In a recent paper1, written jointly with Dr P. P. Laidlaw, methods were described for cultivating Entamoeba histolytica, and various other entozoic amoebae of man and monkeys, from the encysted forms. We there showed that cultures of all the species studied can be readily obtained by merely incubating such cysts in any of the media suitable for the cultivation of the corresponding active forms—provided that certain precautions are taken.

The genus Monospora (= Monosporella renamed) was founded by Metschnikoff in 1884 to designate a parasitic fungus which he discovered in Daphnia magna. This monocellular fungus, of which he described only one species, Monospora bicuspidata, lives free in the body cavity of its host, where it multiplies actively by budding in a yeast-like manner (Fig. I, 1, 2, 3).

One therapeutic oral dose (400 mg/kg) of 153C51 administered to infested mice caused pathological changes in the dorsal region of the tegument of male Schistosoma mansoni during the period 3–24 h after treatment. These changes occurred prior to the ‘hepatic shift’. At the ultrastructural level they consisted of a gradual accumulation in the tegument epidermis of numerous membranous inclusions with the characteristics of residual lysosomes and changes in the localization of acid phosphatase, a lysosomal enzyme. It seemed likely that these changes were due to inhibition or exhaustion of enzyme in the epidermis, followed by re-synthesis of enzyme in the cell bodies and its export to the epidermis. The elimination of hydrolytic activity from lysosomes in the epidermis would explain the accumulation of residual lysosomes. Drug- treated parasites retained their disguise of host red blood cell ghost antigens as shown by indirect fluorescent antibody-labelling and, therefore, it seemed unlikely that immunological factors could be important in producing the tegument pathology.

1. The development of resistance to metachloridine (3-metanilamido-5-chloropyrimidine) was studied in two clones of Plasmodium gallinaceum derived from single erythrocytic parasites and maintained by serial inoculation in young chicks. Resistance developed with equal facility and similarly in the two clones.

2. In thirteen strains of these clones an enhancement of resistance was obtained after three to five courses of treatment, each of seven doses of metachloridine over a period of 3½ days.

3. The effect of the size of the inoculum (2·5 × 107–109 parasites), and of the dose of drug upon the rate of development of resistance was studied. Although the number of courses of treatment required to produce an enhancement of resistance was not always related to the size of the inoculum, with the largest inoculum an enhancement of resistance was observed after the minimum number of courses of treatment (three), whereas with the smallest inoculum no enhancement of resistance was obtained. The rate of the development of resistance did not appear to be related to the size of the dose of drug.

4. In some strains the increase in resistance was sudden, whereas in others it was more gradual. Resistance was retained when parasites of a newly resistant strain were transmitted through Aëdes aegypti or maintained for 43 days in the absence of the drug.

5. A comparison of the development of resistance in populations of normal parasites, and of populations composed of mixtures of known numbers of resistant and normal parasites, indicated that the pattern of the development of resistance in normal populations could be explained by the selections of mutations of a frequency of less than 1 in 5 × 107, or probably less than 1 in 109 parasites.

1. The life cycle of Gigantobilharzia huronensis Najim, 1950, was determined experimentally. The snail, Physa gyrina (Say), served as the intermediate host. The natural definitive hosts were goldfinches (Spinus tristis tristis) and the cardinals (Richmondena cardinalis). Chicks and canaries served as susceptible hosts in the laboratory

2. The shortest time recorded for the hatching of eggs in water is about 20 min. Cercariae were liberated from the snail as early as 24 days after exposure. Eggs were found in the faeces of the definitive host 31 days after the first exposure to the cercariae.

3. Adult worms are elongate and filiform. Females are longer than males, and in both sexes the oral and ventral suckers are absent. The cuticle has no spines. A short gynaecophoric canal is present. Up to 300 testes are present in the male and the cirrus is spiny. Laurer's canal is present and vitelline follicles fill most of the body of the female. Eggs are spherical to very broadly ovoid in shape and averaged 93µ in length by 88µ in width. The uterus contains from one to several eggs at a time, and the genital pore is located shortly posterior to the mouth opening.

