Ephemera strigata (Insecta: Ephemeroptera: Ephemeridae) is the intermediate host of the nematodes Rhabdochona denudata honshuensis and Rhabdochona coronacauda in Japan

Juvenile freshwater parasitic nematodes Rhabdochona denudata honshuensis Moravec and Nagasawa, 1989 and Rhabdochona coronacauda Belouss, 1965 (Spirurida: Thelazioidea: Rhabdochonidae) were found in mayfly nymphs collected in a mountain stream in Japan. Considering the relative density of mayfly nymphs, nematode prevalence, and intensity of parasitism, Ephemera strigata Eaton and Potamanthus formosus Ulmer (Ephemeroptera: Ephemeridae) are frequent natural intermediate hosts for R. d. honshuensis in this locality. The intermediate host of R. coronacauda also is the E. strigata nymph.     

Development and feeding of <Emphasis Type="Italic">Tetraphleps galchanoides</Emphasis>, a predator of the hemlock woolly adelgid

Tetraphleps galchanoides Ghauri (Hemiptera: Anthocoridae) nymphs were collected from hemlock woolly adelgid (HWA) Adelges tsugae Annand (Hemiptera: Adelgidae) infested Tsuga sp. in Baoxing, Sichuan, China. First and second stage nymphs collected from foliage shipped from China; were reared to adults and tested for feeding rates and host preferences. They were reared at 5, 8, 12, and 15 ± 1 °C from November to December, January to March, April, and May to June, respectively, in the quarantine laboratory at Virginia Polytechnic Institute and State University. At 8 °C, development time was 15, 20, and 40 days for the N-III, IV, and V nymphal stages, respectively. Adult males lived 83 days with a range of 21–147 days. A single adult female lived for 21 days. At 5 °C, second stage T. galchanoides nymphs consumed 0.8 HWA nymphs per day, and 2.0 HWA nymphs per day at the N-V stage. At 8 °C, consumption of HWA nymphs ranged from 1.3 to 3.4 nymphs per day for the N-III to N-V stages, respectively. Adult T. galchanoides consumed more HWA eggs than HWA adults, pine bark adelgid (PBA) Pineus strobi (Hartig) (Hemiptera: Adelgidae) adults, and eggs in no-choice tests. In choice tests with HWA eggs and PBA eggs, more HWA eggs were eaten. Adult and nymph body measurements are presented for determination of nymphal instars.     

Redescription and Life History of Contortylenchus brevicomi,a Parasite of the Southern Pine Beetle Dendroctonus frontalis

Larval and adult life stages are described for Contortylenchus brevicomi (Massey) Rühm parasitizing a Mississippi population of Dendroctonus frontalis, the southern pine beetle. Fourth-stage larvae and free-living adult females of this species are identified and described for the first time. The life cycle of C. brevicomi can be reconstructed from this study. The adult female nematode lays eggs in a mature beetle. Larval development progresses within the hemocoel until fourth-stage larvae exit the host. Mating occurs in beetle galleries and only females enter an immature beetle host.     

Onchocerca volvulus: biochemical and morphological characteristics of the surface of third- and fourth-stage larvae

The annulated cuticles of third- and fourth-stage larvae of Onchocerca volvulus have the typical structure of other nematodes but the cuticle of fourth-stage larvae was thinner. The surface of the third-stage larva was wrinkled and fuzzy, while that of the fourth-stage was smooth. Intermediate stages in the formation of the new cuticle and epicuticle beneath the old basal layer and of the separation of the cuticles are shown. Monoclonal antibodies specific to the surface of third-stage larvae did not react with the surface of the fourth-stage larvae. Binding of the monoclonal antibodies to the third-stage larvae was abrogated by treatment of the worms with trypsin and proteinase K, but was unaffected by treatment with periodate or the detergents sodium deoxycholate and SDS. The lectins RCA120 and WGA, but not any of the other lectins tested, bound only to the surface of fourth-stage larvae, and not to that of third-stage larvae. The surfaces of third- and fourth-stage larvae were shown to be different and contained stage-specific surface epitopes.     

Brugia malayi: patterns of expression of surface-associated antigens.

