of the propagules of thermophilic fungi in relation to their special ecological niche is discussed.     

Thermoregulation of Tuvan pastoralists and Western Europeans during cold exposure

Objectives

This study compared the metabolic and vascular responses, to whole-body and finger cold exposure, of a traditional population lifelong exposed to extreme cold winters with Western Europeans.

Methods

Thirteen cold acclimatized Tuvan pastoralist adults (45 ± 9 years; 24.1 ± 3.2 kg/m²) and 13 matched Western European controls (43 ± 15 years; 22.6 ± 1.4 kg/m²) completed a whole-body cold (10°C) air exposure test and a cold-induced vasodilation (CIVD) test, which involved the immersion of the middle finger into ice-water for 30 min.

Results

During the whole-body cold exposure, the durations until the onset of shivering for three monitored skeletal muscles were similar for both groups. Cold exposure increased the Tuvans' energy expenditure by (mean ± SD) 0.9 ± 0.7 kJ min⁻¹ and the Europeans' by 1.3 ± 1.54 kJ min⁻¹; these changes were not significantly different. The forearm-fingertip skin temperature gradient of the Tuvans was lower, indicating less vasoconstriction, than the Europeans during the cold exposure (0 ± 4.5°C vs. 8.8 ± 2.7°C). A CIVD response occurred in 92% of the Tuvans and 36% of the Europeans. In line, finger temperature during the CIVD test was higher in the Tuvans than the Europeans (13.4 ± 3.4°C vs. 3.9 ± 2.3°C).

Conclusion

Cold-induced thermogenesis and the onset of shivering were similar in both populations. However, vasoconstriction at the extremities was reduced in the Tuvans compared to the Europeans. The enhanced blood flow to the extremities could be beneficial for living in an extreme cold environment by improving dexterity, comfort, and reducing the risk of cold-injuries.     

Acute and adaptive responses in humans to exercise in a warm, humid environment

 Acute and repeated exposure for 8–13 consecutive days to exercise in humid heat was studied. Twelve fit subjects exercised at 150 W [45% of maximum O₂ uptake (V.O₂,_max)] in ambient conditions of 35°C and 87% relative humidity which resulted in exhaustion after 45 min. Average core temperature reached 39.9 ± 0.1°C, mean skin temperature (T– _sk) was 37.9 ± 0.1°C and heart rate (HR) 152 ± 6 beats min^–1 at this stage. No effect of the increasing core temperature was seen on cardiac output and leg blood flow (LBF) during acute heat stress. LBF was 5.2 ± 0.3 l min^–1 at 10 min and 5.3 ± 0.4 l min^–1 at exhaustion (n = 6). After acclimation the subjects reached exhaustion after 52 min with a core temperature of 39.9 ± 0.1°C, T– _sk 37.7 ± 0.2°C, HR 146 ± 4 beats min^–1. Acclimation induced physiological adaptations, as shown by an increased resting plasma volume (3918 ± 168 to 4256 ± 270 ml), the lower exercise heart rate at exhaustion, a 26% increase in sweating rate, lower sweat sodium concentration and a 6% reduction in exercise V.O₂. Neither in acute exposure nor after acclimation did the rise of core temperature to near 40°C affect metabolism and substrate utilization. The physiological adaptations were similar to those induced by dry heat acclimation. However, in humid heat the effect of acclimation on performance was small due to physical limitations for evaporative heat loss. Received: 3 July 1996 / Received after revision: 26 September 1996 / Accepted: 7 January 1997     

Improved in vitro cultivation of larvae of the bovine lungworm Dictyocaulus viviparus

