Marine Natural Products as Novel Treatments for Parasitic Diseases.

Parasitic diseases including malaria, leishmaniasis, and trypanosomiasis have received significant attention due to their severe health implications, especially in developing countries. Marine natural products from a vast and diverse range of marine organisms such as sponges, corals, molluscs, and algae have been found to produce unique bioactive compounds that exhibit promising potent properties, including antiparasitic, anti-Plasmodial, anti-Leishmanial, and anti-Trypanosomal activities, providing hope for the development of effective treatments. Furthermore, various techniques and methodologies have been used to investigate the mechanisms of these antiparasitic compounds. Continued efforts in the discovery and development of marine natural products hold significant promise for the future of novel treatments against parasitic diseases.

Nest architecture influences host use by avian brood parasites and is shaped by coevolutionary dynamics.

Brood (social) parasites and their hosts exhibit a wide range of adaptations and counter-adaptations as part of their ongoing coevolutionary arms races. Obligate avian brood parasites are expected to use potential host species with more easily accessible nests, while potential hosts are expected to evade parasitism by building more concealed nests that are difficult for parasites to enter and in which to lay eggs. We used phylogenetically informed comparative analyses, a global database of the world's brood parasites, their host species, and the design of avian host and non-host nests (approx. 6200 bird species) to examine first, whether parasites preferentially target host species that build open nests and, second, whether host species that build enclosed nests are more likely to be targeted by specialist parasites. We found that species building more accessible nests are more likely to serve as hosts, while host species with some of the more inaccessible nests are targeted by more specialist brood parasites. Furthermore, evolutionary-transition analyses demonstrate that host species building enclosed nests frequently evolve to become non-hosts. We conclude that nest architecture and the accessibility of nests for parasitism represent a critical stage of the ongoing coevolutionary arms race between avian brood parasites and their hosts.

Centrosomal protein of 192 kDa (Cep192) fragment possesses bactericidal and parasiticidal activities in Larimichthys crocea.

A novel AMP Lc1773, derived from centrosomal protein of 192 kDa (Cep192), was isolated from Larimichthys crocea using a Bacillus subtilis system. After cDNA libraries construction, repeating selection of B. subtilis system, extraction of extracellular protein, and expression of recombinant protein, we found that B. subtilis 1773, extracellular protein, and rLc1773 had a strong potential to kill Vibrio. parahaemolyticus and V. vulnificus. Further analysis of the antibacterial mechanism revealed that rLc1773 not only disrupted the integrity of bacterial membrane (as confirmed by SEM, TEM, and confocal microscopy observation, and flow cytometry assays), resulting in bacterial cell membrane pore conformation, bacterial rupture, and leakage of cellular contents, but also targeted to block protein synthesis rather than damage nucleic acids (as confirmed by SDS-PAGE, enzyme expression, and gel retardation assays). In addition, rLc1773 had the ability to kill parasite Scuticociliatida in a high rate and low concentration. Critically, the antibacterial activity of rLc1773 had good thermal stability and UV radiation tolerance, but it was affected by pH 9-11 and diverse enzyme to some extent. Lc1773 had neither hemolysis on fish, shrimp, and rabbit erythrocytes，nor significant cytotoxicity. To our knowledge, Cep192 fragment was first demonstrated to possess bactericidal and parasiticidal activities.

DOES PREENING BEHAVIOR REDUCE THE PREVALENCE OF AVIAN FEATHER LICE (PHTHIRAPTERA: ISCHNOCERA)?

