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Identification of specimens belonging to the genus Linguatula (Pentastomida) is relatively easy due to their unique morphology. However, differentiation between species of Linguatula can be challenging for several reasons, including considerable differences between different developmental stages of the parasite within and between species. Currently, 18S rRNA and Cox1 sequences are the only available comparable sequences in GenBank, but recent research has discussed the utility of 28S rRNA for pentastomid phylogenetics. This study presents 28S rRNA gene sequences for two members of the genus Linguatula. Sequences of 28S rRNA were successfully obtained from well-identified samples of L.serrata (collected in Australia) and L.nuttalli (collected in South Africa), with voucher specimens. Phylogenetic analysis of the 28S rRNA region showed 6% difference between L.serrata and L.nuttalli, with low levels of intraspecific variation. In comparison, 18S rRNA and Cox1 sequences from the same specimens showed 0.23% and 13% interspecific differences, respectively. The results of this study show that 28S rRNA has greater genetic diversity to allow for improved differentiation between species of Linguatula than 18S rRNA but is on par with Cox1. Records that do not provide adequate morphological or molecular data to justify independent specific diagnoses must be regarded cautiously, and the need for continued research on species of Linguatula, using a combined morphological and molecular analysis, across a number of different hosts, development stages, geographical regions and molecular markers is highlighted.

Studies on parasite biogeography and host spectrum provide insights into the processes driving parasite diversification. Global geographical distribution and a multi-host spectrum make the tapewormLigula intestinalisa promising model for studying both the vicariant and ecological modes of speciation in parasites. To understand the relative importance of host association and biogeography in the evolutionary history of this tapeworm, we analysed mtDNA and reduced-represented genomic SNP data for a total of 139 specimens collected from 18 fish-host genera across a distribution range representing 21 countries. Our results strongly supported the existence of at least 10 evolutionary lineages and estimated the deepest divergence at approximately 4.99-5.05 Mya, which is much younger than the diversification of the fish host genera and orders. Historical biogeography analyses revealed that the ancestor of the parasite diversified following multiple vicariance events and was widespread throughout the Palearctic, Afrotropical, and Nearctic between the late Miocene and early Pliocene. Cyprinoids were inferred as the ancestral hosts for the parasite. Later, from the late Pliocene to Pleistocene, new lineages emerged following a series of biogeographic dispersal and host-switching events. Although only a few of the currentLigulalineages show narrow host-specificity (to a single host genus), almost no host genera, even those that live in sympatry, overlapped between differentLigulalineages. Our analyses uncovered the impact of historical distribution shifts on host switching and the evolution of host specificity without parallel host-parasite co-speciation. Historical biogeography reconstructions also found that the parasite colonized several areas (Afrotropical and Australasian) much earlier than was suggested by only recent faunistic data.

To date, there have been few published reports of Caracal spp. (or other non-domestic carnivores) preying upon nocturnal strepsirrhine primates anywhere in continental Africa. However, in South Afr...

Fascioliasis is a highly pathogenic disease affecting humans and livestock worldwide. It is caused by the liver flukes

In our previous research on a natural waterbody in South Africa, introduced Australian redclaw crayfish, Cherax quadricarinatus, were found to harbour two species of temnocephalans on their body surfaces and branchial chambers. The temnocephalans were morphologically consistent with Craspedella pedum and Diceratocephala boschmai, both known from this host species and native originally to Australia. In the current study, we report molecular data from the same material. DNA sequences from portions of the gene encoding the nuclear ribosomal 28S RNA indicated a close relationship of these worms with those from Australia. It is highly likely that the worms we found were introduced into South Africa along with the crayfish, following a similar trend elsewhere in the world. There is no evidence that they have been able to establish on native South African hosts.

Studies on parasite biogeography and host spectrum provide insights into the processes driving parasite diversification. Global geographical distribution and a multi-host spectrum make the tapeworm Ligula intestinalis a promising model for studying both the vicariant and ecological modes of speciation in parasites. To understand the relative importance of host association and biogeography in the evolutionary history of this tapeworm, we analysed mtDNA and reduced-represented genomic SNP data for a total of 139 specimens collected from 18 fish-host genera across a distribution range representing 21 countries. Our results strongly supported the existence of at least 10 evolutionary lineages and estimated the deepest divergence at approximately 4.99-5.05 Mya, which is much younger than the diversification of the fish host genera and orders. Historical biogeography analyses revealed that the ancestor of the parasite diversified following multiple vicariance events and was widespread throughout the Palearctic, Afrotropical, and Nearctic between the late Miocene and early Pliocene. Cyprinoids were inferred as the ancestral hosts for the parasite. Later, from the late Pliocene to Pleistocene, new lineages emerged following a series of biogeographic dispersal and host-switching events. Although only a few of the current Ligula lineages show narrow host-specificity (to a single host genus), almost no host genera, even those that live in sympatry, overlapped between different Ligula lineages. Our analyses uncovered the impact of historical distribution shifts on host switching and the evolution of host specificity without parallel host-parasite co-speciation. Historical biogeography reconstructions also found that the parasite colonized several areas (Afrotropical and Australasian) much earlier than was suggested by only recent faunistic data.