4. The miracidium has a peculiar wide gap between the second and third rows of epidermal plates. The body covering consists of twenty-two epidermal plates arranged in four rows of 6:9:4:3. The mother sporocyst has an elongate, sac-like body. A birth pore is present in the mature mother sporocysts. The daughter sporocyst also has an elongate body, but the anterior end is covered with spines. A birth pore is present in the mature daughter sporocysts.

5. The cercaria is an apharyngeal, furcocercous, brevifurcate, distome with pigmented eyespots; the body averaging 240µ in length by 72·5µ in width; the tail stem averaging 268·5µ in length by 30µ, in width; the furca averaging 146·8µ in length by 15µ in width. The furcae have fin folds. The body and tail are covered with spines. There are five pairs of penetration glands and an oral gland. The excretory system is usually 2[(2) + (2+1)]. The cercaria attaches to the surface film of the water with the body parallel to it and the tail hanging downward at different angles. The cercaria is a dermatitis producer in human skin.

A strain of Trypanosoma avium isolated from Corvus frugilegus in England was readily cultivated in vitro in diphasic blood agar, or blood nutrient agar, media at 28 °C.

In both media, small trypanosomes appeared which morphologically resembled the metacyclic forms from the insect vector and which were infective to canaries. They were more numerous, and developed more regularly, in the medium with the nutrient agar base.

At 40–41 °C a few of these small trypanosomes apparently developed into large trypanosomes similar to, but not identical with, those seen in the blood of infected birds.

Experiments were performed to determine at what phase or phases of its parasitic development Boophilus microplus becomes infected with Babesia bigemina. Problems due to Boophilus microplus being a one-host tick were overcome by artificially transferring ticks from one host to a second just prior to a moult. In several experiments, a babesiacide was used to eliminate parasites so that the exposure of ticks to infection could be restricted to one phase of their development. Further information was obtained by correlating the presence or absence of patent parasitaemia in the bovine with the presence or absence of infection in ticks which fed on it.

It was found that ticks were susceptible to infection for approximately the last day of their parasitic life. Infection at other times, that is, during larval, nymphal and the major part of the adult stages was not usual. Four of five attempts to infect larvae and all five attempts to infect nymphs were negative.

Infection of ticks always resulted when a patent parasitaemia coincided with the last day of parasitic life. Ticks were infected in all thirteen instances when these conditions existed but in only seven of eighteen instances when the animal was a carrier but was not showing parasitaemia.

Stage-to-stage transmission did not occur, although suitable conditions for this existed on nine occasions.

The capsule, which surrounds the acanthocephalan parasite Moniliformis dubius in its intermediate host, the cockroach, Periplaneta americana, has been studied in the electron microscope. Some observations were also made with the light microscope.

The capsule surrounding the developing parasite (acanthella) taken from the haemocoel of its host consists largely of closely packed vesicles, which are empty of electron-dense contents.

The vesicles arise from the surface membrane of the insect's haemocytes, which surround the parasite. Long tubular protrusions arise from the haemocyte's surface and, on separating, swell to form rounded vesicles.

The capsule, which surrounds the later resting stage (cystacanth), has apparently been stretched by the growth of the parasite after haemocytic activity has declined. An amorphous, electron–dense component is also evident at this stage.

We are grateful to Mrs A. Baker and Miss R. Jantunen for their technical assistance and to Dr A. C. S. Crossley for his comments on the work.

The transmission of Theileria parva by nymphal and adult Rhipicephalus appendiculatus was compared by the assessment of salivary gland infections in tick batches fed on the same group of infected cattle at the same time. When larval and nymphal R. appendiculatus Muguga ticks were fed concurrently on cattle undergoing acute infection with T. parva Muguga, the resultant nymphae developed a slightly lower prevalence of infection than did the adult ticks. The abundance of infection was 5–20 times higher in the adult ticks than in the nymphae. When larval and nymphal R. appendiculatus Muguga and R. appendiculatus McIlwaine were fed to repletion on cattle infected with T. parva Boleni, a parasite causing subacute infection, resultant adult tick batches had a relatively high prevalence of infection, but infection was not detected in resultant nymphal batches. When cattle that were carriers of 2 stocks of T. parva, Marikebuni and Kiambu 5, were used as the source of infection, the infections developing in adult R. appendiculatus Muguga ticks were much higher than those developing in nymphae. The structure of salivary glands differed between nymphal ticks, adult males and adult females, and this is considered to be an important factor affecting the infection levels. The morphology of the type III acini, the target acini for sporogony, was similar, but the mean numbers of type III acini were different, with 87 in nymphae, 1346 in males and 1736 in females. This difference was correlated with the different infection levels produced in the various instars and sexes. While the process of sporogony in the different tick instars and sexes was similar, the rate of sporogony was fastest in feeding nymphae, taking on average 2–3 days, compared to 3–4 days in females and an irregular period in the males. These results are discussed in relation to the epidemiology of T. parva.

