The Luangwa Valley has a long historical association with Human African Trypanosomiasis (HAT) and is a recognised geographical focus of this disease. It is also internationally acclaimed for its high biodiversity and contains many valuable habitats. Local inhabitants of the valley have developed sustainable land use systems in co-existence with wildlife over centuries, based on non-livestock keeping practices largely due to the threat from African Animal Trypanosomiasis. Historical epidemics of human sleeping sickness have influenced how and where communities have settled and have had a profound impact on development in the Valley. Historical attempts to control trypanosomiasis have also had a negative impact on conservation of biodiversity.Centralised control over wildlife utilisation has marginalised local communities from managing the wildlife resource. To some extent this has been reversed by the implementation of community based natural resource management programmes in the latter half of the 20th century and the Luangwa Valley provides some of the earliest examples of such programmes. More recently, there has been significant uncontrolled migration of people into the mid-Luangwa Valley driven by pressure on resources in the eastern plateau region, encouragement from local chiefs and economic development in the tourist centre of Mfuwe. This has brought changing land-use patterns, most notably agricultural development through livestock keeping and cotton production. These changes threaten to alter the endemically stable patterns of HAT transmission and could have significant impacts on ecosystem health and ecosystem services.In this paper we review the history of HAT in the context of conservation and development and consider the impacts current changes may have on this complex social-ecological system. We conclude that improved understanding is required to identify specific circumstances where win-win trade-offs can be achieved between the conservation of biodiversity and the reduction of disease in the human population.

Background: The European perch, Perca fluviatilis L. is a common paratenic host of dioctophymatid nematodes belonging to the genus Eustrongylides. In this host, once infected oligochaetes, which serve as the first intermediate host, are ingested, Eustrongylides migrates through the intestine and is frequently encountered within the musculature, free within the body cavity, or encapsulated on the viscera. The current study details the first Italian record of Eustrongylides sp. with larvae reported in the muscle of P. fluviatilis. Methods: Uninfected and nematode-infected muscle tissues of perch were fixed and prepared for histological evaluation and electron microscopy. Some sections were subjected to an indirect immunohistochemical method using anti-PCNA, anti-piscidin 3 and anti-piscidin 4 antibodies. Results: A total of 510 P. fluviatilis (TL range 15–25 cm) from Lake Trasimeno, Perugia were post-mortemed; 31 individuals had encysted nematode larvae within their musculature (1–2 worms fish−1). Histologically, larvae were surrounded by a capsule with an evident acute inflammatory reaction. Muscle degeneration and necrosis extending throughout the sarcoplasm, sarcolemmal basal lamina, endomysial connective tissue cells and capillaries was frequently observed. Within the encapsulating reaction, macrophage aggregates (MAs) were seen. Immunohistochemical staining with the proliferating cell nuclear antigen (PCNA) revealed numerous PCNA-positive cells within the thickness of the capsule and in the immediate vicinity surrounding Eustrongylides sp. larvae (i.e. fibroblasts and satellite cells), suggesting a host response had been initiated to repair the nematode-damaged muscle. Mast cells (MCs) staining positively for piscidin 3, were demonstrated for the first time in response to a muscle-infecting nematode. The piscidin 3 positive MC’s were seen principally in the periphery of the capsule surrounding the Eustrongylides sp. larva. Conclusions: A host tissue response to Eustrongylides sp. larvae infecting the musculature of P. fluviatilis was observed. Numerous fibroblasts, MAs and MCs were seen throughout the thick fibroconnectival layer of the capsule enclosing larvae. PCNA positive cells within the capsule suggest that host repair of nematode damaged muscle does occur, while the presence of the antimicrobial peptide piscidin 3 is shown for the first time. This is first report of Eustrongylides sp. in an Italian population of P. fluviatilis.

