We have extracted RNA from Schistosoma mansoni using the lithium chloride–urea method which gives good yields of undegraded RNA. The results of agarose gel electrophoresis of RNA extracted by this procedure suggest that S. mansoni has an in vivo nick in the large rRNA sub-unit. Translation of the RNA in a rabbit reticulocyte lysate gave significant incorporation of [35S]methionine into synthesized proteins. Immuno-precipitation of these translation products using a hyperimmune monkey serum sedimented between 5 and 8% of the radioactivity, which appeared to be present in approximately 13 proteins of molecular weights 18, 20, 21, 22, 23, 35, 40, 54, 60, 70, 74, 78 and 105 K Daltons.

Observations made with the transmission electron microscope are described from the free ovaries of sexually mature female Corynosoma semerme (Palaeacanthocephala) from the ringed seal (Pusa hispida). The structure of the mature ovaries consists of a supporting syncytium, oogonial cells and tissue and appears to be basically similar to that already described for 4 other species of Acanthocephala representing the 3 orders of the phylum. A comparison has been made between the organization of the cytoplasm of the mature oocyte and the zygote with reference to the consequences of fertilization. A detailed description is presented of the heterogeneous inclusions that are observed in the cytoplasm of the mature oocyte but not in the zygote. The fate and possible function of the components of these inclusions is discussed.

The processes which govern the age structure of Fasciola hepatica populations in sheep were investigated by means of a simple mathematical model. The mortality of the metacercariae on the pasture was shown to be a factor of major importance. The mortality rate depends on the micro-climate that prevails at the pasture surface, and thus the age distribution curve of a population of flukes is also determined in part by the weather. This has important implications for the chemotherapy of fascioliasis: flukicides are not equally effective over the whole range of age-classes of parasite and so the factors which govern the age distribution curve of the parasite population within individual hosts ultimately determine the efficiency of chemotherapy.

There are two ways in which trematodes can be used in the control of other trematodes which transmit diseases of medical or veterinary importance. Either they can interfere with the reproductive capacity of the host species of snails, or they can exert an antagonistic effect against the larval stages of the target trematode species inside the snail. Often the two effects act together.

There are six essential criteria for the selection of a suitable species of snail for control: (1) complete and permanent sterilizing effect; (2) clear dominance over target trematodes; (3) strong infectivity to molluscs of all ages; (4) high egg productivity in the definitive host; (5) lack of pathogenicity towards man and domestic animals; (6) ease and low cost of maintenance of the life-cycle.

Various field trials are discussed and it is considered that trematode species producing rediae are usually dominant over those producing only sporocysts (such as the schistosomes). Most species used in control trials to date have belonged to the family Echinostomatidae as they have rediae, and eggs that can be produced in large numbers in a laboratory host such as the rat.

The mathematical basis and feasibility of control schemes are discussed and it is concluded that much more information is necessary before their potential can be evaluated.

The cirrus of a monogenean Diplectanum aequans was isolated by treatment with sodium carbonate solution and studied with a scanning electron microscope. The method may be used in studies of the functional morphology and taxonomy of other organisms.

Eleven gamma-emitting radionuclides (49Sc, 54Mn, 59Fe ( + 2 and + 3), 60Co, 65Zn, 75Se (as selenomethionine, selenocystine, selenite and selenate), 109Cd, 125Sb, 133Ba, 137Cs and 203Hg) were screened as labelling agents for Schistosoma mansoni cercariae by incubation of infected Biomphalaria glabrata snails in radioactive solution according to the technique of Christensen (1977). Only [75Se]methionine yielded satisfactorily labelled cercariae. Multiple regression analysis of volume, number of cercariae and radioactivity from a series of 10 aliquots of unwashed cercarial suspensions yielded estimates of unbound and cercarial-bound radioactivity that were equivalent or superior to estimates based on assay of washed cercariae and eliminated loss of cercariae. Washing of cercarial suspensions over 8 μm pore diameter Millipore filters was found to result in entanglement of 60–90% of the cercariae on the filter disc. Differential external radioassay, a new technique employing partial body shielding within a total body counter, permitted separate estimation of tail and body radioactivity of conscious mice previously exposed by tail immersion to 75Se-labelled cercariae, with measurements repeated as often as desired. Approximately 39% of the 75Se present in emergent cercariae was retained by schistosomula transformed in vitro but this was subject to considerable variation, especially in schistosomula transformed in vivo. Secreted or catabolized label from penetrant cercariae and schistosomula was rapidly removed from the skin by the bloodstream. Numbers of schistosomula in tail skin were directly proportional to the number of reduced silver foci counted on tail autoradiograms; only a very small fraction of tail radioactivity represented unbound (‘spurious’) label. Migration of schistosomula away from skin was 50% complete at 3·8–4·3 days, as determined by probit analysis of autoradiographic data.