An account is given of the bacterial flora isolated from the blood-sucking Reduviid bugs, Triatoma infestans and T. protracta, and the predatory Reduviid, Sinea diadema. The micro-organisms which have been found in Triatoma spp. differ from that found in Rhodnius prolixus (Actinomyces rhodnii), in both specific identity and in their relation to the cells of the mid-gut. The symbiont occurring most frequently in T. infestans has been identified as a diphtheroid organism resembling Coryne-bacterium hofmannii, and sections show this organism inside vacuoles within some of the endothelial cells of the insect's intestine. The organism occurring in R. prolixus, on the other hand, cannot be found in this situation, but remains in the lumen of the gut. While the occurrence of the bacteria found in T. infestans in so intimate a connexion with the cells of its host seems to suggest a more highly organized state of affairs, T. infestans does not show the fixed association with one species of micro-organism which is shown by R. prolixus. Other bacteria occur commonly in T. infestans, although they do not occur in cell-vacuoles; and the range of bacterial types is so great that it seems unlikely that the species recorded in this paper are the only ones that may occur. The presence of these other bacteria may be due either to the anti-biotic action of the Corynebacterium sp. being weaker than that of A. rhodnii, or to masking of the anti-biotic action by the cell wall that encloses the organisms. The situation in R. prolixus shows clearly that a constant symbiotic association with a micro-organism can be maintained without the presence of specialized organs of the mycetome type.

With regard to the predatory species, no general conclusion can be reached on the evidence so far available. Compared with the blood-sucking Reduviids, insects of this kind usually have powerful proteolytic enzymes secreted in the anterior mid-gut, while the same region of the blood-suckers is a simple storage and concentration sac, digestion taking place lower down the intestine. Thus the region of the gut where bacteria are found in blood-suckers might not be a favourable situation for them in predatory bugs. Hence it is perhaps significant that most of the few successful isolations from a predatory type were from the rectum of the insect. It is the usual opinion that insects with a predatory diet do not need bacterial symbionts, but the aim in examining the predatory Reduviidae has been to find out whether they show a condition which might have evolved into that found in their blood-sucking relatives. The search is still in progress for a rather unspecialized type of predatory Reduviid in which the salivary and anterior mid-gut digestion is less strongly developed than in those species which have been available for study up to the present.

On the available data from south Wales and southeast England, the tick Ixodes hexagonus Leach has spring and autumn peaks of activity on hedgehogs, stoats and weasels.

One or more stages of this tick have been recorded on fifteen mammalian hosts. All stages have been collected from hedgehogs, stoats, foxes, polecats and ferrets. The weasel is probably the smallest habitual host. No specimens of I. hexagonus have been found on birds.

Man is an ‘accidental’ host and records of this tick on man in the London area are given. From the present information it seems that ill-effects following the bite of this parasite are rare. In built-up areas the probable sources of infestation are dogs, cats and hedgehogs. Air-raid shelters and similar constructions which have harboured these hosts at one time or another may provide a centre of infestation of man. Our observations indicate that the number of ticks in most hedgehogs' nests is not large. Nothing is known of the tick population in the habitats of other hosts. It is suggested tentatively that the ticks may drop off hedgehogs away from the nest and that after further development (i.e. hatching of eggs and metamorphoses of later stages) they are picked up by such animals as foxes, stoats, weasels.

The records of this tick in Britain are summarized and they indicate that I. hexagonus is a southern species.

In the first part of this paper fifteen species were described, eight of which were from birds. In the present part an additional fifteen are dealt with.

