Parasite Challenge Research Articles

A novel fish meal-free diet formulation supports proper growth and does not impair intestinal parasite susceptibility in gilthead sea bream (Sparus aurata) with a reshape of gut microbiota and tissue-specific gene expression patterns

The exponential growth of the aquaculture sector requires the development of sustainable aquafeeds with less dependence on marine products. The maximized replacement of fish meal (FM) and fish oil (FO) with plant ingredients has been extensively studied in the economically important species gilthead sea bream (Sparus aurata). Recently, major progress has been done with other alternative raw materials, though some non-pathological inflammatory response persisted with feed formulations that increased the circularity of resource utilization. In the present study, we evaluated the effects on growth performance, gene expression, intestinal microbiota and disease resistance of a FM-free diet (NoPAP SANA), based on plant ingredients, aquaculture by-products, algae oil, insect meal and bacterial fermentation biomasses as main dietary oil and protein sources, and supplemented with a commercially available health-promoting feed additive (SANACORE®GM). Juveniles of 21 g initial body weight were fed control or NoPAP SANA diets for 34 days, and head kidney, liver and posterior intestine were collected for gene expression analyses using customized PCR-arrays. Each tissue-specific PCR covered 96 genes in total and included markers of growth performance, lipid and energy metabolism, antioxidant defence, immune system, and intestinal function and integrity. From the same fish, the adherent bacteria of the posterior intestine were studied by Illumina sequencing of the V3-V4 region of the 16S rRNA. The remaining fish were challenged with the intestinal parasite Enteromyxum leei for 78 days and sampled for parasite diagnosis. Both control and NoPAP SANA fish grew efficiently considering gilthead sea bream standards. Before parasite challenge, the NoPAP SANA group showed differential expression of 17, 2 and 4 genes in liver, head kidney and posterior intestine, respectively. The intestinal bacterial composition showed no major differences in diversity or at the phylum level. However, 29 abundant OTUs significantly changed with the diet. From these, 10 OTUs were significantly correlated with differentially expressed genes in the different target tissues. Inferred metagenome analyses revealed that the altered microbiota with NoPAP SANA diet could account for changes in 15 metabolic pathways. The intensity and prevalence of infection after the parasite challenge did not significantly vary between dietary treatments, and infected fish from both groups showed similar disease outcome. Altogether, these results indicate that the NoPAP SANA diet promoted optimal growth and a healthy condition in gilthead sea bream without affecting susceptibility against the tested intestinal parasite, as often observed with alternative diets following current industry formulations.

Leucocytes are Mass‐Invariant in Reptiles Unlike in Endotherms

How do large and small animals defend against parasite challenges given the consequences of body mass on physiology and disease transmission? Functionally equivalent mammalian and avian granulocytes increased disproportionately with mass (i.e., hypermetrically), such that large organisms have higher concentrations than expected by mass alone. However, how ectothermic leucocyte concentrations scale is understudied. We hypothesized that differences exist in the scaling of leucocyte concentrations between endotherms and ectotherms as these groups have substantial differences in their physiology and immunology. Analysis was conducted using MCMCglmm, phylogenetically informed models to determine which existing scaling hypothesis best predicted relationships between mass and lymphocyte and heterophil concentrations among >120 species of reptiles and compared our results with those from birds and mammals. We also compared body mass and life history as a predictor of variation in immune cell concentrations. Reptilian lymphocyte concentrations scaled close to isometrically (i.e., mass‐invariant; phylogenetic MCMCglmm: b(mean, 95% CI) = ‐0.040, [‐0.072, ‐0.007]), and the slope was indistinguishable from the slope found for mammals and birds. In contrast, reptilian heterophil concentrations scaled isometrically (b =0.006, [‐0.028, 0.040]), whereas functionally equivalent mammalian and avian granulocytes scaled hypermetrically. Although reptilian leucocytes were mass‐invariant, the slope was greater than predicted if metabolic rate serves as a pace marker for immune defenses, suggesting that large organisms have greater protection than expected by the Rate of Metabolism Hypothesis. Life history was also a poor predictor of variation in reptilian heterophil and lymphocyte concentrations. Our results provide novel insight into large scale patterns of immune defenses of endothermic and ectothermic vertebrates and have direct implications for modeling the evolution of immune defenses and identifying parasite reservoirs.

