Populations of Fasciola hepatica infecting sheep are regulated by at least 3 density-dependent processes. Parasite deaths due to parasite-induced primary host mortality, fluke fecundity and asexual multiplication in the intermediate snail host, Lymnaea truncatula, vary according to the intensity of infection. Empirical and theoretical evidence is used to assess the extent to which each of these processes in turn affects parasite population numbers.

Guinea pigs acquired resistance to the feeding of immature Amblyomma americanum ticks after a single infestation. The initial larval and nymphal feedings resulted in cumulative mortalities of 27·2 and 28·9%, respectively. The second and third larval infestations yielded cumulative mortalities of 50·5 and 52·4%, compared to 18·9 and 24·4% cumulative mortality from controls. Second and third nymphal feedings resulted in 75·7 and 73·4%, and ticks from controls exhibited 36·6 and 34·7% cumulative mortality. The development of host resistance adversely affected tick feeding and resulted in abnormally coloured ticks and a protracted moulting period with a marked decrease in moulting ability. Histological examination of tick feeding sites on guinea pigs challenged twice revealed greater cavity and lesion sizes in hosts fed upon by nymphs than larvae. The primary cell type within these feeding sites were basophils at 12 h post-attachment, but by 72 h post-attachment eosinophils were dominant. The upper epidermal layer adjacent to both larval and nymphal feeding ticks had accumulations of basophils with a few eosinophils. Lymphocytes may have been involved in the resistance response because local lymph nodes in resistant hosts were much larger than in uninfested and singly infested hosts. Some of these nodes had secondary germinal centres.

A technique is described for the recovery of infective larvae of strongyloid nematodes from sediment resulting from the washing of pasture and soil samples. The process of recovery and counting of larvae is described by a model based on the binomial distribution. Tests of the technique have shown that it will recover approximately 50% of the larvae present in samples of pasture sediment of either 7·5 or 12·5 ml in volume after centrifugation. The efficiency of the technique was independent of the number of larvae present in samples of pasture sediment. The technique was shown to be less efficient and more variable in recovery of larvae from samples of soil sediment. Estimations in efficiency of recovery from soil sediment ranged from 64 to 110·2% and a significant effect of volume of sediment on recovery was detected.

1. Observations on the bionomics of Chrysomyia rufifacies Macq. in India have been recorded.

2. Protein is essential for the maturation of eggs, which is complete in about 5 days after emergence.

3. Mating is necessary for oviposition, though egg formation can proceed independently of mating. A single mating is sufficient for at least three batches of eggs.

4. Eggs hatch in 8–12 hr. after they are laid.

5. The average number of eggs produced is 210 (bred flies).

6. The development from egg to adult occupies about 9½ days in summer.

7. The larva is semi-carnivorous, and the floor of the pharynx is provided with ridges.

8. Females of this species produce offspring of one sex only, either all males or all females. This phenomenon, which is independent of diet and of season, is extremely rare among animals especially as parthenogenesis does not occur in this insect.

A series of observations on the feeding behaviour of a laboratory strain of Rhipicephalus appendiculatus on rabbits and measurements of the fluid consumption during feeding in vitro have been made.

Male and female ticks less than 5 weeks after moulting were less inclined to attach than older ticks.

Female ticks fed slowly during the early stages of attachment until after approximately 6 days of attachment when engorgement proceeded very rapidly and their body weight increased by approximately thirty times. Male ticks fed steadily until they roughly doubled their weight in approximately 7 days.

By an in vitro feeding technique in which whole blood, serum or plasma was offered in capillary tubes placed over the mouthparts, it was possible to measure the volumes absorbed over a period of 24 h. More serum or plasma was consumed than either defibrinated or heparinized blood. After 4 or 5 days’ attachment on a rabbit, females generally consumed greater volumes than males.

Dr G. H. Freeman, Head of the Statistics Division of the East African Agriculture and Forestry Organization, kindly carried out the statistical analysis of the quantitative data on the in vitro feeding.

Acknowledgements are also due to the technical staff who contributed to this work, especially to Mrs D. Backhurst. Mr K. P. Bailey maintained the colony of R. appendiculatus from which the ticks used in this study were derived.

