The whole life cycle of Paramphistomum microbothrium Fischoeder, 1901, found in cattle of Kenya, East Africa, has been established experimentally.

In laboratory conditions, the eggs hatched miracidia on the 14th to 16th day, if they were kept in water at a temperature of 26–28° C.

In a snail, Bulinus alluaudi (Dautzenberg), kept at the temperature of 18–20° C., the miracidium developed into a sporocyst, the elongated body of which, containing young rediae, reached a length of 3·6 mm. in about 2 weeks.

The first rediae which emerged from a sporocyst were observed on the 14th day, and on the 20th day the first generation rediae began to produce second-generation rediae. From the 28th day onward the first-generation parent rediae ceased to produce daughter rediae and began to develop cercariae only. This period of production of cercariae by the redia lasted about 30 days, and when the life of the first-generation rediae drew to its close, the old rediae developed a few daughter rediae again.

Cercariae began to emerge from the first-generation rediae 30 days after exposure of the snail to miracidia. The cercariae left the parent redia in a very immature state and further development occurred in the liver of the snail. The emergence of cercariae from the infected snail began on the 43rd day after exposure to miracidia. Shortly after emerging from the snail, the cercariae attached themselves to vegetation and encysted.

Development of rediae of the second, third, fourth and apparently more successive generations followed an identical course to that outlined for the rediae of the first generation. As a result the successive generations of rediae maintain the infection going in an intermediate host and the infected snails were continually shedding cercariae as long as they lived. In the laboratory experiments the life span of some of the infected snails exceeded a year.

In cattle infected experimentally P. microbothrium reached maturity and began passing out the eggs about a 100 days after the encysted cercariae were fed to the animals.

The distribution of carbohydrates, lipids and enzymes in the daughter sporocyst and contained cercariae of Cercaria bucephalopsis haimaena Lacaze-Duthiers, 1854, is mapped and used as an indication of metabolism. It is concluded that host glucose, glycogen, fatty acids and neutral lipids are absorbed and pass through the syncytial tegument into the cellular sub-tegument. They are then either metabolized or passed into the body cavity and contained cercariae. Glucose seems to be either converted into glycogen and glycoproteins or metabolized anaerobically. The end products of anaerobic respiration are fatty acids and neutral lipids. These pass into the excretory system, which flushes them into the body cavity of the daughter sporocyst, from where they enter the body of the developing cercariae. In the cercariae they are deposited, together with host lipids, possibly as less toxic higher fatty acids or other stored lipids. The stored glycogen and lipids in the cercariae are probably utilized aerobically during the free-living existence. The transport of nutrients and excretory products across the cell membranes may be aided by alkaline phosphatase and esterase activity, as is the metabolism of carbohydrates and lipids. Acid mucopolysaccharide precursors appear to be synthesized in the dense cytoplasm of the mid-tegument of the daughter sporocyst and secreted into the host's visceral haemocoel. Acid phosphatase and esterase found in this region may be involved in the synthesis and also in the breakdown of complex incoming nutrients.

We are grateful to Professor E. W. Knight-Jones for laboratory facilities and to the Science Research Council for a grant to one of us (E. A. B.) which made the work possible.

Malaria and filaria infection rates were determined for anopheline mosquitoes collected whilst biting and resting in village houses in Papua New Guinea. The number of anophelines infected with both parasites was greater than expected from the infection rates of each parasite and this difference was significant in resting collections. The excess of multiply infected mosquitoes is probably a result of a vector population composed of individuals with differing numbers of opportunities to become infected. Malaria-positive, Anopheles punctulatus from resting catches had a significantly greater number of Stage 3 Wuchereria bancroftilarvae than malaria-negative mosquitoes. However, multiply infected mosquitoes appear to suffer greater mortality than non-infected or singly infected mosquitoes when the filarial worm reaches the third stage. Any potential increase in transmission resulting from multiple infections is thereby offset by a greater mortality rate in these mosquitoes.