Plasmodium exposure alters midgut epithelial cell dynamics during the immune memory in Anopheles albimanus

Immunological priming in insects is defined as a previous contact with non-virulent pathogens, which induces protection after a second virulent infection. The mechanism of this process is not well understood. We have observed midgut DNA synthesis (endoreplication) in Plasmodium berghei exposure mosquitoes (primed) and after the immune challenge, which could be an essential component of the priming response in the mosquito. Endoreplication requires cell cycle components re-direction to make multiple DNA copies. Therefore, it is fundamental to understand the role of cell cycle components in priming. Here, we analyzed the expression of the cyclins A, B, E, and AurkA, and the endoreplication components NOTCH and HNT in the mosquito Anopheles albimanus; after priming with non-infective Plasmodium berghei and challenged with an infective P. berghei. The overexpression of cell cycle elements occurred seven days after priming with a quick reduction 24 h after the challenge. Hnt and NOTCH overexpression occurred 24 h after priming. Antimicrobial peptide cecropin is quickly overexpressed after 24 h in primed mosquitoes, then is downregulated at day seven and overexpressed again after parasite challenge. We also found that DNA synthesis occurs in cells with different nuclear sizes, suggesting a change in midgut epithelial dynamics after Plasmodium exposure. Inhibition of DNA synthesis via cisplatin revealed that DNA synthesis is required for priming to limit Plasmodium infection. Our results indicate the importance of cell cycle components on DNA synthesis and Notch pathway during priming response in An. albimanus mosquitoes.

Detrimental Effect ofTrypanosoma brucei bruceiInfection on Memory B Cells and Host Ability to Recall Protective B-cell Responses

Trypanosoma brucei brucei evades host immune responses by multiple means, including the disruption of B-cell homeostasis. This hampers anti-trypanosome vaccine development. Because the cellular mechanism underlying this pathology has never been addressed, our study focuses on the fate of memory B cells (MBCs) in vaccinated mice upon trypanosome challenge. A trypanosome variant surface glycoprotein (VSG) and fluorescent phycoerythrin were used as immunization antigens. Functional and cellular characteristics of antigen-specific MBCs were studied after homologous and heterologous parasite challenge. Immunization with AnTat1.1 VSG triggers a specific antibody response and isotype-switched CD73+CD273+CD80+ MBCs, delivering 90% sterile protection against a homologous parasite challenge. As expected, AnTat1.1 VSG immunization does not protect against infection with heterologous VSG-switched parasites. After successful curative drug treatment, mice were shown to have completely lost their previously induced protective immunity against the homologous parasites, coinciding with the loss of vaccine-induced MBCs. A phycoerythrin immunization approach confirmed that trypanosome infections cause the general loss of antigen-specific splenic and bone marrow MBCs and a reduction in antigen-specific immunoglobulin G. Trypanosomosis induces general immunological memory loss. This benefits the parasites by reducing the stringency for antigenic variation requirements.

Recombinant guanosine-5′-triphosphate (GTP)-binding protein associated with Poloxamer 407-based polymeric micelles protects against Leishmania infantum infection

Leishmania virulence proteins should be considered as vaccine candidates against disease, since they are involved in developing infection in mammalian hosts. In a previous study, a Leishmania guanosine-5′-triphosphate (GTP)-binding protein was identified as a potential parasite virulence factor. In the present work, the gene encoding GTP was cloned and the recombinant protein (rGTP) was evaluated as a vaccine candidate against Leishmania infantum infection. The protein was associated with saponin (rGTP/Sap) or Poloxamer 407-based micelles (rGTP/Mic) as adjuvants, and protective efficacy was investigated in BALB/c mice after parasite challenge. Both rGTP/Sap and rGTP/Mic compositions induced a Th1-type immune response in vaccinated animals, with significantly higher levels of IFN-γ, IL-12, IL-2, TNF-α, GM-CSF, nitrite, specific IgG2a isotype antibody and positive lymphoproliferation, when compared to the control groups. This response was accompanied by significantly lower parasite load in the spleens, livers, bone marrows and draining lymph nodes of the animals. Immunological and parasitological evaluations indicated that rGTP/Mic induced a more polarized Th1-type response and higher reduction in the organ parasitism, and with lower hepatotoxicity, when compared to the use of rGTP/Sap. In conclusion, our preliminary data suggest that rGTP could be considered for further development as a vaccine candidate to protect against VL.