This work was carried out during the assignment of one of us (L.P.J.) by the Food and Agriculture Organization of the United Nations to the East African Veterinary Research Organization.

Dirofilaria immitis is an important filarial parasite of dogs and cats, and a useful model for human filariasis. Current diagnostic tests for heartworm infection in animals rely on the presence of fecund female worms (usually found 6·5 months post-infection or later) and therefore fail to detect pre-patent infections. Putative pepsin inhibitors from 2 filarial parasites of humans namely Onchocerca volvulus (Ov33, Oc3.6, OvDSB) and Brugia malayi (Bm33), have been shown to be useful in diagnosis of onchocerciasis and lymphatic filariasis, respectively. Previous studies have suggested that a homologue exists in D. immitis (DiT33), which may have potential in diagnosis of heartworm infection. In this study, the isolation and characterization of a cDNA clone encoding DiT33 is described.‡ This cDNA contains 12 bases of the nematode-specific 22 nucleotide spliced leader sequence and encodes a 26·4 kDa-protein with a high level of similarity (87–89%) to other filarial members of the family. DJT33 was over-expressed in E. coli as a fusion with the maltose-binding protein and serological analysis was performed using a panel of clinically defined dog sera. The findings of this study indicate that DiT33 is a promising antigen for the early detection of D. immitis and may be a valuable tool in the control and management of heartworm infection.

Teneral male Olossina morsitans centralis, G. austeni, G. palpalis palpalis, G. p. gambiensis, G. fuscipes fuscipes, G. tachinoides and G. brevipalpis were fed on the flanks of Boran calves infected with Trypanosoma vivax stock ILRAD 2241 isolated from a cow in Likoni, Kenya; stock ILRAD 2337 isolated from a cow in Galana, Kenya; stock ILRAD 1392 isolated from a cow in Nigeria; or, stock EATRO 1721 isolated from G. m. submorsitans in Nigeria. The tsetse were fed on the infected hosts for 24 days and were then dissected to determine the infection rates. In G. m. centralis and G. brevipalpis, the mature infection rates of T. vivax from Kenya were 61·1 %, and 75·3% for ILRAD 2241, and 36·2% and 58·2% for ILRAD 2337, respectively. In G. austeni and in the four palpalis group of tsetse, the rates for these two stocks were very low and ranged from 0% in G. p. palpalis to 1·8% in G. austeni for ILRAD 2241 and from 0% in G. f. fuscipes to 5% in G. tachinoides for ILRAD 2337. In contrast, the hypopharyngeal infection rates of T. vivax from Nigeria were quite high in all the 7 tsetse species and sub-species. They ranged from 55·5% in G. austeni to 919% in G. p. gambiensis for ILRAD 1392, and from 71·4% in G. austenei to 97·1 % in G. brevipalpis for EATRO 1721. It is suggested that successful establishment of T. vivax infection in a particular tsetse species could depend on the biochemical characteristics of its attachment sites in the food canal and the efficiency of bloodstream trypomastigotes of a particular T. vivax stock to attach to such sites and undergo complete development to meta-trypanosomes in the hypopharynx of the vector.

An incomplete ring of electric nets was placed around uninfected cattle and cattle infected with Trypanosoma congolense. The numbers of fed and unfed Glossina pallidipes caught on the nets were used to estimate the attractiveness of infected and uninfected cattle to tsetse, and the feeding success of tsetse on the cattle. There was no difference in the attractiveness of infected and uninfected cattle to G. pallidipes. However, the feeding success of G. pallidipes on infected cattle was 75 % greater than on uninfected cattle. This suggests that certain effects of T. congolense on cattle behaviour or physiology act to increase the probability of transmission of the parasite by increasing the feeding success of the vector. The nature of the effects of T. congolense on cattle which caused this result is unknown, but several possibilities are discussed.