Patterns of expression of surface-associated antigens were analyzed in the filarial nematode Brugia malayi immediately prior, and during development in the vertebrate host. Two surface-associated protein molecules, i.e., accessible to surface radioiodination and soluble in aqueous buffers, were investigated: Mrs 29-30,000 and 16,000, both of which are antigenic in infected animals. The Mr 29-30,000 glycoprotein is expressed in a surface-associated manner by adult worms and by fourth-stage larvae, but is not detectable in preparasitic third-stage larvae. The 16,000 component, which appears not to be glycosylated, is surface-associated in adult worms and fourth-stage larvae. In contrast to the 29-30,000 glycoprotein, the 16,000 protein is also expressed both by pre- and postparastic third-stage larvae. However, it becomes surface-associated only after infection. Thus, immediately prior, and during development within the vertebrate host, B. malayi displays at least two different patterns of expression of surface-associated antigens: (i) de novo, intiated either immediately after infection (phase specific) or during genesis of the fourth-stage larva (stage specific); (ii) continuous, but with phase-dependent surface exposure of previously cryptic antigens, during the transition from intermediate to definitive host.     

Development of Dipetalonema viteae third-stage larvae (Nematoda: Filarioidea) in micropore chambers implanted into jirds, hamsters, normal and immunized mice

Development of third-stage larvae of Dipetalonema viteae within subcutaneously implanted micropore chambers proceeded in all hosts tested up to the fourth-stage larvae and occasionally to adolescent worms. In the jird the timing of development was comparable to a natural infection. Although the mouse is an insusceptible host, larval development could take place, but was very slow. Two intraperitoneal inoculations of living third-stage larvae into mice induced the production of antibodies against the larval cuticle and against common antigens. In such immune mice the development of third- and fourth-stage larvae within micropore chambers was significantly inhibited, larval mortality was increased, and the larval motility was impaired.     

Predation of Galendromus helveolus (Acari: Phytoseiidae) on Brevipalpus californicus (Acari: Tenuipalpidae)

The false spider mite, Brevipalpus californicus (Banks), is a major citrus pest in Texas. This phytophagous mite causes damage to fruit, leaves and stems of citrus, and is also a vector of citrus leprosis virus. Galendromus helveolus (Chant) is one of the most prevalent predacious mite species found on Texas citrus. The predation potential of G. helveolus on different stages of B. californicus was evaluated in the laboratory. An individual immature G. helveolus mite consumed an average of 30.7 eggs, 53.6 larvae, or 22.7 nymphs of B. californicus before developing to an adult. An individual adult female G. helveolus consumed an average of 164.8 eggs, 369.6 larvae, or 80.9 nymphs of B. californicus. Both immature and adult G. helveolus never fed on adult stage of B. californicus and never completed development. The development times of the immature stages of G. helveolus were 4.5, 4.1 and 4.6 days when fed on eggs, larvae and nymphs of B. californicus, respectively. When G. helveolus fed on the larval stage of B. californicus, the adults had the longest longevity (18.5 days) and the highest fecundity (14.0 eggs/female).     

Laboratory studies of the biology and food requirements ofMacrorhaphis acuta [Hemiptera: Pentatomidae)

Macrorhaphis acuta were bred in the laboratory (25°C) and fed on larvae ofAscotis selenaria reciprocaria. The incubation period was 8.4 days and the mean hatch was 94.4%. There were 5 nymphal instars which occupied 3.3, 5.0, 5.6, 6.6 and 12.0 days, respectively, from the lst to the 5th. Each nymph consumed an average of 22.8 host larvae to complete development. Adults had a mean longevity of 74.9 days and each consumed an average of 67.5 host larvae. Significant reduction was noted in pupation of 5th instar host larvae sucked by adult predators for periods ranging from 4 to 10 mn. The food requirements of 3rd, 4th, and 5th instar nymphs fed on 3rd instar host larvae were also investigated. The conversion ratio varied with the weight of food consumed during the instar and later instars were more efficient than earlier ones. Predatory value did not vary significantly with successive instars.     

