A profile of research on the parasitic trypanosomatids and the diseases they cause.

The parasitic trypanosomatids cause lethal and debilitating diseases, the leishmaniases, Chagas disease, and the African trypanosomiases, with major impacts on human and animal health. Sustained research has borne fruit by assisting efforts to reduce the burden of disease and by improving our understanding of fundamental molecular and cell biology. But where has the research primarily been conducted, and which research areas have received the most attention? These questions are addressed below using publication and citation data from the past few decades.

Revealing mechanisms of infectious disease spread through empirical contact networks.

The spread of pathogens fundamentally depends on the underlying contacts between individuals. Modeling the dynamics of infectious disease spread through contact networks, however, can be challenging due to limited knowledge of how an infectious disease spreads and its transmission rate. We developed a novel statistical tool, INoDS (Identifying contact Networks of infectious Disease Spread) that estimates the transmission rate of an infectious disease outbreak, establishes epidemiological relevance of a contact network in explaining the observed pattern of infectious disease spread and enables model comparison between different contact network hypotheses. We show that our tool is robust to incomplete data and can be easily applied to datasets where infection timings of individuals are unknown. We tested the reliability of INoDS using simulation experiments of disease spread on a synthetic contact network and find that it is robust to incomplete data and is reliable under different settings of network dynamics and disease contagiousness compared with previous approaches. We demonstrate the applicability of our method in two host-pathogen systems: Crithidia bombi in bumblebee colonies and Salmonella in wild Australian sleepy lizard populations. INoDS thus provides a novel and reliable statistical tool for identifying transmission pathways of infectious disease spread. In addition, application of INoDS extends to understanding the spread of novel or emerging infectious disease, an alternative approach to laboratory transmission experiments, and overcoming common data-collection constraints.

Cryptobia iubilans Infections in Discus Fish in Trinidad and Tobago.

Discus (Symphysodon spp.) are costly and prized specimens in the international ornamental fish trade. The majority of discus submitted to the Aquatic Animal Health Unit at the University of the West Indies School of Veterinary Medicine for necropsy between September 2010 and September 2015 had lesions consistent with Cryptobia iubilans infection, thus prompting this study. To determine the prevalence of the flagellated gastrointestinal protozoan C. iubilans in discus fish, 32 discus were sourced from 10 suppliers, including breeders, importers, and hobbyists across Trinidad. Fish were euthanized, and the internal organs, particularly the stomach and intestine, were observed under a light microscope for characteristic granulomatous lesions and/or live C. iubilans parasites. All wet-mount slides on which granulomas were observed were also Ziehl-Neelsen acid-fast stained to presumptively exclude the presence of Mycobacterium spp., the main differential when diagnosing C. iubilans-associated granulomatous gastritis or to determine the presence of dual infections. Further histological analyses were performed on stomach and intestinal sections, and transmission electron microscopy was used to confirm the parasite in stomach sections. The prevalence of C. iubilans infection was found to be 81.3%, and the prevalence of presumptive dual infections with Mycobacterium spp. was found to be 21.9%. To the best of our knowledge, this is the first documented study of C. iubilans infections in the wider Caribbean region.

Next-Generation Molecular Surveillance of TriTryp Diseases.

Elimination programs targeting TriTryp diseases (Leishmaniasis, Chagas' disease, human African trypanosomiasis) significantly reduced the number of cases. Continued surveillance is crucial to sustain this progress, but parasite molecular surveillance by genotyping is currently lacking. We explain here which epidemiological questions of public health and clinical relevance could be answered by means of molecular surveillance. Whole-genome sequencing (WGS) for molecular surveillance will be an important added value, where we advocate that preference should be given to direct sequencing of the parasite's genome in host tissues instead of analysis of cultivated isolates. The main challenges here, and recent technological advances, are discussed. We conclude with a series of recommendations for implementing whole-genome sequencing for molecular surveillance.

Canine vector-borne protozoa: Molecular and serological investigation for Leishmania spp., Trypanosoma spp., Babesia spp., and Hepatozoon spp. in dogs from Northern Algeria.

