A genomic library of Entamoeba histolytica (pathogenic strain HM-1: IMSS) was screened to detect repetitive DNA clones other than those from the highly abundant ribosomal DNA (rDNA). One such clone (HMc) had a 2·3 kb insert which hybridized with the main genome and not the rDNA circle. Southern hybridization of E. histolytica genomic DNA, digested with EcoR I and probed with HMc, showed multiple bands. The banding pattern was identical in all axenic pathogenic strains tested. Differences, however, existed when the banding pattern of a pathogenic strain was compared with that of a non-pathogenic strain. HMc was present in about 25–30 copies per genome in strain HM-1: IMSS. Nucleotide sequence analysis of HMc revealed a partial open reading frame which hybridized with a 1·35 kb poly A+ transcript in Northern blots. The deduced amino acid sequence did not, however, show significant homology with known proteins. The HMc sequence was found only in E. histolytica as it hybridized with 5 different axenic strains of E. histolytica but did not recognize other closely related species of Entamoeba. It has thus the potential to be used as a species-specific DNA probe.

The surface coat of the infective larvae of the parasitic nematode Trichinella spiralis was characterized with respect to its biophysical properties, morphology and composition. Labelling of larvae with the fluorescent surface probe PKH26 was lost after activation (by incubation in mammalian medium containing trypsin and bile), or following pronase treatment. Electron microscopical examination revealed that pronase treatment resulted in the loss of an amorphous surface layer only, further demonstrating the specificity of PKH26 for the larval surface coat. Surface coat shedding was inhibited by sodium azide and carbonyl cyanide, or by incubation of larvae at 4°C, suggesting the shedding process required metabolic energy. Pre-labelled, unactivated larvae demonstrated continuous slow surface coat shedding and could be re-labelled with PKH26, indicating that the shed coat is replaced in these parasites. However, pre-labelled larvae which were activated failed to re-label with the probe, suggesting that activation provides an irreversible trigger for surface changes. PKH26, therefore, is a useful marker for larval activation. Examination of the shed coat material by scanning electron microscopy revealed 2 types of morphologies; one comprising thin multilaminate sheets and the other of amorphous material with ridges producing a fingerprint-like motif. Western- and lectin-blotting of the shed coat material demonstrated 2 prominent entities; a 90 kDa glycoprotein, which bound Datura stramonium agglutinin and was resistant to N- and O-glycanase treatment and a 47–60 kDa set of protein(s). Analysis of the surface lipids by electrospray mass spectometry revealed the presence of lysophosphatidic acid (lysoPA, C14[ratio ]2) and an unidentifiable component of 339·4 Da. These two lipids constituted 36·9% and 36% by mass of surface coat lipids respectively. The presence of lysoPA was confirmed by thin layer chromatography, which also detected phosphatidic acid (PA). The polar lipids detected in solvent rinses of intact parasites by electrospray mass spectrometry were PI (C48[ratio ]4), PE (C40[ratio ]4 and C38[ratio ]4), PS (C40[ratio ]4), lysoPC (C20[ratio ]2 and C18[ratio ]2) and lysoPA (C14[ratio ]2). These observations are discussed with respect to the role of the surface coat and its shedding in the T. spiralis host–parasite relationship.

Fasciola hepatica is the causative agent of fasciolosis, a zoonosis with significant impact both in human and animal health. Understanding the basic processes of parasite biology, especially those related to interactions with its host, will contribute to control F. hepatica infections and hence liver pathology. Mucins have been described as important mediators for parasite establishment within its host, due to their key roles in immune evasion. In F. hepatica, mucin expression is upregulated in the mammalian invasive newly excysted juvenile (NEJ) stage in comparison with the adult stage. Here, we performed sequencing of mucin cDNAs prepared from NEJ RNA, resulting in six different cDNAs clusters. The differences are due to the presence of a tandem repeated sequence of 66 bp encoded by different exons. Two groups of apomucins one with three and the other with four repeats, with 459 and 393 bp respectively, were identified. These cDNAs have open reading frames encoding Ser-Thr enriched proteins with an N-terminal signal peptide, characteristic of apomucin backbone. We cloned a 4470 bp gene comprising eight exons and seven introns that encodes all the cDNA variants identified in NEJs. By real time polymerase chain reaction and high-resolution melting approaches of individual flukes we infer that fhemuc-1 is a single-copy gene, with at least two different alleles. Our data suggest that both gene polymorphism and alternative splicing might account for apomucin variability in the fhemuc-1 gene that is upregulated in NEJ invasive stage. The relevance of this variation in host–parasite interplay is discussed.