Parasite Challenge: A giant, meaty flower has a fraught relationship with its host plant.

Parasite Challenge: A giant, meaty flower has a fraught relationship with its host plant.

COVID-19 vaccine results might inform malaria vaccine strategies

COVID-19 vaccine results might inform malaria vaccine strategies

Phenomic analysis of the honey bee pathogen-web and its dynamics on colony productivity, health and social immunity behaviors.

Many pathogens and parasites have evolved to overwhelm and suppress their host’s immune system. Nevertheless, the interactive effects of these agents on colony productivity and wintering success have been relatively unexplored, particularly in large-scale phenomic studies. As a defense mechanism, honey bees have evolved remarkable social behaviors to defend against pathogen and parasite challenges, which reduce the impact of disease and improve colony health. To investigate the complex role of pathogens, parasites and social immunity behaviors in relation to colony productivity and outcomes, we extensively studied colonies at several locations across Canada for two years. In 2016 and 2017, colonies founded with 1-year-old queens of diverse genetic origin were evaluated, which represented a generalized subset of the Canadian bee population. During each experimental year (May through April), we collected phenotypic data and sampled colonies for pathogen analysis in a standardized manner. Measures included: colony size and productivity (colony weight, cluster size, honey production, and sealed brood population), social immunity traits (hygienic behavior, instantaneous mite population growth rate, and grooming behavior), as well as quantification of gut parasites (Nosema spp., and Lotmaria passim), viruses (DWV-A, DWV-B, BQCV and SBV) and external parasites (Varroa destructor). Our goal was to examine: 1) correlations between pathogens and colony phenotypes; 2) the dynamics of pathogens and parasites on colony phenotypes and productivity traits; and 3) the effects of social immunity behaviors on colony pathogen load. Our results show that colonies expressing high levels of some social immunity behaviors were associated with low levels of pathogens/parasites, including viruses, Nosema spp., and V. destructor. In addition, we determined that elevated viral and Nosema spp. levels were associated with low levels of colony productivity, and that five out of six pathogenic factors measured were negatively associated with colony size and weight in both fall and spring periods. Finally, this study also provides information about the incidence and abundance of pathogens, colony phenotypes, and further disentangles their inter-correlation, so as to better understand drivers of honey bee colony health and productivity.

Milk production status and associated factors among indigenous dairy cows in Raya Kobo district, north eastern Ethiopia.

BackgroundA cross‐sectional survey study was conducted from September 2020 to April 2021. A total of 217 households were randomly selected. The data collection instruments were structured questionnaires: focus group discussion and key informant interviews. Data were coded, entered and analyzed using Statistical Package for the Social Sciences (SPSS) version 20 software. Ranking indexes as well as binary logistic regression analysis were used to look for the relationship between dependent and independent variables.ResultThe present study showed that season of calving, disease and parasite challenges, housing conditions and shortage of land for forage production with an index value of 0.180, 0.154, 0.153 and 0.126, respectively, were the most important constraints affecting milk production potential. Likewise, foot and mouth disease [adjusted odds ratio (AOR) = 0.001, 95% confidence interval (CI) = (0.000–0.016)], internal parasites [AOR = 0.003, 95% CI = (0.000–0.046)], shortage of grazing land [AOR = 0.017, 95% CI = (0.002–0.148)], summer season of calving [AOR = 0.012, 95% CI = (0.002–0.088)], overall cattle herd composition [AOR = 0.002, 95% CI = (0.000–0.025)], straw shed [AOR = 0.046, 95% CI = (0.006–0.327)] and open yard [AOR = 0.003, 95% CI = (0.000–0.183)] housing conditions were significantly associated with milk production status at p < 0.001 and p < 0.05.ConclusionThe current study indicated that milk production status was poor. Therefore, suitable government policy support and provision of subsidies, genuine participation of dairy producers with governmental and non‐governmental organizations are imperative to improve livestock productivity. Furthermore, future research and development actions should find solutions to decrease the bottlenecks so that the massive potentials of the area could be exploited to its maximum and could advance the livelihood of the community.

