Effects of male-female rations on mating and egg production in Octomyomermis muspratti (Mermithidae: Nematoda)

Octomyomermis muspratti, a mermithid parasite of mosquito larvae, molted to the adult stage in 4–16 days. When male-female ratios were relatively high (1:2), mating and oviposition began soon after maturation, and oviposition was completed in 21–40 days. The number of females developing eggs increased with a corresponding increase in the male-female ratio. About 60% of the females completed oviposition at male-female ratios of 1:20, and about 100% completed oviposition as the male-female ratios were increased to 1:2. No egg development was observed in the absence of males. Male O. muspratti were capable of mating with a number of females and remained reproductively active for at least 50 days after a maturation at 26–28°C.     

Effects of Host Size and Parasite Burden on Sex Ratio in the Mosquito Parasite Octomyomermis muspratti

The ratio of Octomyomermis muspratti to the host mosquito at the time of exposure had little effect on the ratio of male to female parasites that resulted. However, the ratio of males to females increased as the number of parasites/host increased. Hosts with a single nematode produced fewer than 1% males in comparison with hosts with 8 parasites which produced about 40% males; hosts with 10 or more nematodes generally produced more male than female nematodes. Males of O. muspratti usually emerged before females because of the earlier death of multiply-infected mosquitoes. The size of the host at the time of invasion bad no significant influence on nematode sex ratios. Since mating is apparently necessary for reproduction in O. muspratti, the low male to female ratios that occur will be important in developing successful mass production techniques.     

Effect of Aedes aegypti (Diptera:Culicidae) Age on Sex Ratios in Romanomermis culicivorax (Nematoda:Mermithidae)

First, second, third, early and late fourth-instar larvae, and pupae of Aedes aegypti were infected with Romanomerrais culicivorax and reared at 20, 25, and 30 C. An increase in the ratio of male to female nematodes was observed with increase in host age at the time of infection at each temperature. The number of pupal and late fourth-instar infections was low, but R. culicivorax continued to develop in adult A. aegypti. Since male nematodes were recovered from both male and female hosts infected as late fourth instars or pupae, the sex of the host did not influence the sex of the nematode.     

Dispersal and Infectivity of the Entomogenous Nematode,Neoaplectana carpocapsae Weiser (Rhabditida: Steinernematidae), in Sand

Laboratory tests determined the lateral and vertical dispersal patterns of Neoaplectana carpocapsae in sand. In the vertical tests, placement of infective juveniles 15 cm below the sand''s surface resulted in the majority (77%) being recovered above the point of placement after 48 h. Placement of the nematodes on the sand''s surface resulted in the majority (90.4%) remaining within 1 cm of the sand''s surface. Placement of nematodes at depths of 2.5 cm and 5.0 cm below the sand''s surface also resulted in little nematode dispersal. However, vertical hioassay tests showed that juvenile nematodes placed on the sand''s surface dispersed 12 cm down to infect 67% of the Galleria mellonella pupae placed at the depth. Conversely, when nematodes were placed 11 cm below the insect pupae no infection was observed, but 53% infection occurred when nematodes were 7 cm below the site of the insect pupae. In lateral dispersal, 87% of the nematodes rentained within 2 cm of the placement site, although 0.5% were recovered at 12-14 cm away from the point of placement. Lateral bioassay tests indicated that the nematodes were capable of infecting 90, 35, and 5% of the G. mellonella pupae at 7 cm, 10 cm, and 14 cm from the point of placement, respectively.     

Biology of Octomyomermis muspratti,a parasite of mosquitoes as it relates to mass production

The first- and second-instar larvae of Culex pipiens quinquefasciatus were equally susceptible to parasitism by the mermithid nematode Octomyomermis muspratti; the third-instar hosts were significantly less susceptible. The yield of postparasitic nematodes was higher from hosts exposed as second instars then from hosts exposed as first or third instars at comparable levels of parasitism. Mortality of the host prior to emergence of the nematode occurred most frequently among first- and third-instar hosts. About 4% of the infected hosts exposed as first and second instars and 37% of the infected hosts exposed as third instars pupated prior to emergence of the postparasitic nematodes. The exposure of mosquitoes as second instars was the most efficient for mass rearing and resulted in the highest yields of postparasitic O. muspratti.     