Liver samples from 10 Schistosoma mansoni-infected baboons all yielded eggs but neither their egg counts nor duration of infection (< 226 weeks) correlated with the slightly elevated liver collagen levels or minimal histological fibrosis observed. Approximately 10% of inert 9 and 15 μm diameter microspheres injected into the mesenterio veins of 2 baboons with acute, challenge S. mansoni infection reached their lungs (mainly 9 μm microspheres). Lung egg recoveries over 1000 were significantly less common among 175 baboons exposed once to S. monsoni infections than among 56 baboons exposed more than once. Among 221 S. mansoni-and S. haematobium-infected baboons, female worm, total tissue egg and lung egg recoveries were significantly correlated with each other but not with liver or (where available) spleen weights. Liver and spleen weights were strongly correlated with total body weight. Baboons did not develop significant liver fibrosis, even after prolonged schistosome infections. However, some liver ‘leakiness’ developed during acute primary and challenge infections, allowing small inert particles and eggs to pass to the lungs, but this ‘leakiness’ was not associated with resistance to challenge. In contrast to mice, such resistance in baboons cannot, therefore, be explained simply in terms of pathological changes due to previous infections.

Faeces containing eggs of Hyostrongylus rubidus were deposited on grass plots each month throughout 1980. Eggs in faeces deposited on plots from May to October developed into infective larvae but in May development was completed only in artificially moistened faeces, the weather was warm and dry and natural faeces dried out rapidly resulting in the death of eggs and pre-infective larvae. For the remainder of this period the weather was sufficiently warm and wet for development to be completed. More eggs developed into infective larvae on plots with well-grown herbage than on plots with short herbage. In April a few eggs survived and developed into lst-stage larvae but no further development took place. In the laboratory development was completed at temperatures ranging from 10 to 27 °C but no development took place at 4 °C. Out-of-doors infective larvae survived on herbage for up to 10 months while in the laboratory, infective larvae suspended in tap water survived even longer at 10 and 22 °C, but the larvae were rapidly killed by continuous freezing and by desiccation when the relative humidity was less than 95%. The relationship between climatic conditions and the development and survival of the free-living stages is discussed and a comparison made with the free-living stages of Oesophagostomum dentatum.

The severity of the trypanosomiasis problem in a particular location is traditionally assessed in terms of a challenge index – the product of some measure of tsetse abundance and infection rate – which is assumed to be proportional to the force of infection. However, this index masks variation in the force of infection between herds and among individuals within herds. It is also not comparable between sites since the relative abundance of tsetse to hosts may vary. We have studied spatial distribution of herds of cattle in relation to tsetse in The Gambia and calculated an index of challenge based on the ratio of vectors to hosts over the livestock ranging area. This index is strongly correlated with estimates of the force of infection calculated from the incidence of infection in susceptible zebu; and it provides information on heterogeneity in exposure of different herds to tsetse.

Actin depolymerizing factors regulate actin dynamics involved in cellular processes such as morphogenesis, motility, development and infection. Here, a novel actin depolymerizing factor gene (CiADF2) was cloned from the cDNA library of Cryptocaryon irritans, a parasitic ciliate causing cryptocaryonosis. The full-length cDNA of CiADF2 was 531 bp. Its open reading frame (ORF) was 417 bp, encoding a polypeptide of 138 aa with typical features of the ADF/cofilin family. Reverse transcription-PCR suggested that CiADF2 is expressed in all stages of the life cycle. After site-directed mutagenesis of a non-universal genetic code, the ORF was subcloned in Escherichia coli. The bacteria were induced with the addition of isopropylthio-β-D-galactoside to express a fusion protein of recombinant CiADF2 (rCiADF2) with glutathione S transferase. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot confirmed the predicted molecular mass of rCiADF2 of 16·2 kDa. A mouse antibody against rCiADF2 recognized native CiADF2, and rCiADF2 reacted with mouse antisera against C. irritans trophonts. CiADF2 was abundant in the plasma around cytostomes, suggesting that CiADF2 is involved in ciliate movement. Moreover, rCiADF2 showed F-actin binding and depolymerizing activity. This study will help to clarify the pathogenic biology of the parasite and develop effective control measures for cryptocaryonosis.

