In Vitro Parasitism of Rotylenchus robustus by Isolates of Hirsutella rhossiliensis

We tested the hypothesis that isolates of Hirsutella rhossiliensis from host nematodes in the family Hoplolaimidae (Rotylenchus robustus and Hoplolaimus galeatus) would be more virulent to R. robustus than would isolates from host nematodes not in the Hoplolaimidae (Heterodera schachtii and Criconemella xenoplax). Nematodes were touched to 10-20 spores of different isolates and incubated at 20 C in 4.5 mM KC1; the percentage of nematodes colonized (filled with hyphae) was determined after 2, 5, 10, 20, and 30 days. The hypothesis was rejected because isolates from H. schachtii and C. xenoplax were equivalent or better at parasitizing R. robustus than were isolates from R. robustus and H. galeatus. In addition, the R. robustus and H. galeatus isolates were as pathogenic to C. curvata as they were to R. robustus, but produced fewer spores per colonized nematode (H. schachtii) than did the other isolates.     

Predaceous Behavior and Life History of Odontopharynx longicaudata (Diplogasterida)

The behavior of a California isolate of the predaceous nematode, Odontopharynx longicaudata de Man, was studied in water agar culture. When feeding on an Acrobeloides sp. the predator completed its life cycle in 13 to 14 days at 25 C. Optimum temperature for reproduction of the predator was 25 C, few individuals survived at 10 C, and 30 C was lethal. Males were necessary for reproduction, and at 25 C the sex ratio was about 1:1. All postembryonic stages were voracious feeders. A single female predator consumed 30 individuals of another Acrobeloides sp. in 1.5 days. Juveniles must feed in order to complete their development. Three modes of feeding were observed depending on the prey selected. A high degree of prey selectivity occurred; 6 of 17 nematode prey species were readily consumed by the predator, but there was little or no feeding on the remaining 11 species. Predation percentage varied with prey species. Consumption of Anguina pacificae and the two Acrobeloides spp. was almost 100%, consumption of A. amsinckiae, Pratylenchus vulnus, and Trichodorus sp. was 70-78%. Difference in final predator population densities was obtained after feeding on the two species of Acrobeloides. Final predator population densities increased linearly with increasing inoculum levels of the first Acrobeloides sp.     

Anhydrobiosis in Five Species of Plant Associated Nematodes

Five species of nematodes - Hemicriconemoides pseudobrachyurum, Hemicycliophora conida, Macroposthonia ornata, Aphelenchoides ritzemabosi, and Psilenchus hilarulus -were desiccated to study their capacity to survive anhydrobiotically. Results indicate that the ability of the sheath to shrink quickly and its relatively loose attachment with the nematode body allow H. conida to survive longer than H. pseudobrachyurum; the survival of M. ornata was intermediate, A. ritzemabosi and P. hilarulus survived immersion in paraffin oil for 12 and 17 days, respectively. Both of these nematodes possess multiple contraction ability; i.e., coiling coupled with transverse and longitudinal folding of the cuticle. P. hilarulus is a new addition to the list of anhydrobiotic nematodes.     

Atomic Force Microscopy of Plant-Parasitic Nematodes

A simple method for atomic force microscopy (AFM) of nematode cuticle was developed to visualize the external topography of Helicotylenchus lobus, Meloidogyne javanica, M. incognita, and Xiphinema diversicaudatum. Endospores of two isolates of the nematode parasite, Pasteuria penetrans, adhering to M. incognita and X. diversicaudatum were also visualized and measured by this technique. Scanning procedures were applied to specimens killed and dehydrated in air or dehydrated and stored in glycerol. Atomic force microscopy scanning of nematodes in constant height mode yielded replicated high-resolution images of the cuticle showing anatomical details such as annulations and lateral fields. Submicrometer scale images allowed the identification of planar regions for further higher resolution scans.     

Evaluation of Hot Water Treatments for Management of Ditylenchus dipsaci and Fungi in Daffodil Bulbs

