Accurate survey methods are required for any wildlife research to yield reliable population data. This constraint finds significance in amphibian research that involves a highly threatened group of animals with a large proportion of cryptic species not easily detected by conventional survey methods. Across a growing spectrum of zoology research, survey outcomes are benefitting from the efficacy of scent detection dogs in assisting with species detection. We investigated the ability of a scent detection dog to locate and identify traces of giant bullfrog, Pyxicephalus adspersus scent and investigate methods of preserving frog scent for use in subsequent conditioning training of dogs. The scent detection dog was able to detect 100,000 times diluted scent with 87% sensitivity and 84% efficacy. High specificity (98,6%) was also achieved while presented with the challenge of detecting P. adspersus scent amid that of other frog species. Detection sensitivity was negatively correlated with scent preservation time but yielded the highest sensitivity for samples that were preserved as skin swabs stored at 4 °C and diluted shortly before use. Conservationists, scientists, and customs officials alike can benefit from scent detection dog detection of amphibians through enhanced sample acquisition rates with reduced collection biases.
The sicklefin redhorse, Moxostoma sp. (Cypriniformes: Catostomidae), is an innominate imperiled catostomid endemic to the Hiwassee and Little Tennessee river basins, which has been restricted to a few tributaries of these systems by impoundments. During collections to propagate sicklefin redhorse for reintroduction, a myxozoan, described herein, was observed infecting sicklefin redhorse in the Little Tennessee River Basin, North Carolina. Myxobolus naylori Ksepka et Bullard sp. n. infects the stratum spongiosum covering the scales of sicklefin redhorse. Myxospores of the new species differ from all congeners by the combination of having a mucous envelope, intercapsular process, and sutural markings as well as lacking an iodinophilic vacuole in the sporoplasm. and A phylogenetic analysis of the 18S rDNA gene recovered the new species in a polytomy with Myxobolus marumotoi Li et Sato, 2014 and a clade comprised of species of Myxobolus Bütschli, 1882; Thelohanellus Kudo, 1933, and Dicauda Hoffman et Walker, 1973. Histological sections of infected sicklefin redhorse skin revealed myxospores within a plasmodium in the stratum spongiosum dorsal to scales, encapsulated in collagen fibres, and associated with focal erosion of scales directly beneath the plasmodium; in some instances, the scale was perforated by the plasmodium. The specificity of the new species to sicklefin redhorse may make it a useful biological tag to differentiate sicklefin redhorse from morphologically similar species. The new species is the first parasite reported from sicklefin redhorse, a species of concern to the United States Fish and Wildlife Service. No species of Myxobolus has been reported from species of Moxostoma in the Southeast United States. As it was observed that Myxobolus minutus Rosser, Griffin, Quiniou, Alberson, Woodyard, Mischker, Greenway, Wise et Pote, 2016 is a primary junior homonym of Myxobolus minutus Nemeczek, 1911, we propose the replacement name Myxobolus diminutus (Rosser, Griffin, Quiniou, Alberson, Woodyard, Mischker, Greenway, Wise et Pote, 2016).
In the Pyrenees, brown bear population abundance is estimated from non-invasive genetic analyses of scat and hair samples. Although such analyses are highly beneficial for population monitoring and research, it can be especially difficult for humans to locate bear scats in the field. To address this, we have incorporated a dog (trained from an early age to detect bear scats) into these efforts since 2014. Here, we compared the effectiveness of the scat-detection dog/handler and human-only teams to locate bear scats using our work in the Pyrenees as a case study. A species validation was systematically carried out, either genetically or visually using a microscope, based on the presence of bear hair, for all scats collected from 2010 to 2019. From 2014 to 2019, the use of the dog/handler team in addition to human-only teams increased the average number of bear scats collected annually by four times in comparison with the 2010-2013 period when only humans were searching for scats. This temporal augmentation could not be explained by the increase in bear population size. From 2014 to 2019, the annual percentage of outings during which at least one bear scat was found was 17 times higher for the dog than for humans. The use of the dog also resulted indirectly in a better genotyping success and genetic identification of more individuals due to a larger choice of viable samples that could be sent to the molecular laboratory, as well as a larger number of cub scats detected by the dog. We found that even the use of a single scat-detection dog can greatly improve the efficiency of detecting target scats in challenging monitoring conditions.