Transfer of methyl-branched hydrocarbons from the parasitoid, Eretmocerus mundus, to silverleaf whitefly nymphs during oviposition

The parasitic wasp Eretmocerus mundus (Hymenoptera: Aphelinidae), a natural enemy of the silverleaf whitefly Bemisia argentifolii (Homoptera: Aleyrodidae), deposits eggs beneath nymphs and not within them. Experiments were designed to establish whether ovipositing E. mundus females leave marking chemicals on nymphs to enable searching females to discriminate parasitized from unparasitized hosts. Cuticular lipids from three experimental treatment groups were characterized: parasitoid-exposed nymphs that had a parasitoid egg between the nymph and leaf; control nymphs not exposed to E. mundus; and a third treatment condition of parasitized nymphs, held for 10 days after wasp exposure. Lipids were solvent-extracted from the nymphal cuticles of the various treatment groups and the lipid components were characterized and quantified by gas chromatography and mass spectrometry. Results indicated the presence of quantities of C31 and C33 dimethylalkanes only from parasitoid-exposed groups of nymphs and not in the extracts from control nymphs or the parasitized nymphs after 10-day exposure. Furthermore, the C31 and C33 dimethylalkanes were shown to be major lipid components of the hexane extracts from E. mundus females. Since the lipids were removed from parasitoid-exposed nymphs before interaction with hatched parasitoid larvae, the findings indicated that the dimethylalkanes were transferred onto nymphal cuticles by ovipositing E. mundus females.     

Population biology of Contracaecum rudolphii sensu lato (Nematoda) in the great cormorant (Phalacrocorax carbo) from northeastern Poland

Parameters related to the occurrence, aggregation, and population structure of the nematode Contracaecum rudolphii Hartwich, 1964 , in the great cormorant ( Phalacrocorax carbo ) from northeastern Poland were analyzed. A total of 491 birds of different ages (adults, immatures, and nestlings) was examined; the cormorants studied were taken from both fresh- and brackish water habitats. Contracaecum rudolphii were found in stomachs of 454 birds (92.5%); the 46,244 nematodes included third- (L3) and fourth-stage larvae (L4), and sub-adult and adult females and males. The distribution of nematodes in the host population were highly aggregated. The occurrence of C. rudolphii was significantly dependent on the host's age and habitat, as well as on season; the proportion of development stages differed significantly depending on birds' age and season (the latter only in adult birds). The infrapopulations of C. rudolphii in the adult cormorants showed distinct density-dependent correlations: that is, as the infrapopulation size increased, the number of adult females C. rudolphii decreased, and the proportion of larvae and sub-adult females increased. A higher proportion of larvae and sub-adult females characterized the component population structure of the nematodes in the cormorant nestlings, compared with adult birds, probably because of immune system deficiency in the immature birds, coupled with the development of the nematode population. Seasonal changes in the C. rudolphii population, observed in the adult cormorants, were not related to seasonality of the L3 occurrence in food; instead, the changes are believed to have resulted from independent processes of elimination of the oldest nematodes and their replacement by larval stages that subsequently matured.     

九香虫形态及生物学特征研究

通过建立网室近自然实验种群,结合野生个体观察记述了云南玉溪九香虫Coridius chinensis (Dallas)种群的各虫期形态特征、在云南玉溪红塔区的年生活史、寄主范围、取食偏好、成虫及若虫的行为习性,为该虫的保育和规模化养殖提供依据。结果显示,九香虫卵有绿色和粉色两种生态型,若虫有5个龄期;在云南玉溪红塔区1年仅发生1代,仅以成虫态越冬;红塔区九香虫野生个体主要以佛手瓜和南瓜等葫芦科植物为食;取食偏好选择试验表明该虫最喜取食葫芦,冬瓜和南瓜次之;雌雄成虫可多次交配多次产卵,单雌单次平均产卵量为16.3±6.0粒。研究表明,云南玉溪红塔区的九香虫种群为典型的一化性昆虫,休眠期可达8个月;该虫食性较广,可取食多科植物,但以葫芦科瓜类为主;羽化成虫每年9月中旬至10月上旬陆续从佛手瓜、南瓜等寄主植物上迁至枯树皮下、柴堆等隐蔽场所越冬,翌年4月下旬陆续解除休眠,迁至佛手瓜、南瓜等作物上取食繁殖。     