Viable eggs produced weekly per infective stage was used as a measure of the reproductive success of Echinostoma friedi during the first 12 weeks of infection in hamsters. The weekly reproductive success was not constant during the experiment in relation to the egg output and the proportion of viable eggs produced. The egg release started during week 2 post-inoculation, attaining a maximum during week 3. A decline in egg output was observed from week 9. Viable eggs were only produced from week 3 post-inoculation and a maximum was attained at week 4 of the experiment. A decline in egg viability was observed from week 9. Considering together the egg output and the egg viability, the maximum weekly reproductive success was obtained during week 4 post-inoculation. The changes in the weekly reproductive success were not reflected in variations in worm numbers and body sizes during the course of the infection. The humoral immune response of golden hamsters during the infection with E. friedi was determined. Increases of IgG levels against somatic and excretory/secretory products of E. friedi were detected coinciding with the reduction in the reproductive success.

Larvae of the monogeneans Pseudodiplorchis americanus and Neodiplorchis scaphiopodis develop within egg capsules retained in the uterus of the parent parasite. This study reveals adaptations for the storage and nutrition of infective stages which have no known precedent amongst other platyhelminths. The vitelline system is greatly reduced and appears to contribute little to embryo nutrition. An electron-dense ‘shell’ which encloses the newly formed embryos is subsequently replaced by a thin flexible sac composed of concentric layers of membranes which are derived from the uterus wall. Further membranes are added to this multi-laminate structure as the encapsulated embryos pass along the uterus, and this enables the sac to expand continuously as the larva inside it grows. The capsule lining extends into fine cytoplasmic processes which ‘plumb in’ to the larval tegument between ciliated cells. These connexions are packed with glycogen and appear to perform a placenta-like function. The unique adaptations correlate with the demands of the life-cycle. Large numbers of oncomiracidia (up to 300) can be packed, in membranous sacs, within the storage capacity of the uterus; direct transfer of nutrients from parent to offspring enables resources to be supplied over an extended period; and infective larvae can be maintained in readiness for an unpredictable opportunity for transmission.

A striking feature of toxoplasmic seroconversion is the prominent and early IgM response to a low molecular weight antigen of 4–5 kDa. Two different monoclonal antibodies directed against the 4–5 kDa antigen have been generated and used to characterize this molecule. Using these monoclonal antibodies, we could demonstrate the surface localization of the low Mr antigen by immunofluorescence and immuno-electron microscopy assays. By immunoblotting, we observed that one of the monoclonal antibodies was unable to recognize the 4–5 kDa antigen in tachyzoites propagated in cell culture, indicating an epitope variability between Toxoplasma gondii tachyzoites grown in vivo and in vitro. We discuss the implications of this latter finding in the design of diagnostic reagents.

Administration of the immunostimulants Corynebacterium parvum, Bacillus Calmette-Guérin (BCG) or Bordetella pertussis prior to, or at the same time as, challenge with Trypanosoma congolense significantly increased survival times in mice, both of trypano-susceptible (A/J) and trypano-resistant (C57Bl) strains. The increased survival time was associated with significant alterations in parasitaemia, which included lengthening of the pre-patent period, a delay in the time taken to reach the first peak of parasitaemia and a reduction in the level of parasitaemia. Similar results were obtained when these strains of mice were challenged with Trypanosoma brucei following pre-treatment with C. parvum. Thus, by the use of immunostimulants it was possible to reduce the susceptibility of mice to trypanosomiasis and the hope is that this can also be achieved with domestic livestock.

The global increase of the human parasite, the common bed bug Cimex lectularius, calls for specific pest control target sites. The bed bug is also a model species for sexual conflict theory which suggests that seminal fluids may be highly diverse. The species has a highly unusual sperm biology and seminal proteins may have unique functions. One-dimensional PAGE gels showed 40–50% band sharing between C. lectularius and another cimicid species, Afrocimex constrictus. However, adult, sexually rested C. lectularius males were found to store 5–7 μg of seminal protein and with only 60 μg of protein we obtained informative 2-D PAGE gels. These showed 79% shared protein spots between 2 laboratory populations, and more than half of the shared protein spots were detected in the mated female. Further analysis using liquid chromatography electrospray ionization tandem mass spectrometry revealed that 26·5% of the proteins had matches among arthropods in databases and 14·5% matched Drosophila proteins. These included ubiquitous proteins but also those more closely associated with reproduction such as moj 29, ubiquitin, the stress-related elongation factor EF-1alpha, a protein disulfide isomerase and an antioxidant, Peroxiredoxin 6.

