In the early 2000s, experimental rearing of spotted wolffish, Anarhichas minor, was started in Iceland. Health surveillance, carried out at regular intervals during the rearing period, revealed persistent and highly prevalent Kudoa infections of fish muscles which caused great financial losses due to post mortem myoliquefaction. In addition, during the traditional process of drying and smoking wild Atlantic lumpfish, Cyclopterus lumpus, the muscles from some fish almost completely disappear and the fish have to be discarded. To describe the etiological agent responsible for these conditions, spotted wolffish, Atlantic wolffish Anarhichas lupus, northern wolffish Anarhichas denticulatus and Atlantic lumpfish were caught off the Icelandic coast and examined for the presence of Kudoa. We describe a novel myxosporean, Kudoa islandica n. sp., using morphological and molecular data, and show with histopathology that it causes extensive myoliquefaction in three different wild fish hosts, which all are commercially valuable species in Iceland. Although some spore dimensions varied significantly between fish species, the molecular analyses showed that the same parasite was responsible for infection in all fish. The northern wolffish was not found to be infected. Although robustly placed in the Kudoa clade in phylogenetic analyses, K. islandica was phylogenetically distinct from other kudoids. A single myxosporean, K. islandica, is responsible for the infections in the somatic muscles of lumpfish and wolffish, causing extensive post mortem myoliquefaction. This myxosporean is likely to infect other fish species and it is important to study its life cycle in order to evaluate any threat to salmonid culture via the use of lumpfish as a biocontrol for sea lice.
Malaria parasites in the phylum Apicomplexa (Order: Haemosporida) infect diverse vertebrates and invertebrate hosts. At least seven genera of haemosporidian parasites have been described to exclusively infect bats. Most of these parasites remain enigmatic with a poorly known host range. Here, we investigated 271 bats belonging to 21 species and seven families from six provinces of Thailand. Overall, 124 out of 271 bats (45.8%) were positive for haemosporidian parasites, while none had Plasmodium, based on microscopic examination of blood smears and PCR amplification. We obtained 19 distinct cytochrome b (cytb) nucleotide haplotypes of Hepatocystis from seven bat species (families: Craseonycteridae, Hipposideridae, Pteropodidae, and Rhinolophidae). Nycteria was found in four bat species (Craseonycteridae, Emballonuridae, Megadermatidae, and Pteropodidae) and Polychromophilus in two species (Emballonuridae, Vespertilionidae). Phylogenetic analysis inferred from cytb sequences placed Hepatocystis into 2 different clades. Most Hepatocystis infections were found in insectivorous bats and clustered together with a sequence from Hipposideros larvatus in Cambodia (in subclade 1a). A single sequence of Hepatocystis obtained from a frugivorous bat, Cynopterus brachyotis, was placed in the same clade with Hepatocystis from the same bat species previously reported in Malaysia (clade 2). Nycteria in these Thai bats were clearly separated from the African isolates previously reported in bats in the family Rhinolophidae. Polychromophilus murinus from Myotis siligorensis was placed in a distinct clade (clade 2) from Polychromophilus melanipherus isolated from Taphozous melanopogon (clade 1). These results confirmed that at least two distinct species of Polychromophilus are found in Thailand. Collectively, Hepatocystis presented no host specificity. Although Megaderma spasma seemed to be infected by only Nycteria, its respective parasite does not show specificity to only a single bat host. Polychromophilus murinus and P. melanipherus seem to infect a narrower host range or are somehow restricted to bats in the families Vespertilionidae and Emballonuridae, respectively.