Dogs are competent reservoirs/hosts of several protozoan pathogens transmitted by blood-feeding arthropods. Throughout their long history of domestication, they have served as a link for the exchange of parasites among livestock, wildlife, and humans and therefore remain an important source of emerging and re-emerging diseases. In Algeria, while canine leishmaniosis (CanL) is well known to be endemic, no data are available on other vector-borne protozoans. Here, we investigate the occurrence and diversity of trypanosomes, piroplasms and Hepatozoon spp. and update the epidemiological status of CanL in dogs from Kabylia, northern Algeria. A total of 227 dogs from three regions of Kabylia were enrolled, including 77 dogs with clinical signs. Dogs were clinically examined and were tested for L. infantum antibodies using a Rapid Immuno-Migration (RIM™) and a quantitative indirect Immunofluorescence Antibody Test (IFAT). PCR screening and sequencing were performed for vector-borne protozoa. Sixty two percent (141/227) of dogs presented at least one infection, whereas 26% (59/227) were co-infected. L. infantum antibodies were detected in 35.7% (81/227) of dogs including 88.7% (68/77) of sick dogs. Molecular investigation revealed prevalence of: 6.6% (15/227), 13.2% (30/227), 41% (93/227) for Trypanosoma spp., B. vogeli and H. canis, respectively. T. evansi (3.1%) and potential new subspecies of T. congolense had been identified. Dog''s clinical status correlates positively with L. infantum antibody titers and the presence of co-infections. Susceptibility to CanL varied according to the dog's aptitude and guard dogs were more infected (51%) (P-value = .001). B. vogeli infection was more frequent in juveniles than adults (32% vs 9%, P-value < .001) and in females than males (21% vs 10%, P-value = .02). To the authors' knowledge, this is the first report on vector-borne protozoa infected dogs in Algeria. Current results are important not only for animal health, but also to avoid serious public health and livestock problems.

Non-Leishmania Parasite in Fatal Visceral Leishmaniasis-Like Disease, Brazil.

Through whole-genome sequencing analysis, we identified non-Leishmania parasites isolated from a man with a fatal visceral leishmaniasis-like illness in Brazil. The parasites infected mice and reproduced the patient's clinical manifestations. Molecular epidemiologic studies are needed to ascertain whether a new infectious disease is emerging that can be confused with leishmaniasis.

Ichthyobodo spp. Infection in Meagre (Argyrosomus regius) from Turkey: Parasitological and Pathological Findings.

OBJECTIVE: The aim of this study is to describe the first report of Ichthyobodo spp. infection in meagre (Argyrosomus regius) fry in a marine aquaculture facility in Turkey. METHODS: The material of the study was composed of 30 meagre A. regius in 2-3 g weight taken from the fry adaptation unit of a fish farm in the Aegean Sea. In this study, parasitological and pathological examinations were performed on the meagre. Ichthyobodo spp. was determined on the body surfaces and gills. RESULTS: Pathological examination revealed grayish mucous and erosions between the pin head and lentin over the skin of the examined specimens. Microscopic examinations revealed significant spongiosis, vacuolar degeneration, and hyperplasia in epidermal malpighian cells and hyperplasia in goblet cells. CONCLUSION: In the present study, Ichthyobodo spp. infection was for the first time determined in an alternative cultured meagre in Turkey.

Seasonal variation in Azumiobodo hoyamushi infection among benthic organisms in the southern coast of Korea.

BACKGROUND: Recent studies have reported that soft tunic syndrome (STS) in the edible ascidian Halocynthia roretzi is caused by the kinetoplastid parasite Azumiobodo hoyamushi. In this study, we attempted to detect and quantify the pathogen in benthic animals. METHODS: Four species of ascidians, three species of echinoderms, two species of bivalves, one species each of sponge and algae, as well as seawater, were collected in 2014 and 2015 from an ascidian farm on the southern coast of Korea by SCUBA diving. Samples were collected from ascidian hanging culture ropes or the sea bottom. Inhalent siphons were excised for the analysis of ascidians, and soft body tissues were excised from the other species. Membrane filters were used to filter collected seawater. Tissues and membrane filters were analysed using culture testing, PCR testing, and qPCR diagnoses. RESULTS: Only organisms belonging to Ascidiacea are susceptible to A. hoyamushi infection. The infection rate (% infected of the total number collected) and infection intensity (number of cells infected/g tissue wet weight) varied depending on the seasonal variation in seawater temperatures. Most ascidians examined were infected with A. hoyamushi and showed higher infection intensity in cold water seasons (April 2014 and February 2015), followed by a dramatic drop during warm water seasons (August and November, 2014). In addition, infection intensity of A. hoyamushi during the warm water period was higher in ascidians from the sea bottom than those from the hanging culture rope. CONCLUSIONS: Among benthic organisms that inhabit the southern coast of Korea, most ascidians are susceptible to A. hoyamushi infection. Seasonal cycle of infection rates and intensities of the pathogen correspond well with the STS disappearance and onset cycle observed in ascidian farms. The high intensity of A. hoyamushi infection in the ascidians on the sea bottom of ascidian farms during summer suggest further studies on the role of the pathogen in resumption of STS occurrence in late fall or early winter in the southern coast of Korea.