The oyster Crassostrea gasar is a species widely used as food and a source of income for the local population of the estuaries of Northeast Brazil. Perkinsus marinus and Perkinsus olseni are deleterious parasites for oyster farming and were recently detected in Brazil. In this study, a histopathologic survey of the oyster C. gasar cultured in the estuary of the River Mamanguape (Paraíba State) was performed. Adult oysters were collected in December 2011 and March, May, August and October 2012 and processed for histology and Perkinsus sp. identification by molecular analyses. Histopathological analysis revealed the presence of parasitic organisms including viral gametocytic hypertrophy, prokaryote-like colonies, protozoans (Perkinsus sp. and Nematopsis sp.) and metazoans (Tylocephalum sp. and cestodes). Other commensal organisms were also detected (the protozoan Ancistrocoma sp. and the turbellarian Urastoma sp.). The protozoan parasite Perkinsus sp. had the highest overall prevalence among the symbiotic organisms studied (48·9%), followed by Nematopsis sp. (36·3%). The other organisms were only sporadically observed. Only the protozoan Perkinsus sp. caused alterations in the oysters’ infected organs. Molecular analyses confirmed the presence of P. marinus, P. olseni and Perkinsus beihaiensis infecting the oyster C. gasar. This is the first report of P. beihaiensis in this oyster species.

Notoedric mange, caused by obligately parasitic sarcoptiform Notoedres mites, is associated with potentially fatal dermatitis with secondary systemic disease in small mammals, felids and procyonids among others, as well as an occasional zoonosis. We describe clinical spectra in non-chiropteran hosts, review risk factors and summarize ecological and epidemiological studies. The genus is disproportionately represented on rodents. Disease in felids and procyonids ranges from very mild to death. Knowledge of the geographical distribution of the mites is highly inadequate, with focal hot spots known for Notoedres cati in domestic cats and bobcats. Predisposing genetic and immunological factors are not known, except that co-infection with other parasites and anticoagulant rodenticide toxicoses may contribute to severe disease. Treatment of individual animals is typically successful with macrocytic lactones such as selamectin, but herd or wildlife population treatment has not been undertaken. Transmission requires close contact and typically is within a host species. Notoedric mange can kill half all individuals in a population and regulate host population below non-diseased density for decades, consistent with frequency-dependent transmission or spillover from other hosts. Epidemics are increasingly identified in various hosts, suggesting global change in suitable environmental conditions or increased reporting bias.

Understanding the frequency distribution of parasites and parasite stages among hosts is essential for efficient experimental design and statistical analysis, and is also required for the development of sustainable methods of controlling infection. Nematodirus battus is one of the most important organisms that infect sheep but the distribution of parasites among hosts is unknown. An initial analysis indicated a high frequency of animals without N. battus and with zero egg counts, suggesting the possibility of a zero-inflated distribution. We developed a Bayesian analysis using Markov chain Monte Carlo methods to estimate the parameters of the zero-inflated negative binomial distribution. The analysis of 3000 simulated data sets indicated that this method out-performed the maximum likelihood procedure. Application of this technique to faecal egg counts from lambs in a commercial upland flock indicated that N. battus counts were indeed zero-inflated. Estimating the extent of zero-inflation is important for effective statistical analysis and for the accurate identification of genetically resistant animals.