Influence of breed and parasite challenge on the immune response to naturally acquired intestinal nematode infection in sheep.

Natural infection by intestinal nematodes was assessed in Santa Ines and Ile de France sheep breeds, over a five-month grazing period, with emphasis on the development of the immune response in lambs under two anthelmintic treatment regimens. Nineteen Santa Ines and 19 Ile de France male lambs were allocated into two treatment groups: animals under suppressive treatment with anthelmintics; and animals under targeted selective treatment (TST). The nematodes Trichostrongylus colubriformis and Strongyloides papillosus showed the highest infection intensity in the TST animals in both breeds. Parasite-specific immunoglobulin G levels were significantly higher and more rapidly induced in Santa Ines lambs. Additionally, these lambs had higher levels of parasite-specific immunoglobulin A in intestinal mucus. Animals under TST had higher means of mast cells and globule leukocytes in the intestinal mucosa due to longer and greater parasite challenge in comparison with the suppressive group. A breed effect (P < 0.05) was recorded for mast cells, with Santa Ines lambs presenting the highest counts. Although Santa Ines lambs had lower intestinal nematode counts than Ile de France lambs, they had shown a large variation in T. colubriformis burden, with most of them presenting a marked worm burden, despite showing an earlier and more intense immune response to intestinal nematode infection.

Trypanosoma cruzi Affects Rhodnius prolixus Lipid Metabolism During Acute Infection

The interaction between Rhodnius prolixus and Trypanosoma cruzi has huge medical importance because it responds to the transmission of Chagas disease, a neglected tropical disease that affects about eight million people worldwide. It is known that trypanosomatid pathogens depend on active lipid endocytosis from the insect host to meet growth and differentiation requirements. However, until now, knowledge on how the parasite affects the lipid physiology of individual insect organs was largely unknown. Herein, the biochemical and molecular dynamics of the triatomine R. prolixus lipid metabolism in response to T. cruzi acute infection were investigated. A qRT-PCR approach was used to determine the expression profile of 12 protein-coding genes involved in R. prolixus lipid physiology. In addition, microscopic and biochemical assays revealed the lipid droplet profile and the levels of the different identified lipid classes. Finally, spectrometry analyses were used to determine fatty acid and sterol composition and their modulation towards the infection. T. cruzi infection downregulated the transcript levels of protein-coding genes for lipid biosynthetic and degrading pathways in individual triatomine organs. On the other hand, upregulation of lipid receptor transcripts indicates an attempt to capture more lipids from hemolymphatic lipoproteins. Consequently, several lipid classes (such as monoacylglycerol, diacylglycerol, triacylglycerol, cholesteryl ester, phosphatidylcholine, and phosphatidylethanolamine) were involved in the response to the parasite challenge, although modulating only the insect fat body. T. cruzi never leaves the insect gut and yet it modulates non-infected tissues, suggesting that the association between the parasite and the vector organs is reached by cell signaling molecules. This hypothesis raises several intriguing issues to inspire future studies in the parasite-vector interaction field.