Cysteine protease is a superfamily of widespread proteolytic enzymes and plays a major role in larval invasion, migration, exsheathing, survival and immune evasion in parasites. In the present study, the gene coding cysteine proteinase of the nematode Trichinella spiralis (Owen, 1835) was cloned into pQE-80L and subsequently expressed in E. coli JM109. The rTsCP was purified and its antigenicity was identified by Western blot and ELISA. Using anti-rTsCP serum the native TsCP was identified in muscle larval crude proteins. The results of quantitative real-time PCR and immunofluorescence test demonstrated that the TsCP was expressed in all stages of T. spiralis and located mainly in cuticle, stichosome and reproductive organs. The immunisation of mice with rTsCP elicited Th2-predominant immune responses. Anti-rTsCP antibodies could partially inhibit the in vitro larval invasion of intestinal epithelial cells and kill the newborn larvae by an antibody-dependent cell-mediated dose-dependent cytotoxicity. The vaccinated mice exhibited a 54% reduction of adults and a 33% reduction of muscle larvae following challenge infection. The results suggested that the TsCP might be an indispensable protein in Trichinella invasion, development and survival of T. spiralis in hosts, and could be a potential vaccine target against infection., Yan Yan Song, Li Ang Wang, Hua Na Ren, Xin Qi, Ge Ge Sun, Ruo Dan Liu, Peng Jiang, Xi Zhang, Jing Cui, Zhong Quan Wang., and Obsahuje bibliografii
The Natal multimammate mouse, Mastomys natalensis, occurs throughout sub-Saharan Africa. Mitochondrial phylogenetics indicate this species was fragmented during the Pleistocene, forming six matrilineage phylogroups: A-I, A-II, A-III, B-IV, B-V, B-VI with distinct ranges. All except the A-III lineage are identified as natural reservoirs of mammarenaviruses. M. natalensis A-III is found in western Ethiopia and is the only lineage reported in the country. While screening 203 small mammal samples from Dhati Welel National Park for mammarenaviruses, we detected mammarenavirus RNA in nine samples, eight from M. natalensis and one from M. awashensis. A sequence similarity search and phylogenetic analysis confirmed the M. natalensis mitochondrial DNA belongs to the A-III lineage. We characterised the complete virus genome, which showed typical mammarenavirus organisation. Phylogenetic analysis indicated it clusters with Gairo virus found in M. natalensis B-IV in Tanzania, while showing sufficient divergence from other mammarenaviruses to be considered as a new species, for which we proposed the name Dhati Welel. Additional sampling in the M. natalensis A-III phylogeographic range should help determine whether the detection of the virus in M. awashensis represents a local spill-over or if the virus circulates in both Mastomys species.
Black rats (Rattus rattus) are native to the Indian subcontinent but have now colonized most continents and islands following human movements and international trade. They are involved in the circulation and transmission to humans of many zoonotic agents as well as in massive damage to food stocks and native biodiversity in the regions they have settled. This study investigates the genetic diversity and possible origins of black rats from Benin, West Africa. We sequenced the complete mitochondrial cytochrome b gene in 90 individuals from nine localities in Benin. These sequences were subsequently compared to 390 other cytochrome b haplotypes from individuals from various European, Asian, American and African localities. Nucleotide polymorphism analysis, haplotype network and maximum likelihood phylogenetic tree reconstructions showed low mitochondrial diversity in black rats from Benin. Our results also suggest at least two distinct introduction events: one introduction probably occurred during the spice trade (15th-17th century) through the Indies Road connecting Europe to Asia. Other introduction events could have occurred more recently following the intensification of globalized trade from the eighteenth century, and onwards.
The elastase, which belongs to the serine protease family, hydrolyses various proteins and may be involved in the parasite invasion. In this study, complete sequence of elastase-1 (TsE) the nematode Trichinella spiralis (Owen, 1835) was cloned into the plasmid pcDNA3.1 as TsE DNA vaccine. After intramuscular vaccination, serum anti-Trichinella antibodies (IgG and subclass IgG1/IgG2a, and IgA), total and specific intestinal mucosal sIgA in mice vaccinated with pcDNA3.1/TsE were measured by ELISA. The results showed that vaccination with pcDNA3.1/TsE induced a systemic humoral immune response (high levels of serum IgG and subclass IgG1/IgG2a and IgA) and local intestinal mucosal immune responses (high levels of TsE-specific sIgA). Vaccination of mice with TsE DNA vaccine also triggered a systemic and local concomitant Th1/Th2 response, as demonstrated by significant elevation of Th1 (IFN-γ and IL-2) / Th2 (IL-4 and IL-10) cytokine levels after the spleen, mesenteric lymph node and Peyer's patch cells from vaccinated mice were stimulated with recombinant TsE (rTsE). The vaccination of mice with pcDNA3.1/TsE displayed a 17% reduction of intestinal adult worms and a 39% reduction of muscle larvae. Our results indicated that TsE DNA vaccine elicited a systemic concomitant Th1/Th2 response and an enteral local sIgA response, and produced a partial protection against infection with T. spiralis. The TsE may be regarded as a potential candidate vaccine target against Trichinella infection. The oral polyvalent vaccines should be developed to improve the protective efficacy of anti-Trichinella vaccines.