Implications of co-infection of Leptomonas in visceral leishmaniasis in India.

Protozoan parasites Leishmania donovani (family: Trypanosomatidae) cause fatal visceral leishmaniasis (VL) and the infection relapses in apparently cured population as post kala-azar dermal leishmaniasis (PKDL) in the Indian subcontinent. In recent years co-infection of another Trypanosomatid parasite Leptomonas with L. donovani during VL/PKDL in this region has become prominent. The observation of clinically lesser-known insect parasite, Leptomonas in leishmaniasis is intriguing to researchers. The presence of Leishmania look alike Leptomonas in the cultures of clinical isolates of Leishmania has been worrisome to those, who prefer to work with pure Leishmania cultures for drug and vaccine development or immune response studies. The exact implications of such a co-habitation, which might lead to a delay in the diagnostics of VL and elevate mortality, need a thorough investigation. Also whether Leptomonas is involved in leishmaniasis manifestation needs to be ascertained. Thus we are currently witnessing a new paradigm of a parasitic co-infection in VL/PKDL cases in India and this review outlines various opportunities for further research in understanding such emerging co-infection.

Host specificity, pathogenicity, and mixed infections of trypanoplasms from freshwater fishes.

This work summarizes the results of the 8-year study focused on Trypanoplasma sp. parasitizing freshwater fishes in the vicinity of Kyiv, Ukraine. Out of 570 fish specimens of 2 different species analyzed, 440 individuals were found to be infected. The prevalence of infection ranged from 24 % in Abramis brama Linnaeus (freshwater bream) to 100 % in Cobitis taenia Linnaeus (spined loach). The level of parasitemia also varied between moderate in freshwater bream and very high in spined loach. Interestingly, no clinical manifestations of trypanoplasmosis were observed even in extremely heavily infected C. taenia. We hypothesize that different species may differ in evolutionary timing allowing for reciprocal adaptation of the members of the "host-parasite" system. Molecular analysis of the 18S rRNA sequences revealed that several specimens were simultaneously infected with at least two different trypanoplasm species. To the best of our knowledge, this is the first report of the mixed infection with fish trypanoplasms.

Notes on the occurrence of Trypanosoma sp. (Kinetoplastida: Trypanosomatidae) in freshwater fishes from South Africa.

A total of 257 fishes from four families, Clariidae, Cichlidae, Cyprinidae and Schilbeidae were collected from three localities: the Sand River Dam, Swaziland; the Nylsvlei Nature Reserve, South Africa and the Vaal Dam and Vaal River Barrage, South Africa. Only fishes (n= 154) from Clariidae and Cichlidae were found to be infected with trypanosomes. A total of 221 Clarias gariepinus (Burchell 1822) were collected from the Vaal Dam and Vaal Barrage area, South Africa. Of these, 74%(89/121) were infected with trypanosomes from the Vaal Dam and 63%(63/100) from the Vaal River Barrage, with no seasonal infection pattern. A prevalence of 25%(1/4) was found in C. gariepinus from the Sand River Dam, Swaziland, and a 50% (1/2) prevalence was found in Tilapia sparrmanii from the Nylsvlei Nature Reserve, South Africa. Standard measurements conformed closely to the morphometric and morphological descriptions of Trypanosoma mukasai. This article provides new locality records for T. mukasai from the Vaal Dam, Vaal River Barrage and Nylsvlei Nature Reserve (South Africa) and the Sand River Dam (Swaziland). Tilapia sparrmanii collected in the Sand River Dam in Swaziland is also noted as a new host record.