15 Differentially Expressed Long Non-coding RNA Associated with Gastrointestinal Nematode Infection in Divergent Immune Response Sheep

Abstract Gastrointestinal nematode infection is one of the major production problems for sheep producers worldwide due its high incidence, morbidity, and mortality in affected flocks. The study of long non-coding RNA (lncRNA) in liver tissue of high (HIR) and low immune responder (LIR) sheep to GINs using RNA-Sequencing technology may provide a better understanding of the gene regulation mechanism associated with the host response to the infection. The aim of this study was to identify differentially expressed (DE) lncRNA between HIR and LIR natural infested sheep and control group. Liver tissue samples from the 13 divergent animals (out of a population of 211) based on their immunoglobulin G levels after vaccination using Hen Egg White (HEW) Lysozyme, and immature abomasum worm counts [HIR (&amp;gt; 4000) (n = 5), LIR (&amp;lt; 1500) (n=5) and control (no parasite challenge) (n=4) groups] were used to perform transcriptomic analysis using RNA-Sequencing. The “Large Gap read mapping “and “Transcript Discovery” tools from CLC Genomics Workbench 20.0.4 (CLC Bio, Aarhus, Denmark), were used to map reads to a reference genome (Oar_rambouillet_v1.0) and transcript discovery, respectively. The FEELnc software was used to identify, from predicted transcript model, potential lncRNAs and classify those transcripts into intro putative lncRNAs and protein coding RNAs. As preliminary results, 8 and 48 DE lncRNAs for HIR and LIR compared to control group were identified, respectively using an adjusted p-value False Discovery Rate (FDR) &amp;lt; 0.05 and Fold change (FC) abs &amp;gt; 2. Functional analyses using the list of DE lncRNAs identified metabolic pathways related to immune function. In depth analysis will help to better understand the physiological mechanisms of resilience of high immune sheep.

Canine Leishmaniosis in Greece: An Updated Countrywide Serological Study and Associated Risk Factors.

Canine leishmaniosis (Leishmania infantum) is a zoonotic disease that affects dogs worldwide. Greece is enzootic for this disease, and updated data for its current distribution are of major importance. The aim of this cross-sectional serological study was primarily to update the current knowledge of Leishmania infantum seropositivity status within the asymptomatic Greek canine population and, furthermore, to assess the possible climatological and other risk factors. In total, sera of 1265 asymptomatic dogs were collected from all prefectures of the country. A questionnaire that included all individual dog information was completed for all animals. The Speed Leish K® canine Leishmania antibody test kit (BVT Groupe Virbac, France) was employed. Potential risk factors were evaluated utilizing logistic regression models. Overall, 13.8% (n = 175) of the sampled dogs were seropositive to Leishmania infantum originating from all geographical departments of the country, whereas most prefectures had at least one seropositive animal. Outdoor living, high mean humidity, low mean wind speed and high total annual rainfall were found to increase the seropositivity status against the parasite. Conclusively, Leishmania infantum remains a common parasite challenge in the asymptomatic canine population of Greece, and therefore, its early diagnosis and effective prevention are significant in the country.

Molecular Characterization and Expression Analysis of Intercellular Adhesion Molecule-1 (ICAM-1) Genes in Rainbow Trout (Oncorhynchus mykiss) in Response to Viral, Bacterial and Parasitic Challenge.

The intercellular adhesion molecule-1 (ICAM-1), known as CD54, is a transmembrane cell surface glycoprotein that interacts with two integrins (i.e., LFA-1 and Mac-l) important for trans-endothelial migration of leukocytes. The level of ICAM-1 expression is upregulated in response to some inflammatory stimulations, including pathogen infection and proinflammatory cytokines. Yet, to date, our knowledge regarding the functional role of ICAM-1 in teleost fish remains largely unknown. In this study, we cloned and characterized the sequence of ICAM-1 in rainbow trout (Oncorhynchus mykiss) for the first time, which exhibited that the molecular features of ICAM-1 in fishes were relatively conserved compared with human ICAM-1. The transcriptional level of ICAM-1 was detected in 12 different tissues, and we found high expression of this gene in the head kidney, spleen, gills, skin, nose, and pharynx. Moreover, upon stimulation with infectious hematopoietic necrosis virus (IHNV), Flavobacterium columnare G4 (F. columnare), and Ichthyophthirius multifiliis (Ich) in rainbow trout, the morphological changes were observed in the skin and gills, and enhanced expression of ICAM-1 mRNA was detected both in the systemic and mucosal tissues. These results indicate that ICAM-1 may be implicated in the mucosal immune responses to viral, bacterial, and parasitic infections in teleost fish, meaning that ICAM-1 emerges as a master regulator of mucosal immune responses against pathogen infections in teleost fish.