Treatment of daffodil (Narcissus pseudonarcissus) bulbs in a 0.37% formaldehyde water solution at 44 C for 240 minutes is a standard practice in California for management of the stem and bulb nematode, Ditylenchus dipsaci. Recent concern over the safety of formaldehyde and growers'' requests for a shorter treatment time prompted a reevaluation of the procedure. The time (Y, in minutes) required to raise the temperature at the bulb center from 25 to 44 C was related to bulb circumference (X, in cm) and is described by the linear regression Y = -15 + 3.4X. The time required for 100% mortality of D. dipsaci in vitro without formaldehyde was 150, 60, and 15 minutes at 44, 46, and 48 C, respectively. Hot water treatment (HWT) with 0.37% formaldehyde at 44 C for 150 minutes controlled D. dipsaci and did not have a detrimental effect on plant growth and flower production. Shorter formaldehyde-HWT of 90, 45, and 30 minutes at 46, 48, and 50 C, respectively, controlled D. dipsaci but suppressed plant growth and flower production. Fungal genera commonly isolated from the bulbs in association with D. dipsaci were Penicillium sp., Fusarium oxysporum f. sp. narcissi, and Mucor plumbeus, representing 60, 25, and 5%, respectively, of the total fungi isolated. These fungi caused severe necrosis in daffodil bulbs. HWT at 44 C for 240 minutes reduced the number of colonies recovered from bulbs. The effects of formaldehyde, glutaraldehyde, and sodium hypochlorite in reducing the population of fungi within bulbs were variable. Satisfactory control of D. dipsaci within bulbs can be achieved with HWT of bulbs at 44 C for 150 minutes with 0.37% formaldehyde or at 44 C for 240 minutes without chemicals.     

Caenorhabditis elegans: Effects of 5-hydroxytryptophan and dopamine on behavior and development

5-Hydroxytryptophan (5-HTP) was introduced iontophoretically into the vulva region of Caenorhabditis elegans to examine behavioral responses to this putative neurotransmitter. Responses in esophageal basal bulb pulsation and/or vulval contractions occurred. Little relation was observed between dosage and behavioral response. Similar behavioral responses followed topical applications of 5-HTP. An inverse relationship between the rates of esophageal pumping and vulval contraction was recorded following both iontophoretic injection and topical application. Following iontophoretic application, young nematodes resumed body movement sooner than old nematodes did. Growth significantly increased when 5-HTP or dopamine was added to the culture medium, but neither chemical influenced fecundity or life span.     

Host-Parasite Relationships of Meloidogyne arenaria and M. incognita on Susceptible Soybean

Pathogenicity and reproduction of single and combined populations of Meloidogyne arenaria and M. incognita on a susceptible soybean (Glycine max cv. Davis) were investigated. Significant galling and egg mass production were observed on roots of greenhouse-grown soybean inoculated with M. arenaria and M. incognita, in combination and individually. M. arenaria produced more galls and egg masses than M. incognita, whereas in combined inoculation with both nematode species, gall and egg production was intermediate. In growth chamber tests, inoculations with M. arenaria and M. incognita, singly or in combination, produced more galls and egg masses at 30 C than at 25 C. At 25 C, M. arenaria alone produced significantly more galls and egg masses than the combined M. arenaria plus M. incognita, while M. incognita produced the fewest. At 30 C, numbers of egg masses produced by M. arenaria did not differ significantly from combined M. arenaria and M. incognita. In temperature tank tests, M. incognita produced more galls and egg masses at 28 C than at 24 C soil temperature. In contrast, numbers of galls, egg masses, and eggs of M. arenaria were slightly higher at 24 C than at 28 C. Combined inoculum of both nematode species produced greater numbers of galls at 24 C than at 28 C.     

Dermal gland secretion improves the heat tolerance of the brown dog tick, Rhipicephalus sanguineus, allowing for their prolonged exposure to host body temperature

Dermal glands (sensilla sagittiformia) secrete when brown dog ticks, Rhipicephalus sanguineus, are mechanically disturbed, presumably as a defensive mechanism. Recently, we observed that these glands secrete due to the pressure stimulation of engorgement. In this study, we examine how dermal gland secretion alters the physiology of R. sanguineus, particularly if this secretion is an important mechanism during blood feeding. The ability of ticks to retain water was not modified by dermal gland secretion, but heat tolerance was enhanced. Short-term heat shock was improved slightly (1 h at 50 °C to 1 h at 56 °C) and featured reduced injury responses and greater recovery after heat shock. When exposed to their host body temperature (37 °C) for prolonged periods, individuals that had secreted survived over 1 week longer than individuals that did not secrete. Dorsal application of squalene, the main component of dermal gland secretion, did not increase temperature tolerance, suggesting that the act of secreting rather than the physical properties of the secretion itself is responsible for the increase in heat tolerance. Based on our results, dermal gland secretion may be an essential mechanism in certain tick species (Amblyomma, Dermacentor, Hyalomma, Rhipicephalus, but not Ixodes) for tolerating body temperature and not succumbing to heat stress during the extended time periods of feeding on a mammalian host, serving as a mechanism to prevent heat damage from the host during feeding.     