Molecular tools for the detection and identification of Ichthyobodo spp. (Kinetoplastida), important fish parasites.

Ichthyobodo spp. are ectoparasitic flagellates of fish that may cause disease (ichthyobodosis), a common problem affecting the aquaculture industry worldwide. Ichthyobodosis in farmed fish is often associated with a range of other infectious agents and diagnosis in for example gill disease may be difficult. Sensitive and effective methods for detection and identification of Ichthyobodo spp. are needed to aid diagnosis of ichthyobodosis and epizootiological studies on Ichthyobodo spp. We have designed a specific quantitative real-time PCR assay targeting SSU rDNA for the detection of Ichthyobodo spp. infections. Also, several novel primer sets are presented for use in identification of Ichthyobodo spp. through PCR and sequencing. These PCR methods have been optimized and tested on samples from wild caught and farmed fish from different geographical areas in Norway. The real-time PCR assay has been tested for sensitivity and efficiency, and we present data demonstrating its use for absolute quantification of Ichthyobodo salmonis in tissue samples through RT-qPCR and qPCR. We demonstrate the use of the described set of molecular tools for the detection and sequencing of Ichthyobodo spp. from farmed and wild fish, and also show that they may aid the discovery of new Ichthyobodo species. The detection of light Ichthyobodo spp. infections through microscopy is time consuming and less sensitive compared to PCR methods. Initial real-time PCR testing and subsequent sequencing of positive samples is a powerful method that will increase diagnostic precision, aid carrier detection and promote species discoveries in the Ichthyobodonidae. Our preliminary observations indicate a high Ichthyobodo spp. diversity.

Interspecific geographic distribution and variation of the pathogens Nosema bombi and Crithidia species in United States bumble bee populations.

Several bumble bee (Bombus) species in North America have undergone range reductions and rapid declines in relative abundance. Pathogens have been suggested as causal factors, however, baseline data on pathogen distributions in a large number of bumble bee species have not been available to test this hypothesis. In a nationwide survey of the US, nearly 10,000 specimens of 36 bumble bee species collected at 284 sites were evaluated for the presence and prevalence of two known Bombus pathogens, the microsporidium Nosema bombi and trypanosomes in the genus Crithidia. Prevalence of Crithidia was ≤10% for all host species examined but was recorded from 21% of surveyed sites. Crithidia was isolated from 15 of the 36 Bombus species screened, and were most commonly recovered from Bombus bifarius, Bombus bimaculatus, Bombus impatiens and Bombus mixtus. Nosema bombi was isolated from 22 of the 36 US Bombus species collected. Only one species with more than 50 sampled bees, Bombus appositus, was free of the pathogen; whereas, prevalence was highest in Bombus occidentalis and Bombus pensylvanicus, two species that are reportedly undergoing population declines in North America. A variant of a tetranucleotide repeat in the internal transcribed spacer (ITS) of the N. bombi rRNA gene, thus far not reported from European isolates, was isolated from ten US Bombus hosts, appearing in varying ratios in different host species. Given the genetic similarity of the rRNA gene of N. bombi sampled in Europe and North America to date, the presence of a unique isolate in US bumble could reveal one or more native North American strains and indicate that N. bombi is enzootic across the Holarctic Region, exhibiting some genetic isolation.

Prevalence of Colacium vesiculosum (Colaciales: Euglenophyceae) on planktonic crustaceans in a subtropical shallow lake of Argentina.

Colacium vesiculosum (Euglenophyceae) is an epibiont common on planktonic microcrustaceans of continental waters. The interaction between epibionts and substrate organisms is not very well known, particularly in subtropical environments of South America. In the present work, we analyzed the prevalence, density, biomass and attachment sites of C. vesiculosum on planktonic microcrustaceans from Paiva Lake, a subtropical lake of Argentina. With the aim to evaluate whether epibionts affect the filtering rates of Notodiaptomus spiniger, the dominant planktonic crustacean, we carried out bioassays using phytoplankton < 53 microm. Crustaceans were sampled using a PVC tube (1.2m long and 10cm in diameter), filtering 50L of water through a 53 microm-mesh. Microcrustaceans were counted in Bogorov chambers under a stereoscopic microscope. The infested organisms were separated and observed with a photonic microscope to determine density and biovolume of epibionts, by analyzing their distribution on the exoskeleton. The prevalence of C. vesiculosum was higher in adult crustaceans than in their larvae and juveniles. The most infested group was that of calanoid copepods, related to their high density. The attachment sites on the exoskeleton were found to be the portions of the body which have a higher probability of encounter with epibionts during locomotion and feeding, i.e., antennae and thoracic legs in copepods, and thoracic legs and postabdomen in cladocerans. The similar values found in the filtering rate of infested and uninfested individuals of N. spiniger and the constant prevalence (< 40%) of epibiont algae, suggest that C. vesiculosum does not condition the life of planktonic crustaceans of Paiva Lake.