Three species of nematodes from the Camallanidae that are known to infect Xenopus laevis Daudin (Anura: Pipidae) were collected from several localities across South Africa. New data on morphology, partial 28S and cox1 genes, infection levels and distribution are presented herein. The most common species, Batrachocamallanus slomei Southwell et Kirshner, 1937, from the stomach and less often oesophagus, was found in eight localities. Camallanus kaapstaadi Southwell et Kirshner, 1937, also from the oesophagus, was found in two localities and C. xenopodis Jackson et Tinsley, 1995, from the intestine, at a single locality. New localities for both C. kaapstaadi and C. xenopodis provide a geographical range extension. Males of C. xenopodis are described for the first time herein. The existence of a left spicule in the males of both the species of Camallanus Railliet and Henry, 1915 is confirmed and measurements are provided. Although C. xenopodis is distinguished from C. mazabukae Kung, 1948 in the present study, we suggest greater sampling effort in other African amphibians to confirm the species status of the latter taxon. Finally, the new molecular data showed distant relationships between collected species of Camallanus and species parasitising fish and freshwater turtles., Roman Svitin, Anneke L. Schoeman, Louis H. du Preez., and Obsahuje bibliografii
Polystomatid monogeneans have a wide diversity of life cycles correlated with the varied ecology and behaviour of their aquatic vertebrate hosts. Typically, transmission involves a swimming infective larva but most hosts are amphibious and invasion is interrupted when hosts leave water. A key life cycle adaptation involves a uterus that, in the most specialised cases, may contain several hundred fully-developed larvae prepared for instant host-to-host transmission. By contrast, one subfamily of the Polystomatidae - the Polystomoidinae, specific to chelonians (freshwater turtles) - has a simplified reproductive system without a uterus. Recently, Polystomoides nelsoni Du Preez et Van Rooyen, 2015 has been described with a uterus containing multiple eggs. The present study explores the exceptional interest of this parasite - for the functional biology of egg production, for the evolution of a reproductive system unique amongst ca 60 species in the subfamily, and for systematic relationships. A new genus is proposed, Uteropolystomoides gen. n., separate from the four currently-recognised genera Polystomoides Ward, 1917, Uropolystomoides Tinsley et Tinsley, 2016, Neopolystoma Price, 1939 and Polystomoidella Price, 1939 which lack a uterus. In addition, U. nelsoni (Du Preez et Van Rooyen, 2015) comb. n. has a suite of distinctive copulatory stuctures: a massive genital bulb with an exceptionally large number of very long genital spines and hyper-development of the vaginal openings. These characters set U. nelsoni apart from all other polystomoidines worldwide except Polystomoides multifalx Stunkard, 1924 and P. stunkardi Harwood, 1931. Missing data for these latter species preclude definitive assessment of inter-relationships but the distinguishing characters of U. nelsoni, especially the unique occurrence of the uterus, suggest a novel evolutionary pathway isolated from other lineages of polystomatids infecting chelonians., Richard C. Tinsley., and Obsahuje bibliografii
A comparative study of the scoleces of monozoic tapeworms (Cestoda: Caryophyllidea), parasites of catostomid and cyprinid fishes (Teleostei: Cypriniformes) in the Nearctic Region, was carried out using light and scanning electron microscopy. Scoleces of 22 genera of North American caryophyllideans were characterised and their importance for taxonomy, classification and phylogenetic studies was critically reviewed. Nearctic genera exhibit a much higher variation in the shape and form of scoleces compared with taxa in other biogeographical regions. The following basic scolex types can be recognised in Nearctic caryophyllideans: monobothriate (Promonobothrium Mackiewicz, 1968), loculotruncate (Promonobothrium, Dieffluvium Williams, 1978), bothrioloculodiscate (Archigetes Leuckart, 1878, Janiszewskella Mackiewicz et Deutsch, 1976, Penarchigetes Mackiewicz, 1969, Pseudoglaridacris Oros, Uhrovič et Scholz, 2018), fixomegabothriate (Capingens Hunter, 1927), bulbate and bulboacuminate (Atractolytocestus Anthony, 1958), cuneiloculate (Hypocaryophyllaeus Hunter, 1927, Rowardleus Mackiewicz et Deutsch, 1976, Spartoides Hunter, 1929), biacetabulate, bulboloculate, bothrioloculodiscate (Biacetabulum Hunter, 1927), tholate (Hunterella Mackiewicz et McCrae, 1962), cuneifimbriate (Khawia Hsü, 1935), cuneiform (Calentinella Mackiewicz, 1974, Caryophyllaeides Nybelin, 1922, Edlintonia Mackiewicz, 1970), hastate (Pseudolytocestus Hunter, 1929), loculotholate (Bialovarium Fischthal, 1953, Pliovitellaria Fischthal, 1951), and cuneiformoloculate (Glaridacris Cooper, 1920, Isoglaridacris Mackiewicz, 1965). The same type of scolex may be shared by species of different genera or families and species of the same genus can have a scolex of conspicuously different morphology, e.g. in Promonobothrium. Scolex morphology may be therefore of limited use in generic designation.