No group of ticks presents a more interesting biological study than the genus Dermacentor. The diversity of habits of these ticks is remarkable. These differences in habits, especially when we find them correlated with certain structural characteristics, would lead some to consider splitting the genus into two or more genera or subgenera. This we do not think well advised.

Tick distribution depends on the drop-off rhythm of the previous stage and on the suitability for tick survival of the environment where they are disseminated. Studies were implemented in Burkina Faso to assess detachment pattern of engorged Amblyomma variegatum nymphs. Experiments were carried out with naturally infested cattle kept in a paddock or monitored when grazing community pasture. In the pasture, 80% of the nymphs detached between 14.00 h and 17.00 h while less than 25% did so in the paddock. Further investigation was implemented to assess whether the density of adult ticks might be modified by herd management. During the early dry season, zebus grazed in 4 plots fenced in natural savannahs and fallows. Two of the plots were used in the morning and the two others in the afternoon. Six months later, zebus were put in these plots, in turn, on 9 occasions. The number of A. variegatum adults picked up by the cattle in each plot was highly variable: they captured more ticks in the plots installed on good lush savannah and 3-fold more ticks in those where the herd had grazed in the afternoon during the previous dry season. An integrated tick control strategy taking these results into account is proposed.

The examination of the rabbits of the Aberystwyth area has shown heavy infection with adult cestodes. Of 516 small intestines of rabbits examined 358, or 66 per cent., have harboured adult cestodes. Two species occur—Cittotaenia pectinata (Goeze 1782) and Cittotaenia denticulata (Rudolphi 1804). Neither appear to have been recorded for the British Isles before. Theobald (1910) refers to reports of the prevalence of tapeworms in rabbits from near Winchester and from Devonshire. The known distribution of C. denticulata is Germany and France, and of C. pectinata, Germany and France and a variety, var. americana, in North America. C. pectinata is found in that part of the small intestine nearest the stomach. C. denticulata occurs in the lower part of the small intestine. On no occasion have the two species been found in one rabbit.

Establishing the putative links between sylvatic and domestic transmission cycles of Trypanosoma cruzi, the etiological agent of Chagas disease, is of public health relevance. We conducted three surveys to assess T. cruzi infection in wild mammals from a rural and a preserved area in Misiones Province, Northeastern Argentina, which had recently been declared free of vector- and blood-borne transmission of human T. cruzi infection. A total of 200 wild mammals were examined by xenodiagnosis (XD) and/or polymerase chain reaction (PCR) amplification of the hyper-variable region of kinetoplast DNA minicircles of T. cruzi (kDNA-PCR). The overall prevalence of T. cruzi infection was 8%. Nine (16%) of 57 Didelphis albiventris opossums and two (7%) of 29 Desmodus rotundus vampire bats were positive by both XD and kDNA-PCR. Additionally, one D. rotundus positive for T. cruzi by kDNA-PCR tested positive by satellite-DNA-PCR (SAT-DNA-PCR). The T. cruzi-infected bats were captured indoors and in the yard of a vacant dwelling. All D. albiventris were infected with TcI and both XD-positive D. rotundus by TcII. Fifty-five opossum cubs within the marsupium were negative by XD. The mean infectiousness to the vector was 62% in D. albiventris and 50% in D. rotundus. Mice experimentally infected with a parasite isolate from a vampire bat displayed lesions typically caused by T. cruzi. Our study documents the presence of the genotype TcII in a sylvatic host for the first time in Argentina, and the occurrence of two transmission cycles of T. cruzi in a district free of domestic vector-borne transmission.