Pasture plots in a warm coastal region were separately contaminated in the autumn, winter and spring with eggs of a coastal strain of Ostertagia ostertagi derived from dairy calves. At the same times, a parallel set of plots was contaminated by a strain transferred from within a beef cattle management system in a cooler tablelands region. A reciprocal transfer of the coastal strain was also made to the tablelands, where pasture plots were contaminated with eggs of both strains of O. ostertagi in winter. Successive groups of parasite-free calves were grazed on the plots for 10–14 days at 4-week intervals and then killed for worm counts 14 days after their removal from pasture. In both environments, irrespective of the season of pasture contamination, inhibition of development reached a maximum in spring and the two strains differed only in the proportion of the population inhibited, which was significantly greater for the table- lands strain. There was no strain difference for either numbers of infective larvae on pasture or total O. ostertagi numbers. This suggested that the difference between coastal and tablelands populations in the extent of inhibition is genetically determined. If an environmental stimulus was implicated in the spring occurrence of inhibition, the stimulus was provided equally in coastal and tablelands environments and it cannot be exposure to low temperatures.

A simple and robust method for the isolation of gastro-intestinal nematode eggs from faeces is described that uses both salt- and sugar solutions for flotation. Application of this ‘salt–sugar’; isolation method to large numbers of faecal samples of adult dairy cows indicates a 3- or 4-fold reduction in the proportion of e.p.g.-negative cows relative to studies that used other techniques for egg isolation. The procedure detects more eggs than the Wisconsin flotation method in replicate samples and in spiked egg-free faeces. The number of recovered eggs in spiked faecal samples is linear over a range of egg concentrations, and the transparent faecal preparations that result from the protocol can be stored as digital images which can be used as input for an efficient automated egg-counting procedure. The increased rate of processing of faeces combined with the large reduction of the percentage of e.p.g.-negative cows allows more accurate analysis of large numbers of adult or resistant animals for studies of nematode parasitism.

An AFLP approach was established to investigate genetic diversity within Oesophagostomum bifurcum (order Strongylida) from human and non-human primates. Evaluation of different combinations of restriction enzymes (n=8) and primers (n=29) demonstrated that the use of HindIII/BglII digested templates and primers with the selective nucleotides +AG/+AC, respectively, was the most effective for the analysis of O. bifurcum DNA. A total of 63 O. bifurcum adults from human, Patas monkey, Mona monkey and Olive baboon hosts from different geographical regions in Ghana were subjected to analysis using this method. Cluster analysis revealed 4 genetically distinct groups, namely O. bifurcum from the Patas monkey (I), from the Mona monkey (II), from humans (III) and from the Olive baboon (IV). These findings were concordant with those achieved previously using RAPD analysis and supports population genetic substructuring within O. bifurcum according to host species. The results demonstrated the effectiveness of the present AFLP method for establishing genetic variation within O. bifurcum, and indicates its applicability to other parasitic nematodes of human and/or veterinary health importance.

The variability inherent in different isolates of Fasciola hepatica has been evident from reports in the literature but to date there has been no systematic examination of the relationship between these differences and the fecundity of the parasite. In this study we have attempted to remedy this situation by comparing the relative efficiencies with which 2 well-characterized isolates of the liver fluke (Oberon and Fairhurst) progress through both their definitive and intermediate hosts. We did not observe a reduction in fitness in the Oberon isolate which has been reported to be triclabendazole-resistant, compared to the triclabendazole-susceptible Fairhurst isolate, but considerable inter- and intra-isolate variability at different life-cycle stages was recorded. Thus the Oberon isolate gave 4-fold the number of cercariae when 100 snails were each challenged with a single miracidium and was more successful in establishing productive infections in rats. Fairhurst metacercariae excysted at a higher rate than those from the Oberon isolate and Fairhurst flukes produced 4-fold more eggs. The extent of the intra- and inter-isolate variability revealed in this work will provide a basis for the development of models of population dynamics aimed at predicting the response of the liver fluke to changing environmental conditions such as the use of anthelmintics or climatic change.

Although Phthirus pubis occurs in Europe, Asia, Africa, North America and Australia, and is found on negroes as well as whites, as herein recorded, further data are required relating to the geographical distribution of the species and the races of man it infests.

The crab-louse occurs less frequently on man than does Pediculus and appears to be parasitic chiefly on persons leading an active sexual life. It has, so far, only been twice recorded on another host than man, i.e. the dog.