Amphistomiasis is caused by different species of trematodes in the family paramphistomatidae and is one of the neglected parasitic diseases of livestock. Amphistomiasis has a wide geographic distribution, especially in subtropical and tropical regions. Although adult stage of most of the amphistomes are seen in the rumen and reticulum of ruminants, but sometimes can be seen in bile ducts too. A few parasites were collected from the bile ducts of a native cattle from the Kerman slaughterhouse, Kerman Province, southeastern Iran, and samples were studied by histomorphological as well as, Scanning Electron Microscopy. The worms were identified as Explanatum explanatum using identification keys and SEM. This trematode has been identified in livestock from only two other provinces of Iran but this is the first report of it in Kerman Province. Specific epidemiological studies with exact slaughter inspections are needed to recognize the extent of the parasite prevalence in native animals of Iran.

The Proterodiplostomidae Dubois, 1936 is a relatively small family of diplostomoidean digeneans parasitising the intestines of reptilian hosts associated with freshwater environments in tropical and subtropical regions. The greatest diversity of proterodiplostomids is found in crocodilians, although some parasitise snakes and turtles. According to the most recent revision, the Proterodiplostomidae included 17 genera within 5 subfamilies. Despite the complex taxonomic structure of the family, availability of testable morphology-based phylogenetic hypotheses and ancient hosts, molecular phylogenetic analyses of the group were practically lacking. Herein, we use novel DNA sequence data of the nuclear lsrRNA gene and mitochondrial cox1 gene from a broad range of proterodiplostomid taxa obtained from crocodilian, fish, and snake hosts on four continents to test the monophyly of the family and evaluate the present morphology-based classification system of the Proterodiplostomidae in comparison with the molecular phylogeny. This first detailed phylogeny for the Proterodiplostomidae challenges the current systematic framework. Combination of molecular phylogenetic data with examination of freshly collected quality specimens and re-evaluation of morphological criteria resulted in a number of systematic and nomenclatural changes along with a new phylogeny-based classification of the Proterodiplostomidae. As the result of our molecular and morphological analyses: (i) the current subfamily structure of the Proterodiplostomidae is abolished; (ii) three new genera, Paraproterodiplostomum n. g., Neocrocodilicola n. g. and Proteroduboisia n. g., are described and Pseudoneodiplostomoides Yamaguti, 1954 is restored and elevated from subgenus to genus level; (iii) two new species, Paraproterodiplostomum currani n. g., n. sp. and Archaeodiplostomum overstreeti n. sp., are described from the American alligator in Mississippi, USA. Comparison of the structure of terminal ducts of the reproductive system in all proterodiplostomid genera did not support the use of these structures for differentiation among subfamilies (or major clades) within the family, although they proved to be useful for distinguishing among genera and species. Our study includes the first report of proterodiplostomids from Australia and the first evidence of a snake acting as a paratenic host for a proterodiplostomid. A key to proterodiplostomid genera is provided. Questions of proterodiplostomid-host associations parasitic in crocodilians are discussed in connection with their historical biogeography. Our molecular phylogeny of the Proterodiplostomidae closely matches the current molecular phylogeny of crocodilians. Directions for future studies of the Proterodiplostomidae are outlined. Electronic supplementary material The online version of this article (10.1007/s11230-020-09928-2) contains supplementary material, which is available to authorized users.

