Morphological and molecular characterization of a new species of leech (Glossiphoniidae,Hirudinida): Implications for the health of its imperiled amphibian host (Cryptobranchus alleganiensis)

The hellbender (Cryptobranchus alleganiensis) is among the most intriguing and imperiled amphibians in North America. Since the 1970s and 80s, western populations of the Ozark and eastern subspecies in Missouri have declined by nearly 80%. As a result of population declines, the Ozark hellbender was recently federally protected as an endangered species, and the eastern subspecies was granted protection under CITES. Although habitat degradation is probably the biggest threat to hellbender populations, recent evidence suggests that pathogens including chytrid fungus and “flesh-eating” bacteria may also contribute to declines in Ozark hellbenders. Leeches, which are very common on Ozark hellbenders, have recently been implicated as possible vectors of disease among Ozark hellbenders but have not been described in eastern hellbenders or outside of Missouri and Arkansas. We discovered a population of leeches on eastern hellbenders in southwest Virginia and confirmed that the species of leech is within the genus Placobdella, but is morphologically and genetically distinct from all previously described leech species. We named the new species Placobdella appalachiensis sp. n. Moser and Hopkins, based on the mountainous region in which it was discovered. Our surveys over a three consecutive year period suggested that this leech species may be patchily distributed and/or have a narrow geographic range. We consistently detected leeches at one site (mean prevalence in 80 hellbenders = 27.5%; median intensity = 3.0 leeches per parasitized hellbender [range 1 – >250 leeches]) during three years of surveys, but we never found leeches in any of our other seven study sites in two streams (mean prevalence in 139 hellbenders = 0%). We found a significant positive relationship between hellbender body size and the intensity of parasitism, and we suggest the possibility that the behavioral ecology of adults leading up to reproduction may increase their encounter rates with parasites. We discuss the potential conservation implications of discovery of leeches in this stream, and make recommendations for future mitigation and monitoring efforts.     

A new species of Orobdella (Hirudinida, Arhynchobdellida, Orobdellidae) from Taipei, Taiwan

A new quadrannulate species of Orobdella, Orobdella ketagalansp. n., from Taipei, Taiwan, is described. This is the first record of Orobdella and the family Orobdellidae from Taiwan. This new species possesses small, paired sperm duct bulbs in the male reproductive system. In addition to these bulbs, the following combination of characters distinguishes this new species from other quadrannulate species: somite IV uniannulate, male gonopore at XI b6, female gonopore at XIII a1, 1/2 + 4 + 1/2 between gonopores, simple tubular gastroporal duct, lacking epididymides, and undeveloped atrial cornua. Phylogenetic analyses using nuclear 18S rDNA and histone H3 as well as mitochondrial COI, 12S rDNA, tRNA(Val), and 16S rDNA markers showed that Orobdella ketagalan is related to the two Ryukyu Archipelago species Orobdella dolichopharynx Nakano, 2011 and Orobdella shimadae Nakano, 2011.     

A new sexannulate species of Orobdella (Hirudinida, Arhynchobdellida, Orobdellidae) from Yakushima Island, Japan

A new sexannulate species of the genus Orobdella Oka, 1895, Orobdella mononokesp. n., is described on the basis of five specimens collected from Yakushima Island, Japan. Orobdella mononokesp. n. differs from other sexannulate Orobdella species in its possessing the following combination of characters: dorsal surface bicolor in life, I-XIII, XXVII and caudal sucker grayish purple, XIV-XXVI amber, male gonopore at XI c11/c12, female gonopore at XIII b2, 8 + 1/2 between gonopores, tubular but bulbous at junction with crop gastroporal duct, epididymides in XV-XIX, and atrial cornua ovate. Phylogenetic analyses using nuclear 18S rDNA and histone H3, and mitochondrial COI, tRNA(Cys), tRNA(Met), 12S rDNA, tRNA(Val) and 16S rDNA markers show that Orobdella mononokesp. n. is closely related to Orobdella esulcata Nakano, 2010 from Kyushu, Japan, and two species, Orobdella dolichopharynx Nakano, 2011 and Orobdella shimadae Nakano, 2011, from the Ryukyu Archipelago, Japan.     