Animals defend themselves against parasites in many ways. Defenses, such as physiological immune responses, are capable of clearing some infections. External parasites that do not feed on blood, however, are not controlled by the physiological immune system. Instead, ectoparasites like feather-feeding lice (Phthiraptera: Ischnocera) are primarily controlled by behavioral defenses such as preening. Here we test the hypothesis that birds able to preen are capable of clearing infestations of feather lice. We experimentally manipulated preening ability in a captive population of rock pigeons (Columba livia) that were infested with identical numbers of feather lice (Columbicola columbae or Campanulotes compar or both). We then monitored the feather louse infestations for 42 wk. Birds with impaired preening remained infested throughout the experiment; in contrast, the prevalence of lice on birds that could preen normally decreased by 50%. These data indicate that it is indeed possible for birds to clear themselves of feather lice, and perhaps other ectoparasites, by preening. We note, however, that captive birds spend more time preening than wild birds, and that they are less likely to be reinfested than wild birds. Thus, additional studies are necessary to determine under what circumstances wild birds can clear themselves of ectoparasites by preening.

Transcriptomic and biochemical analysis from the venom gland of the neotropical ant Odontomachus chelifer.

The genus Odontomachus is widely distributed in neotropical areas throughout Central and South America. It is a stinging ant that subdues its prey (insects) by injecting them a cocktail of toxic molecules (venom). Ant venoms are generally composed of formic acid, alkaloids, hydrocarbons, amines, peptides, and proteins. Odontomachus chelifer is an ant that inhabits neotropical regions from Mexico to Argentina. Unlike the venom of other animals such as scorpions, spiders and snakes, this ant venom has seldom been analyzed comprehensively, and their compositions are not yet completely known. In the present study, we performed a partial investigation of enzymatic and functional activities of O. chelifer ant venom, and we provide a global insight on the transcripts expressed in the venom gland to better understand their properties. The crude venom showed phospholipase A2 and antiparasitic activities. RNA sequencing (Illumina platform) of the venom gland of O. chelifer generated 61, 422, 898 reads and de novo assembly Trinity generated 50,220 contigs. BUSCO analysis against Arthropoda_db10 showed that 92.89% of the BUSCO groups have complete gene representation (single-copy or duplicated), while 4.05% are only partially recovered, and 3.06% are missing. The 30 most expressed genes in O. chelifer venom gland transcriptome included important transcripts involved in venom function such as U-poneritoxin (01)-Om1a-like (pilosulin), chitinase 2, venom allergen 3, chymotrypsin 1 and 2 and glutathione S-transferase. Analysis of the molecular function revealed that the largest number of transcripts were related to catalytic activity, including phospholipases. These data emphasize the potential of O. chelifer venom for prospection of molecules with biotechnological application.

Female Cuckoo Calls Deceive Their Hosts by Evoking Nest-Leaving Behavior: Variation under Different Levels of Parasitism.

The common cuckoo (Cuculus canorus) is an obligate brood parasite that has evolved a series of strategies to trick its hosts. The female cuckoo has been hypothesized to mimic the appearance and sounds of several raptors to deceive the hosts into exhibiting anti-predator behavior. Such behavior would relax the protection of the host nest and thus allow the female cuckoo to approach the host nest unopposed. Many anti-parasite strategies have been found to vary among geographical populations due to different parasitic pressures from cuckoos. However, the effect of female cuckoo calls related to different levels of parasitic pressure has not been examined. Here, we studied the effect of female cuckoo calls on the oriental reed warbler (Acrocephalus orientalis), one of the major hosts of the common cuckoo, in two geographical populations experiencing different levels of parasite pressure. Four kinds of sounds were played back to the hosts: the calls from female common cuckoos, male common cuckoos, sparrowhawks (Accipiter nisus), and oriental turtle doves (Streptopelia orientalis). The results showed that the female cuckoo calls induced the hosts to leave their nests more frequently than the male cuckoo or dove calls in both populations, and two populations of the hosts reacted similarly to the female cuckoo calls, implying that the function of female cuckoo calls would not be affected by the difference in parasitism rate. This study indicates that female cuckoo calls function to distract the hosts' attention from protecting their nests. However, we propose that such a deception by the female cuckoo call may not be due to the mimicry of sparrowhawk calls, but rather that the rapid cadence of the call that causes a sense of anxiety in the hosts.