Transovarially-transmitted intracellular microorganisms in adult and larval stages of Brugia malayi

Observation of intracellular organisms in the lateral chords of Brugia malayi adults initiated further studies to determine the prevalence of these organisms within the tissues of adult worms and of larvae. The organisms were found in the lateral chords of adult males and females, microfilariae, first-, second-, third-, and fourth-stage larvae. In the females, they were present in the oogonia, oocytes, and developing eggs, suggesting transovarial transmission within the life cycle of the filarid. The organisms may have a developmental cycle consisting of more than one stage, including a small spheroidal stage up to 0.6 micrometer in size and a larger form up to 1.5 micrometer in length, all of which occur in the cytoplasm within a vesicle formed of host membrane. Each stage lacks a definite cell wall, being bound by 2 trilaminate membranes. The bacterial entities in B. malayi resemble both in morphology and development the organisms found in other filarids, but whether they affect the vertebrate host in any way remains to be determined. Their presence within certain cells of the developing eggs could be exploited as intracellular markers for the organogenesis of the lateral chords and the ovary.     

The life cycle of Paraquimperia tenerrima: a parasite of the European eel Anguilla anguilla

Previous studies on the life history of the nematode eel specialist Paraquimperia tenerrima (Nematoda: Quimperiidae) have failed to determine whether an intermediate host is required in the life cycle. In the laboratory, eggs failed to hatch below 10 degrees C, hatching occurring only at temperatures between 11 and 30 degrees C. Survival of the free-living second stage larvae (L2) was also temperature dependent, with maximal survival between 10 and 20 degrees C. Total survival of the free-living stages (eggs and L2) is unlikely to exceed a month at normal summer water temperatures, confirming that parasite could not survive the 6 month gap between shedding of eggs in spring and infection of eels in early winter outside of a host. Eels could not be infected directly with L2, nor could a range of common freshwater invertebrate species. Third stage larvae (L3) resembling P. tenerrima were found frequently and abundantly in the swimbladder of minnows Phoxinus phoxinus from several localities throughout the year and were able to survive in this host in the laboratory for at least 6 months. Third stage larvae identical to these larvae were recovered from minnows experimentally fed L2 of P. tenerrima, and eels infected experimentally with naturally and experimentally infected minnows were found to harbour fourth stage larvae (L4) and juvenile P. tenerrima in their intestines. Finally, the whole life cycle from eggs to adult was completed in the laboratory, confirming that minnows are an obligate intermediate host for P. tenerrima.     

Development of Ascarophis sp. (Nematoda: Cystidicolidae) to maturity in Gammarus deubeni (Amphipoda)

Experimentally transmitted Ascarophis sp. (Spirurida) developed to adult worms in the invertebrate host, Gammarus deubeni (Amphipoda), collected in the intertidal zone in Passamaquoddy Bay, New Brunswick, Canada. The morphological development and growth of larval stages is very similar to other cystidicolids, which are found as adults in fish. Unlike virtually all other Spirurida, which require a vertebrate definitive host, infective larvae of Ascarophis sp. migrate from the invertebrate host musculature into the hemocoel where they molt twice to become adults. Gravid females appear at 80 days and 69 days post-infection at 10-12 C and 18-20 C, respectively. While there is little evident host reaction to the parasite within the muscle tissue, within the hemocoel there is hemocytic reaction to shed nematode cuticles, released eggs, and sometimes the worm itself, including some melanization. The worms are morphologically similar to Ascarophis sp. from G. oceanicus in the Baltic and White seas and among Ascarophis species from fish is most similar to A. arctica. It is suggested that Ascarophis sp. no longer requires a vertebrate host and is transmitted between amphipods either through death and disintegration of infected amphipods and dispersal of the nematode eggs, or more likely through cannibalism or necrophagy.     