Lemuricola (Protenterobius) nycticebi is the only pinworm species known to infect strepsirrhine primates outside Africa, and the only pinworm species yet described in slow lorises. Here, we provided a detailed morphological comparison of female and male worms, and a first description of fourth-stage larvae collected from free-living slow lorises (Nycticebus menagensis) in Sabah, Malaysian Borneo. Using mitochondrial and nuclear markers, we also reconstructed the species' phylogenetic relationship with other pinworms infecting primates. Both morphological and molecular results indicated a distinct association between L. (P.) nycticebi and its host. However, while taxonomy identified this species as a member of the Lemuricola clade and grouped pinworms infecting lemurs and slow lorises together, phylogenetic reconstruction split them, placing L. (P.) nycticebi within the Enterobius clade. Our results suggest that L. (P.) nycticebi may represent a different taxon altogether, and that it is more closely related to pinworm species infecting Old World primates outside Madagascar. Pongobius pongoi (Foitová et al., 2008) n. comb. is also proposed.
The tropical rainforests of Sundaland are a global biodiversity hotspot increasingly threatened by human activities. While parasitic insects are an important component of the ecosystem, their diversity and parasite-host relations are poorly understood in the tropics. We investigated parasites of passerine birds, the chewing lice of the speciose genus MyrsideaWaterston, 1915 (Phthiraptera: Menoponidae) in a natural rainforest community of Malaysian Borneo. Based on morphology, we registered 10 species of lice from 14 bird species of six different host families. This indicated a high degree of host specificity and that the complexity of the system could be underestimated with the potential for cryptic lineages/species to be present. We tested the species boundaries by combining morphological, genetic and host speciation diversity. The phylogenetic relationships of lice were investigated by analyzing the partial mitochondrial cytochrome oxidase I (COI) and the nuclear elongation factor alpha (EF-1α) genes sequences of the species. This revealed a monophyletic group of Myrsidea lineages from seven hosts of the avian family Pycnonotidae, one host of Timaliidae and one host of Pellorneidae. However, species delimitation methods supported the species boundaries hypothesized by morphological studies and confirmed that four species of Myrsidea are not single host specific. Cophylogenetic analysis by both distance-based test ParaFit and event-based method Jane confirmed overall congruence between the phylogenies of Myrsidea and their hosts. In total we recorded three cospeciation events for 14 host-parasite associations. However only one host-parasite link (M. carmenae and their hosts Terpsiphone affinis and Hypothymis azurea) was significant after the multiple testing correction in ParaFit. Four new species are described: Myrsidea carmenaesp.n. ex Hypothymis azurea and Terpsiphone affinis, Myrsidea franciscaesp.n. ex Rhipidura javanica, Myrsidea ramonisp.n. ex Copsychus malabaricus stricklandii, and Myrsidea victoriaesp.n. ex. Turdinus sepiarius.
Due to their unusual life cycle that includes parasitic larval and free living adult stages, gnathiid isopods are typically overlooked in biodiversity surveys, even those that focus on parasites. While the Philippines sits within the region of highest marine biodiversity in the world, the coral triangle, no gnathiid species have been identified or described from that region. Here we present the first records of two gnathiid species collected from the Visayas, central Philippines: Gnathia malaysiensis Müller, 1993, previously described from Malaysia, and G. camuripenis Tanaka, 2004, previously described from southern Japan. This paper provides detailed morphological redescriptions, drawings and scanning electron microscope images as well as the first molecular characterisation of both species, Furthermore, a summary of the Central-Indo Pacific Gnathia species is provided.
Haemosporidians infect a wide diversity of bat genera and species, yet little is known about their transmission cycles or epidemiology. Though several recent studies have focused on the genus Hepatocystis, an Old World parasite primarily infecting bats, monkeys, and squirrels, this group is still understudied with little known about its transmission and molecular ecology. These parasites lack an asexual erythrocytic stage, making them unique from the Plasmodium vertebrate life cycle. In this study, we detected a prevalence of 31% of Hepatocystis in short-nosed fruit bats (Cynopterus brachyotis) in Singapore. Phylogenetic reconstruction with a partial cytochrome b sequence revealed a monophyletic group of Hepatocystis from C. brachyotis in Malaysia, Singapore, and Thailand. There was no relationship with infection and bat age, sex, location, body condition or monsoon season. The absence of this parasite in the five other bat species sampled in Singapore indicates this Hepatocystis species may be host restricted.