Using biochemical genetic methods, we have distinguished 2 sibling species in the complex Bothrimonus nylandicus (Schneider, 1902), which infest 2 congeneric species of sole (Solea lascaris and Solea impar) on European coasts (Atlantic and Mediterranean). Neither of the parasite species is specific for either of the sole species, but one of them is present all year round, whereas the other is absent in the autumn and winter and only appears in the spring, subsequently disappearing at the end of the summer. Only S. impar lives in the Mediterranean, and is equally infested by both cestodes, whereas both species occur in the Atlantic and each of them is preferentially infested by 1 species of cestode. The shortness of the adult stage of the parasite in the definitive host and the presence of 2 life-cycles associated with competition between the 2 hosts in the Atlantic could be responsible for the biological differences observed and for maintaining the sibling species in sympatry.

Morphological features of the bladder wall of Cysticercus fasciolaris and carbohydrate analysis of the contained fluid are described. The bladder fluid was found to have a higher concentration of glucose than the plasma of the respective host animal. The bladder wall has a prominent duct system with numerous cytoplasmic extensions and interconnections. It is suggested that the bladder may function in the nutrition of the developing cysticercus by providing a reservoir of raw material which may be utilized by the growing strobila.

The authors wish to thank Dr Theodor von Brand for many helpful discussions, and Dr Harley G. Sheffield and Mr Ned M. Etherington for their advice and technical assistance in the preparation of the electron micrographs.

The diversity and prevalence of malaria parasites of the genera Plasmodium and Haemoproteus were determined in the globally-threatened Aquatic Warbler Acrocephalus paludicola. Birds were sampled during migration in Portugal and at the wintering quarters in Senegal and parasites were detected using molecular methods. Only three generalist parasite lineages (Plasmodium) were found. There were no significant differences in the prevalence of parasites between sexes in Europe, but adults had higher prevalence than first-year birds, and birds in Europe had higher prevalence than those captured in Africa. When comparing with other Acrocephalus species and taking sample size into account, Aquatic Warblers had the lowest prevalence and, together with another threatened species, the Seychelles Warbler Acrocephalus sechellensis, the lowest diversity of malaria parasites. We hypothesize that the low diversity of parasites and absence of specialist lineages of Aquatic Warblers are caused by its small population size and fragmented distribution. Furthermore, Aquatic Warblers’ extreme habitat specialization may decrease their exposure to malaria parasites, but other explanations such as high mortality (which would constraint the sampling of infected birds) or, in contrast, very efficient immunological system in clearing the infections cannot be ruled out. This study contributes to explain variation in prevalence and diversity of malaria parasites among hosts.

We have identified and partially sequenced 8 ABC transporters from an EST dataset of Sarcoptes scabiei var. hominis, the causative agent of scabies. Analysis confirmed that most of the known ABC subfamilies are represented in the EST dataset including several members of the multidrug resistance protein subfamily (ABC-C). Although P-glycoprotein (ABC-B) sequences were not found in the EST dataset, a partial P-glycoprotein sequence was subsequently obtained using a degenerate PCR strategy and library screening. Thus a total of 9 potential S. scabiei ABC transporters representing the subfamilies A, B, C, E, F and H have been identified. Ivermectin is currently used in the treatment of hyper-infested (crusted) scabies, and has also been identified as a potentially effective acaricide for mass treatment programmes in scabies-endemic communities. The observation of clinical and in vitro ivermectin resistance in 2 crusted scabies patients who received multiple treatments has raised serious concerns regarding the sustainability of such programmes. One possible mechanism for ivermectin resistance is through ABC transporters such as P-glycoprotein. This work forms an important foundation for further studies to elucidate the potential role of ABC transporters in ivermectin resistance of S. scabiei.