Phylogenetic Relationships and Genetic Variation in Longidorus and Xiphinema Species (Nematoda: Longidoridae) Using ITS1 Sequences of Nuclear Ribosomal DNA

Genetic analyses using DNA sequences of nuclear ribosomal DNA ITS1 were conducted to determine the extent of genetic variation within and among Longidorus and Xiphinema species. DNA sequences were obtained from samples collected from Arkansas, California and Australia as well as 4 Xiphinema DNA sequences from GenBank. The sequences of the ITS1 region including the 3'' end of the 18S rDNA gene and the 5'' end of the 5.8S rDNA gene ranged from 1020 bp to 1244 bp for the 9 Longidorus species, and from 870 bp to 1354 bp for the 7 Xiphinema species. Nucleotide frequencies were: A = 25.5%, C = 21.0%, G = 26.4%, and T = 27.1%. Genetic variation between the two genera had a maximum divergence of 38.6% between X. chambersi and L. crassus. Genetic variation among Xiphinema species ranged from 3.8% between X. diversicaudatum and X. bakeri to 29.9% between X. chambersi and X. italiae. Within Longidorus, genetic variation ranged from 8.9% between L. crassus and L. grandis to 32.4% between L. fragilis and L. diadecturus. Intraspecific genetic variation in X. americanum sensu lato ranged from 0.3% to 1.9%, while genetic variation in L. diadecturus had 0.8% and L. biformis ranged from 0.6% to 10.9%. Identical sequences were obtained between the two populations of L. grandis, and between the two populations of X. bakeri. Phylogenetic analyses based on the ITS1 DNA sequence data were conducted on each genus separately using both maximum parsimony and maximum likelihood analysis. Among the Longidorus taxa, 4 subgroups are supported: L. grandis, L. crassus, and L. elongatus are in one cluster; L. biformis and L. paralongicaudatus are in a second cluster; L. fragilis and L. breviannulatus are in a third cluster; and L. diadecturus is in a fourth cluster. Among the Xiphinema taxa, 3 subgroups are supported: X. americanum with X. chambersi, X. bakeri with X. diversicaudatum, and X. italiae and X. vuittenezi forming a sister group with X. index. The relationships observed in this study correspond to previous genera and species defined by morphology.     

Population Response to Temperature in the Subfamily Tylenchorhynchinae

The effects of temperature on population development of 11 species of stunt nematodes in the subfamily Tylenchorhynchinae were compared on red clover or Kentucky bluegrass in constant-temperature tanks at 5-degree intervals from 10 to 35 C. The optimum temperature for population increase on red clover in 90 days was 30 C for Tylenchorhynchus agri, T. nudus, T. martini, and T. clarus, 25 C for T. sylvaticus and T. dubius, and 20 C for T. canalis, Merlinius brevidens, and Quinisulcius capitatus. The optimum was 30 C for T. robustoides and 25 C for T. maximus on Kentucky bluegrass. The temperature range for population increase was 20-35 C for T. agri, T. nudus, T. martini, and T. clarus, 20-30 C for T. sylvaticus and T. robustoides, 15-25 C for T. maximus, 10-25 C for T. dubius, and 10-20 C for M. brevidens and Q. capitatus. T. canalis increased only at 20 C. All species were recovered in numbers near their inoculum level at 10 C. There was no survival of T. sylvaticus, T. dubius, T. canalis, T. robustoides, T. maximus, M. brevidens, and Q. capitatus at 35 C, or of the last three of these species at 30 C. Temperature had no effect on sex ratio in final populations. Population increase was greatest in T. martini and least in T. canalis.     

The enigmatic molar from Gondolin, South Africa: Implications for Paranthropus paleobiology

The specific attribution of the large hominin M₂ (GDA-2) from Gondolin has significant implications for the paleobiology of Paranthropus. If it is a specimen of Paranthropus robustus it impacts that species' size range, and if it belongs to Paranthropus boisei it has important biogeographic implications. We evaluate crown size, cusp proportions and the likelihood of encountering a large-bodied mammal species in both East and South Africa in the Early Pleistocene. The tooth falls well outside the P. robustus sample range, and comfortably within that for penecontemporaneous P. boisei. Analyses of sample range, distribution and variability suggest that it is possible, albeit unlikely to find a M₂ of this size in the current P. robustus sample. However, taphonomic agents - carnivore (particularly leopard) feeding behaviors - have likely skewed the size distribution of the Swartkrans and Drimolen P. robustus assemblage. In particular, assemblages of large-bodied mammals accumulated by leopards typically display high proportions of juveniles and smaller adults. The skew in the P. robustus sample is consistent with this type of assemblage. Morphological evidence in the form of cusp proportions is congruent with GDA-2 representing P. robustus rather than P. boisei. The comparatively small number of large-bodied mammal species common to both South and East Africa in the Early Pleistocene suggests a low probability of encountering an herbivorous australopith in both. Our results are most consistent with the interpretation of the Gondolin molar as a very large specimen of P. robustus. This, in turn, suggests that large, presumptive male, specimens are rare, and that the levels of size variation (sexual dimorphism) previously ascribed to this species are likely to be gross underestimates.     