Sarcocystis scandentiborneensis sp. nov. was discovered in histological sections of striated musculature of treeshrews (Tupaia minor, T. tana) from Northern Borneo. Sarcocysts were cigar-shaped, 102 μm-545 μm long, and on average 53 μm in diameter. The striated cyst wall varied in thickness (2-10 μm), depending on whether the finger-like, villous protrusions (VP) were bent. Ultrastructurally, sarcocysts were similar to wall type 12 but basal microtubules extended into VPs that tapered off with a unique U-shaped, electron-dense apical structure. In phylogenetic trees of the nuclear 18S rRNA gene, S. scandentiborneensis formed a distinct branch within a monophyletic subclade of Sarcocystis spp. with (colubrid) snake-rodent life cycle. We mapped all intraspecific (two haplotypes) and interspecific nucleotide substitutions to the secondary structure of the 18S rRNA gene: in both cases, the highest variability occurred within helices V2 and V4 but intraspecific variability mostly related to transitions, while transition/transversion ratios between S. scandentiborneensis, S. zuoi, and S. clethrionomyelaphis were skewed towards transversions. Lack of relevant sequences restricted phylogenetic analysis of the mitochondrial Cytochrome C oxidase subunit I (COI) gene to include only one species of Sarcocystis recovered from a snake host (S. pantherophisi) with which the new species formed a sister relationship. We confirm the presence of the functionally important elements of the COI barcode amino acid sequence of S. scandentiborneensis, whereby the frequency of functionally important amino acids (Alanine, Serine) was markedly different to other taxa of the Sarcocystidae. We regard S. scandentiborneensis a new species, highlighting that structurally or functionally important aspects of the 18S rRNA and COI could expand their utility for delineation of species. We also address the question why treeshrews, believed to be close to primates, carry a parasite that is genetically close to a Sarcocystis lineage preferably developing in the Rodentia as intermediate hosts.
Thelohanellus nikolskii, Achmerov, 1955 is a well-known myxozoan parasite of the common carp (Cyprinus carpio L.). Infection regularly manifests in numerous macroscopic cysts on the fins of two to three month-old pond-cultured carp fingerlings in July and August. However, a Thelohanellus infection is also common on the scales of two to three year-old common carp in ponds and natural waters in May and June. Based on myxospore morphology and tissue specificity, infection at both sites seems to be caused by the same species, namely T. nikolskii. This presumption was tested with molecular biological methods: SSU rDNA sequences of myxospores from fins of fingerlings and scales of older common carp were analysed and compared with each other and with related species available in GenBank. Sequence data revealed that the spores from the fins and scales represent the same species, T. nikolskii. Our study revealed a dichotomy in both infection site and time in T. nikolskii-infections: the fins of young carp are infected in Summer and Autumn, whereas the scales of older carp are infected in Spring. Myxosporean development of the species is well studied, little is known, however about the actinosporean stage of T. nikolskii. A previous experimental study suggests that aurantiactinomyxon actinospores of this species develop in Tubifex tubifex, Müller, 1774. The description included spore morphology but no genetic sequence data (Székely et al., 1998). We examined >9000 oligochaetes from Lake Balaton and Kis-Balaton Water Reservoire searching for the intraoligochaete developmental stage of myxozoans. Five oligochaete species were examined, Isochaetides michaelseni Lastochin, 1936, Branchiura sowerbyi Beddard, 1892, Nais sp., Müller, 1774, Dero sp. Müller, 1774 and Aelosoma sp. Ehrenberg, 1828. Morphometrics and SSU rDNA sequences were obtained for the released actinospores. Among them, from a single Nais sp., the sequence of an aurantiactinomyxon isolate corresponded to the myxospore sequences of T. nikolskii.