Coinfection with human visceral leishmaniasis (HVL) and human immunodeficiency virus (HIV) has become an emerging public health problem in several parts of the world, with high morbidity and mortality rates. A systematic review was carried out in the literature available in PubMed, Scielo and Lilacs related to HVL associated with HIV coinfection, seeking to analyze epidemiological, clinical and laboratory aspects. Of the 265 articles found, 15 articles were included in the qualitative analysis, which referred to the results of HVL treatment in patients coinfected with HIV. In the published articles between 2007 and 2015, 1171 cases of HVL/HIV coinfection were identified, 86% males, average age 34 years, liposomal amphotericin B was the most commonly used drug, cure rates 68 and 20% relapses and 19% deaths, five different countries, bone marrow was used in 10/15 manuscripts. HVL/HIV coinfection is a major challenge for public health, mainly due to the difficulty in establishing an accurate diagnosis, low response to treatment with high relapse rates and evolution to death. In addition, these two pathogens act concomitantly for the depletion of the immune system, contributing to worsening the clinical picture of these diseases, which requires effective surveillance and epidemiological control measures.

The distribution of genetic differentiation in a population of the parasitic nematode Strongyloides ratti divided between rat hosts was determined. We applied population genetic theory to these data to determine the source of new infections. We estimate the rate at which a rat acquires a new infection from (a) the existing subpopulation of parasites within that rat (‘self-reinfection’) versus (b) the wider environment (‘immigration’). We find that the observed levels of genetic diversity and differentiation in the study population are consistent with low to moderate rates of self-reinfection and inconsistent with high rates of self-reinfection.

Current theories on disease-diversity relationships predict a strong influence of host richness on disease transmission. In addition, identity effect, caused by the occurrence of particular species, can also modify disease risk. We tested the richness effect and the identity effects of mammal species on bovine tuberculosis (bTB), based on the regional bTB outbreak data in cattle from 2005–2010 in Africa. Besides, we also tested which other factors were associated with the regional bTB persistence and recurrence in cattle. Our results suggested a dilution effect, where higher mammal species richness (MSR) was associated with reduced probabilities of bTB persistence and recurrence in interaction with cattle density. African buffalo had a positive effect on bTB recurrence and a positive interaction effect with cattle density on bTB persistence, indicating an additive positive identity effect of buffalo. The presence of greater kudu had no effect on bTB recurrence or bTB persistence. Climatic variables only act as risk factors for bTB persistence. In summary, our study identified both a dilution effect and identity effect of wildlife and showed that bTB persistence and recurrence were correlated with different sets of risk factors. These results are relevant for more effective control strategies and better targeted surveillance measures in bTB.

DNA barcoding is now a common tool in parasitology and epidemiology, which require good methods for identification not only of parasites and pathogens but vectors and reservoirs. This special issue presents some advances and challenges in barcoding of microbes, parasites, and their vectors and reservoirs. DNA barcoding found new applications in disease ecology, conservation parasitology, environmental parasitology and in paleoparasitology. New technologies such as next-generation sequencing and matrix-assisted laser desorption–ionization time-of-flight have made it now possible to investigate large samples of specimens. By allowing the investigation of parasites at the interface between environment, biodiversity, animal and human health, barcoding and biobanking have important policy outcomes as well as ethics and legal implications. The special issue ‘Advances and challenges in the barcoding of parasites, vectors and reservoirs’ illustrates some recent advances and proposes new avenues for research in barcoding in parasitology.

Entamoeba histolytica trophozoites recovered from the host-parasite interface during abscess development obtain different stimuli compared with long-term cultured cells. In order to have a better understanding about the mechanisms in which the 140 kDa fibronectin (FN)-binding molecule (EhFNR) is involved during the invasive process, we decided to compare the regulation process of this molecule among long-term cultured trophozoites, FN-stimulated trophozoites, and trophozoites recently recovered from a liver abscess. A cDNA clone (5A) containing a fragment of the EhFNR that shows identity to the C-terminal region of the intermediate galactose lectin subunit Igl, was selected with a mAb (3C10). Identity of EhFNR with Igl was confirmed by immunoprecipitation with 3C10 and EH3015 (against the Gal/GalNAc intermediate subunit) mAbs. The 3C10 mAb was used as a tool to explore the modulation of the amoebic receptor (EhFNR). Our results showed specific regulation of the EhFNR in FN-interacted amoebas, as well as in trophozoites recovered at different stages of abscess development. This regulation involved mobilization of the receptor molecule from internal vesicles to the plasma membrane. Therefore, we suggest that in the host-parasite interface, the EhFNR (Igl) plays an important role in the adhesion process during abscess development.