This review highlights published research on the principal entomogenous nematodes that have potential as biological control agents of insects. The life-cycles and status of promising members of the families Allantonematidae, Diplogasteridae, Heterorhabditidae, Mermithidae, Neotylenchidae, Rhabditidae, Sphaerulariidae, Steinernematidae and Tetradonematidae are discussed. Emphasis is placed on attempts to control insect populations with these nematodes. Mass propagation of the Steinernematidae and Mermithidae are also discussed, including current in vivo and in vitro rearing systems. A number of these nematodes show promise as biological control agents but commercial development has been slow because of environmental limitations, host specificity and inherent problems associated with manipulation of living organisms. Although some of these nematodes have been considered for commercial preparation, it appears that they will remain ‘on the shelf’ until the need for such biological control provides small businesses with the incentive to make them available for general use.

Various biological mixtures commonly used as microbiological nutrients were added to a chemically defined medium, containing 0·2% (w/v) albumin, in order to assess their ability to support maturation of eggs deposited by adult Schistosoma mansoni in vitro. Of the supplements tested, only lactalbumin hydrolysate and casein hydrolysate stimulated egg deposition and subsequent egg development in vitro. Upon addition of foetal calf serum, further stimulation was noted only in casein hydrolysate-containing media. The addition of mouse red blood cells did not significantly improve the system. Some miracidia which hatched from eggs derived from casein hydrolysate-fortified cultures displayed normal behavioural characteristics, penetrated susceptible snails and established patent infections. These results provide an experimental system for exploring the nutritional requirements of developing miracidia, and the physiological determinants that influence miracidial penetrative capacity and subsequent ability to develop in a susceptible intermediate host.

There are many species of fungi attacking terrestrial and aquatic insects of agricultural and medical importance. Of these, few species have received much attention with a view to their use as biological control agents. The mechanisms of infection have been comparatively well studied, although many questions remain to be answered. The knowledge so far obtained has shed some light on the reasons for host specificity. Temperature, above all relative humidity, and their interactions are the most important physical factors influencing infection of terrestrial insects by entomopathogenic fungi and these are discussed in relation to epizootiological events. In aquatic environments, temperature, salinity and organic pollution are the important factors. In addition to these physical factors, numerous biotic factors at the level of the insect and the fungal pathogen influence both infection and spread of disease in insect populations. The complexity of the interactions of the biotic and abiotic factors makes it extremely difficult to study the influence of any one of these. Virulence of fungal pathogens and its measurement are discussed, together with the shortcomings of present bioassay systems; virtually no laboratory bioassay system exists which has been designed to yield data meaningful in the field. Mass-production techniques are described as are their inherent problems and those of formulation and storage. Finally, the achievements of fungal control of insects in the field are reviewed. Thus far, several species are mass-produced and are in widespread use, two of which, Verticillium lecanii and Hirsutella thompsonii, have been commercialized. More studies, ecological, fundamental and developmental are required in this field to realize fully the potential of other candidate fungi.

The weight gain and growth rate of Trichoplusia ni larvae were severely depressed following parasitization by the insect parasite, Hyposoter exiguae. The development of the host was also delayed and juvenile hormone esterase activity present at the normal time of moulting was absent in parasitized insects. Similar effects on growth and development were achieved by rearing host insects on nutrient-deficient media. Accompanying the above effects of parasitization were alterations in dry/wet weight ratio and host haemolymph osmolality. Parasitization resulted in immediate increases in the concentrations of haemolymph non-glycogen carbohydrate (composed of trehalose, maltose and glucose) and protein. The concentration of fat body glycogen also increased following parasitization. In contrast, the concentration of haemolymph non-glycogen carbohydrate and protein, as well as fat body glycogen, decreased in nutrient-deprived larvae. The significance of the above results as they relate to the insect parasite–host relationship is discussed.

Monoclonal antibodies have been produced after fusion of NS-1 murine myeloma cells with spleen cells from mice immunized either by chronic primary infection or with irradiated cercariae: in both cases, animals were challenged with live cercariae 7 days before fusion. The initial cultures were screened for anti-schistosomular antibodies both by a radioimmunoassay with whole schistosomulum extracts and by immunofluorescence. There was no correlation between the two techniques and subsequent screening was carried out by immunofluorescence. Cloning was carried out in soft agar and 7 cloned cell lines, from 5 initial cultures, were selected for detailed study. Products of 6 of these 7 lines were monoclonal, as judged by isoelectricfocusing of [35S]methionine-labelled supernatant fluids, and their binding to live schistosomula was specific. None of the antibodies showed detectable activity in mediating eosinophil- or complement-dependent damage to schistosomula in vitro. However, 2 antibodies were successfully used to isolate surface proteins with an apparent molecular weight of 24000 on SDS-polyacrylamide gel electrophoresis.