Prevalence of Colacium vesiculosum (Colaciales: Euglenophyceae) on planktonic crustaceans in a subtropical shallow lake of Argentina

Colacium vesiculosum (Euglenophyceae) is an epibiont common on planktonic microcrustaceans of continental waters. The interaction between epibionts and substrate organisms is not very well known, particularly in subtropical environments of South America. In the present work, we analyzed the prevalence, density, biomass and attachment sites of C. vesiculosum on planktonic microcrustaceans from Paiva Lake, a subtropical lake of Argentina. With the aim to evaluate whether epibionts affect the filtering rates of Notodiaptomus spiniger, the dominant planktonic crustacean, we carried out bioassays using phytoplankton <53µm. Crustaceans were sampled using a PVC tube (1.2m long and 10cm in diameter), filtering 50L of water through a 53µm-mesh. Microcrustaceans were counted in Bogorov chambers under a stereoscopic microscope. The infested organisms were separated and observed with a photonic microscope to determine density and biovolume of epibionts, by analyzing their distribution on the exoskeleton. The prevalence of C. vesiculosum was higher in adult crustaceans than in their larvae and juveniles. The most infested group was that of calanoid copepods, related to their high density. The attachment sites on the exoskeleton were found to be the portions of the body which have a higher probability of encounter with epibionts during locomotion and feeding, i.e., antennae and thoracic legs in copepods, and thoracic legs and postabdomen in cladocerans. The similar values found in the filtering rate of infested and uninfested individuals of N. spiniger and the constant prevalence (<40%) of epibiont algae, suggest that C. vesiculosum does not condition the life of planktonic crustaceans of Paiva Lake. Rev. Biol. Trop. 59 (3): 1295-1306. Epub 2011 September 01.

Colacium vesiculosum Ehrenberg (Euglenophyceae) es un epibionte común en microcrustáceos del plancton de aguas continentales. La interacción epibiontes y organismo substrato es poco conocida, particularmente en ambientes subtropicales de América del Sur. Se analiza la prevalencia, densidad, biomasa y sitios de adhesión de C. vesiculosum en microcrustáceos planctónicos de un lago subtropical de Argentina. Con el fin de evaluar si los epibiontes afectan la tasa de filtración de Notodiaptomus spiniger Brian, crustáceo dominante del zooplancton, se realizaron bioensayos utilizando el fitoplancton <53µm. Los crustáceos fueron muestreados con un tubo de PVC (1m de largo, 10cm de diámetro) filtrándose 50L de agua a través de una red de 53µm de abertura de malla. El recuento de los microcrustáceos se realizó en cámaras de Bogorov bajo microscopio estereoscópico. Los organismos infectados fueron separados y observados al microscopio fotónico para determinaciones de densidad y biovolumen de los epibiontes, y su distribución en el exoesqueleto. La prevalencia de C. vesiculosum fue mayor en crustáceos adultos que en estadíos larvales y juveniles. El grupo más infestado fue el de los copépodos calanoideos en relación con su alta densidad. Los sitios de adhesión sobre el exoesqueleto fueron las regiones del cuerpo que durante la locomoción y alimentación tienen mayor probabilidad de encuentro con los epibiontes: antenas y patas toráxicas en copépodos, patas toráxicas y postabdomen en cladóceros. Valores similares en la tasa de filtración entre individuos infestados y no infestados de N. spiniger y la constante prevalencia (<40%) del alga epibionte sugieren que C. vesiculosum no condicionaría la vida de los microcrustáceos planctónicos del lago Paiva.