Demographics of the semi-slug Parmarion martensi, an intermediate host for Angiostrongylus cantonensis in Hawai‘i, during laboratory rearing

The semi-slug, Parmarion martensi, is an intermediate host of the zoonotic nematode, Angiostrongylus cantonensis, the aetiological agent of neuroangiostrongyliasis or rat lungworm disease in humans. Rearing methods were developed for P. martensi to facilitate studies on nematode transmission and control. Parmarion martensi exhibited high survivorship when reared on a diet of dog food and fresh fruits and vegetables in temperature-controlled cabinets at 21.4°C, 98% relative humidity and 12:12 L:D cycle. Rearing containers were lined with moist paper towels for substrate and plastic pots were provided for hiding/resting and egg-laying. Under these conditions, time to first reproduction was 165.3 ± 12.3 days, fecundity was approximately 34.5 ± 7.8 eggs per adult, and hatch rate was 52.7 ± 3.2%. Survivorship post egg hatch was 86.2 ± 2.9% at 30 days (neonates had a mortality rate of about 14%) and 99% thereafter for up to a year. The demographics of laboratory-reared and wild-caught P. martensi were similar except for the weight of reproductive adults, which was significantly higher in laboratory-reared adults (4.0 ± 0.2 g) than in field-collected adults (1.5 ± 0.1 g).

From a Movement-Deficient Grapevine Fanleaf Virus to the Identification of a New Viral Determinant of Nematode Transmission

Grapevine fanleaf virus (GFLV) and arabis mosaic virus (ArMV) are nepoviruses responsible for grapevine degeneration. They are specifically transmitted from grapevine to grapevine by two distinct ectoparasitic dagger nematodes of the genus Xiphinema. GFLV and ArMV move from cell to cell as virions through tubules formed into plasmodesmata by the self-assembly of the viral movement protein. Five surface-exposed regions in the coat protein called R1 to R5, which differ between the two viruses, were previously defined and exchanged to test their involvement in virus transmission, leading to the identification of region R2 as a transmission determinant. Region R4 (amino acids 258 to 264) could not be tested in transmission due to its requirement for plant systemic infection. Here, we present a fine-tuning mutagenesis of the GFLV coat protein in and around region R4 that restored the virus movement and allowed its evaluation in transmission. We show that residues T258, M260, D261, and R301 play a crucial role in virus transmission, thus representing a new viral determinant of nematode transmission.

Endemic infection reduces transmission potential of an epidemic parasite during co-infection

Endemic, low-virulence parasitic infections are common in nature. Such infections may deplete host resources, which in turn could affect the reproduction of other parasites during co-infection. We aimed to determine whether the reproduction, and therefore transmission potential, of an epidemic parasite was limited by energy costs imposed on the host by an endemic infection. Total lipids, triacylglycerols (TAG) and polar lipids were measured in cockroaches ( Blattella germanica ) that were fed ad libitum, starved or infected with an endemic parasite, Gregarina blattarum. Reproductive output of an epidemic parasite, Steinernema carpocapsae , was then assessed by counting the number of infective stages emerging from these three host groups. We found both starvation and gregarine infection reduced cockroach lipids, mainly through depletion of TAG. Further, both starvation and G. blattarum infection resulted in reduced emergence of nematode transmission stages. This is, to our knowledge, the first study to demonstrate directly that host resource depletion caused by endemic infection could affect epidemic disease transmission. In view of the ubiquity of endemic infections in nature, future studies of epidemic transmission should take greater account of endemic co-infections.

A Stretch of 11 Amino Acids in the βB-βC Loop of the Coat Protein of Grapevine Fanleaf Virus Is Essential for Transmission by the Nematode Xiphinema index

ABSTRACT Grapevine fanleaf virus (GFLV) and Arabis mosaic virus (ArMV) from the genus Nepovirus, family Secoviridae, cause a severe degeneration of grapevines. GFLV and ArMV have a bipartite RNA genome and are transmitted specifically by the ectoparasitic nematodes Xiphinema index and Xiphinema diversicaudatum, respectively. The transmission specificity of both viruses maps to their respective RNA2-encoded coat protein (CP). To further delineate the GFLV CP determinants of transmission specificity, three-dimensional (3D) homology structure models of virions and CP subunits were constructed based on the crystal structure of Tobacco ringspot virus, the type member of the genus Nepovirus. The 3D models were examined to predict amino acids that are exposed at the external virion surface, highly conserved among GFLV isolates but divergent between GFLV and ArMV. Five short amino acid stretches that matched these topographical and sequence conservation criteria were selected and substituted in single and multiple combinations by their ArMV counterparts in a GFLV RNA2 cDNA clone. Among the 21 chimeric RNA2 molecules engineered, transcripts of only three of them induced systemic plant infection in the presence of GFLV RNA1. Nematode transmission assays of the three viable recombinant viruses showed that swapping a stretch of (i) 11 residues in the βB-βC loop near the icosahedral 3-fold axis abolished transmission by X. index but was insufficient to restore transmission by X. diversicaudatum and (ii) 7 residues in the βE-αB loop did not interfere with transmission by the two Xiphinema species. This study provides new insights into GFLV CP determinants of nematode transmission.