Within host communities, related species are more likely to share common parasitic agents, and as a result, morphological similarities have led researchers to conclude that parasites infecting closely related hosts within a community represent a single species. However, genetic diversity within parasite genera and host range remain poorly investigated in most systems. Strongyloides is a genus of soil-transmitted nematode that has been reported from several primate species in Africa and Asia, and has been estimated to infect hundreds of millions of people worldwide, although no precise estimates are available. Here we describe a case of infection with a cryptic species of Strongyloides in a Bornean (Philippine) slow loris (Nycticebus menagensis) living within a diverse community of several primate species in the Lower Kinabatangan Wildlife Sanctuary, Malaysian Borneo. Fresh fecal samples were collected from five primate species and nematode larvae cultured from these samples were selected for phylogenetic analyses. Sequences obtained for most larvae were identified as S. fuelleborni, grouping into three different clusters and showing no aggregation within specific hosts or geographic location. In contrast, a set of parasite sequences obtained from a slow loris clustered closely with S. stercoralis into a different group, being genetically distinct to sequences reported from other primate hosts, humans included. Our results suggest that although S. fuelleborni infects all haplorrhines sampled in this primate community, a different species might be infecting the slow loris, the only strepsirrhine in Borneo and one of the least studied primates in the region. Although more data are needed to support this conclusion, we propose that Strongyloides species in primates might be more diverse than previously thought, with potential implications for ecological and evolutionary host-parasite associations, as well as epidemiological dynamics.
Bat flies are highly-specialized, hematophagous arthropods that are globally ubiquitous. There is little published research on bat flies (Diptera: Nycteribiidae) in Singapore and understanding the diversity of nycteribiids, host association and infestation rates can provide insight into this host-ectoparasite relationship. Nycteribiids were collected from bats trapped in Singapore (2011-2016) and identified using morphological keys. Host-ectoparasite relationships were investigated with logistic regression and Bayesian poisson regression. Nycteribiids were found to be monoxenously associated with their host bat species and host age, sex, species, and BBCI appear to contribute to differences in prevalence and intensity. Differences in host specificity between bat fly species in Singapore and their conspecifics in less disturbed habitats with higher bat biodiversity, such as Malaysia, Philippines and Thailand, suggest that the high host specificity in Singapore derives from the paucity of suitable hosts and abundance of single species roosts and not from their coevolved restrictions to them.
Gastropod-nematode associations are underreported worldwide. In the present study, juvenile and adult nematodes were found in the freshwater gastropod Burnupia stenochorias (Melvill & Ponsonby, 1903), from the Vaal River, South Africa. The nematodes were confirmed to belong to the genus Daubaylia chitwood & chitwood, 1934 (Daubayliidae). This is the first report of Daubaylia from a snail belonging to the family Burnupiidae, and the first report of this nematode taxon in southern Africa. Like D. pearsoni and D. malayanum from Australia and Malaysia respectively, adult females of the current species possess multiple well-developed eggs in the uteri, with larvae developing in utero. Morphological and molecular characteristics showed that the nematodes are distinct from all the described species of Daubaylia. Thus, they are considered a new species, Daubaylia burnupiae n. sp. The species differs from its congeners based on spicule shape, the short tail of the male, an anal cuticular knob-like protrusion on the female, and oesophagi with short isthmi and short glandular basal bulbs in both sexes. Three club-shaped pharyngeal lobes, extending slightly above the surface of the cephalic lips in both sexes and a pre-cloacal median papilla on the male were described using scanning electron microscopy, the first of such observations for the daubayliids. Genetic analyses showed that partial sequences of D. burnupiae n. sp. differed from species for which genetic data are available, by at least 26 and 9 base pair differences for 28S and 18S rDNA, respectively. Our results show that low prevalence and abundance of nematodes in the snails, corresponded with increased pollution in the river. We suspect that exposure to pollutants reduces the viability of the infective gravid female nematode during transmission. Therefore, the nematode is a potential bioindicator for aquatic pollution.