Whilst the crab-louse is usually found confined to the pubic and perianal region, it frequently spreads upward upon the abdomen and breast, and may infest the axillae severely, or it may spread downward along the thighs. Generalized infestation is rare, in such cases only the hands and feet are spared, although the head and neck are as a rule also free from parasites. The crab-louse has occasionally been found localized upon the head in infants, being either confined entirely to the eyelids, the nits being encountered on the eyelashes, or occurring also upon the eyebrows and hairy portions of the scalp. In adults, the parasite occurs much more rarely in these situations; a case is recorded of an Arab in whom, in addition, the beard and moustache were infested. Such cases are rare in children, very rare indeed in adults.

Apparently the head is so rarely infested because it is less suitable as a habitat. As Waldeyer points out, the head is probably not a suitable habitat because the scalp-hairs are crowded close together and finer than on the pubis and in the axillae. The majority of the active stages are found clinging to two hairs on the regions of the body they most infest, and where the hairs are sparser than on the head. In these parts, the hairs are 2 mm. or more apart. The reach between the extended two hind leg-pairs of the adult insect is about 2 mm., these legs being the ones that are used for grasping the hair.

Whilst infestation usually takes place through coitus, there are many exceptions to the rule as proved by the occurrence of crab-lice on infants. The latter may become infested by their parents or other adults. The insect may pass from one infant to another through their sharing a cradle, or it may pass from soldier to soldier when crowding occurs in barracks or billets. Scratching of their persons by infested individuals no doubt aids in the spread of the parasite upon them and the dissemination of the insect further afield. Both the act of coitus and scratching promote the shedding of hairs, and these, when bearing nits or young larvae, may be of considerable importance in disseminating the parasite. Such hairs, and no doubt occasionally active stages of the louse, are shed on clothing, bedding, the seat of the privy, etc., and readily become entangled with the pubic or other hair of clean persons who may come in contact therewith. A detached louse promptly clings to any hair with which it comes in contact. Therefore, whilst Pthirus is commonly conveyed directly, it may also be acquired indirectly. It is a helpless creature when removed from the hair to which it clings continuously upon the body, where it moves about by shifting from hair to hair; it is therefore much more likely to be conveyed passively from host to host than is Pediculus.

Unless disturbed, the parasite remains confined throughout its life to a limited area upon the host's skin as I have demonstrated experimentally.

The females appear to preponderate over the males in number when upon the host, their relative proportion being about 3:2. In copulation, the male does not seize the female as in Pediculus; he seizes the hairs to which the female clings, using these as a support. Oviposition occurs as in Pediculus. A female that was raised experimentally, laid up to three eggs per day, laying a total of 26 eggs. The hatching period (on the leg) lasted 7–8 days. Like Pediculus, the crab-louse passes through three moults (there being as many larval stages) before it attains sexual maturity. When the adults are about 10 days old they exhibit to a well-marked degree the greenish coloration of the fat-body which is seen by transparency at the sides of the abdomen in the living insect. The life-cycle, from egg to egg, is completed in 22–27 days (on the leg).

Young unfed larvae usually die within ten hours of emergence. When removed from man, they survive longer at 16–20° C. than at 30° C. and die much more rapidly in a dry than in a moist atmosphere; none of the numerous lice of all stages that were tested were found to survive up to 42½ hours, when maintained under different conditions. Living on man a male survived 22 days, a female 17 days, but the insects can doubtless live longer.

The feeding habits of Pthirus recall those of Ixodidae in being practically continuous, and, like Boophilus, they stop feeding to moult upon the host. After moulting they promptly proceed to feed again, shifting but slightly to a fresh feeding ground. On emerging, the young larva clasps a single hair and feeds at its base, and after a day or more it may clasp two hairs as the later stages do usually. The habit of continuous feeding explains why the insects die so quickly when removed from the host.

Whilst a brief exposure to light renders the insects restless but does not cause them to wander away whilst feeding, a warm body brought within their sphere promptly produces great activity in Pthirus when it is removed from man.

1. The male rat is more susceptible to infections of Nematospiroides dubius than the female. As the rat grows older the resistance of the female rat to infection increases at a greater rate than that of the male.

