Ichthyophthirius multifiliis (Ciliophora) Exit from Gill Epithelium1

The first change in the sequence of morphological events occurring as fully developed Ichthyophthirius multifiliis trophonts spontaneously left gill epithelium or as younger trophonts departed, following experimentally induced death of the fish, was the separation of parasites from overlying host cells. Discharge of contractile vacuoles may have played a role in this process. Spaces then appeared between host cells, and individual epithelial cells became vacuolated. Finally, the epithelium ruptured and the parasites swam free. In induced exit after three days residence in the host, departure of the trophont was evident only after autolysis of epithelium had occurred. Induced departure of trophonts after four days residence was more rapid, suggesting an active role for the parasite in exit. Changes in parasite and epithelium observed in induced exit were similar to those in spontaneous departure after five days residence.     

Ichthyophthirius multifiliis (Ciliophora) Development in Gill Epithelium1

ABSTRACT. The development of Ichthyophthirius multifiliis trophonts in gill epithelium of channel catfish was studied on the first five days post-exposure (PE) by light and electron microscopy. Trophonts increased in average diameter from 48 μm at one day PE to 248 μm at five days PE. Although theronts invaded any part of the gill epithelium, at three days PE most parasites were found adjacent to major blood vessels, particularly the afferent vessel. From one to three days PE, 70–90% of trophonts were immediately adjacent to gill epithelium, but at four and five days PE only 50% of trophonts were closely apposed by host epithelial cells. Notable ultrastructural changes in the trophonts over the five-day period occurred in the mucocysts and lipid inclusions, both of which increased markedly in number. The few crystalline mucocysts present at one day PE were near the cell periphery but rarely attached to the plasma membrane. At three days PE, crystalline mucocysts were significantly more abundant, and at five days PE, they occurred in large numbers throughout the cytoplasm as well as at the periphery. At three days PE, secretory mucocysts were first observed. Contractile vacuoles were more prominent at five days PE than earlier in development. Development of mucocysts and lipid reserves is probably essential to survival and reproduction of the ciliate after it leaves the host.     

Ichthyophthirius multifiliis as a potential vector of Edwardsiella ictaluri in channel catfish

There is limited information on whether parasites act as vectors to transmit bacteria in fish. In this trial, we used Ichthyophthirius multifiliis and fluorescent Edwardsiella ictaluri as a model to study the interaction between parasite, bacterium, and fish. The percentage (23-39%) of theronts fluorescing after exposure to E.?ictaluri was significantly higher than control theronts (~?6%) using flow cytometry. Theronts exposed to E.?ictaluri at 4?×?10(7) CFU?mL(-1) showed a higher percentage (~?60%) of fluorescent theronts compared to those (42%) exposed to 4?×?10(3) CFU?mL(-1) at 4?h. All tomonts (100%) carried the bacterium after exposure to E.?ictaluri. Edwardsiella ictaluri survived and replicated during tomont division. Confocal microscopy demonstrated that E.?ictaluri was associated with the tomont surface. Among theronts released from tomonts exposed to E.?ictaluri, 31-66% were observed with attached E.?ictaluri. Sixty percent of fish exposed to theronts treated with 5?×?10(7) E.?ictaluri?mL(-1) were positive for E.?ictaluri at 4?h as determined by qPCR or fluorescent microscopy. Fluorescent E.?ictaluri were observed on trophonts in skin and gill wet mounts of dead fish. This study demonstrated that Ich could vector E.?ictaluri to channel catfish.     

Ichthyophthirius (Ciliophora): Population Studies Suggest Reproduction in Host Epithelium1

Evidence that Ichthyophthirius multifiliis trophonts may reproduce within the epithelium of the host was obtained from experimental infections of channel catfish. Mean number of parasites spontaneously leaving the fish increased from 0 on day 3 postexposure (PE) to 66.5 per fish on day 7 PE. Mean population density in fin, however, increased five-fold from day 3 to day 5 PE in the absence of opportunity for reinfection. At day 3 PE, 100% of parasite loci in fin and gill arches contained solitary trophonts. At day 4 PE, 10% of loci in fin contained clusters of two trophonts; at day 7 PE, 69% contained clusters of two or more trophonts. The first clusters of four trophonts in fin were observed day 5 PE and of eight trophonts, day 8 PE. Trophonts in clusters were flattened against one another.     