In 1945 I received a large collection of Streblidae from the late Prof. A. M. Adamson, of the Imperial College of Tropical Agriculture. This collection was made by a post-graduate student, Mr T. S. Jones, and Dr E. C. Humphries, in Trinidad. Later on Mr Jones sent me another three collections, which together with the first comprised three hundred and sixty specimens, the largest number of Streblidae which has ever been collected in America. In addition, I received seven specimens from Mr P. C. J. Brunet, of Keble College, Oxford, which were also collected in Trinidad. I take this opportunity to express my thanks to all these gentlemen, and especially to Mr Jones, who also supplied me with the list of the bats of Trinidad, and some useful information regarding their roosting habits. My thanks are also due to Dr T. C. S. Morrison-Scott and Mr R. W. Hayman of the British Museum (Natural History) for their help in checking the names of several species of bats. The present record is based on the abovementioned five collections and also includes all those records which had been known from Trinidad before.

Evidence for the existence of 2 first intermediate host-specific sibling species of the 45 collar-spined echinostome Echinoparyphium recurvatum is presented. Experimental studies on their life-cycles were carried out under controlled laboratory conditions. The two entities were found to be morphologically indistinguishable in all major respects, yet they exhibited distinct biological characteristics. One of the sibling species utilizes the freshwater lymnaeid pulmonate snail Lymnaea peregra as first intermediate host, and the adults occur in the anterior small intestine of the wildfowl experimental definitive host Anas platyrhynchos. The other utilizes the freshwater mesogastropod prosobranch snail Valvata piscinalis as first intermediate host, and the adults occur in the posterior small intestine and rectum of A. platyrhynchos. The existence of the two sibling species in sympatry may be explained by the fact that the life-cycles of the two forms represent two distinct cycles of transmission serving to reduce excessive competition between them.

Immunity to Hymenolepis nana was adoptively transferred to recipient mice of 4 strains using mesenteric lymph node cells (MLNC), spleen cells (SC), and/or serum taken from infected donor mice. Donor cells capable of transferring immunity appeared in the mesenteric lymph nodes and spleen at an early stage (5 days) after infection. Immunity was most effectively transferred with MLNC and immune serum did not enhance this effect. It is suggested that effective immunity to H. nana involves both antibody and T cell-mediated inflammatory events.

Ocular, peroral, intraperitoneal, intramuscular, and subcutaneous inoculation of severe combined immunodeficient (SCID) mice with spores of the human isolate (CDC: V404) of Brachiola algerae (syn. Nosema algerae) (Phylum Microspora) revealed that the microsporidium develops in viscera of the immunodeficient mouse host, but only after the ocular administration of spores. It is hypothesized that the physico-chemical milieu of the conjunctiva and cornea helped to adapt the originally ‘poikilothermic microsporidian’ to the conditions within the homoiothermic organism. Ocular application of spores caused no clinical signs of disease at the application site. However, severe infection in the liver was found 60 days after infection, manifested as hepatosplenomegaly and multifocal miliary necroses and granulomas containing parasites. No microsporidia were found in any other tissues. Transmission electron microscopy revealed characteristic tubulovesicular ‘secretory materials’ on the plasma membrane of all developmental stages of B. algerae except sporoblasts and spores. These formations increase the parasite surface and allow more efficient metabolic communication of the parasite with the host cell. It is hypothesized that the presence of these structures is a factor helping the parasite to grow in a variety of hosts and tissues. Ultrastructural characters support the likelihood that B. algerae and B. vesicularum are conspecific, and that there exists a relationship between species of the genera Brachiola and Anncaliia.

A key issue relating to developing multi-component anti-malarial vaccines, lies in studying Plasmodium vivax surface proteins' genetic variation. The present work was aimed at amplifying, cloning and sequencing the gene encoding P. vivax merozoite surface protein 5 (PvMSP5) in samples obtained from infected patients from Colombian areas having varying malaria transmission rates.Nucleotide sequence data reported in this paper are available in the GenBank, EMBL and DDBJ databases under Accessions numbers DQ341586 to DQ341601. Our results have revealed that PvMSP5 is one of the P. vivax surface proteins having greater polymorphism, this being restricted to specific protein regions. The intron and exon II (which includes the GPI anchor and EGF-like domain) were both highly conserved when compared to exon I; exon I displayed the greatest variation and most of the recombination events occurred within it. No geographical grouping was observed. The Nei-Gojobori test revealed significant positive selection in the samples analysed here, whereas Tajima and Fu and Li tests presented a neutral selection pattern. The results reflected a localized variation pattern, recombination between PvMSP5 alleles and also functional and immune pressures, where stronger selective forces might be acting on exon I than on exon II, suggesting that the latter could be an important region to be included in an anti-malarial vaccine.