Major Molecular Factors Related to Leishmania Pathogenicity.

Leishmaniasis is a major health problem with 600k - 1M new cases worldwide and 1 billion at risk. It involves a wide range of clinical forms ranging from self-healing cutaneous lesions to systemic diseases that are fatal if not treated, depending on the species of Leishmania. Leishmania sp. are digenetic parasites that have two different morphological stages. Leishmania parasites possess a number of invasive/evasive and pathoantigenic determinants that seem to have critical roles in Leishmania infection of macrophages which leads to successful intracellular parasitism in the parasitophorous vacuoles. These determinants are traditionally known as "virulence factors", and are considered to be good targets for developing specific inhibitors to attenuate virulence of Leishmania by gene deletions or modifications, thus causing infective, but non-pathogenic mutants for vaccination. Pathway of biosynthesis is critical for keeping the parasite viable and is important for drug designing against these parasites. These drugs are aimed to target enzymes that control these pathways. Accordingly, maintaining low level of parasitic infection and in some cases as a weapon to eradicate infection completely. The current paper focuses on several virulence factors as determinants of Leishmania pathogenicity, as well as the metabolites produced by Leishmania to secure its survival in the host.

Effects of neonicotinoids on honey bee autogrooming behavior against the tracheal mite Acarapis woodi.

The European honey bee, Apis mellifera, is the most common and important pollinator of crops worldwide. Honey bees are damaged by destructive parasitic mites, but they also have evolved a behavioral immune system to remove them. Exposures to neonicotinoids, however, can cause significant behavioral effects because these compounds alter the central role of nicotinic acetylcholine receptor in insect brains. In this study, we assessed the effects of three neonicotinoids that have a high toxicity to bees-imidacloprid, thiamethoxam, and clothianidin-on the behavioral immune system of honey bees. We used A. mellifera and the endoparasitic mite Acarapis woodi as a behavioral immune system model because A. mellifera can effectively remove the mite by autogrooming. Our results did not demonstrate an effect of neonicotinoid application on whether bees show autogrooming or on mite removal, but the time to initial autogrooming became shorter and the number of autogrooming attempts increased. As opposed to previous studies, our findings indicate that the honey bee response to parasitic mites becomes more sensitive after exposure to neonicotinoids.Clinical Trials Registration: Not applicable.

Inhibiting parasite proliferation using a rationally designed anti-tubulin agent.

Infectious diseases caused by apicomplexan parasites remain a global public health threat. The presence of multiple ligand-binding sites in tubulin makes this protein an attractive target for anti-parasite drug discovery. However, despite remarkable successes as anti-cancer agents, the rational development of protozoan parasite-specific tubulin drugs has been hindered by a lack of structural and biochemical information on protozoan tubulins. Here, we present atomic structures for a protozoan tubulin and microtubule and delineate the architectures of apicomplexan tubulin drug-binding sites. Based on this information, we rationally designed the parasite-specific tubulin inhibitor parabulin and show that it inhibits growth of parasites while displaying no effects on human cells. Our work presents for the first time the rational design of a species-specific tubulin drug providing a framework to exploit structural differences between human and protozoa tubulin variants enabling the development of much-needed, novel parasite inhibitors.

Environmental and individual determinants of fecal avoidance in semi-free ranging woolly monkeys (Lagothrix lagotricha poeppigii).