A comparison is given of the ultrastructure of the vitelline cell in Schistosoma mansoni, S. haematobium, S. japonicum and S. mattheei. Four stages in development of the vitelline cell have been categorized as follows: Stage 1, the undifferentiated cell; Stage 2, the developing cell showing the beginning of synthetic activity; Stage 3, the developing cell showing active protein synthesis; Stage 4, the fully mature vitelline cell. These stages in development have been defined morphologically and Stages 1, 2 and 3 are very similar in all 4 species. Lipid is present in the Stage 4 cells of all species but appears earlier at Stage 3 in S. haematobium and S. mattheei. There are several differences as to the intercellular inclusions of the Stage 4 cells, the most marked of these being the absence of calcareous corpuscles from S. japonicum as compared with the other 3 species.

Investigations were conducted on the infection rates and transmission charecteristics of sterile male Glossina morsitans morsitans for Trypanosoma vivax, T. congolense and T. brucei. Pupae were irradiated after the first flush of female emergences with 7, 10 or 13 krad. in a 137caesium radiation source under ambient conditions. The emergent males were fed on a T. viuax-infected goat at peak parasitaemia. They were subsequently mated at 8 days old with 3-day-old normal, virgin females. A batch of sexually fertile males which had been infected as tenerals on the goat at the same time were similarly mated to serve as controls. All the tsetse were maintained on rabbits' ears; the survival and reproductive performance were monitored for seven age-group periods of 9 days each. The sterility rates of the males were 73, 91 and 98% for the 7, 10 and 13 krad. doses of radiation; the percentage of T. vivax infection rates were 78·3, 82·3 and 74·4, respectively. For the control males, the fecundity of the mated females was 90% and the infection rate was 75·7%. In all other experiments, the 10 krad. dose was selected since both the survival and the sterility of the irradiated males were good. The T. vivax-challenged cows and goats died with symptoms of anaemia. There was no difference in the transmission characteristics between sterile and fertile vectors. There was also no significant difference between the sterile and normal males in the infection rates and the transmission frequency of T. congolense and T. brucei to cattle, goats and mice. These results indicate that release of sterile male tsetse in tsetse control programmes will potentially increase the trypanosomiasis risk in the affected area. Consideration must therefore be given to integrating adequate surveillance and chemotherapeutic measures for protection of man and his domestic livestock in the region during the period of such tsetse control campaigns.

This review begins with a description of the parasitoid life-style and the ecological and evolutionary factors which generate the remarkable diversity of insect parasitoids. We then describe the various ways that parasitoids have been used in the biological control of insect pests, and survey their success to date. The use of parasitoids remains largely an art, aided by past experience of success and failure. A more fundamental approach, involving basic research and theory, has not as yet contributed significantly to practical biological control. We explore the potential for such a science of parasitoid use and review basic research on parasitoid ecology and evolution which is of particular relevance to biological control. Mathematical models are used to identify and examine those parasitoid and host attributes which lead to successful biological control. Factors such as parasitoid foraging behaviour, fecundity, larval survival and sex ratio are shown to be important in influencing the depression of host populations and/or the stability of host–parasitoid interactions after depression. Multiple release is discussed and a model for inundative release of parasitoids is explored.

The design and operation of a modular, bacteriological continuous-flow culture system have been adapted for the growth of Trypanosoma cruzi epimastigotes in simple monophasic media. The system was designed to achieve a minimum of 200 days of continuous culture and provision was made for the continuous supply of medium and collection of parasites under sterile conditions. The system provides large quantities of epimastigotes with homogeneous morphology and uniform viability. The system also lends itself tothe analysis of the factors which affect parasite growth. We have examined the effects of changes in environmental parameters on epimastigote growth rate. Optimal growth was observed at 27 °C. The rate ofstirring of the culture had a small but definable effect on the growth rate, which was greatest at 80 r.p.m. Growth was only slightly affected by the level of dissolved oxygen between 10 and 50% saturation, but was inhibited at higher concentrations. Growth was slower at extreme values of pH but showed a broad optimum around pH 7·4.