Rodent species, such as Rattus rattus diardii and Rattus norvegicus are invasive species of wild rats that serve as potential reservoirs of important human's pathogens. Parasitic zoonosis accounts for over 60% of all human infectious diseases worldwide. This situation arises from the recent changes in the global climate and ecosystem composition, which led to the spread of rodents and rodent-borne pathogens globally. The aim of this study was to determine the occurrence of rodent's parasites and their zoonotic potentials in some selected areas in UPM. Rodents were captured using live-traps and euthanised for helminths and protozoan recovery. Intestinal parasites were detected and identified from stool samples using formalin ethyl-acetate concentration technique (FECT), while tissue parasites were identified by histopathological examination of selected tissue sections of the liver, brain, lungs, and muscle. In this study, a total of 89 wild rats were captured. Twelve species of intestinal and tissue parasites were recorded, of which, Taenia taeniaeformis accounts for the highest infection recorded (28%) followed by Hymenolepis nana (19.5%) and Capillaria hepatica (19.1%), while Toxoplasma gondii was the least parasite (6.7%) identified. Furthermore, other parasites species observed include, Cryptosporidium spp. (21.3%), Entamoeba histolytica/Entamoeba dispar and Moniliformis moniliformis (17.9%), Angiostrongylus cantonensis (16.8%), Hymenolepis diminuta (16.1%), Giardia spp. (14.6%), Trichuris spp. (12.3%), and Sarcocystis spp. (6.74). Based on the results obtained in the present study, 17.1% and 15.4% of the rodents captured were confirmed positive for at least one species of intestinal or tissue parasites, respectively. The presence of these zoonotic parasites in the wild rats suggests the potential risk of rodent-borne zoonotic disease transmission to humans. Hence, the need to improved rats control intervention and public health awareness among the populace.
We investigated the morphology and phylogenetic relationships of novel and previously recognized Sarcocystis spp. infecting small mammals and colubrid snakes in Asia. The nuclear 18S rRNA and mitochondrial cox1 of Sarcocystis sp.1 from mangrove snakes (Boiga dendrophila) in Thailand and Sarcocystis sp.2 from a ricefield rat (Rattus argentiventer) in Sumatra were partially sequenced. Sporocysts of Sarcocystis sp.1 induced development of sarcocysts in experimentally infected rats, which showed a unique ultrastructure that was observed previously by S.P. Kan in rats from Malaysia; therefore, we describe this species as Sarcocystis kani sp. nov. Its integration into the 18S rRNA phylogeny of Sarcocystis spp. cycling between small mammals and colubrid snakes helped clarify relationships among the so-called S. zuoi-complex of molecularly cryptic species: Sarcocystis kani sp. nov., S. sp.2, S. attenuati, S. scandentiborneensis, and S. zuoi were all included in this clade. Tree topology was resolved into dichotomies congruent with the morphological disparities between the taxa. However, cox1 gene sequencing (including newly sequenced S. singaporensis and S. zamani) revealed that Sarcocystis kani, S. attenuati, and S. scandentiborneensis were identical suggesting a recent, common ancestry. To identify other distinctive features, lineage-specific molecular patterns within both genes were examined revealing that all 18S rRNA sequences of the S. zuoi - complex possess a unique, 7-nt long motif in helix 38 of domain V7 that was different in S. clethrionomyelaphis which branched off basally from the complex. Three-dimensional homology modelling of COX1 protein structure identified amino acid substitutions within the barcode area specific for the S. zuoi-complex and substantial divergence in structurally important amino acids between Sarcocystis species of snakes as definitive hosts and other lineages of the Sarcocystidae. We discuss the utility of selected genes for species delimitation of the Sarcocystis spp. under investigation, which probably evolved during recent radiations of their intermediate and definitive hosts.