Biology of Microplitis kewleyi (Hymenoptera: Braconidae): A parasite of Agrotis ipsilon (Lepidoptera: Noctuidae) larvae

Microplitis kewleyi Muesebeck is a gregarious internal parasite of larvae of the black cutworm Agrotis ipsilon (Hufnagel). Studies of the biology of the parasite revealed that there was an inverse relationship between host instar and parasite preference. Duration of development from egg to pupa ranged from 18 days at 27°C to 68.7 days at 16°C. Development from egg to pupa took 13.5–21.6 days when fourth and first instar host larvae, respectively, were parasitized. A larger number of parasites emerged from hosts parasitized in the fourth instar (22.4) than the first instar (11.5). Parasite pupation occurred when the host was in the fifth/sixth instar, depending on the instar parasitized. Thirty‐nine per cent of host larvae exposed as first instars to parasites died before parasite emergence. This decreased to 0% for host larvae exposed as fourth instars. The sex ratio was 1:1.2 (M:F). Thirty‐seven per cent of hosts exposed diurnally were stung, compared to 24% exposed nocturnally. Mean daily progeny was highest (12) on the first day, decreasing to zero after 20 days. Percent host parasitism was also highest on the first day (35%) decreasing to nearly 0% after 18 days. There appear to be three parasite larval instars. Host larvae often remained alive after parasite emergence.     

Ichthyophthiriasis in the mirror carp Cyprinus carpio (L.) V. Acquired immunity*

Mirror carp (Cyprinus carpio L.) infected with sublethal doses of Ichthyophthirius multifiliis, were free of parasites 21 days after infection. Fish remained free of parasites for at least 8 months when maintained in an infective environment. Such fish were refractory to reinfection with numbers of parasites that killed all normal, previously unexposed fish. Serum from fish recovered from previous infections with sublethal doses of parasites, immobilized free-swimming stages of I. multifiliis to a dilution of up to 1: 1024. The rise in serum-immobilization titre occurred between the 10th and 22nd days of infection, the period during which parasites disappeared from the body surface of the fish. Infective stages of the parasite were unable to penetrate the mucus body covering of resistant fish.     

Ichthyophthirius multifiliis (Ciliophora) exit from gill epithelium

The first change in the sequence of morphological events occurring as fully developed Ichthyophthirius multifiliis trophonts spontaneously left gill epithelium or as younger trophonts departed, following experimentally induced death of the fish, was the separation of parasites from overlying host cells. Discharge of contractile vacuoles may have played a role in this process. Spaces then appeared between host cells, and individual epithelial cells became vacuolated. Finally, the epithelium ruptured and the parasites swam free. In induced exit after three days residence in the host, departure of the trophont was evident only after autolysis of epithelium had occurred. Induced departure of trophonts after four days residence was more rapid, suggesting an active role for the parasite in exit. Changes in parasite and epithelium observed in induced exit were similar to those in spontaneous departure after five days residence.     

Consequences of divergent temperature optima in a host–parasite system

It was suggested that parasite infections become more severe with rising temperature, as expected during global warming. In ectothermic systems, the growth of a parasite and therefore its reproductive capacity is expected to increase with temperature. However, the outcome of the interaction depends on the temperature optima of both host and parasite. Here we used experimental infections of three‐spined stickleback fish Gasterosteus aculeatus with its specific tapeworm parasite Schistocephalus solidus to investigate in detail the temperature optima for both host and parasite. We analyzed the fitness consequences thereof, focusing on growth and immunity of the host, and growth and offspring production of the parasite as fitness correlates. We checked for potential differences among populations, using the offspring of hosts and parasites derived from four study sites in Iceland, Germany and Spain that differ in average annual temperature ranging between 4.8°C and 18.4°C. We found differences in temperature optima of host and parasites that were quite consistent across the populations: while sticklebacks grew faster and had higher immune activity at low temperatures, the parasites did not even grow fast enough to reach sexual maturity in these conditions. By contrast, with increasing temperatures, parasite growth, egg production and offspring hatching increased strongly while host immunity and growth were impaired. Our results show that divergent temperature optima of hosts and parasites can have drastic fitness consequences and support the expectation that some parasites will benefit from global warming.     