Lifecycle of Rhipicephalus haemaphysaloides under laboratory conditions

Rhipicephalus haemaphysaloides Supino, 1897 (Ixodida: Ixodidae) is a three‐host hard tick widely distributed in Sri Lanka. The lifecycle of the R. haemaphysaloides population in Sri Lanka was studied under laboratory conditions using fully engorged females collected from dogs. Larvae, nymphs and adults were reared on New Zealand rabbits. Engorged females weighed a mean ± standard deviation (SD) of 470.1 ± 106.5 mg and laid a mean ± SD of 4067.4 ± 3136.2 eggs within 16.1 ± 3.7 days. The mean ± SD preoviposition period was 6.5 ± 1.1 days. The mean ± SD reproductive efficiency index was 8.5 ± 4.1, and was higher in heavier females (Pearson correlation, r = 0.8; P = 0.001). The incubation period of eggs was shorter at higher temperatures. The majority (86.9%) of larvae successfully moulted after a mean ± SD premoulting period of 7.9 ± 0.7 days following a parasitic period of 3.2 ± 0.5 days. Unfed larvae survived a mean ± SD of 103.4 ± 19.8 days. Nymphs completed the bloodmeal after 3.7 ± 0.9 days, weighed 5.7 ± 1.7 mg and demonstrated a premoulting period of 15.1 ± 1.9 days. Unfed nymphs survived a mean ± SD of 45.8 ± 3.8 days. Heavier nymphs moulted into females, whereas lighter nymphs moulted into males (two‐sample t‐test, P < 0.001). The bodyweight of adult females increased more than 100‐fold after feeding. The lifecycle of R. haemaphysaloides was successfully completed on rabbits within a mean of 91.3 days (range: 69–117 days) under laboratory conditions.     

Cytosolic glutathione S-transferases of Oesophagostomum dentatum

Oesophagostomum dentatum stages were investigated for glutathione S-transferase (GST) expression at the protein and mRNA levels. GST activity was detected in all stages (infectious and parasitic stages including third- and fourth-stage larvae of different ages as well as males and females) and could be dose-dependently inhibited with sulfobromophthalein (SBP). Addition of SBP to in vitro larval cultures reversibly inhibited development from third- to fourth-stage larvae. Two glutathione-affinity purified proteins (23 and 25 kDa) were detected in lysates of exsheathed third-stage larvae by SDS-PAGE. PCR-primers were designed based on peptide sequences and conserved GST sequences of other nematodes for complete cDNA sequences (621 and 624 nt) of 2 isoforms, Od-GST1 and Od-GST2, with 72% nucleotide similarity and 75% for the deduced proteins. Genomic sequences consisted of 7 exons and 6 introns spanning 1296 bp for Od-GST1 and 1579 and 1606 bp for Od-GST2. Quantitative real-time-PCR revealed considerably elevated levels of Od-GST1 in the early parasitic stages and slightly reduced levels of Od-GST2 in male worms. Both Od-GSTs were most similar to GST of Ancylostoma caninum (nucleotides: 73 and 70%; amino acids: 80 and 73%). The first three exons (75 amino acids) corresponded to a synthetic prostaglandin D2 synthase (53% similarity). O. dentatum GSTs might be involved in intrinsic metabolic pathways which could play a role both in nematode physiology and in host-parasite interactions.     

The life history of Plagioporus shawi (McIntosh 1939) (Trematoda:Opecoelida), an intestinal parasite of salmonid fishes.

Podocotyle shawi is transferred to the genus Plagioporus. The miracidium develops and hatches in 11 to 14 days at 22 to 25 C. The operculate snail, Lithoglyphus virens, serves as first intermediate host. Mother and daughter sporocysts develop, the latter producing cotylomicrocercous cercariae within 90 to 130 days. Cercariae encyst in the amphipod, Hyalella azteca; stonefly nymphs of the genus Arcynopteryx; mayfly nymphs of the genera Heptagenia and Paraleptophlebia; chironomid larvae of the genera Chironomus, Polypedilum, Phaenopsectra, Ablabesmyia, and Psectrocladius and caddis larvae of the genera Brachycentrus, Limnephilus, and Hydropsyche. Metacercariae 60 to 100 days old were fed to young steelhead trout, Salmo gairdneri, and partially developed specimens of P. shawi were recovered 26 days later. Stonefly nymphs are believed to be the most important second intermediate host in the natural environment. The parasite seems to be restricted to the Pacific Northwest of the United States and has been reported from the silver salmon, Oncorhynchus kisutch; kokanee, O. nerka; cutthroat trout, Salmo clarki; steelhead trout, S. gairdneri; and the mountain whitefish, Prosopium williamsoni.     