Effect of Soil Texture on the Distribution and Infectivity of Neoaplectana carpocapsae (Nematoda: Steinernematidae)

The vertical migration of N. carpocapsae infective juveniles applied to the soil surface or introduced 14 cm below the soil surface was studied in four different soil types (pure silica sand, coarse sandy loam, silty clay loam, and clay). The percentage of juveniles able to migrate and infect wax moth pupae placed in the soil decreased as the percentage of clay and silt increased. Most nematodes placed on the soil surface remained within 2 cm of the surface, but some penetrated to a depth of 10 cm in pure silica sand and coarse sandy loam to infect pupae. Some pupae at the same depth were also infected with nematodes in silty clay loam soil. In pure silica sand and coarse sandy loam, nematodes introduced 14 cm below the soil surface were able to infect wax moth pupae located between 4 and 24 cm. Movement was least in clay soil and limited in silty clay loam. Nematodes showed a tendency to disperse upwards from the point of application. In all cases the number of migrating nematodes was greatest when wax moth pupae were present.     

Rhabditis terricola: An opportunistic nematode parasite of earthworm cocoons

During a study on the breeding rate of the earthworms Lumbricus rubellus and Eisenia foetida, nematodes identified as Rhabditis terricola were found consistently in large numbers in earthworm cocoons that failed to hatch after an adequate incubation period. This nematode species, which was previously known as a saprophyte, was found to invade earthworm cocoons and reproduce within, causing extensive productivity losses in earthworm cultures. In this study, R. terricola was effectively eradicated from earthworm cultures by rinsing the earthworms in tap water and transferring them repeatedly to sterile bedding.     

Catenaria anguillulae in the mermithid nematode Romanomermis culicivorax

Disease epizootics in laboratory cultures of the mermithid nematode Romanomermis culicivorax were caused by a chytridiomycetous fungal parasite, Catenaria anguillulae. Techniques used for mass rearing the nematode provided ideal conditions for dispersal of zoospores and multiplication of the fungus; up to 90% of the postparasites of R. culicivorax were parasitized. The fungus was isolated and grown on yeast extract-soluble starch agar, and the effects of different temperature and pH regimes on the fungus were investigated as possible methods for prevention or control. A temperature suitable for mass rearing the nematodes but unsuitable for the fungus could not be found, as C. anguillulae had a broad temperature range. Adjustment of pH or a short period of chilling (0°C) were more promising control measures. At acid pH (<5), zoospore motility and germination and infection of postparasites were suppressed. The disease was controlled by rearing nematodes in water adjusted to a pH of 4.5.     

Growth and Development of Romanomermis culicivorax In Vitro

Various combinations of vertebrate and invertebrate tissue culture and microbiological media were utilized in an attempt to culture Romanomermis culicivorax (Mermithidae: Nematoda) in vitro. Most media were unsuitable and caused nematodes to become lumpy, vacuolated, and granular. Slow and limited growth and development of internal structures of the nematodes were obtained with variously supplemented Grace''s tissue culture and Schneider''s Drosophila media. In an enriched Grace''s medium, development attained by the nematodes after 3-4 wk was comparable to 4-5-day-old parasites grown in vivo in the mosquito host, Culex pipiens. Two molts were observed in vitro. Maximum dimensions in vitro were 7.0-mm length and 87-μm width at the widest point. The stichosome, stichocytes, and trophosome developed prominently. A filiform tail and highly cuticularized tube persisted throughout the culture period in vitro.     

Biological Control of Mosquitoes with Mermithids

Mermithid nematodes parasitizing mosquitoes have substantial potential for vector control. Studies on the physiological ecology of Romanomermis culicivorax have defined some of the general requirements of mermithid nematodes and produced general guidelines for the experimental release of mermithids in biological control. Experimental field studies have established the biological control potential of R. culicivorax, but further development and ulilization of this parasite will require a substantial commitlnent of scientific man-years and ftmds.     