The impact of food type, temperature and starvation on larval development of Balanus amphitrite Darwin (Cirripedia: Thoracica)

The impact of diatom food species (Chaetoceros calcitrans and Skeletonema costatum), temperature and starvation on the larval development of Balanus amphitrite was evaluated. Starvation threshold levels for different ages of larvae (0- to 5-day-old) fed with C. calcitrans and S. costatum and then starved at 5, 15 and 25 °C temperature were estimated as ultimate recovery hour (URH; denoting the starvation point in hours at the end of which larvae can recover and continue development). Effect of temperature on starvation threshold varied significantly with larval age and food species. The URH declined with larval age at 5 °C, but not at 15 and 25 °C. The URH and grazing rates were high for early instars fed on C. calcitrans, and for advanced instars fed on S. costatum. Carbon gain through feeding was maximum for 2-day-old larvae when fed with C. calcitrans and decreased with larval age. However, when fed with S. costatum carbon gain increased with larval age. This confirms that with development the utility of food types changes. The differences in the carbon gain can be attributed to differences in grazing rate due to variations in the size of the diatom cells, larval intersetular distance, diatom sinking rate and the photo-taxic behavior of larvae. Molting was observed at times when larvae were undergoing starvation and this could be viewed as stress-induced molting, and it differed with the larval age and food organisms.     

Effect of Temperature on Attachment,Development, and Interactions of Pasteuria penetrans on Meloidogyne incognita

The effect of temperature (10, 20, 25, 30, and 35 C) on attachment and development of Pasteuria penetrans on Meloidogyne arenaria race 1 was elevated in growth chambers. The greatest attachment rate of endospores of P. penetrans occurred on second-stage juveniles at 30 C. The bacterium developed more quickly within its host at 30 and 35 C than at 25 C or below. The development of the bacterium within the nematode female was divided into nine recognizable life stages, which ranged from early vegetative thalli to mature sporangia. Mature sporangium was the predominant life stage observed after 35, 40, 81, and 116 days at 35, 30, 25, and 20 C, respectively. The body width and length of M. arenaria females infected with P. penetrans were smaller initially than the same dimensions in uninfected females, but became considerably larger over time at 25, 30, and 35 C. This isolate of P. penetrans also parasitized and completed its life cycle in males of M. arenaria.     

Effect of Time,Temperature, and Inoculum Density on Reproduction of Pratylenchus thornei in Carrot Disk Cultures

Reproduction of Pratylenchus thornei on carrot disk cultures at different time periods after inoculation, temperature, and initial inoculum density was studied. At 25 C and with an initial inoculum of 25 females per disk, the final nematode population increased with increasing time after inoculation, although the populations after 25 and 50 days were not different. Nematode numbers increased by 1,255-fold and 3,619-fold at 75 and 100 days, respectively. Over 35 days incubation at 15, 20, 25, and 30 C, the nematode multiplied 1.8, 8.4, 10.5, and 0.4 times, respectively. The optimum temperature for reproduction was between 20 and 25 C, and the nematode life cycle was completed in about 25-35 days. Increasing nematode inoculum (25, 50, 100, 500, 1,000 nematodes per disk) increased the final nematode population but did not increase reproduction rate, the highest being 25.3 at an initial inoculum density of 100 nematodes per disk.     

Influence of Temperature on Population Development of Eight Species of Pratylenchus on Soybean

In a soil temperature study, population increase on ''Clark 63'' soybeatt was most rapid at 30 C in Pratylenchus alleni, P. brachyurus, P. cofleae, P. neglectus, P. scribneri, and P. zeae and at 25 C in P. penetrans and P. vulnus. The last two were the only species that reproduced at 15 C. Populations of all species increased over the range of 20-30 C, except those of P. neglectus at 20 C and P. coffeae, which was not tested below 25 C. Only P. brachyurus, P. neglectus, P. scribneri and P. zeae reproduced at 35 C. At their optimum temperatures, P. scribneri exhibited the greatest population increase, 1248-fold, and P. penetrans the least, 32-fold. This is the first report of soybean as a host for P. vulnus.     