The ovaries of the largescale yellowfish, Labeobarbus marequensis (Teleostei: Cypriniformes: Cyprinidae), are made up of the germinal epithelium, nests of late chromatin nucleolus stage oocytes, and ovarian follicles. Each follicle is composed of a single oocyte, which is surrounded by somatic follicular cells and a basal lamina covered by thecal cells. We describe polarization and ultrastructure of oocytes during the primary growth stage. The oocyte nucleus contains lampbrush chromosomes, nuclear bodies and fibrillar material in which multiple nucleoli arise. Nuage aggregations composed of material of a nuclear origin are present in the perinuclear cytoplasm. The Balbiani body (Bb) contains aggregations of nuage, rough endoplasmic reticulum, individual mitochondria and complexes of mitochondria with nuage (cement). Some mitochondria in the Bb come into close contact with endoplasmic reticulum cisternae and vesicles that contain granular material. At the start of primary growth, the Bb is present in the cytoplasm close to the nucleus. Next, it expands towards the oocyte plasma membrane. In these oocytes, a spherical structure, the so-called yolk nucleus, arises in the Bb. It consists of granular nuage in which mitochondria and vesicles containing granular material are immersed. Later, the Bb becomes fragmented and a fully grown yolk nucleus is present in the vegetal region. It contains numerous threads composed of granular nuage, mitochondria, lysosome-like organelles and autophagosomes. We discuss the formation of autophagosomes in the cytoplasm of primary growth oocytes. During the final step of primary growth, the cortical alveoli arise in the cytoplasm and are distributed evenly. The eggshell is deposited on the external surface of the oocyte plasma membrane and is made up of two egg envelopes that are pierced by numerous pore canals. The external egg envelope is covered in protuberances. During primary growth no lipid droplets are synthesized or stored in the oocytes.

Background IgE to galactose alpha-1,3 galactose (alpha-gal) causes alpha-gal syndrome (delayed anaphylaxis after ingestion of mammalian meat). Development of sensitization has been attributed to tick bites; however, the possible role of other parasites has not been well studied. Objective Our aims were to assess the presence, relative abundances, and site of localization of alpha-gal–containing proteins in common ectoparasites and endoparasites endemic in an area of high prevalence of alpha-gal syndrome, as well as to investigate the ability of ascaris antigens to elicit a reaction in a humanized rat basophil in vitro sensitization model. Methods Levels of total IgE, Ascaris-specific IgE, and alpha-gal IgE were measured in sera from patients with challenge-proven alpha-gal syndrome and from controls without allergy. The presence, concentration, and localization of alpha-gal in parasites were assessed by ELISA, Western blotting, and immunohistochemistry. The ability of Ascaris lumbricoides antigen to elicit IgE-dependent reactivity was demonstrated by using the RS-ATL8 basophil reporter system. Results Alpha-gal IgE level correlated with A lumbricoides–specific IgE level. Alpha-gal protein at 70 to 130 kDa was detected in A lumbricoides at concentrations higher than those found in Rhipicephalus evertsi and Amblyomma hebraeum ticks. Immunohistochemistry was used to localize alpha-gal in tick salivary acini and the helminth gut. Non–alpha-gal–containing A lumbricoides antigens activated RS-ATL8 basophils primed with serum from subjects with alpha-gal syndrome. Conclusion We demonstrated the presence, relative abundances, and site of localization of alpha-gal–containing proteins in parasites. The activation of RS-ATL8 IgE reporter cells primed with serum from subjects with alpha-gal syndrome on exposure to non–alpha-gal–containing A lumbricoides proteins indicates a possible role of exposure to A lumbricoides in alpha-gal sensitization and clinical reactivity.

Smales, Lesley, Halajian, Ali, Rampedi, Kgethedi Michael (2021): A new species of Mediorhynchus Van Cleave, 1916 (Acanthocephala: Gigantorhynchidae) from Burhinus capensis (Aves: Charadriiformes) and a report of Mediorhynchus africanus Amin et al. 2013 from Guttera edouardi (Aves: Galliformes) from South Africa. Zootaxa 5005 (3): 395-400, DOI: https://doi.org/10.11646/zootaxa.5005.3.15

The genus Clinostomoides Dollfus, 1950 was erected to accommodate a single worm from Ardea goliath sampled in the Belgian Congo. The specimen was distinguished from other clinostomids by its large size and posterior genitalia. In the following years, metacercariae of Clinostomoides brieni, have been described in Clarias spp. in southern and western Africa. A few authors have referred to Clinostomum brieni, but all such usages appear to be lapsus calami, and the validity of Clinostomoides remains widely accepted. In this study our aim was: position C. brieni among the growing clinostomids molecular database, and redescribe the species with emphasis on characters that have emerged as important in recent work. We sequenced two nuclear (partial 18S and ITS) and one mitochondrial marker (partial cytochrome c oxidase I) and studied morphology in metacercariae from hosts and localities likely to harbour the type species (Clarias spp., Democratic Republic of the Congo, South Africa). Phylogenetic analysis shows C. brieni belongs within Clinostomum Leidy, 1856. We therefore transfer C. brieni to Clinostomum, amend the diagnosis for the genus Clinostomum and provide a critical analysis of other species in Clinostomoides, all of which we consider species inquirendae, as they rest on comparisons of different developmental stages.