Extensive insecticide use has led to the resistance of mosquitoes to these insecticides, posing a major barrier to mosquito control. Previous Solexa high-throughput sequencing of Culex pipiens pallens in the laboratory has revealed that the abundance of a novel microRNA (miRNA), miR-13664, was higher in a deltamethrin-sensitive (DS) strain than a deltamethrin-resistant (DR) strain. Real-time quantitative PCR revealed that the miR-13664 transcript level was lower in the DR strain than in the DS strain. MiR-13664 oversupply in the DR strain increased the susceptibility of these mosquitoes to deltamethrin, whereas inhibition of miR-13664 made the DS strain more resistant to deltamethrin. Results of bioinformatic analysis, quantitative reverse-transcriptase polymerase chain reaction, luciferase assay and miR mimic/inhibitor microinjection revealed CpCYP314A1 to be a target of miR-13664. In addition, downregulation of CpCYP314A1 expression in the DR strain reduced the resistance of mosquitoes to deltamethrin. Taken together, our results indicate that miR-13664 could regulate deltamethrin resistance by interacting with CpCYP314A1, providing new insights into mosquito resistance mechanisms.

Midgut proteases contribute to the success or failure of Plasmodium infection of the mosquito. This paper examines the reciprocal effect of Plasmodium yoelii nigeriensis on midgut trypsin, chymotrypsin, aminopeptidase and carboxypeptidase in the mosquito Anopheles stephensi. The total protein ingested and the rate of protein digestion were unaffected by the parasite, but more protein was ingested at the first than the second bloodmeal. All peptidases were unaffected by the presence of the parasite during the first gonotrophic cycle, when ookinetes were penetrating the midgut. In the second gonotrophic cycle, trypsin and chymotrypsin were unaffected by growing oocysts, but aminopeptidase activity was reduced in the midguts of infected mosquitoes. Chymotrypsin activity was depressed and aminopeptidase activity elevated during the second gonotrophic cycle. Plasmodium infection has a negligible effect on bloodmeal digestion and does not limit the availability of the protein for egg production. The significance of changes in aminopeptidase activity when oocysts are present is discussed.

Finch trichomonosis, caused by Trichomonas gallinae, emerged in the Canadian Maritime provinces in 2007 and has since caused ongoing mortality in regional purple finch (Carpodacus purpureus) and American goldfinch (Carduelis tristis) populations. Trichomonas gallinae was isolated from (1) finches and rock pigeons (Columbia livia) submitted for post-mortem or live-captured at bird feeding sites experiencing trichomonosis mortality; (2) bird seed at these same sites; and (3) rock pigeons live-captured at known roosts or humanely killed. Isolates were characterized using internal transcribed spacer (ITS) region and iron hydrogenase (Fe-hyd) gene sequences. Two distinct ITS types were found. Type A was identical to the UK finch epidemic strain and was isolated from finches and a rock pigeon with trichomonosis; apparently healthy rock pigeons and finches; and bird seed at an outbreak site. Type B was obtained from apparently healthy rock pigeons. Fe-hyd sequencing revealed six distinct subtypes. The predominant subtype in both finches and the rock pigeon with trichomonosis was identical to the UK finch epidemic strain A1. Single nucleotide polymorphisms in Fe-hyd sequences suggest there is fine-scale variation amongst isolates and that finch trichomonosis emergence in this region may not have been caused by a single spill-over event.