Ophryoglena hemophaga n. sp. (Ciliophora: Ophryoglenidae): a parasite of the digestive gland of zebra mussels Dreissena polymorpha

Ophryoglena hemophaga n. sp. is described from a freshwater Dreissena polymorpha population in the Rhine delta of the Netherlands. This is the first ophryoglenine species (order Hymenostomatida, suborder Ophryoglenina) recorded as a molluscan parasite. As is typical of ciliates in the suborder Ophryoglenina, O. hemophaga exhibits a polymorphic life history with cystment and reproduction by palintomy. Trophonts were observed within digestive gland lumina, and zebra mussel hemocytes were present in some of their digestive vacuoles. The presence of a single, longitudinal tract of multiple contractile vacuoles represents its most unique feature and distinguishes it from all other described Ophryoglena spp. The number of somatic kineties of O. hemophaga (range 100 to 124) is also distinguishing as it is one of the lowest for [corrected] an Ophryoglena sp. Other characteristics of this species include: ovoid to elongate trophonts 96 to 288 microm in length, with an elongate macronucleus 41 to 65 microm in length; tomonts 50 to 150 microm in diameter producing a clear mucous cyst envelope, whose thickness is approximately half of the tomont diameter; elongated theronts 96 to 131 microm in length which emerge after 1 to 3 cell divisions taking 36 to 48 h at 20 +/- 3 degrees C. Protomonts and theronts are, respectively, negatively and positively phototactic--characteristics that likely aid in maintenance of infection in zebra mussel populations.     

Biological studies withChrysocharis parksi [Hym.: Eulophidae] a parasite ofLiriomyza spp. [Dipt.: Agromyzidae]

Biological studies withChrysocharis parksi Crawford, a parasite of agromyzid leafminers, were conducted.C. parksi successfully parasitized the following species in the genusLiriomyza Mik.,L. huidobrensis (Blanchard),L. sativae Blanchard,L. trifolii (Burgess), andL. trifoliearum Spencer. Successful parasite development was recorded from 8 plant families and 16 genera. Host plants which were suitable for leafminer development to the adult stage were also suitable for parasite development. Mean immature developmental period (egg to adult, usingL. trifolii as the host) at 21.1°, 26.7°, and 32.2° C was (for both sexes) 23, 14, and 14 days, respectively. Longevity of females provided only water was inversely related to temperature; significantly longer survival occurred at 21.1° C (5.0 days) compared to 26.7° C and 32.2° C (3.2 and 2.1 days, respectively). The addition of honey to the diet significantly improved longevity of both sexes at all temperatures. Adult female parasites which were provided an average of 33.0L. trifolii larvae per day produced an average of 135 offspring at a constant 26.7° C.C. parksi host-fed on ca. 3.7 leafminer larvae per day over an 11-day adult lifespan.      

Note sur la biologie deDiatraeophaga striatalis Townsend tachinaire parasite deProceras sacchariphagus Boj

Summary The author describes the life cycle ofDiatraeophaga striatalis, a tachinid parasite ofProceras sacchariphagus, under conditions of laboratory rearing. At 27°C and 80% relative humidity the average length of endoparasitic life and of nymphosis is of 10 days. Embryogenesis occurs in the uterus of the fertilized females which laid their first planidia at least 8 days after mating. At 18°C the endoparasitic life lasts from 20 days to 32 days. At 15°C the parasite larva stop developping and dies in the host; the nymph cannot tolerate this temperature for over 15 days. The oviposition process is inhibited at 15°C but females survive to this treatment for less than a week. Superparasitism does not involve the delayed development of larvae and has no effect on the size and the weight of the pupae obtained if the size of the host caterpillar is large enough and if the two parasites are not fixed on the same trachea.      