Five Cape fur seals, Arctocephalus pusillus pusillus, 1 dusky kob, Argyrosomus coronus, and 55 silver kob, A. inodorus, collected from Cape Cross, Henties Bay, and Skeleton Coast Park, Namibia, were examined for acanthocephalans. Adults of Bolbosoma vasculosum and Corynosoma australe were recovered from 5 Cape fur seals, and cystacanths of C. australe from the single dusky kob and 3 of the silver kobs that were examined. This is the first record of B. vasculosum and the second of C. australe from the Namibian coast. Scanning electron microscopy of specimens of C. australe revealed features such as bursal papillae and micropores both on the proboscis hook surfaces and across the epidermal surface of the trunk. X-ray analyses of gallium cut hook, egg, and spines of C. australe are presented with analyses of spines of Corynosoma strumosum for comparison.

Digeneans are an important class of trematodes which infect variety of animals ranging from vertebrates (mainly final hosts) to invertebrates (mainly as intermediate hosts). Additionally, some of these parasites are zoonotic and one health importance. Diversity of digeneans in Iran and reports of several zoonotic cases show the importance of these parasites in this country. This is the checklist of digenean parasites reported from Iran representing a total of 118 species, from 105 genera, 43 families, 21 subfamilies, and 3 orders that are reported from 139 species of different hosts including humans. Some of the most important final number of families, genera, and species reported in Iran fauna include: Fasciolidae, Genus: Fasciola, Fasciola gigantica, Fasciola hepatica, Family: Dicrocoeliidae, Genus: Dicrocoelium, Dicrocoelium dendriticum, Family: Diplostomidae, Genus: Diplostomum, Diplostomum spathaceum, Family: Heterophyidae, Genus: Haplorchis, Haplorchis taichui, Family: Notocotylidae, Genus: Notocotylu, Notocotylus aegyptiacus, Family: Bucephalidae, Genus: Rhipidocotyle, Rhipidocotyle illense.

Genetic diversity within partial 18S rRNA sequences from Hepatozoon protozoan parasites from wild felines in South Africa was assessed and compared with data from domestic cats to assess patterns of host specificity. Lions, leopards, servals, a caracal and an African wildcat were all positive for parasites of the Hepatozoon felis-complex. However, haplotypes were not species-specific, and potential mixed infections were widespread. Additional genetic markers are needed to untangle the extremely complex situation of these parasites in both domestic cats and wild felines in South Africa.

All species of hard ticks associated with reptiles as hosts throughout their life cycle, are currently assigned to genera including Amblyomma and Africaniella. Among these species, based on literature data, Africaniella transversale has never been investigated for the presence of tick-borne pathogens. In this study, seven DNA extracts (two from A. transversale and five from Amblyomma exornatum) were screened for the presence of important tick-borne protozoa (piroplasms) and bacteria (Anaplasmataceae and Rickettsiaceae) with conventional PCRs and sequencing. A new heat shock protein chaperonin (groEL) gene-specific PCR was also developed to identify Occidentia spp. in these samples. In A. transversale, Occidentia massiliensis (previously detected in rodent-associated soft ticks) and Rickettsia hoogstraalii were present. While the latter was molecularly identical with formerly reported sequences of this rickettsia, the genotype of O. massiliensis was new based on sequence and phylogenetic analyses of its groEL gene. In A. exornatum, a Rickettsia genotype closely related to R. tamurae and R. monacensis, was detected. The ompA sequence of this genotype was identical to that of Rickettsia sp. Ae-8 reported from A. exornatum in a reptile breeding facility in the USA. These results show that A. transversale might carry O. massiliensis which (unless having a symbiotic nature in ticks) may originate either from the reptile host of this hard tick species or the rodent prey of reptiles. This is also the first detection of the reptile tick-associated Rickettsia sp. Ae-8 (phylogenetically aligning with R. tamurae, R. monacensis) in Africa, i.e. within the original geographical range of A. exornatum.

A total of 509 mammalian vertebrates, belonging to 76 species, were examined for infection with pentastomid parasites. These animals were from 8 of the 9 provinces in South Africa. Linguatulid pentastomes were found only in 7 animals, specifically the African Lion (n = 3) and African Buffalo (n = 4). Adult parasites were found in the lion but nymphs, of various stages, were found in the buffalo. A detailed morphological examination of adult parasites using both light and scanning electron microscopy techniques suggested the specimens were Linguatula nuttalli Sambon1922. Sequences of 18S ribosomal DNA and Cox1 regions obtained from both adult and nymph stages suggested they belong to the one species. Phylogenetic analyses of Linguatula spp. based on the 18S and Cox1 sequences available in GenBank and obtained in the present study showed a clear distinction between L. nuttalli, L. arctica and L. serrata (from Europe and Australia). Several specimens from the Palearctic region which were previously assumed to be L. serrata formed a distinct group in the phylogenetic tree suggesting they probably belong to a different, and as of yet, unknown species., Graphical abstract Image 1, Highlights • Genetic and morphological characterisation of Linguatula nuttalli in South Africa. • Occurrence of Linguatula nuttalli in 509 mammalian vertebrates, belonging to 76 species. • Phylogenetic analyses of Linguatula spp. from different continents.