The benzodiazepine Ro 11-3128 (methyl-clonazepam) presents several similarities with praziquantel with regard to its anti-schistosomal mode of action, since both drugs cause spastic paralysis, calcium influx and tegumental disruption in the parasites. In order to know whether the two compounds share the same binding sites in the schistosomes, we performed in vivo and in vitro competition experiments. We took advantage of the fact that Ro 11-3128 is active against immature Schistosoma mansoni (whereas praziquantel is inactive), and praziquantel is active against S. japonicum (which is insensitive to Ro 11-3128). An excess of praziquantel did not inhibit the activity of Ro 11-3128 against immature S. mansoni and an excess of Ro 11-3128 did not inhibit the activity of praziquantel against S. japonicum, suggesting that the schistosome binding sites of the two drugs are different. On the other hand, cytochalasin D, an agent known to perturb – among other things – calcium channel function, was capable of inhibiting the schistosomicidal activity of both praziquantel and Ro 11-3128, thus adding another element of similarity between the two anti-schistosomal agents. A similar, albeit partial, inhibition of the schistosomicidal activity of the two drugs was exerted by some of the classical calcium channel blockers. Taken together, these results suggest that praziquantel and Ro 11-3128, although binding to different schistosome receptor sites, may use the same basic anti-schistosomal effector mechanisms.

The literature contains many examples of changes induced by parasites in the behaviour and/or other phenotypic traits of insects. From an evolutionary perspective, the nature of these changes is usually difficult to assess. Parasite-induced changes in host behaviour can be adaptations of either host or parasite, or they can be mere pathological consequences of infection. Of the many criteria and experimental tests necessary to distinguish between adaptations and non-selected consequences, two are particularly important: the demonstration of fitness benefits for either host or parasite associated with the behavioural change, and the elucidation of the proximate mechanism responsible for the behavioural change. Another approach can serve to identify adaptive changes in behaviour: mapping specific behavioural alterations on a phylogeny of either hosts or parasites. The usefulness of this approach is illustrated with two examples, acanthocephalan- cockroach associations and insect-fungus associations. The adaptive nature of parasite-induced behavioural changes will always be difficult to evaluate because they are the product of two distinct but interacting genotypes. However, experimental and phylogenetic approaches can provide valuable insights into the evolutionary history of insect-parasite interactions.

Toxoplasma gondii and Sarcocystis neurona are protozoan parasites with terrestrial definitive hosts, and both pathogens can cause fatal disease in a wide range of marine animals. Close monitoring of threatened southern sea otters (Enhydra lutris nereis) in California allowed for the diagnosis of dual transplacental transmission of T. gondii and S. neurona in a wild female otter that was chronically infected with both parasites. Congenital infection resulted in late-term abortion due to disseminated toxoplasmosis. Toxoplasma gondii and S. neurona DNA was amplified from placental tissue culture, as well as from fetal lung tissue. Molecular characterization of T. gondii revealed a Type X genotype in isolates derived from placenta and fetal brain, as well as in all tested fetal organs (brain, lung, spleen, liver and thymus). This report provides the first evidence for transplacental transmission of T. gondii in a chronically infected wild sea otter, and the first molecular and immunohistochemical confirmation of concurrent transplacental transmission of T. gondii and S. neurona in any species. Repeated fetal and/or neonatal losses in the sea otter dam also suggested that T. gondii has the potential to reduce fecundity in chronically infected marine mammals through parasite recrudescence and repeated fetal infection.

Psoroptic sheep scab and psoroptic otoacariasis of domestic rabbits occur in the same geographical regions of Switzerland. To address the question as to whether Psoroptes mites are naturally transmitted between sheep and rabbits, we determined the sequences of the rDNA second internal transcribed spacer (ITS-2) from psoroptic mites from sheep (5 field isolates from Switzerland and 2 laboratory isolates from Ireland and the UK) and from rabbits (8 field isolates from Switzerland). The ITS-2 sequences for all Psoroptes mites originating from sheep were identical and differed at 1 nucleotide position from all the sequences of the rabbit-derived isolates. A statistically significant difference between a rabbit-derived isolate and isolates originating from sheep was also obtained by morphometric analysis of the lengths of the outer opisthosomal setae. For comparative purposes, the ITS-2 sequences from Chorioptes and Sarcoptes collected in Switzerland were also determined. No intraspecies variation was found in 6 sarcoptic isolates from red foxes, with a sequence identity of 41% as compared to Psoroptes. The ITS-2 sequences of 3 chorioptic isolates differed by 24–29% from the Psoroptes sequence. Identical sequences were found for the Chorioptes isolates from sheep and a camel, which differed by 18% from the sequence of an isolate from a cow. These genetic data of psoroptic mites originating from sheep and rabbits from the same geographical area suggest the existence of epidemiologically separated populations.