Is selection for resistance and resilience to Haemonchus contortus possible in Morada Nova sheep?

Gastrointestinal nematodes (GIN) are a major constraint for sheep production. Selection of animals resistant to GIN infection is an effective and sustainable control strategy. In this study, the possibility of selection for resistance and resilience, as well as potential correlated responses, were evaluated. A total of 256 lambs and 123 ewes were submitted to two consecutive independent artificial infections with 4,000 infective larvae (L3) of Haemonchus contortus. Records of faecal egg count (FEC), packed cell volume (PCV), and body weight (BW) were taken serially until day 42 after infection in both challenges. Estimates of genetic parameters were obtained for individual records (in lambs) and for overall traits (in lambs and ewes) using mixed models. Phenotypic and genetic correlations were estimated between traits in lambs, with genetic correlations being based on estimated breeding values (EBVs). In addition, correlations between FEC in lambs and FEC in ewes, as well as PCV in lambs and ewes, were calculated. Heritability estimates for FEC in the first and second parasite challenge were, respectively, 0.25 ± 0.18 and 0.46 ± 0.19 for lambs, and 0.00 ± 0.09 and 0.20 ± 0.16 for ewes. For PCV, heritability estimates were 0.23 ± 0.14 and 0.32 ± 0.16 for lambs and 0.13 ± 0.11 and 0.37 ± 0.18 for ewes. For the overall daily weight gain (DWG), the heritability estimate was 0.70 ± 0.21. No significant genetic correlation was found between DWG and the other traits, while there was a negative genetic correlation between FEC and PCV (-0.70 ± 0.03). Genetic correlations of FEC and PCV between lambs and ewes were 0.36 ± 0.08 and 0.42 ± 0.08, respectively. The results of the present study suggest that selection for low FEC, high PCV and high BW/DWG is possible in Morada Nova lambs. Furthermore, selection for low FEC should have a correlated response on PCV (leading to higher PCV), while no correlated response is expected on DWG. Selection for low FEC in lambs would, in future generations, lead to higher resistance of lambs and also of ewes. Simultaneous selection for all three traits is possible in Morada Nova lambs, which would increase performance, decrease losses due to parasite infection and reduce the need for anthelmintic treatments, with beneficial consequences regarding selection pressure on parasite populations and productivity.

Monoclonal Antibodies Targeting an Opisthorchis viverrini Extracellular Vesicle Tetraspanin Protect Hamsters against Challenge Infection.

Opisthorchis viverrini causes severe pathology in the bile ducts of infected human hosts, and chronic infection can culminate in bile duct cancer. The prevention of infection by vaccination would decrease opisthorchiasis-induced morbidity and mortality. The tetraspanin protein, Ov-TSP-2, is located on the membrane of secreted extracellular vesicles (EVs), and is a candidate antigen for inclusion in a subunit vaccine. To address the role of anti-Ov-TSP-2 antibodies in protection, we assessed the protective capacity of anti-Ov-TSP-2 monoclonal antibodies (mAbs) against opisthorchiasis. Two anti-TSP-2 IgM mAbs, 1D6 and 3F5, and an isotype control were passively transferred to hamsters, followed by parasite challenge one day later. Hamsters that received 3F5 had 74.5% fewer adult flukes and 67.4% fewer eggs per gram of feces compared to hamsters that received the control IgM. Both 1D6 and 3F5 (but not the control IgM) blocked the uptake of fluke EVs by human bile duct epithelial cells in vitro. This is the first report of passive immunization against human liver fluke infection, and the findings portend the feasibility of antibody-directed therapies for liver fluke infection, bolstering the selection of TSPs as components of a subunit vaccine for opisthorchiasis and fluke infections generally.

Epidemiology of Trypanosomiasis in Wildlife-Implications for Humans at the Wildlife Interface in Africa.