Maupasina weissi (Seurat, 1913), is redescribed from the eastern rock sengi, Elephantulus myurus Thomas & Schwann, from Limpopo Province and compared with material collected from Free State and North West Provinces, South Africa, as well as the description of the type-material from South Tunisia. Distinguishing features of the species include a corona radiata of 12 pointed leaflike elements, a complex bipartite buccal capsule with 3 large bicuspid denticular lobes and 4 smaller club-shaped lobes and 11 (occasionally 10) pairs of cloacal papillae in the male. The complex taxonomic history of the genus Maupasina Seurat, 1917 is discussed. The probable misidentification of Macroscelides proboscideus Shaw, the short eared sengi, from North West Province, South Africa, as a host of M. weissi is explained, indicating that M. weissi occurs only in species of the genus Elephantulus Thomas & Schwann. The widely separated geographical regions, stretching the length of the African continent, in which M. weissi has been found are indicative of a conservative species with a broad geographical distribution. Tenebrionid beetles, up to 50% of stomach contents of E. myurus in Limpopo Province may act as intermediate hosts in the life-cycle of M. weissi.

Introduced alien fish species and their associated parasites may result in a serious threat to indigenous biodiversity. Furthermore, this may have negative impacts on cultured fish as well as on native parasitic fauna. In the present study, the invasive Asian nematode, Camallanus cotti Fujita, 1927 (Nematoda: Camallanidae), is reported from the guppy (Poecilia reticulata) for the first time in Africa. This parasite is assumed to be introduced into Africa along with the introduction of exotic poeciliid fishes, which are known to be the most common hosts of C. cotti in ornamental fish industry worldwide.The presence of this parasite in both aquarium-cultured fish as well as fish from natural waterbodies is evidence of the introduction of the alien organisms due to insufficient prophylactic veterinary control during transfer of non-native hosts between countries and the spread of them by the anthropogenic introduction to natural systems.

Background Bats are regarded as the primary (ancestral) hosts of bugs of the family Cimicidae. The historically and economically most important species in the family is the common bedbug (Cimex lectularius), because of its worldwide occurrence and association with humans. This molecular-phylogenetic study was initiated in order to expand the knowledge on the phylogeny of cimicid bugs of bats, by investigating samples from Hungary, Romania (representing central-eastern Europe) and two further countries (South Africa and Vietnam). Results Altogether 216 cimicid bugs were collected (73 Ci. lectularius, 133 Ci. pipistrelli, nine Cacodmus ignotus and one Ca. sparsilis). Members of the Cimex lectularius species group were found both in the environment of bats (only Myotis emarginatus, which is a cave/attic-dwelling species) and on three crevice-dwelling bat species (two pipistrelloid bats and M. bechsteinii). On the other hand, Ci. pipistrelli always occurred off-host (near M. myotis/blythii, which are cave/attic-dwelling species). In addition, two Cacodmus spp. were collected from Pipistrellus hesperidus. The morphological characters of these specimens are illustrated with high resolution pictures. Analysis of cytochrome c oxidase subunit 1 (cox1) sequences generated from 38 samples indicated relative genetic homogeneity of Ci. pipistrelli, while the Ci. lectularius group had two haplotypes (collected from pipistrelloid bats in Hungary and Vietnam) highly divergent from other members of this species group. These results were confirmed with molecular and phylogenetic analyses based on the internal transcribed spacer 2 (ITS2). Bat-associated bugs morphologically identified as Ca. ignotus and Ca. sparsilis were different in their cox1, but identical in their ITS2 sequences. Conclusions Molecular evidence is provided here on the existence of two new genotypes, most likely new species, within the Ci. lectularius species group. The relevant specimens (unlike the others) were collected from pipistrelloid bats, therefore the association of Ci. lectularius with different bat host species (pipistrelloid vs myotine bats) should be evaluated further as a possible background factor of this genetic divergence. In addition, Ca. ignotus is reported for the first time in South Africa.