The aim of the present study was to investigate the prevalence and genotyping of Toxoplasma gondii in Iranian human immunodeficiency virus (HIV)-positive patients using multilocus-nested polymerase chain reaction restriction fragment length polymorphism (Mn-PCR-RFLP). A total of 102 serum samples obtained from infected patients were collected from the laboratory centres in northern Iran. Anti-T. gondii antibodies and deoxyribonucleic acid (DNA) detection were accomplished by an enzyme-linked immunosorbent assay and PCR. The Mn-PCR-RFLP method was used for the genotyping of T. gondii. Overall, 68.6% (70/102) and 11.7% (12/102) of the individuals were tested positive for anti-T. gondii immunoglobulin G and T. gondii DNA, respectively. Complete genotyping was performed on 10/12 (83.3%) PCR-positive samples. Accordingly, the samples were classified as genotype #1 (type II clonal; n = 3, 30%), genotype #2 (type III clonal; n = 2, 20%), genotype #10 (type I clonal; n = 2, 20%), genotype #27 (type I variant; n = 1, 10%), genotype #35 (type I variant; n = 1, 10%) and genotype #48 (type III variant; n = 1, 10%). The results were indicative of the high frequency of the type I and type I variant of T. gondii strains in HIV-positive patients in northern Iran. Given the high prevalence of T. gondii and frequency of pathogenic types (pathogen in laboratory mice) in the patients, special measures should be taken to prevent the possible increased incidence of encephalitis by T. gondii.

We investigated whether parasites or hosts benefit from reduced reproduction in infected hosts. When parasites castrate their hosts, the regain of host reproduction is necessary for castration to be a host adaptation. When infecting Daphnia magna with Pasteuria ramosa, in a lake water based medium, 49 2% of the castrated females regained reproduction. We investigated the relationship between castration level, and parasite and host fitness proxies to determine the adaptive value of host castration. Hosts which regained reproduction contained less spores and had a higher lifetime reproduction than permanently castrated hosts. We also found a negative correlation between parasite and host lifetime reproduction. For hosts which regained reproduction we found no optimal level of castration associated with lifetime reproduction. These results support the view that host castration only is adaptive to the parasite in this system. In addition, we suggest that permanent castration might not be the norm under natural conditions in this system. Finally, we argue that a reduction in host reproduction is more likely to evolve as a property favouring parasites rather than hosts. To our knowledge this is the only experimental study to investigate the adaptive value of reduced host reproduction when castrated hosts can regain reproduction.

Interactions between adult Hymenolepis diminuta and rat C-reactive protein (CRP) were investigated in vivo and in vitro. Using an ELISA technique, serum levels of CRP were monitored in rats infected with 100 cysticercoids. Although infection increased the level of this protein in the early stages of parasitization, the increase was not significant until 35 days post-infection (p.i.). Secondary infections did not enhance the response. When H. diminuta was cultured in the presence of CRP, reduced worm motility and opaque areas were observed and electron microscopical studies revealed shedding of microtriches and lysis of the tegument. Initially, damage was restricted to the strobila which correlated with the regional distribution of phosphorylcholine as visualized using immunofluorescence.

Mice which have recovered from infections with A. garnhami do not succumb to; challenge with the piroplasms B. rodhaini or B. microti and over half such animals are protected against the malaria parasites P. v. vinckei and P. v. chabaudi. This protection does not extend to the malaria parasites P. b. berghei or P. b. yoelii. Mice which have recovered from infections with the piroplasms are protected against A. garnhami, as are those which have recovered from infections with P. v. vinckei or P. v. chabaudi. About half the mice which have recovered from infections with P. b. berghei or P. b. yoelii are also protected against A. garnhami. This interaction between intra-erythrocytic protozoa may influence the incidence of these parasites in the wild. A. garnhami is unlikely to be of any use in the immunization of humans against malaria but could be of use in the protection of cattle against piroplasmosis.