Two hybridomas secreting monoclonal IgM antibody to Schistosoma mansoni have been isolated following fusion of spleen cells from Balb/c mice immunized with living S. mansoni and NS1 myeloma cells. One monoclonal IgM antibody (WP66.4) mediated about the same level of passive protection against a challenge infection as immune serum from mice with a chronic S. mansoni infection. The other monoclonal antibody (WP66.2) did not give a significant level of passive protection. This result indicates that the effective monoclonal antibody recognizes an antigen which may be a valuable candidate for experimental vaccination. In vitro one monoclonal antibody (WP66.4) caused a much higher level of complement-dependent cytotoxicity than the other (WP66.2), suggesting a possible mechanism for the effect observed in vivo. With indirect immunofluorescence both monoclonal antibodies reacted with surface determinants on living S. mansoni schistosomula, adult worms and miracidia but these determinants were not detected on cercariae or lung schistosomula. Neither monoclonal antibody cross-reacted with S. haematobium schistosomula or Fasciola hepatica metacercariae, indicating a possible use for these reagents in differential diagnosis of S. mansoni infections.

Mice infected with 25 to 35 unsexed Schistosoma mansoni cercariae became resistant to homologous challenge by 8 weeks, and the degree of resistance that was acquired correlated with the size of the worm burden and the tissue egg burden. Mice given 100 male or 100 female cercariae alone failed to become resistant even after 30–40 weeks of infection. When a super-infection of 100 male or 100–300 female cercariae was given to mice with 25 unsexed cercariae, the degree of resistance that was acquired was generally lower than in mice given 25 unsexed cercariae alone. In the super-infected mice there was no correlation between the size of the worm burden (mainly unpaired male or female worms) and the degree of acquired resistance. In mice super-infected with male worms there was an inverse correlation between the number of male worms and the number of eggs deposited in the liver and intestine by each mature worm pair. A positive correlation was, however, still found between the degree of acquired resistance and total tissue egg counts in mice super-infected with single sex cercariae. Mice injected intraperitoneally or subcutaneously with intact S. mansoni eggs failed to become resistant to S. mansoni challenge. Transfer of lymphoid cells or serum, separately or together, from heavily infected and demonstrably resistant donor mice to naive normal or T-cell deprived recipients failed to render the recipients resistant to S. mansoni challenge. The results indicate that resistance to re-infection in mice with recently patent S. mansoni infections is dependent on the number of S. mansoni eggs entrapped in the tissues, and that excessive numbers of unpaired S. mansoni male or female worms can inhibit the fecundity of established worm pairs. The apparent acquired resistance to S. mansoni that has been observed in this model system may be largely a consequence of non-specific factors interfering with normal patterns of challenge worm migration and maturation.

The effects of minocycline alone or in combination with sulphadiazine were evaluated against acute murine toxoplasmosis and compared with the efficacy of pyrimethamine and sulphadiazine. The data presented show that mice could be protected from acute toxoplasmosis by minocycline, in spite of using a virulent strain of Toxoplasma gondii, a large inoculum, and a delay in the administration of the drug. Acute toxoplasmic infection in mice could be eradicated by a minocycline–sulphadiazine combination, which was found to be as effective as pyrimethamine–sulphadiazine combination therapy.

Haemoproteus kopki (De Mello, 1916) is reported from the geckoes Teratoscincus scincus and T. microlepis collected in three localities on the Afghan border of Pakistan. A detailed description of H. kopki is provided in correlation with the courses of an initial, acute infection and a relapsed, chronic infection. Haemoproteus kopki is characterized by large round or oval gametocytes which show sexual dimorphism in distribution of pigment: macrogametocytes show pigment dispersed in dark granules throughout the cytoplasm, while pigment in microgametocytes is aggregated into large, round golden masses within vacuoles in one, or rarely two foci. Other sexual differences exist in gametocyte dimensions. Gametocytes produced in the relapse infection grew much more slowly than did those of the initial infection, and never attained a comparable size despite being observed for twice the length of time. Despite a maximum parasitaemia of 53·8% in the initial infection, increased erythropoiesis did not occur. No asexual stages were found in tissues examined. The presence of mixed infections of H. kopki, Leishmania sp. and microfilariae in a harsh sand dune habitat with presumably limited dipteran fauna suggests their having a common vector in intimate association with the burrows inhabited by the geckoes.

A comprehensive account is given of the evolution of registration guidelines and safety testing procedures for microbial insecticidal agents. Particular emphasis is given to the use of viruses for pest control and the various guidelines developed to assess their possible hazards. The likely gains and risks associated with using viruses are discussed. Several meetings have been held in the last 10 years to assess the hazards of virus insecticides. Some of these meetings have produced recommendations, some have developed guidelines for safety testing. These meetings are reviewed. The various guidelines developed for safety testing are critically evaluated and the UK Registration Criteria for Biological Agents used as Pesticides are reproduced in full. Examples of viruses that have been safety tested and registered for use are given and the criteria used are described. Conclusions are drawn on the merits of the guidelines presently available and the likely future development of safety testing schemes is considered.