In the present work, we carried out a morphological, biometrical and molecular study of whipworms Trichuris Roederer, 1761 (Nematoda: Trichuridae) parasitizing Papio ursinus Keer 1792 (Chacma baboon). Biometrical and molecular data suggest a new species of Trichuris parasitizing baboons. In addition of main morphological features (spicule, spicule sheath, spicule tube, proximal cloacal tube, distal cloacal tube, vulva, vagina), the mean values of individual variables between Trichuris colobae, Trichuris suis, Trichuris trichiura examined by Student's t tests suggest that T. ursinus n. sp. constitutes a new species. The combined analysis of three markers (cox1, cob and ITS2) revealed a sister relationships between T. colobae and T. ursinus n. sp. Mitochondrial sequences revealed a higher inter-specific similarity between T. ursinus n. sp., T. suis and T. colobae. Phylogenetic hypotheses for both mitochondrial genes strongly supported distinct genetic lineages corresponding to different species of the genus Trichuris associated with certain hosts. Thus, T. suis, T. colobae and T. ursinus n. sp. appeared as a sister group and separated from Trichuris spp. from humans and other species of primates.

Single juvenile specimens of a species of Centrorhynchus were collected from the small intestines of 3 stray dogs in Qom, Qom Province, Iran, constituting the third report of a centrorhynchids from a dog. The proboscis morphology and armature, 24–28 longitudinal rows of 9–10 hooks, were most similar to juveniles of Centrorhynchus mysentri from the Indian bullfrog, Hoplobatrachus tigerinus, and adults of Centrorhynchus globirostris from the pheasant crow Centropus sinensis, which are described from India and Pakistan, respectively. The dogs may have become infected when eating infected frogs.

In the Afrotropic region, the genus Clinostomum is represented by four accepted and four unnamed species distinguished using molecular data. Here, we describe one of the four unnamed species as Clinostomum ukolii n. sp. based on metacercariae from siluriform fishes (Synodontis batensoda, Schilbe intermedius) collected in Nigeria and South Africa. The new species is distinguished by molecular data (39 new sequences of partial cytochrome c oxidase I≥ 6.7% divergent from those of other species) and morphological differences from named and unnamed species in the same region. Metacercariae of C. ukolii n. sp. can be distinguished based on size, tegumental spines, and various aspects of the genital complex, including its position, lobation of the anterior testis, and the disposition and shape of the cirrus pouch. Although descriptions of new species of digeneans are typically based on the morphology of adults, we argue that in cases where data are available from metacercariae from regionally known species, new species can be described based on metacercariae, particularly when supported by molecular data, as here. Moreover, sub-adult reproductive structures can be clearly visualized in metacercaria of Clinostomum. Considering metacercariae as potential types for new species could advance clinostome systematics more rapidly, because metacercariae are encountered much more often than adults in avian definitive hosts.

Background Despite the increasingly recognized eco-epidemiological significance of bats, data from molecular analyses of vector-borne bacteria in bat ectoparasites are lacking from several regions of the Old and New Worlds. Methods During this study, six species of ticks (630 specimens) were collected from bats in Hungary, Romania, Italy, Kenya, South Africa, China, Vietnam and Mexico. DNA was extracted from these ticks and analyzed for vector-borne bacteria with real-time PCRs (screening), as well as conventional PCRs and sequencing (for pathogen identification), based on the amplification of various genetic markers. Results In the screening assays, Rickettsia DNA was only detected in bat soft ticks, whereas Anaplasma phagocytophilum and haemoplasma DNA were present exclusively in hard ticks. Bartonella DNA was significantly more frequently amplified from hard ticks than from soft ticks of bats. In addition to Rickettsia helvetica detected by a species-specific PCR, sequencing identified four Rickettsia species in soft ticks, including a Rickettsia africae-like genotype (in association with a bat species, which is not known to migrate to Africa), three haemotropic Mycoplasma genotypes in Ixodes simplex, and Bartonella genotypes in I. ariadnae and I. vespertilionis. Conclusions Rickettsiae (from both the spotted fever and the R. felis groups) appear to be associated with soft rather than hard ticks of bats, as opposed to bartonellae. Two tick-borne zoonotic pathogens (R. helvetica and A. phagocytophilum) have been detected for the first time in bat ticks. The present findings add Asia (China) to the geographical range of R. lusitaniae, as well as indicate the occurrence of R. hoogstraalii in South Africa. This is also the first molecular evidence for the autochthonous occurrence of a R. africae-like genotype in Europe. Bat haemoplasmas, which are closely related to haemoplasmas previously identified in bats in Spain and to “Candidatus Mycoplasma haemohominis”, are reported here for the first time from Central Europe and from any bat tick. Electronic supplementary material The online version of this article (10.1186/s13071-019-3303-4) contains supplementary material, which is available to authorized users.