2. The course of the infection is modified by the sex of the host.

3. More larvae penetrated the intestinal mucosa to encyst in the male than in the female. More larvae, however, formed cysts in the female than in the male rat by the fifth day.

4. The male harboured more adult worms than the female rat, although this difference was not significant in the immature animals.

5. The sex resistance of the rat to N. dubius infections was removed by bilateral gonadectomy. Castration decreased the susceptibility of the male rat, while spaying increased it in the female compared with the susceptibility in the respective normal hosts.

6. Subsequent replacement of the homologous sex hormone in the gonadectomized rat restores the sex resistance, and may even increase it (particularly in the immature animals). Oestradiol increased the resistance of the spayed female rat, while testosterone increased the susceptibility of the castrate male rat to infection.

7. Oestradiol implanted in castrate male rats increased the resistance of these hosts to a greater level than was shown in the normal male rat.

8. The rat shows a marked age resistance over which the sex resistance is superimposed.

9. The relationship between the sex of the host and its resistance to infection is discussed.

This work was done during the tenure of a Department of Scientific and Industrial Research Studentship. My thanks are due to Dr E. T. B. Francis for his helpful and critical supervision and to Professor I. Chester Jones, in whose department the work was done, for the facilities he provided.

There are strong biological, evolutionary and immunological arguments for predicting extensive polymorphism among helminth parasites, but relatively little data and few instances from which the selective forces acting on parasite diversity can be discerned. The paucity of information on intraspecific variation stands in contrast to the fine detail with which helminth species have been delineated by morphological techniques, accentuating a trend towards considering laboratory strains as representative of a relatively invariant organism. However, in the fast-moving evolutionary race between host and parasite one would predict a monomorphic species would be driven to extinction. We review the arena of intraspecific variation for the major helminth parasites, ranging from biological properties such as host or vector preference, to biochemical and immunological characteristics, as well as molecular markers such as DNA sequence variants. These data are summarized, before focusing in more detail on polymorphisms within protein-coding genes of potential relevance to the host-parasite relationship, such as vaccine candidates. In particular, we discuss the available data on a number of major antigens from the filarial nematode Brugia malayi. Information is currently too sparse to answer the question of whether there is antigenic variation in filariasis, but the indications are that proteins from the blood-borne microfilarial stage show significant intraspecific variability. Future work will define whether polymorphisms in these antigens may be driven by exposure to the host immune response or reflect some other facet of parasite biology.

Programmed cell death (PCD) has been observed in many unicellular eukaryotes; however, in very few cases have the pathways been described. Recently the early divergent amitochondrial eukaryote Giardia has been included in this group. In this paper we investigate the processes of PCD in Giardia. We performed a bioinformatics survey of Giardia genomes to identify genes associated with PCD alongside traditional methods for studying apoptosis and autophagy. Analysis of Giardia genomes failed to highlight any genes involved in apoptotic-like PCD; however, we were able to induce apoptotic-like morphological changes in response to oxidative stress (H2O2) and drugs (metronidazole). In addition we did not detect caspase activity in induced cells. Interestingly, we did observe changes resembling autophagy when cells were starved (staining with MDC) and genome analysis revealed some key genes associated with autophagy such as TOR, ATG1 and ATG 16. In organisms such as Trichomonas vaginalis, Entamoeba histolytica and Blastocystis similar observations have been made but no genes have been identified. We propose that Giardia possess a pathway of autophagy and a form of apoptosis very different from the classical known mechanism; this may represent an early form of programmed cell death.

The range of interactions between blood-sucking arthropods and their hosts is enormous and their importance to man would be difficult to overestimate. From the tundra to the tropics man, and animals in which he has a vital nutritional or economic interest, represent to the blood-sucking arthropod a source of animal protein. By their activities in obtaining a blood meal the host may be subjected to minimal inconvenience, acute irritation, or suffer severely from blood loss or from the effects of toxins introduced during feeding. Superimposed upon these situations is an enormous variety of disease agents which may be transmitted to the host by the parasite during or after feeding.