Background: Insecticides are widely used to control malaria vectors and have significantly contributed to the reduction of malaria-caused mortality. In addition, the same classes of insecticides were widely introduced and used in agriculture in Benin since 1980s. These factors probably contributed to the selection of insecticide resistance in malaria vector populations reported in several localities in Benin. This insecticide resistance represents a threat to vector control tool and should be monitored. The present study reveals observed insecticide resistance trends in Benin to help for a better management of insecticide resistance. Methods: Mosquito larvae were collected in eight sites and reared in laboratory. Bioassays were conducted on the adult mosquitoes upon the four types of insecticide currently used in public health in Benin. Knock-down resistance, insensitive acetylcholinesterase-1 resistance, and metabolic resistance analysis were performed in the mosquito populations based on molecular and biochemical analysis. The data were mapped using Geographical Information Systems (GIS) with Arcgis software. Results: Mortalities observed with Deltamethrin (pyrethroid class) were less than 90% in 5 locations, between 90-97% in 2 locations, and over 98% in one location. Bendiocarb (carbamate class) showed mortalities ranged 90-97% in 2 locations and were over 98% in the others locations. A complete susceptibility to Pirimiphos methyl and Fenitrothion (organophosphate class) was observed in all locations with 98-100% mortalities. Knock-down resistance frequencies were high (0.78-0.96) and similar between Anopheles coluzzii, Anopheles gambiae, Anopheles arabiensis, and Anopheles melas. Insensitive acetylcholinesterase-1 was rare (0.002-0.1) and only detected in Anopheles gambiae in concomitance with Knock-down resistance mutation. The maps showed a large distribution of Deltamethrin resistance, Knock-down mutation and metabolic resistance throughout the country, a suspected resistance to Bendiocarb and detection of insensitive acetylcholinesterase-1 from northern Benin, and a wide distribution of susceptible vectors to Pirimiphos methyl and Fenitrothion. Conclusion: This study showed a widespread resistance of malaria vectors to pyrethroid previously located in southern Benin, an early emergence of carbamates resistance from northern Benin and a full susceptibility to organophosphates. Several resistance mechanisms were detected in vectors with a potential cross resistance to pyrethroids through Knock-down and metabolic resistance mechanisms.

Background: Members of the Anopheles gambiae complex are the main transmitters of malaria. Anopheles merus is a member of the complex found along the Kenyan coast because it breeds in saline waters. An entomological study was conducted in Garithe Malindi District, to investigate the physicochemical and environmental factors affecting the distribution of An. merus. Methods: Field and laboratory studies were used to investigate the breeding habitats of the subspecies. Mosquito larvae were sampled using standard dipping technique from small pockets of pools, ponds, hoof prints, road drain, wells and mangrove swamps found in Garithe. All 3rd and 4th instars of Anopheles larvae sampled were identified microscopically into species. A representative of Anopheles gambiae complex was then identified to specific sibling species using r-DNA PCR technique.The habitats were characterized based on temperature, conductivity, salinity, dissolved oxygen, total dissolved solids, pH, size, distance to nearest house, canopy coverage, surface debris, presence of algae, emergent plants, turbidity and habitat types. Results: A total of 159 morphologically identified late stage instar Anopheles gambiae s.l larvae were selected for r-DNA analysis by PCR. Out of these, 60.4% (n = 96) were Anopheles merus, 8.8% (n = 14) were Anopheles arabiensis, 18.2% (n = 29) were Anopheles gambiae s.s and 12.6% (n = 20) were unknown.Using paired t-test (t (121) = −3.331, P = 0.001) a significantly high proportion of An. merus was observed in all habitats compared to An. arabiensis, and An. gambiae s. s.In habitat characterization, Pearson’s correlation analysis test showed different parameters being associated with the occurrence of An. merus larvae in the different habitats sampled. Six out of the 55 correlation coefficients (10.9%) were statistically significant, suggesting non-random association between some pairs of variables. Those that had a significantly high positive correlation with An. merus included temperature, salinity, conductivity, total dissolved solids and algae. Conclusions: Different physicochemical parameters and environmental parameters affect the occurrence of An. merus. In this study, higher temperatures accelerate the growth of the larvae and aids in growth of micro-organisms and algae which are food sources for the larvae. Saline waters favour the growth and development of An. merus larvae; they are also able to develop in a range of saline waters. Conductivity, total dissolved solids and canopy coverage are among the important factors influencing the development and abundance of An. merus larvae in their habitats. Habitat type also influences the abundance of An. merus larvae. They mainly prefer to breed in pools and ponds, but not swamps, hoof prints and wells.