In vitro culture technique for Cryptocaryon irritans, a parasitic ciliate of marine teleosts

A medium for the in vitro culture of Cryptocaryon irritans, which is an obligatorily parasitic ciliate of marine teleosts and causes 'white spot disease', was developed. The medium consisted of a layer of cultured fish cells (FHM), with an agarose gel layer covering the cell layer. The agarose gel contained 0.22% agarose, 10% fetal calf serum, 100 I.U. ml(-1) Penicillin G potassium and 100 microg ml(-1) streptomycin sulphate. Theronts of C. irritans transformed to trophonts and grew to 180 microm in mean length in the medium, although they gradually decreased in number. When trophonts fully developed in medium were transferred into seawater 4 d after inoculation, approximately 70% of them transformed to encysted tomonts and released theronts. When fish were challenged with theronts obtained from in vitro-raised parasites, approximately 40% of the theronts were recovered from fish, indicating comparative infectivity of in vitro-raised theronts to those of in vivo-raised theronts. This is the first report that C. irritans fully developed in vitro and its entire life cycle was completed without a host fish.     

Salinitätstoleranz des parasitischen CiliatenIchthyophthirius multifiliis

Eels,Anguilla anguilla (L.), infested withIchthyophthirius multifiliis Fouquet, were exposed to different salinity levels (8, 15, and 30 ‰). Following exposure to 15° C for 2 to 4 weeks the parasite was able to survive on 20 cm long, young eels. This demonstrates thatI. multifiliis is not being damaged below the epidermis of its host; in form of free cysts or as swarmers, it is, however, no longer able to cause infection. In a second experiment, 2 eels of 40 cm length were infested withI. multifiliis at 15° C in 8 ‰ S; however, in 15 ‰ S, the parasite disappeared after 4 weeks.     

Comparison of serum antibody responses and host protection against parasite Ichthyophthirius multifiliis between channel catfish and channel × blue hybrid catfish

There is limited information available on the immune protection of channel catfish Ictalurus punctatus × blue catfish I. furcatus (CB) hybrid against the fish parasite Ichthyophthirius multifiliis (Ich). The objective of this study was to compare serum antibody response and host protection between channel catfish and CB hybrid catfish using a cohabitation model. Channel catfish and CB hybrid catfish were immunized with live theronts by immersion or by IP injection at the dose of 10,000–20,000 theronts per fish in two trials. The fish were then challenged with theronts to compare serum antibody response and protection against the parasite between channel catfish and CB hybrid catfish. The immunized channel catfish and CB hybrid catfish showed a significantly higher (p < 0.05) serum anti-Ich antibody (titer > 1120) compared to non-immunized controls (titer = 0). After being challenged with live theronts, the immunized channel catfish and CB hybrid catfish had none or a low number of the parasites (<50 trophonts per fish) and showed a significantly higher (p < 0.05) survival (90–100%) than non-immunized controls (0%). Overall results indicated that there was no statistical (p > 0.05) difference on serum anti-Ich antibody, parasite infection and fish survival between immunized channel catfish and CB hybrid catfish.     

Experimental warming drives a seasonal shift in the timing of host‐parasite dynamics with consequences for disease risk

Multi‐species experiments are critical for identifying the mechanisms through which climate change influences population dynamics and community interactions within ecological systems, including infectious diseases. Using a host–parasite system involving freshwater snails, amphibians and trematode parasites, we conducted a year‐long, outdoor experiment to evaluate how warming affected net parasite production, the timing of infection and the resultant pathology. Warming of 3 °C caused snail intermediate hosts to release parasites 9 months earlier and increased infected snail mortality by fourfold, leading to decreased overlap between amphibians and parasites. As a result, warming halved amphibian infection loads and reduced pathology by 67%, despite comparable total parasite production across temperature treatments. These results demonstrate that climate–disease theory should be expanded to account for predicted changes in host and parasite phenology, which may often be more important than changes in total parasite output for predicting climate‐driven changes in disease risk.     