Vertical Distribution of Three Nematode Genera in a Bentgrass Putting Green in Central Illinois

A study was conducted to determine the vertical distribution of Tylenchorhynchus nudus, Criconemella curvata, and Helicotylynchus cornurus in the upper 5 cm of bentgrass (Agrostis palustris cv. Penncross) putting green turf. The effect of fenamiphos on the vertical distribution of these species also was examined. Experimental design was a split-plot in which whole-plots were fenamiphos treated (0.11 kg a.i./100 m²) or untreated, and sub-plots were two strata (depths of 0-2.5 crn and 2.5-5.0 cm). Soil samples were collected during the growing season for 2 years after treatment to determine root weight and number of nematodes. Root weight was greater in the upper stratum on all sampling dates in both years. When differences between strata in population density were observed, T. nudus, C. curvata, and H. cornurus were more concentrated in the upper stratum. Vertical distribution of T. nudus, C. curvata, and H. cornurus was similar to the distribution of root weight. The difference in population density of H. cornurus between upper and lower strata was affected by fenamiphos on some dates, whereas differences between strata were unaffected for T. nudus and C. curvata. Double arcsine transformed proportions of the total populations of T. nudus, C. curvata, and H. cornurus in the upper stratum on each sampling date indicated no differences between fenamiphos treated and untreated plots in 1989 or 1990.     

Temperature response of photosynthesis in different drug and fiber varieties of Cannabis sativa L.

The temperature response on gas and water vapour exchange characteristics of three medicinal drug type (HP Mexican, MX and W1) and four industrial fiber type (Felinq 34, Kompolty, Zolo 11 and Zolo 15) varieties of Cannabis sativa, originally from different agro-climatic zones worldwide, were studied. Among the drug type varieties, optimum temperature for photosynthesis (T_opt) was observed in the range of 30–35 °C in high potency Mexican HPM whereas, it was in the range of 25–30 °C in W1. A comparatively lower value (25 °C) for T_opt was observed in MX. Among fiber type varieties, T_opt was around 30 °C in Zolo 11 and Zolo 15 whereas, it was near 25 °C in Felinq 34 and Kompolty. Varieties having higher maximum photosynthesis (P_{N max}) had higher chlorophyll content as compared to those having lower P_{N max}. Differences in water use efficiency (WUE) were also observed within and among the drug and fiber type plants. However, differences became less pronounced at higher temperatures. Both stomatal and mesophyll components seem to be responsible for the temperature dependence of photosynthesis (P_N) in this species, however, their magnitude varied with the variety. In general, a two fold increase in dark respiration with increase in temperature (from 20 °C to 40 °C) was observed in all the varieties. However, a greater increase was associated with the variety having higher rate of photosynthesis, indicating a strong association between photosynthetic and respiratory rates. The results provide a valuable indication regarding variations in temperature dependence of P_N in different varieties of Cannabis sativa L.     

Penetration of Celery and Alfalfa Roots by Pratylenchus penetrans as Affected by Temperature

A greater percentage of females than juveniles or males of P. penetrans penetrated celery roots grown in infested soil at 5, 18, or 30 C; the difference was greatest at 5 C. The time of initial penetration of alfalfa seedlings inoculated with single nematodes on water agar varied with temperature. Females penetrated the seedlings earlier and over a wider range of temperatures than did males or juveniles. The rate of penetration was highest for females. After initial penetration, the penetration rate decreased with time. At 13-28 C, approximately 80% of roots were penetrated by females and only 25-30% by males and juveniles by the end of the experiment.     

Effect of dietary history and algal traits on feeding rate and food preference in the green sea urchin Strongylocentrotus droebachiensis

Feeding behaviour is influenced by a variety of factors, including nutritional requirements, the quality of available foods, and environmental conditions. We examined the effect of two factors, food morphology and dietary history, on the feeding rate and preference of the sea urchin Strongylocentrotus droebachiensis. Standardizing food shape and structure did not alter urchins' expected preference for the native kelp Laminaria longicruris over the invasive alga Codium fragile ssp. tomentosoides. However, when foods containing L. longicuris were shaped to mimic the algae, the C. fragile mimic was consumed more rapidly than the kelp mimic. Dietary history had no effect on single diet feeding rate. Urchins feeding on C. fragile consistently consumed twice as much (by mass) as those fed kelp, regardless of their previous diet. Despite higher feeding rates on C. fragile, urchins feeding on this alga were unable to compensate for its low energetic content and ingested less energy. Dietary history had a short-term effect on food preference, with urchins tending to prefer less familiar foods. Our findings suggest that urchins feed on C. fragile at a high rate, due to ease of handling and/or compensatory feeding, and that they do not a have strict preference hierarchy. Rather, food choice appears to reflect active maintenance of a mixed diet.