Background: In Ethiopia, urinary schistosomiasis caused by Schistosoma haematobium has been known to be endemic in several lowland areas of the country where it causes considerable public health problems, mainly among school-age children. However, information on recent magnitude and risk factors of the disease is lacking, particularly for Gambella area. Therefore, this study aimed to assess the prevalence of urinary schistosomiasis and associated risk factors among Abobo Primary School children in Gambella, southwestern Ethiopia. Methods: A cross-sectional study involving 304 school children was conducted in Abobo Primary School, Gambella Regional State, southwestern Ethiopia, from February to June 2014. Ten ml of urine sample was collected from each study participant and processed for microscopic examination by the urine filtration method; egg load for positive individuals was determined per 10 ml of urine. Data on socio-demographic characteristics and risk factors were collected using an interview-based questionnaire. The data were entered into and analyzed with SPSS version 20. Logistic regression and odds ratio were used to measure association and strength between variables, respectively. P-value < 0.05 at 95% CI was considered as statistically significant. Results: The prevalence of urinary schistosomiasis was 35.9% (109/ 304) with a mean egg intensity of 8.76 per 10 ml of urine. Being male [AOR (95%CI) = 2.15(1.31, 3.52)], having father as a farmer [AOR (95%CI) = 1.96(1.19, 3.22)] and children living apart from parents [AOR (95% CI): 3.09 (1.14, 8.4)] were significantly associated with urinary schistosomiasis. Conclusion: The present study area in Gambella Regional State, southwestern Ethiopia, represents moderate-risk community for urinary schistosomiasis. Sex, father’s occupation and living apart from parents were found to be associated with infection. Treatment of all school-age children and fishermen is required once every 2 years until the prevalence of infection falls below the level of public health importance. It is also recommended to complement praziquantel treatment with supplementary measures such as provision of sanitation facilities and health education.

T. T. Ao et al.

Background: Parvicapsula pseudobranchicola (Myxozoa) causes widespread infections in farmed Atlantic salmon in northern Norway. Heavily infected salmon become runts, probably due to vision impairment or blindness. The salmon are likely infected by waterborne actinospores, released by an alternating annelid host, but the life cycle of P. pseudobranchicola is unknown. Seatrout and Arctic charr have been considered possible hosts for the parasite, but firm evidence has been lacking.FindingsWe show for the first time the presence of mature spores of P. pseudobranchicola in seatrout. The seatrout were infected with high intensities of P. pseudobranchicola in the pseudobranchs in early April. The presence of mature spores in early spring suggests that the fish had been infected late the previous year, a pattern of infection similar to that observed for farmed salmon stocked in autumn. Although heavily infected, the fish did not display any symptoms consistent with parvicapsulosis. The results suggest that the life cycle of P. pseudobranchicola is more adapted to seatrout, rather than to Atlantic salmon. Conclusions: The presence of mature spores of P. pseudobranchicola in seatrout confirms that seatrout is a natural host for this myxosporean and this is also the first record of these spores in the pseudobranch of a wild salmonid. Furthermore, wild trout from non-farming areas may become heavily infected with P. pseudobranchicola, developing pseudobranch pathology resembling that of farmed Atlantic salmon suffering from parvicapsulosis.

Background: Toxoplasma gondii is an obligate, intracellular protozoan that infects almost all warm-blooded animals, including humans, domesticated and wild animals. Recent studies of Toxoplasma gondii isolates from animals in different regions of China have shown a limited genetic diversity with the dominance of the ToxoDB PCR-RFLP genotype #9 named as “Chinese 1”. However, there is not much published information regarding its prevalence in domestic animals from Guizhou province, a subtropical region in Southwest China. The objectives of this study were to determine seroprevalence and genetic diversity of T .gondii in pigs, dogs and cats in Guizhou province, Southwest China.FindingsThe anti-T. gondii IgG were detected in 70.0%(49/70) pigs, 20.56%(22/107) dogs and 63.16(12/19) cats. The anti-T. gondii IgM were found in 0.93%(1/107) dogs, 21.53%(4/19) cats, but not in pigs. In addition, the toxoplasma circulating antigen (CAG) were detected in 16.9%18/70)pigs, 13.1% (14/107) dogs and 10.5%(2/19) cats. The T. gondii DNA were detected in 31.5%(22/70) pigs, 3.7%(4/107) dogs and 52.63%(10/19) cats. Five T. gondii isolates were obtained(3 from pigs and 2 from cats). The genotype of these five isolates belonged to the predominant genotype “Chinese 1”. Conclusions: The high prevalence of T. gondii infection in pigs,cats and dogs indicated that the T. gondii infection is common in Guizhou province. Additionally, the T. gondii genotype “Chinese 1” was dominant in Southwest China.