OBJECTIVES: Parasite selection pressures have driven the evolution of numerous behavioral defenses in host species, but recent studies revealed individual variation in their expression. As little is known about the factors causing heterogeneity among individuals in infection-avoidance behaviors, we investigated in woolly monkeys (Lagothrix lagotricha poeppigii) the influence of several environmental and individual characteristics on the tendency to avoid food contaminated by soil and by their own and conspecifics' feces. MATERIALS AND METHODS: We conducted feeding tests on 40 semi-free ranging individuals rescued from the pet trade. Using generalized linear mixed models, we investigated the effect of season, sex, age, dominance rank, and exposure to non-natural living conditions on feeding decisions. RESULTS: Woolly monkeys did not avoid soil-contaminated food and equally avoided food contaminated by their own and conspecifics' feces. Individuals varied greatly in their level of fecal avoidance. Only females exhibited strong avoidance of fecally contaminated food, but adapted their behavior to food availability, highlighting the trade-off between nutritional intake and parasite avoidance. Additionally, low-ranking females, less competitive over food resources, exhibited lower avoidance than dominant ones. Juveniles were more cautious than adults, possibly to compensate for a higher parasite susceptibility. Finally, we reported an unknown effect of exposure to non-natural living conditions on behavioral defenses, as animals kept as household pets for an extended period apparently lost their ability to avoid fecally contaminated food. CONCLUSION: We argue that striving to understand variation in infection-avoidance behaviors in natural populations is crucial to predict disease spread and inform conservation policies.

Evaluation of selected plant extracts for in vitro anti-marine leech (Zeylanicobdella arugamensis) activity

@#The aqueous extracts of thirty-four (34) tropical plants were tested in vitro for potential anti-marine leech (Zeylanicobdella arugamensis) activity. The anti-leech activity was determined by exposing 8 adult leeches (Z. arugamensis) (9.3 ± 1.5 mm, aged 15 days) to 20 μl of plant extract (0.5 g/ml) for 5 min in a 24-well plate. After 5 min of exposure, the leeches were rinsed and transferred into seawater, to enable them to revive from the effect of the extract. Leech movements were observed from time to time and the numbers of paralyzed or dead leeches were recorded at 5, 20, 30 and 240 min. The efficacy of the plant extract in killing the adult Z. arugamensis during the 5 min exposure is reflected on the anti-leech property of the extract. The anti-leech property of positive plant extracts was also determined at different exposure time (1, 3, 5 min) and dilutions (1/2 (0.25 g/ml), 1/5 (0.1 g/ml) and 1/10 (0.05 g/ml). The extracts of 4 plants (Melastoma malabathricum, Piper betle, Tetracera indica and Etlingera coccinea) demonstrated anti-leech activity. The effects of M. malabathricum, P. betle and E. coccinea extracts on the leeches were very rapid causing death as early as a few seconds upon exposure. However, all four positive plant extracts were found not effective in killing the leeches at 1/10 dilution (0.05 g/ml). A Scanning Electron Microscopy examination on leeches exposed to the positive plant extracts exhibited effects such as protruding proboscis and shrunken or swollen bodies.

COVID-19: The Immune Responses and Clinical Therapy Candidates.

The pandemic of coronavirus disease 2019 (COVID-19), with rising numbers of patients worldwide, presents an urgent need for effective treatments. To date, there are no therapies or vaccines that are proven to be effective against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Several potential candidates or repurposed drugs are under investigation, including drugs that inhibit SARS-CoV-2 replication and block infection. The most promising therapy to date is remdesivir, which is US Food and Drug Administration (FDA) approved for emergency use in adults and children hospitalized with severe suspected or laboratory-confirmed COVID-19. Herein we summarize the general features of SARS-CoV-2's molecular and immune pathogenesis and discuss available pharmacological strategies, based on our present understanding of SARS-CoV and Middle East respiratory syndrome coronavirus (MERS-CoV) infections. Finally, we outline clinical trials currently in progress to investigate the efficacy of potential therapies for COVID-19.

Is Allopreening a Stimulus-Driven Defense Against Ectoparasites?