Parasitic larvae of Dictyocaulus viviparus are of major importance for the development of immunity in cattle. The conditions for in vitro cultivation of D. viviparus larvae as well as their morphology have thus far been only poorly investigated. Exsheathed larvae were cultivated in vitro in RPMI-1640 (Gibco-BRL, pH 7.2) containing 20% newborn Calf serum, 200 U Moronal/ml, and 200 U penicillin/streptomycin/ml. Incubation was performed at 39.5 °C at 0, 5%, 10%, and 20% CO₂. Average development rates to third-moult (3 M) or fourth-stage (L4) larvae at 5% CO₂ incubation were 8.33% (SD ± 7.76%), 22.52% (SD ± 13.09%) at 10% CO₂, and 38.01% (SD ± 15.63%) at 20% CO₂. These differences were statistically significant. Some morphological features of these larvae are described. Received: 28 March 1998 / Accepted: 17 April 1998     

Effects of temperature, ionic strength and pH on the function of skeletal muscle myosin from a eurythermal fish, Fundulus heteroclitus

The mummichog, Fundulus heteroclitus, is an intertidal fish that exhibits little change in swimming ability despite large and rapid variations in environmental parameters. We therefore tested the hypothesis that this nearly constant function is due to Fundulus myosin being intrinsically insensitive to changes of temperature, ionic strength and pH. In vitro motility assays were used to quantify the speed of unregulated actin filaments on myosin purified from F. heteroclitus glycolytic skeletal muscle. Filament speed was 2.07±0.17 μm s⁻¹ at 26°C, ionic strength (Γ/2) of 0.08 M Γ/2 and pH 7.4. Speed increased as temperature increased over the range of 5–36°C with an activation energy (E _a) of 94.0±7.0 kJ mol⁻¹) and an enthalpy (ΔH ^‡) of 91.5±7.0 kJ mol⁻¹ at 20°C. A linear relationship between temperature and ATPase activity was also obtained with actin-activated myosin Mg²⁺-ATPase assays over the temperature range 5–35°C with E _a=59.9±2.4 kJ mol⁻¹ and ΔH ^‡=57.4±2.4 kJ mol⁻¹ at 20°C. There was little or no effect of ionic strength on filament speed over the range 0.19 M Γ/2–0.54 M Γ/2. Speed increased significantly at lower ionic strengths and was 7.9-fold higher at 0.08 M Γ/2 than at 0.19 M Γ/2. Speed increased with pH with a 16-fold increase between pH 6.7 and 7.4. These results indicate that changes in physiological parameters that include temperature, pH and ionic strength affect the function of unregulated F. heteroclitus myosin, and thus other factors must be responsible for the mummichog’s swimming performance being comparatively insensitive to environmental variation.     

In vitro predatory activity of nematophagous fungi and after passing through gastrointestinal tract of equine on infective larvae of Strongyloides westeri

Three isolates of predator fungi Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34), and Arthrobotrys robusta (I-31) were assessed in in vitro test regarding the capacity of prey infective larvae (L₃) Strongyloides westeri. Compared to control, without fungus, there was a significant decrease (P < 0.01) of 80.4%, 67.9%, and 72.8% in means of infective larvae S. westeri recovered from treatments with isolates AC001, NF34, and I-31, respectively. All tested isolates were efficient in the capture of S. westeri (P > 0.01) in vitro test. Linear regression coefficients of treated and control groups were −0.21 for control, −0.32 for D. flagrans, −0.34 for M. thaumasium, and −0.22 for A. robusta. In the following, isolates AC001 and NF34 were assessed in vivo regarding the capacity of supporting the passage through equine gastrointestinal tract without loss of ability of preying infective larvae S. westeri. Fungal isolates survived the passage and were efficient in preying L₃ since the first 12 h of collection (P < 0.01) in relation to the control group (without fungus). Compared to control, there was a significant decrease (P < 0.01) of 76.4% and 76.7% (12 h), 86.4% and 85.9% (24 h), 88.3% and 87.7% (48 h), and 89.9% and 87.2% (72 h) in means of infective larvae S. westeri recovered from treatments with isolates AC001 and NF34, respectively. Linear regression coefficients of L₃ of recovered S. westeri regarding the collections due to time were 1.93 for control, −3.52 for AC001, and −2.64 for NF34. Fungi D. flagrans and M. thaumasium (NF34) have demonstrated to be promising for use in the biological control of equine parasite S. westeri.