Background: In eukaryotes, histone arginine methylation associates with both active and repressed chromatin states depending on the residues involved and the status of methylation. Even when the amino-terminus of Entamoeba histolytica histones diverge from metazoan sequences, these regions contain arginine residues that are potential targets for methylation. However, histone arginine methylation as well as the activity of arginine methyltransferases (PRMTs) has not been studied in this parasite. The aim of this work was to examine the dimethylation of arginine 3 of H4 histone (H4R3me2) and to identify the parasite PRMT that could be responsible for this modification (EhPRMT1). Methods: To examine the presence of H4R3me2 in E histolytica, we performed Western blot and immunofluorescence assays on trophozoites using an antibody against this epigenetic mark. To recognize the PRMT1 enzyme of this parasite that possibly perform that modification, we first performed a phylogenetic analysis of E. histolytica and human PRMTs. RT-PCR assays were carried out to analyze the expression of the putative PRMT1 genes. One of these genes was cloned and expressed in Escherichia coli. The recombinant protein was tested by its recognition by an antibody against human PRMT1 and in its ability to form homodimers and to methylate commercial histones. Results: The arginine 3 of human H4, which is subjected to post translational methylation, was aligned with the arginine 8 of E. histolytica H4, suggesting that this residue could be methylated. The recognition of an 18 kDa nuclear protein of E. histolytica by an antibody against H4R3me2 confirmed this assumption. We found that this parasite expresses three phylogenetic and structural proteins related to PRMT1. Antibodies against the human PRMT1 detected E. histolytica proteins in cytoplasm and nuclei and recognized a recombinant PRMT1 of this parasite. The recombinant protein was able to form homodimers and homotetramers and displayed methyltransferase activity on arginine 3 of chicken H4. Conclusion: All these results suggest that E. histolytica contains as a minimum one structural and functional protein ortholog to PRMT1, enzyme that potentially dimethylates H4R8. This modification may play an important role in the gene expression regulation of this microorganism.

Background: Theileria annae is a tick-transmitted small piroplasmid that infects dogs and foxes in North America and Europe. Due to disagreement on its placement in the Theileria or Babesia genera, several synonyms have been used for this parasite, including Babesia Spanish dog isolate, Babesia microti-like, Babesia (Theileria) annae, and Babesia cf. microti. Infections by this parasite cause anemia, thrombocytopenia, and azotemia in dogs but are mostly subclinical in red foxes (Vulpes vulpes). Furthermore, high infection rates have been detected among red fox populations in distant regions strongly suggesting that these canines act as the parasite’s natural host. This study aims to reassess and harmonize the phylogenetic placement and binomen of T. annae within the order Piroplasmida. Methods: Four molecular phylogenetic trees were constructed using a maximum likelihood algorithm based on DNA alignments of: (i) near-complete 18S rRNA gene sequences (n = 76 and n = 93), (ii) near-complete and incomplete 18S rRNA gene sequences (n = 92), and (iii) tubulin-beta gene sequences (n = 32) from B. microti and B. microti-related parasites including those detected in dogs and foxes. Results: All phylogenetic trees demonstrate that T. annae and its synonyms are not Theileria parasites but are most closely related with B. microti. The phylogenetic tree based on the 18S rRNA gene forms two separate branches with high bootstrap value, of which one branch corresponds to Babesia species infecting rodents, humans, and macaques, while the other corresponds to species exclusively infecting carnivores. Within the carnivore group, T. annae and its synonyms from distant regions segregate into a single clade with a highly significant bootstrap value corroborating their separate species identity. Conclusion: Phylogenetic analysis clearly shows that T. annae and its synonyms do not pertain to Theileria and can be clearly defined as a separate species. Based on the facts that T. annae and its synonyms have not been shown to have a leukocyte stage, as expected in Theileria, do not infect humans and rodents as B. microti, and cluster phylogenetically as a separate species, this study proposes to name this parasite Babesia vulpes sp. nov., after its natural host, the red fox V. vulpes.

H. K. Hassan et al.

E. Castro-Sánchez et al.

L. Breakwell et al.

E. Schneider et al.

E. de Vries et al.

P. Dickmann et al.

M. Nemoto et al.

C. Petrosian-Husa

D. Tappe et al.

J. Reiche et al.