The polymerase chain reaction (PCR) method was used to characterize trypanosome infections in tsetse flies from 3 sleeping sickness foci in Cameroon. The predominant tsetse species found was Glossina palpalis palpalis. An average infection rate of 12·1% was revealed by microscopical examination of 888 non-teneral tsetse flies. PCR amplification analyses for trypanosome identification were carried out on 467 flies, with primer sets specific for Trypanosoma (Trypanozoon) brucei s.l., T. (Duttonella) vivax, T. (Nannomonas) simiae and forest type T. (Nannomonas) congolense. Of 467 flies 93 were positive by microscopical analysis while PCR succeeded in identifying 89 positive flies. Of the PCR-positive flies 34 (38·2%) were negative by microscopical examination. PCR amplification, when compared to the parasitological technique, gave a higher estimate of infection rate of trypanosomes in natural tsetse populations. The PCR technique did, however, fail to identify 40·9% (38/93) of the parasitologically positive flies. The reasons for this failure are discussed. The overall prevalence of mixed infections, assessed by PCR, was 37·1%; the majority (72·7%) involved T. brucei and forest type T. congolense.

Drugs that kill or inhibit the sexual stages of Plasmodium in order to prevent transmission are important components of malaria control programmes. Reducing gametocyte carriage is central to the control of Plasmodium falciparum transmission as infection can result in extended periods of gametocytaemia. Unfortunately the number of drugs with activity against gametocytes is limited. Primaquine is currently the only licensed drug with activity against the sexual stages of malaria parasites and its use is hampered by safety concerns. This shortcoming is likely the result of the technical challenges associated with gametocyte studies together with the focus of previous drug discovery campaigns on asexual parasite stages. However recent emphasis on malaria eradication has resulted in an upsurge of interest in identifying compounds with activity against gametocytes. This review examines the gametocytocidal properties of currently available drugs as well as those in the development pipeline and examines the prospects for discovery of new anti-gametocyte compounds.

The host recognition and invasion process of Myxobolus cerebralis actinospores (triactinomyxon, TAM) was studied on a genetic level. A small-scale in vitro assay was developed to activate a large number of TAMs simultaneously, and to monitor the host invasion in the absence of live fish. The transcriptomes of non-activated and in vitro-activated TAMs were compared by suppressive subtractive hybridization (SSH) to identify parasite genes involved in the host invasion process. Differential screening and a subsequent BLAST search revealed 15 of 452 SSH-library clones expressed differently in activated TAMs. None of the 15 transcripts obtained has previously been identified from M. cerebralis. Quantitative real-time PCR was used to examine the relative expression profile of 8 selected transcripts upon TAM activation and after penetration of the host. Four of these were found to be up-regulated in activated TAMs, while expression was relatively low in non-activated TAMs and in infected fish tissue, indicating that they are relevant genes during host recognition or subsequent host invasion of M. cerebralis TAMs.

The diversity and biology of Cryptosporidium that is specific for rats (Rattus spp.) are not well studied. We examined the occurrence and genetic diversity of Cryptosporidium spp. in wild brown rats (Rattus norvegicus) by microscopy and polymerase chain reaction (PCR)/sequencing targeting the small subunit rDNA (SSU), actin and HSP70 genes. Out of 343 faecal samples tested, none were positive by microscopy and 55 were positive by PCR. Sequence analysis of SSU gene revealed the presence of Cryptosporidium muris (n = 4), C. andersoni (n = 3), C. ryanae (n = 1), C. occultus (n = 3), Cryptosporidium rat genotype I (n = 23), Cryptosporidium rat genotype IV (n = 16) and novel Cryptosporidium rat genotype V (n = 5). Spherical oocysts of Cryptosporidium rat genotype I obtained from naturally-infected rats, measuring 4.4–5.4 μm × 4.3–5.1 μm, were infectious to the laboratory rats, but not to the BALB/c mice (Mus musculus) nor Mongolian gerbils (Meriones unguiculatus). The prepatent period was 3 days post infection and the patent period was longer than 30 days. Naturally- and experimentally-infected rats showed no clinical signs of disease. Percentage of nucleotide similarities at the SSU, actin, HSP70 loci between C. ratti n. sp. and the rat derived C. occultus and Cryptosporidium rat genotype II, III, IV, and V ranged from 91.0 to 98.1%. These genetic variations were similar or greater than that observed between closely related species, i.e. C. parvum and C. erinacei (93.2–99.5%). Our morphological, genetic and biological data support the establishment of Cryptosporidium rat genotype I as a new species, Cryptosporidium ratti n. sp.