While both human and animal trypanosomiasis continue to present as major human and animal public health constraints globally, detailed analyses of trypanosome wildlife reservoir hosts remain sparse. African animal trypanosomiasis (AAT) affects both livestock and wildlife carrying a significant risk of spillover and cross-transmission of species and strains between populations. Increased human activity together with pressure on land resources is increasing wildlife–livestock–human infections. Increasing proximity between human settlements and grazing lands to wildlife reserves and game parks only serves to exacerbate zoonotic risk. Communities living and maintaining livestock on the fringes of wildlife-rich ecosystems require to have in place methods of vector control for prevention of AAT transmission and for the treatment of their livestock. Major Trypanosoma spp. include Trypanosoma brucei rhodesiense, Trypanosoma brucei gambiense, and Trypanosoma cruzi, pathogenic for humans, and Trypanosoma vivax, Trypanosoma congolense, Trypanosoma evansi, Trypanosoma brucei brucei, Trypanosoma dionisii, Trypanosoma thomasbancrofti, Trypanosma elephantis, Trypanosoma vegrandis, Trypanosoma copemani, Trypanosoma irwini, Trypanosoma copemani, Trypanosoma gilletti, Trypanosoma theileri, Trypanosoma godfreyi, Trypansoma simiae, and Trypanosoma (Megatrypanum) pestanai. Wildlife hosts for the trypansomatidae include subfamilies of Bovinae, Suidae, Pantherinae, Equidae, Alcephinae, Cercopithecinae, Crocodilinae, Pteropodidae, Peramelidae, Sigmodontidae, and Meliphagidae. Wildlife species are generally considered tolerant to trypanosome infection following centuries of coexistence of vectors and wildlife hosts. Tolerance is influenced by age, sex, species, and physiological condition and parasite challenge. Cyclic transmission through Glossina species occurs for T. congolense, T. simiae, T. vivax, T. brucei, and T. b. rhodesiense, T. b. gambiense, and within Reduviid bugs for T. cruzi. T. evansi is mechanically transmitted, and T. vixax is also commonly transmitted by biting flies including tsetse. Wildlife animal species serve as long-term reservoirs of infection, but the delicate acquired balance between trypanotolerance and trypanosome challenge can be disrupted by an increase in challenge and/or the introduction of new more virulent species into the ecosystem. There is a need to protect wildlife, animal, and human populations from the infectious consequences of encroachment to preserve and protect these populations. In this review, we explore the ecology and epidemiology of Trypanosoma spp. in wildlife.

MPLA and AddaVax® Adjuvants Fail to Promote Intramuscular LaAg Vaccine Protectiveness against Experimental Cutaneous Leishmaniasis.

There is so far no vaccine approved for human leishmaniasis, mainly because of the lack of appropriate adjuvants. This study aimed to evaluate in mice the capacity of a mixture of monophosphoryl lipid A (MPLA) and AddaVax® adjuvants in enhancing the efficacy of a Leishvacin®-like vaccine comprised of Leishmania amazonensis whole antigens (LaAg). For that, mice were immunized with LaAg plus MPLA/AddaVax® by the intramuscular route (i.m.) prior to challenge with 2 × 105 and 2 × 106 living parasites. Immunization with LaAg alone reduced the lesion growth of the 2 × 105-challenged mice only in the peak of infection, but that was not accompanied by reduced parasite load, and thus not considered protective. Mice given a 2 × 106 -challenge were not protected by LaAg. The association of LaAg with MPLA/AddaVax® was able to enhance the cutaneous hypersensitivity response compared with LaAg alone. Despite this, there was no difference in proliferative cell response to antigen ex vivo. Moreover, regardless of the parasite challenge, association of LaAg with MPL/AddaVax® did not significantly enhance protection in comparison with LaAg alone. This work demonstrated that MPL/AddaVax® is not effective in improving the efficacy of i.m. LaAg vaccine against cutaneous leishmaniasis.