Effect of Soil Texture on the Distribution and Infectivity of Neoaplectana glaseri (Nematoda: Steinernematidae)

The vertical migration of infective juveniles of Neoaplectana glaseri applied to the soil surface or introduced 16 cm below the soil surface was studied in pure silica sand, coarse sandy loam, silty clay loam, and clay. The number of juveniles that migrated and infected wax moth pupae placed in the soil decreased as the proportion of clay and silt increased. The majority of nematodes moved downwards 2-6 cm from the surface, but some penetrated to a depth of 14 cm in pure silica sand and coarse sandy loam. In pure silica sand and coarse sandy loam, nematodes introduced 16 cm below the soil surface were able to infect wax moth pupae located at depths of 0-4 cm and 28-32 cm. Nematodes showed a greater tendency to disperse downwards from the point of application. Movement of the nematode was least in clay soil and limited in silty clay loam soil. The number of migrating nematodes was greatest when wax moth pupae were present.     

Field Efficacy of Furfural as a Nematicide on Turf

A commercial formulation of furfural was recently launched in the United States as a turfgrass nematicide. Three field trials evaluated efficacy of this commercial formulation on dwarf bermudagrass putting greens infested primarily with Belonolaimus longicaudatus, Meloidogyne graminis, or both these nematodes, and in some cases with Mesocriconema ornatum or Helicotylenchus pseudorobustus. In all these trials, furfural improved turf health but did not reduce population densities of B. longicaudatus, M. graminis, or the other plant-parasitic nematodes present. In two additional field trials, efficacy of furfural at increasing depths in the soil profile (0 to 5 cm, 5 to 10 cm, and 10 to 15 cm) against B. longicaudatus on bermudagrass was evaluated. Reduction in population density of B. longicaudatus was observed in furfural-treated plots for depths below 5 cm on several dates during both trials. However, no differences in population densities of B. longicaudatus were observed between the furfural-treated plots and the untreated control for soil depth of 0 to 5 cm during either trial. These results indicate that furfural applications can improve health of nematode-infested turf and can reduce population density of plant-parasitic nematodes in turf systems. Although the degree to which turf improvement is directly caused by nematicidal effects is still unclear, furfural does appear to be a useful nematode management tool for turf.     

Effect of the Entomogenous Nematode Nemplectana carpocapsae on the Tachinid Parasite Compsilura concinnata (Diptera: Tachinidae)

The entomogenous nematode Neoaplectana carpocapsae and its associated bacterium, Xenorhabdus nematophilus, could not infect the pupal stage of the tachinid Compsilura concinnata through the puparium. N. carpocapsae had an adverse effect on 1-, 2- and 3-day-old C. concinnata larvae within the armyworm host in petri dish tests. All 1-day-old larvae treated with nematodes died in their hosts, whereas 61% and 69% of 2- and 3-day-old larvae treated with nematodes, respectively, died. However, the survivors developed to adults. Nine to thirty-seven percent of adult tachinids which emerged from nematode-treated soil (50 nematodes/cm²) were infected with N. carpocapsae. The nematode adversely affects C. concinnata directly by the frank infection of the tachinid and indirectly by causing the premature death of the host which results in tachinid death.     

THE RELATION BETWEEN THE ELECTRICAL CONDUCTIVITY OF THE EXTERNAL MEDIUM AND THE RATE OF CELL DIVISION IN SEA URCHIN EGGS

Dilution of sea water with isotonic sugar solution leaves the rate of cleavage of Arbacia eggs almost unchanged until the proportion of sea water is decreased to 20 or 25 volumes per cent. From this point cleavage becomes progressively slower with further dilution. Many eggs fail to cleave at dilutions of 5 to 6 volumes per cent. No cleavage occurs in 2 volumes per cent sea water or in pure sugar solution. Eggs returned from these media to sea water resume cleavage and development. There is thus no relation between the rate of cleavage and the electrical conductivity of the medium, except possibly within the range of dilutions from 20 to 5 volumes per cent sea water. In this range cleavage rate decreases as conductivity decreases, but the relation is not a linear one.     

Hemolymph pH of the larvae of three species of mosquitoes,and the effect of Romanomermis culicivorax parasitism on the blood pH of Culex pipiens

The hemolymph pHs of late fourth-instar Culex pipiens, Aedes taeniorhynchus, and Toxorhynchites amboinensis were measured with pH-sensitive glass microelectrodes and were slightly alkaline; pH 7.51, 7.62, and 7.37, respectively. The hemolymph pH remained relatively constant during the development of C. pipiens larvae through the third and early and late fourth instars. The hemolymph pH in C. pipiens larvae parasitized with the mermithid nematode Romanomermis culicivorax was unaltered. These measurements provide an approximate pH (ca 7.4) which is normal for the hemolymph of larval mosquitoes, and should be useful for further development of a culture medium for R. culicivorax.     