Allopreening occurs when 1 bird preens another bird. The behavior is normally directed at the head and neck of the recipient, i.e., regions that the bird cannot self-preen. Studies of penguins, pigeons, and other groups of birds suggest that allopreening plays a role in the control of ectoparasites, such as ticks and feather lice. However, it is not known whether allopreening increases in response to increases in parasite load, or whether it is a programmed response that occurs independently of parasite load. We conducted a laboratory experiment using wild-caught rock pigeons (Columba livia) to test the relationship between ectoparasite load and allopreening rate. We added feather lice (Columbicola columbae) to captive pigeons and tested for changes in allopreening rates compared to control birds with no lice added. Allopreening rates did not change in response to the addition of lice. Interestingly, however, our data revealed a negative correlation between allopreening and self-preening rates.

Unconditional care from close maternal kin in the face of parasites.

Several species mitigate relationships according to their conspecifics' parasite status. Yet, this defence strategy comes with the costs of depriving individuals from valuable social bonds. Animals therefore face a trade-off between the costs of pathogen exposure and the benefits of social relationships. According to the models of social evolution, social bonds are highly kin-biased. However, whether kinship mitigates social avoidance of contagious individuals has never been tested so far. Here, we build on previous research to demonstrate that mandrills (Mandrillus sphinx) modulate social avoidance of contagious individuals according to kinship: individuals do not avoid grooming their close maternal kin when contagious (parasitized with oro-faecally transmitted protozoa), although they do for more distant or non-kin. While individuals' parasite status has seldom been considered as a trait impacting social relationships in animals, this study goes a step beyond by showing that kinship balances the effect of health status on social behaviour in a non-human primate.

Variation in Tolerance to Parasites Affects Vectorial Capacity of Natural Asian Tiger Mosquito Populations.

Globally, diseases transmitted by arthropod vectors, such as mosquitoes, remain a major cause of morbidity and mortality [1]. The defense responses of mosquito and other arthropod vectors against parasites are important for understanding disease transmission dynamics and for the development of novel disease-control strategies. Consequently, the mechanisms by which mosquitoes resist parasitic infection (e.g., immune-mediated killing) have long been studied [2, 3]. However, the ability of mosquitoes to ameliorate the negative fitness consequences of infection through tolerance mechanisms (e.g., tissue repair) has been virtually ignored (but see [4, 5]). Ignoring parasite tolerance is especially taxing in vector biology because unlike resistance, which typically reduces vectorial capacity, tolerance is expected to increase vectorial capacity by reducing parasite-mediated mortality without killing parasites [6], contributing to the recurrent emergence of vector-borne diseases and its stabilization and exacerbation. Despite its importance, there is currently no evidence for the evolution of tolerance in natural mosquito populations. Here, we use a common-garden experimental framework to measure variation in resistance and tolerance to dog heartworm (Dirofilaria immitis) between eight natural Aedes albopictus mosquito populations representing areas of low and high transmission intensity. We find significant inter-population variation in tolerance and elevated tolerance where transmission intensity is high. Additionally, as expected, we find that increased tolerance is associated with higher vectorial capacity. Consequently, our results indicate that high transmission intensity can lead to the evolution of more competent disease vectors, which can feed back to impact disease risk.

Trypanothione synthetase confers growth, survival advantage and resistance to anti-protozoal drugs in Trypanosoma cruzi.