Aquatic molluscs as auxiliary hosts for terrestrial nematode parasites: implications for pathogen transmission in a changing climate

SUMMARYNematodes are common parasites of molluscs but are often overlooked. Both metastrongyloid and rhabditoid species dominate the fauna within land snail and slug populations. Nevertheless, a key characteristic of many laboratory studies is the ability of these terrestrial nematodes to utilize aquatic molluscs as auxiliary hosts. The significance of this to the ecology of the parasite has never been evaluated. There is increasing concern as to the impact of climate change on the epidemiology of many parasitic diseases. In particular, it has been suggested that host switching may increase under the pressure of extreme climatic conditions. It is therefore timely to assess the role that aquatic molluscs may play in transmitting terrestrial nematodes, which include species of medical and veterinary importance such as Angiostrongylus cantonensis, A. vasorum, and Muellerius capillaris. The present review assesses the mechanisms of terrestrial nematode transmission through aquatic molluscs focusing on metastrongyloid and rhabditoid species, the importance of variable susceptibility of molluscan hosts, field studies on natural occurrence within aquatic habitats, and the impact of extreme climatic events (floods and droughts) that may increase in frequency under climate change.

Transmission of the pine wood nematode Bursaphelenchus xylophilus through oviposition activity of Monochamus galloprovincialis (Coleoptera: Cerambycidae)

Transmission of Bursaphelenchus xylophilus (PWN) by the oviposition of Monochamus galloprovincialis was studied in Portugal. Female insects laid eggs on 77% of pine bolts on the laboratory, of which 37% became infected with PWN, with a mean of 290 nematodes. Inside the PWN affected zone 50 P. pinaster trap-trees were created between May and September 2001 and 2002, of which 29 were colonised by M. galloprovincialis and only four became infected by the nematode. The low transmission efficiency detected both on laboratory and field suggests that nematode transmission through the vector’s oviposition activity may be a secondary component of the pine wilt disease epidemiology in Portugal.

Characteristics of the emergence of Monochamus alternatus, the vector of Bursaphelenchus xylophilus (Nematoda: Aphelenchoididae), from Pinus thunbergii logs in Nanjing, China, and of the transmission of the nematodes through feeding wounds

The pine wood nematode (PWN), Bursaphelenchus xylophilus, which in China and East Asia is vectored by Monochamus alternatus, is the causal agent of pine wilt disease of Japanese black pine, Pinus thunbergii. The vector emergence and the transmission of nematodes through vector feeding on pine twigs were monitored during 2004 and 2005 in Nanjing, China. The emergence started from late April to the end of June and peaked from late May to early June. There were 438 and 927 adults collected in 2004 and 2005, respectively, and approximately 70% of the beetles emerged during the peak period. Visual estimation of the nematode burden on vectors by observation of the atrium of the first abdominal spiracle gave unreliable information. The percentage of beetles carrying PWN of the total number of emerged beetles was between 20 and 30%. Seventy percent of the nematodes were released from beetle cadavers after 3 days of extraction. The sex and longevity of the beetle had no significant relationship with the number of nematodes that remained in cadavers. Transmission of nematodes into pine twigs through beetle feeding started 10 days after emergence of the vector. The period of nematode transmission could last for up to 79 days after beetle emergence. Two types of nematode-transmission curve were found by measuring the number of nematodes transmitted into pine twigs per day. The unimodal pattern peaked between 3 and 6 weeks after adult emergence; the bimodal pattern had two transmission peaks: one between 2 and 3 weeks, and another between 5 to 7 weeks after beetle emergence.

Mild energy restriction alters mouse–nematode transmission dynamics in free-running indoor arenas

Energy restriction reduces Heligmosomoides polygyrus (Dujardin, 1845) (Nematoda) infection by reducing transmission-related behaviours but prolongs parasite survival by suppressing immune responses in individually housed mice. To determine the relative importance of these two processes in accumulation of worms in mouse populations, 10 female CD1 mice were housed in each of eight indoor arenas with ad libitum access to either an energy-sufficient (ES) diet or an energy-restricted (ER) diet with 20% less metabolizable energy (four arenas per diet). After 3 weeks, H. polygyrus transmission was initiated by introducing larvae onto damp peat trays. Mice adapted to the ER diet through increased food intake and nesting and reduced overall activity; after 6 weeks, nutritional and immunological measures were comparable between diet groups. With continuing exposure to parasite larvae, mice in both ER and ES arenas developed resistance to the incoming larvae; however, mice in the ER arenas accumulated lower worm burdens than mice in the ES arenas despite their increased contact with peat. We suggest that the comparable immunocompetence of mice in the ER and ES arenas enabled the ER mice exposed to higher transmission rates to more rapidly reject the parasites, leading to lower final worm numbers, a pattern frequently observed in other helminth infections.