Genetic diversity within partial 18S rRNA sequences from Hepatozoon protozoan parasites from domestic cats in South Africa was assessed and compared against published data to assess global biogeographic patterns. Multiple distinct haplotypes of Hepatozoon felis were identified, as well as an unrelated Hepatozoon lineage. Hepatozoon felis genetic diversity globally is very high, indicating a likely complex of species. The recently described Hepatozoon apri from wild boars is closely related to some lineages of H. felis. Sarcocystis and Babesia parasites were also detected. Since Hepatozoon felis is apparently a species complex, potential differences between genetically distinct forms need to be assessed. The finding of an unrelated Hepatozoon indicates that felids can be infected by more species of Hepatozoon than currently known, and that trophic interactions may increase the number of Hepatozoon species found in carnivores. Genetic screening again is demonstrated to identify previously unrecognised parasites from vertebrate hosts.

The ovaries of the largemouth bass Micropterus salmoides, an alien and invasive species in South Africa, contain a germinal epithelium which consists of germline and somatic cells, as well as previtellogenic and late vitellogenic ovarian follicles. The ovarian follicle consists of an oocyte surrounded by follicular cells and a basal lamina; thecal cells adjacent to this lamina are covered by an extracellular matrix. In this article, we describe the Balbiani body and the polarization and ultrastructure of the cytoplasm (ooplasm) in previtellogenic oocytes. The nucleoplasm in all examined oocytes contains lampbrush chromosomes, nuclear bodies and several nucleoli near the nuclear envelope. The ultrastructure of the nucleoli is described. Numerous nuage aggregations are present in the perinuclear cytoplasm in germline cells as well as in the ooplasm. Possible roles of these aggregations are discussed. The ooplasm contains the Balbiani body, which defines the future vegetal region in early previtellogenic oocytes. It is comprised of nuage aggregations, rough endoplasmic reticulum, Golgi apparatus, mitochondria, complexes of mitochondria with nuage-like material, and lysosome-like organelles. In mid-previtellogenic oocytes, the Balbiani body surrounds the nucleus and later disperses in the ooplasm. The lysosome-like organelles fuse and transform into vesicles containing material which is highly electron dense. As a result of the fusion of the vesicles of Golgi and rough endoplasmic reticulum, the cortical alveoli arise and distribute uniformly throughout the ooplasm of late previtellogenic oocytes. During this stage, the deposition of the eggshell (zona radiata) begins. The eggshell is penetrated by canals containing microvilli and consists of the following: the internal and the external egg envelope. In the external envelope three sublayers can be distinguished.

International audience; The phylogeny of the hard tick genus Amblyomma Koch, 1844 deserves special attention, because several poorly studied tick species associated with reptiles still bear the name of this genus, although they may not belong to it. This study focuses on the phylogeny of two such species with uncertain taxonomic status, i.e., Amblyomma transversale (Lucas, 1845) and Amblyomma exornatum Koch, 1844, analyzing two mitochondrial (cytochrome c oxidase subunit 1 and 16S rRNA) and two nuclear (18S and 28S rRNA) genes.In the cox1 phylogenetic analysis, both Am. transversale and Am. exornatum were part of a sister group to all other Metastriata, whereas in the 16S rRNA gene analysis, Am. transversale belonged to a sister group to three subfamilies (Amblyomminae Neumann, 1911; Haemaphysalinae Hoogstraal and Aeschlimann, 1982; Bothriocrotoninae Klompen, Dobson and Baker, 2002), and Am. exornatum formed a sister group to other Amblyomminae. However, based on the 18S and 28S rRNA genes, Am. transversale belonged to a sister group of either Bothriocrotoninae alone or of both Bothriocrotoninae and Haemaphysalinae, respectively. In the latter two phylogenetic analyses Am. exornatum always clustered within Amblyomminae. Morphological comparisons revealed that Am. transversale has at least four unique characters and shares a high number of traits with the genera Robertsicus Barker and Burger, 2018 and Archaeocroton Barker and Burger, 2018, as well as with the subgenus Alloceraea Schulze, 1918 (represented by Haemaphysalis inermis Birula, 1895). These results justify that the genus Africaniella Travassos Dias, 1974 should be reinstated, and the species name of Am. transversale should be used as Africaniella transversale (Lucas, 1845).