BACKGROUND: Chagas cardiomyopathy, caused by Trypanosoma cruzi infection, continues to be a neglected illness, and has a major impact on global health. The parasite undergoes several stages of morphological and biochemical changes during its life cycle, and utilizes an elaborated antioxidant network to overcome the oxidants barrier and establish infection in vector and mammalian hosts. Trypanothione synthetase (TryS) catalyzes the biosynthesis of glutathione-spermidine adduct trypanothione (T(SH)2) that is the principal intracellular thiol-redox metabolite in trypanosomatids. METHODS AND RESULTS: We utilized genetic overexpression (TryShi) and pharmacological inhibition approaches to examine the role of TryS in T. cruzi proliferation, tolerance to oxidative stress and resistance to anti-protozoal drugs. Our data showed the expression and activity of TryS was increased in all morphological stages of TryShi (vs. control) parasites. In comparison to controls, the TryShi epimastigotes (insect stage) recorded shorter doubling time, and both epimastigotes and infective trypomastigotes of TryShi exhibited 36-71% higher resistance to H2O2 (50-1000 µM) and heavy metal (1-500 µM) toxicity. Treatment with TryS inhibitors (5-30 µM) abolished the proliferation and survival advantages against H2O2 pressure in a dose-dependent manner in both TryShi and control parasites. Further, epimastigote and trypomastigote forms of TryShi (vs. control) T. cruzi tolerated higher doses of benznidazole and nifurtimox, the drugs currently administered for acute Chagas disease treatment. CONCLUSIONS: TryS is essential for proliferation and survival of T. cruzi under normal and oxidant stress conditions, and provides an advantage to the parasite to develop resistance against currently used anti-trypanosomal drugs. TryS indispensability has been chemically validated with inhibitors that may be useful for drug combination therapy against Chagas disease.

Functional significance of cuckoo Cuculus canorus calls: responses of conspecifics, hosts and non-hosts.

Male cuckoos Cuculus canorus produce calls that differ in number of syllables depending on environmental conditions and presence of male and female conspecifics. Why different males produce so repeatable calls that vary greatly in duration among males remains an open question. We used playback of cuckoo calls with few or many syllables (hereafter short and long calls), and woodpigeon calls (a control that also produces few or many syllables), predicting that playback of longer cuckoo calls should attract more male cuckoos (if males with such calls are dominant and successfully out-compete other males due to intraspecific competition), and attract more hosts mobbing male cuckoos (cuckoos with such calls and their females attract more hosts because of an increased risk of parasitism). Because cuckoos differentially parasitize hosts away from human habitation, we also tested whether the number of syllables in cuckoo calls differed with distance from buildings. Playback showed significant effects of number of syllables in cuckoo calls, but not woodpigeon Columba palumbus calls, with an additional effect of distance from human habitation decreasing the response to playback. These findings are consistent with the hypothesis that longer cuckoo calls, especially played back near human habitation, attract more conspecifics and hosts than shorter calls. To the best of knowledge this is the first study showing that cuckoo call response modified both other cuckoo individuals, as well as hosts response.

Age, exposure and immunity.

The acquisition of immunity to malaria by an individual depends on their age and the number of infectious mosquito bites they have received.

Quantification of anti-parasite and anti-disease immunity to malaria as a function of age and exposure.

Fundamental gaps remain in our understanding of how immunity to malaria develops. We used detailed clinical and entomological data from parallel cohort studies conducted across the malaria transmission spectrum in Uganda to quantify the development of immunity against symptomatic P. falciparum as a function of age and transmission intensity. We focus on: anti-parasite immunity (i.e. ability to control parasite densities) and anti-disease immunity (i.e. ability to tolerate higher parasite densities without fever). Our findings suggest a strong effect of age on both types of immunity, not explained by cumulative-exposure. They also show an independent effect of exposure, where children living in moderate/high transmission settings develop immunity faster as transmission increases. Surprisingly, children in the lowest transmission setting appear to develop immunity more efficiently than those living in moderate transmission settings. Anti-parasite and anti-disease immunity develop in parallel, reducing the probability of experiencing symptomatic malaria upon each subsequent P. falciparum infection.

Inevitability of the emergence and persistence of genetic parasites caused by evolutionary instability of parasite-free states.

Genetic parasites, including viruses and mobile genetic elements, are ubiquitous among cellular life forms, and moreover, are the most abundant biological entities on earth that harbor the bulk of the genetic diversity. Here we examine simple thought experiments to demonstrate that both the emergence of parasites in simple replicator systems and their persistence in evolving life forms are inevitable because the putative parasite-free states are evolutionarily unstable. REVIEWERS: This article has been reviewed by Yitzhak Pilpel, Bojan Zagrovic, and Eric van Nimwegen.