Ticks on reptiles and amphibians in Central Amazonia,with notes on rickettsial infections

Reptiles and amphibians are exceptional hosts for different ectoparasites, including mites and ticks. In this study, we investigated tick infestations on reptiles and amphibians trapped in Central Amazonia, and also assessed the presence of rickettsial infections in the collected ticks. From September 2016 to September 2019, 385 reptiles (350 lizards, 20 snakes, 12 tortoises, and three caimans) and 120 amphibians (119 anurans and one caecilian) were captured and examined for ectoparasites. Overall, 35 (10%) lizards, three (25%) tortoises and one (0.8%) toad were parasitized by ticks (124 larvae, 32 nymphs, and 22 adults). In lizards, tick infestation varied significantly according to landscape category and age group. Based on combined morphological and molecular analyses, these ticks were identified as Amblyomma humerale (14 larvae, 12 nymphs, 19 males, and one female), Amblyomma nodosum (three larvae, one nymph, and one female), and Amblyomma rotundatum (four larvae, three nymphs, and one female), and Amblyomma spp. (103 larvae and 16 nymphs). Our study presents the first records of A. nodosum in the Amazonas state and suggests that teiid lizards are important hosts for larvae and nymphs of A. humerale in Central Amazonia. Moreover, a nymph of A. humerale collected from a common tegu (Tupinambis teguixin) was found positive for Rickettsia amblyommatis, which agrees with previous reports, suggesting that the A. humerale-R. amblyommatis relationship may be more common than currently recognized.

     

Seasonal patterns of abundance and host relationships of the Australian paralysis tick,Ixodes holocyclus Neumann (Acarina: Ixodidae), in southeastern Queensland

Estimates of seasonal abundance of larvae, nymphs and females of the paralysis tick, Ixodes holocyclus, were obtained by collecting ticks that engorged on small mammals and birds trapped in two localities in southeastern Queensland: Brisbane (wet sclerophyll forest) and Tamborine Mountain (cleared rain forest). The long-tailed short-nosed bandicoot, Isoodon macrourus, was the most common mammal trapped but small numbers of other marsupials, rodents and ground frequenting birds were also captured. Small numbers of five other tick species were also collected. In both habitats there was clearly one dominant generation of I. holocyclus;7er year, although the presence of all stages at most times of the year indicated overlapping of smaller cohorts. Females were most abundant in spring and early summer, larvae in summer-autumn, and nymphs in autumn-winter. I. holocyclus was abundant on I. macrourus and rare on most other mammals and birds captured. At the peak of abundance of each instar, each bandicoot dropped from 500 to 2000 engorged larvae, 100 to 200 engorged nymphs, and four to six engorged females. Life tables were compiled for the tick in both habitats and these indicate that there was relatively high survival from engorged larva to engorged nymph and thence to engorged female and that most mortality occurred between detaching of the engorged female and the detaching of the engorged larva. The tick was more abundant on bandicoots from cleared rain forest and rain forest edge, than on those from sclerophyll woodland. The survival of engorged larvae and nymphs of I. holocyclus and the larval productivity of engorged females were examined in a warm moist climate where the tick was abundant (Tamborine Mountain) and in a hotter dryer climate where the tick was rare or absent (Amberley). In both localities engorged larvae and nymphs survived to the next instar in all seasons of the year. On most occasions engorged females produced eggs which hatched. Mature bandicoots from tick infested areas showed little or no resistance to infestation with larvae or nymphs of I. holocyclus, whereas other small mammals from the same area showed an appreciable degree of resistance to the immature stages of the tick. Feeding larvae and nymphs exposed to normal light-dark cycles in the laboratory detached during the afternoon and early evening. This behaviour and host resistance are discussed in relation to the daily activity cycles of host species, their habitat preferences, and their role as hosts for I. holocyclus.