In the present work, a comparative molecular study of Ctenocephalides felis and Ctenocephalides canis isolated from dogs (Canis lupus familiaris) from different geographical regions (Spain, Iran and South Africa) was carried out. We found morphological variations in C. felis that do not correspond with molecular differences. The Internal Transcribed Spacers 1 and 2 (ITS1, ITS2) and 18S rRNA partial gene, and cytochrome c-oxidase 1 (cox1) mtDNA partial gene sequences were determined to clarify the taxonomic status of these two species, and to assess interpopulation variation and interspecific sequence differences. In addition, a comparative phylogenetic study with other species of fleas using Bayesian, Maximum Parsimony and Maximum Likelihood analysis was performed. The 18S rRNA partial gene fragment was useful neither to discriminate C. canis and C. felis nor to infer phylogenetic relationships at this level, whereas ITS1 and ITS2 assessed for specific determination in the genus Ctenocephalides. The cox1 mtDNA sequences of C. felis revealed three main haplotypes and we suggest that there has been introgression of C. canis cox1 mtDNA into C. felis by Wolbachia pipientis. Based on cox1 sequences, restriction mapping identified many endonucleases that could be used to delineate different haplotypes of C. felis and to differentiate C. felis and C. canis.

The study of the primate microbiome is critical in understanding the role of the microbial community in the host organism. To be able to isolate the main factors responsible for the differences observed in microbiomes within and between individuals, confounding factors due to technical variations need to be removed. To determine whether alterations due to preservatives outweigh differences due to factors such as host population, host species, body site, and habitat, we tested three methods (no preservative, 96% ethanol, and RNAlater) for preserving wild chimpanzee (fecal), wild lemur (fecal), wild vervet monkey (rectal, oral, nasal, otic, vaginal, and penile), and captive vervet monkey (rectal) samples. All samples were stored below - 20°C (short term) at the end of the field day and then at - 80°C until DNA extraction. Using 16S rRNA gene sequencing, we show a significant preservative effect on microbiota composition and diversity. Samples stored in ethanol and RNAlater appear to be less different compared with samples not stored in any preservative (none). Our differential analysis revealed significantly higher amounts of Enterococcaceae and Family XI in no preservative samples, Prevotellaceae and Spirochaetaceae in ethanol and RNAlater preserved samples, Oligosphaeraceae in ethanol-preserved samples, and Defluviitaleaceae in RNAlater preserved samples. While these preservative effects on the microbiome are not large enough to remove or outweigh the differences arising from biological factors (e.g., host species, body site, and habitat differences) they may promote misleading interpretations if they have large enough effect sizes compared to the biological factors (e.g., host population).

The cat flea (Ctenocephalides felis) is the most common parasite of domestic cats and dogs worldwide. Due to the morphological ambiguity of C. felis and a lack of - particularly largescale - phylogenetic data, we do not know whether global C. felis populations are morphologically and genetically conserved, or whether human-mediated migration of domestic cats and dogs has resulted in homogenous global populations. To determine the ancestral origin of the species and to understand the level of global pervasion of the cat flea and related taxa, our study aimed to document the distribution and phylogenetic relationships of Ctenocephalides fleas found on cats and dogs worldwide. We investigated the potential drivers behind the establishment of regional cat flea populations using a global collection of fleas from cats and dogs across six continents. We morphologically and molecularly evaluated six out of the 14 known taxa comprising genus Ctenocephalides, including the four original C. felis subspecies (Ctenocephalides felis felis, Ctenocephalides felis strongylus, Ctenocephalides felis orientis and Ctenocephalides felis damarensis), the cosmopolitan species Ctenocephalides canis and the African species Ctenocephalides connatus. We confirm the ubiquity of the cat flea, representing 85% of all fleas collected (4357/5123). Using a multigene approach combining two mitochondrial (cox1 and cox2) and two nuclear (Histone H3 and EF-1α) gene markers, as well as a cox1 survey of 516 fleas across 56 countries, we demonstrate out-of-Africa origins for the genus Ctenocephalides and high levels of genetic diversity within C. felis. We define four bioclimatically limited C. felis clusters (Temperate, Tropical I, Tropical II and African) using maximum entropy modelling. This study defines the global distribution, African origin and phylogenetic relationships of global Ctenocephalides fleas, whilst resolving the taxonomy of the C. felis subspecies and related taxa. We show that humans have inadvertently precipitated the expansion of C. felis throughout the world, promoting diverse population structure and bioclimatic plasticity. By demonstrating the link between the global cat flea communities and their affinity for specific bioclimatic niches, we reveal the drivers behind the establishment and success of the cat flea as a global parasite.

Tissue samples from wildlife from South Africa were opportunistically collected and screened for haemoprotozoan parasites using nonspecific PCR primers. Samples of 127 individuals were tested, comprising over 50 different species. Haemogregarines were the most commonly identified parasites, but sarcocystids and piroplasmids were also detected. Phylogenetic analyses estimated from the 18S rDNA marker highlighted the occurrence of several novel parasite forms and the detection of parasites in novel hosts. Phylogenetic relationships, which have been recently reviewed, appear to be much more complex than previously considered. Our study highlights the high diversity of parasites circulating in wildlife in this biodiverse region, and the need for further studies to resolve taxonomic issues.