Mass Production of the Mosquito Parasite Romanomermis culicivorax: Effect of Density

When numbers of Romanomermis culicivorax Ross and Smith were varied in containers with a constant surface area and depth of sand, densities of 12-24 nematodes per cm² yielded significantly more preparasites than higher densities. When container surface area and numbers of nematodes were constant and sand volume and depth were varied, yields did not differ significantly. When numbers of nematodes and sand volume were constant and surface area and sand depths were varied, yields were significantly higher for a density of 24 nematodes per cm². Yields of preparasites were tripled by simply setting up three cultures, each containing 5 g of nematodes, instead of a single culture containing 15 g.     

Reproduction of Criconemoides simile, Helicotylenchus pseudorobustus,and Paratylenchus projectus on Soybean

Reproduction of Criconernoides simile, Paratylenchus projectus, and Helicotylenchus pseudorobustus, alone and in certain combinations, was characterized during 50 days on the soybean cultivars Custer and Hood. Custer was the more suitable host for all three nematodes and C. simile was the most vigorous parasite on both cultivars. C. simile and H. pseudorobustus reproduced as well in combination as they did alone; addition of P. projectus to this combination did not alter results. Reproduction by P. projectus was repressed on both cultivars when it was combined with C. simile and H. pseudorobustus. The relative importance of time was assessed, as it relates both to the establishment of parasitism by applied nematodes and to the rate of development of larvae in the various host-parasite combinations. The necessity of characterizing as well as enumerating populations when evaluating population development of ectoparasitic nematodes was demonstrated.     

Development and Fecundity of Reesimermis nielseni,a Nematode Parasite of Mosquitoes

Maturation of the mermithid nematode Reesimermis nielseni to the adult stage began by the tenth day after emergence of the nematodes from their hosts at ambient temperatures (24-27 C). Most postparasitic males and females reached the adult stage after 50 and 70 days, respectively. The first females exhibiting egg development and oviposition were observed 25-30 days after emergence, but some oviposition was still taking place 150 days later. Reesimermis nielseni laid an average 2,480 eggs per female over an 18-day oviposition period. A majority of the mature eggs hatched within 7 h after the cultures were flooded. The preparasites are short-lived, and only a few were able to infect exposed hosts after 72 h.     

Control of Larval Northern Corn Rootworm. (Diabrotica barberi) with Two Steinernematid Nematode Species

The entomogenous nematodes Steinerema feltiae and S. bibionis did not penetrate the roots of corn, Zea mays, to infect larval northern corn rootworm (NCR), Diabrotica barberi, feeding within. Laboratory bioassays against first instar NCR indicated that S. feltiae, Mexican strain (LD₅₀ = 49 nematodes/insect) is more virulent than S. bibionis (LD₅₀ = 100). Numbers of NCR larvae in a grain corn crop were reduced by both nematode species applied at corn seeding time at the rate of 10,000 infective-stage juveniles per linear meter of corn row. The chemical insecticide fonofos provided significantly better control than either nematode species.     

Plant-Parasitic Nematodes of South Viet Nam

Between 1974 and 1978, 2,842 identifications of plant-parasitic nematodes were made from more than 1,700 soil and plant samples collected in eight provinces of South Viet Nam. Species in nine genera—Helicotylenchus, Criconemoides, Meloidogyne, Pratylenchus, Tylenchorhynchus, Hoplolaimus, Hirschmanniella, Xiphinema, and Rotylenchulus—comprised 96.1% of the identifications; the remaining 3.9% were species of 11 genera. Fourteen genera were associated with rice which was grown on about 2,500,000 ha in 1970. Of these, Ditylenchus, Hirschmanniella, and Meloidogyne were most important. Ditylenchus angustus caused severe damage to about 50,000 ha of flooded rice in the Mekong Delta in 1976. Hirschmanniella spp. were found in all samples examined from flooded rice fields. Meloidogyne spp. were common in rice seedbeds, upland rice, and rice not kept flooded continuously. Meloidogyne and Pratylenchus spp. were found in roots of 22 of the 32 crop plants sampled. Little or no attempt was made in South Viet Nam to control nematodes.