Dietary and Plasmodium challenge effects on the cuticular hydrocarbon profile of Anopheles albimanus

The cuticular hydrocarbon (CHC) profile reflects the insects’ physiological states. These include age, sex, reproductive stage, and gravidity. Environmental factors such as diet, relative humidity or exposure to insecticides also affect the CHC composition in mosquitoes. In this work, the CHC profile was analyzed in two Anopheles albimanus phenotypes with different degrees of susceptibility to Plasmodium, the susceptible-White and resistant-Brown phenotypes, in response to the two dietary regimes of mosquitoes: a carbon-rich diet (sugar) and a protein-rich diet (blood) alone or containing Plasmodium ookinetes. The CHCs were analyzed by gas chromatography coupled to mass spectrometry or flame ionization detection, identifying 19 CHCs with chain lengths ranging from 20 to 37 carbons. Qualitative and quantitative changes in CHCs composition were dependent on diet, a parasite challenge, and, to a lesser extent, the phenotype. Blood-feeding caused up to a 40% reduction in the total CHC content compared to sugar-feeding. If blood contained ookinetes, further changes in the CHC profile were observed depending on the Plasmodium susceptibility of the phenotypes. Higher infection prevalence caused greater changes in the CHC profile. These dietary and infection-associated modifications in the CHCs could have multiple effects on mosquito fitness, impacts on disease transmission, and tolerance to insecticides.

Thrombospondin‐1 Plays an Essential Role in Hippo Signaling During the Early Phase of Trypanosoma cruzi Infection in Heart Endothelial Cells

Trypanosoma cruzi is an obligate intracellular hemoflagellate parasite which cause Chagas disease. This neglected tropical disease causes severe morbidity and mortality in endemic regions. 30% of chronically infected people suffer from cardiac disorder. Chagas heart diseasecan lead to sudden death due to cardiac arrhythmias or progressive heart failure. Invasive trypomastigotes released from infected cells can be carried in vascular endothelial system to infect neighboring and distant cells. During the process of cellular infection, the parasite induces host cells to increase the levels of host thrombospondin-1 (TSP-1) to enable the process of infection. TSP-1 plays important roles in the functioning of vascular cells including vascular endothelial cells with important implications in cardiovascular health. Hippo signaling components, including yes-associated protein 1 (YAP)/transcriptional coactivator with PDZ-binding motif (TAZ), which are upstream of TSP-1 have been linked to the pathophysiology of heart disease. The molecular mechanisms by which T. cruzi signals and eventually infect heart endothelial cells remain unknown. To evaluate the importance of TSP-1 expression in heart endothelial cells during the process of T. cruzi infection, we challenge heart endothelial cells prepared from Wild Type and TSP-1 Knockout mouse (MHEC) to invasive T. cruzi trypomastigotes at different time points and evaluated changes in the Hippo signaling cascade using immunoblotting and immunofluorescence assays. We found that the parasite turned off the hippo signaling pathway in TSP-1KO heart endothelial cells. The levels of SAV1 and MOB1A increased to a maximum of 2.70±0.23 and 5.74±1.45 fold at 3 and 6 h, respectively, in TSP-1KO MHEC compared to WT MHEC following parasite challenge. This was accompanied by a significant continuous increase in the nuclear translocation of downstream effector molecule YAP to a maximum mean nuclear fluorescence intensity of 10.14±0.40 at 6 h compared to wild type cells. Besides this, we found that increased nuclear translocated YAP significantly colocalized with the transcription co-activator molecule pan-TEAD with a maximum Pearson's correlation coefficient of 0.51±0.06 at 6 h compared to YAP-Pan-TEAD colocalization in the WT MHEC, which decreased significantly with to a minimum Pearson's correlation coefficient of 0.30±0.01 at 6 h. Our finding suggests that upregulated TSP-1 is essential for the regulation of the hippo signaling pathway during the early phase of infection. These studies advance our understanding of the Host-Parasite molecular interactions occurring between heart endothelial cells and T. cruzi in the presence and absence of TSP-1. Taken together, this further supports our concept that an increase in the level of TSP-1 induced by the parasite is essential for T. cruzi infection and pathogenesis.