Non-human primates of South-East Asia remain under-studied concerning parasite epidemiology and co-infection patterns. Simultaneously, efforts in conservation demand knowledge of parasite abundance and biodiversity in threatened species. The Endangered proboscis monkey, Nasalis larvatus, a primate flagship species for conservation in Borneo, was investigated in the present study. Habitat loss and fragmentation are among the greatest threats to bachelor and harem groups of this folivorous colobine. Designed as a follow-up study, prevalence and co-infection status of intestinal parasites from N. larvatus in a protected area in Malaysian Borneo were analyzed from fecal samples using a flotation method. For the first time, the intestinal parasite co-infection patterns were examined using quantitative analyses. Overall, 92.3% of fecal samples (N = 652) were positive for helminth eggs. Five helminth groups were detected: (1) trichurids (82.7% prevalence) including Trichuris spp. (82.1%) and Anatrichosoma spp. (1.4%), (2) strongyles (58.9%) including Trichostrongylus spp. (48.5%) and Oesophagostomum/Ternidens spp. (22.8%), (3) Strongyloides fuelleborni (32.7%), (4) Ascaris lumbricoides (8.6%), and (5) Enterobius spp. (5.5%). On average, an individual was co-infected with two different groups. Significant positive associations were found for co-infections of trichurids with strongyles and S. fuelleborni as well as S. fuelleborni with A. lumbricoides and strongyles. This study shows a high prevalence of various gastrointestinal helminths with potential transmission pathways primarily related to soil and with zoonotic relevance in wild proboscis monkeys in their remaining natural habitats. Observed positive associations of trichurids with strongyles and Strongyloides spp. may result from the high prevalence of trichurids. Similarly, positive associations between Strongyloides and Ascaris were found, both of which typically occur predominantly in juvenile hosts. These findings should be considered when proposing conservation actions in altered habitats nearby human settlements and when managing captive populations.
Borrelia burgdorferi sensu lato (s.l.) DNA was detected by PCR in Ixodes persulcatus Schulze, 1930, Haemaphysalis concinna Koch, 1844, Haemaphysalis japonica douglasi Nuttall et Warburton, 1915 and Dermacentor silvarum Olenev, 1932 ticks collected in the Amur region, the Jewish Autonomous region, the Sakhalin region and on the Khabarovsk territory. Infection rate of I. persulcatus with B. burgdorferi s.l. 10-69% exceeded the corresponding values of three other tick species in all examined regions during 1999-2014 despite different tick abundance and dominance structure. Bacterial loads estimated on the base of quantitative real time PCR varied from 102 to 109 genome-equivalents per a tick with maximal values for I. persulcatus and H. japonica. Phylogenetic analysis of 16S rRNA gene and 5S-23S rRNA intergenic spacer nucleotide sequences revealed two species: 1) Borrelia garinii of Asian type NT29 with several isolates of European type 20047; 2) Borrelia afzelii with identical sequences of the majority of studied isolates and VS461 reference strain in all regions except the Sakhalin Island where B. afzelii was not found. Borrelia miyamotoi of the relapsing fever group was detected as monoinfection or in combination with B. burgdorferi s.l. in 4.0 ± 0.9% and 4.8 ± 0.9% I. persulcatus ticks, respectively. Multiple locus sequence analysis of three fragments of 16S rRNA, glpQ and p66 genes proved that all the Far Eastern B. miyamotoi isolates belonged to the Asian type identical to FR64b strain (GenBank CP004217) from Japan. Wide distribution of Borrelia DNA in ticks, relative genetic homogeneity with similar sequences of the coding regions and the intergenic spacer of Borrelia wild isolates and temporal stability with high homology levels of the Far Eastern isolates of B. garinii, B. afzelii and B. miyamotoi with previously described spirochetes from the surrounding regions of Russia, China and Japan allowed us to suggest multiple ecological niches as the stability factor of the parasitic system.