The effect of host age ofLymantria dispar larvae [Lep.: Lymantriidae] on the development ofGlyptapanteles liparidis [Hym.: Braconidae]

The endoparasitic development ofG. liparidis was examined in 3 different host stages of gypsy moth larvae. Hatching ofG. liparidis-larvae occurred 3 to 5 days after oviposition in hosts parasitized during their premoulting period, and after 5 to 7 days in those parasitized in the 3^rd midinstar state. The parasites generally moulted to the 2^nd larval instar between the 11^th and 13^th day in the first group, and between the 13^th and 15^th day in the latter, when they had reached a volume of 0.04–0.05 mm³. The positive correlation between host ecdysis and the ecdysis of 1^st stadium larvae to L₂ suggested that host moulting influenced the development of the parasitoid larvae. Emergence from the host larvae occurred at 20°C after 27 days on average, and coincided with the parasites moulting to the 3^rd instar. Five to 7 days after spinning their cocoons near the developmentally arrested host larva, the male, and 1 to 2 days later the female wasps eclosed. Due to the variation in the number of parasites per host, no difference was observed between the hosts parasitized at various stages; however, a tendency for later parasitized hosts to contain more parasite larvae was evident. The nutritional conditions of the moth parental generation influenced both host and parasite development. On the other hand no influence of host age was observed on emergence dates of larvae and wasps.      

Diversity of parasites in Cichlasoma amazonarum Kullander, 1983 during rainy and dry seasons in eastern Amazon (Brazil)

This study investigated the influence of the rainy and dry seasons on the parasite communities of Cichlasoma amazonarum (Kullander, 1983) in a tributary of the Amazon River system, northern Brazil. Of 112 fish examined, 95.5% were parasitized by Ichthyophthirius multifiliis, Piscinoodinium pillulare, Gussevia disparoides, Posthodiplostomum sp., Procamallanus (Spirocamallanus) inopinatus, and Echinorhynchus paranensis, as well as by the Glossiphoniidae leech. Ichthyophthirius multifiliis and P. pillulare were the dominant parasite species, with I. multifiliis the most prevalent and abundant; the leech (Glossiphoniidae) was the least prevalent and least abundant. Parasites presented an aggregate dispersion pattern and seasonal variations in infestation levels, influenced by the environmental conditions in the rainy season. Variation in the parasite dynamics created overall changes in the parasite communities, characterized by greater diversity, species richness and evenness during the rainy season. However, there was a high similarity (99.8%) in the parasite community structure between the rainy and dry seasons. During the dry season the hosts had predominantly 1–3 parasite species compared to 3–4 parasites in the rainy season. Some parasites in the eastern Amazon undergo population changes relating to seasonality. These results thus indicate the correct season to apply adequate prophylactic measures to reduce negative impacts of parasites in this wild ornamental fish when captured.     

Behavior,net reproduction,longevity, and mummy-stage survival ofAphidius matricariae [Hym. Aphidiidae]

The courtship, mating and ovipositional behavior ofA. matricariae Haliday were studied. UsingMyzus persicae (Sulzer) as the host, the production of progeny per female parasite and survival from mummy stage to the adult were studied at constant temperatures of 10°, 12.8°, 15.6°, 18.3°, 21°, 24°, 26.7°, 29.5° and 32°C. The longevity of male and female parasites was determined at temperatures of 7°, 10°, 15.6°, 21°, 26.7°, 29.5° and 32°C. The greatest number of progeny (392) was produced at 21°C. The optimal temperatures for production of progeny and survival of the parasites during the mummy stage were from 12.8°C to 21°C. The longevity of male and female adult parasites decreased as temperatures increased and male parasites lived significantly (P<0.05) longer than females at 10° and 15.6°C.