Biocatalytic surface modification of knitted fabrics made of poly (ethylene terephthalate) with hydrolytic enzymes from Thermobifida fusca KW3b

Knitted fabrics made from poly(ethylene terephthalate) fibres were treated with an enzyme preparation from Thermobifida fusca KW3b showing hydrolytic activity on p-nitrophenyl butyrate. The fabrics were also treated with NaOH and the results were compared. Both enzyme- and alkaline-treated fabrics showed an increase in reactive dye uptake, vertical wicking height and water absorption capacity of the fabrics indicating an increased surface hydrophilicity. However, X-ray photoelectron (XPS) and energy-dispersive spectroscopy (EDS) did not give conclusive results on the presence of newly introduced hydrophilic groups on the surface of the fibres. Analysis of the enzyme-treated poly(ethylene terephthalate) fibres by scanning electron microscopy (SEM) and by atomic force microscopy (AFM) indicated an increase in surface roughness of the fibres, which may contribute to the observed increased hydrophilicity of the PET fabrics. However, much longer treatment times (24 h) were required to obtain these effects with the enzymes compared to the chemical treatment (1 h).     

On the classification, evolution and biogeography of terrestrial haemadipsoid leeches (Hirudinida: Arhynchobdellida: Hirudiniformes)

A scourge of tropical and subtropical jungles, bloodfeeding terrestrial leeches of Haemadipsidae have long confused systematists and defied sensible biogeographic interpretation. The family Haemadipsidae usually includes problematic taxa that neither fit the typical IndoPacific distribution of the group, nor properly match diagnostic characters used to define the family. Historically, four additional families-Xerobdellidae, Diestecostomatidae Mesobdellidae and Nesophilaemonidae-have occasionally been recognized for New World and European representatives, though agreement on the composition of those families has not been consistent. Here, we expand the phylogenetic sampling of non-IndoPacific (among other) genera to include Meso American Diestecostoma species and Nesophilaemon skottsbergii from the Juan Fernandez Archipelago in order to critically assess prior hypotheses in a molecular phylogenetic analysis of arhynchobdellid leeches. The result, based on nuclear 18S rDNA and 28S rDNA and mitochondrial COI indicates that there are two distantly related lineages of bloodfeeding terrestrial leeches. The otherwise monophyletic family Haemadipsidae is found to exclude species of Xerobdella, Mesobdella and Diestecostoma. Xerobdellidae is formally resurrected to accommodate species of those three genera. Morphological characteristics corroborate the distinction of Haemadipsidae and Xerobdellidae on the basis of sexual and nephridial characters. Idiobdella seychellensis belongs in Haemadipsidae notwithstanding its lacking respiratory auricles. Nesophilaemon skottsbergii too is in Haemadipsidae notwithstanding its geographic proximity to the xerobdellid Mesobdella gemmata. The characters used to define haemadipsoid families are reevaluated. Feeding preferences and biogeographic patterns are also examined.     

Holotype redescription of Mimobdella japonica (Hirudinida, Arhynchobdellida, Erpobdelliformes) and taxonomic status of the genus Mimobdella

Mimobdella japonica Blanchard, 1897, the type species of the genus Mimobdella Blanchard, 1897, is redescribed based on the holotype. This species is characterized by the following characteristics: mid-body somites novem-annulate, two post-anal annuli, male gonopore in XI/XII, female gonopore in XII/XIII, 9 annuli (one full somite) between gonopores, strepsilaematous pharynx and three myognaths with stylets, possessing post-crop caeca in pairs, ovisacs reaching to XXI a2. The genus Mimobdella is placed under the family Salifidae, not Gastrostomobdellidae or Erpobdellidae, according to its possessing three myognaths bearing pharyngeal stylets.     

Two new species of Austrobdella (Hirudinida: Piscicolidae) from Chile

Austrobdella coliumicus n. sp. is described from Coliumo Bay, Chile. It is characterized by a continuous, external coelomic canal (= marginal lacuna), 5 pairs of testisacs, accessory gland cells, a body not distinctly divided into trachelosome and urosome, 2 pairs of dorsal ocelli on the trachelosome, dorsal and ventral segmental ocelli present on the urosome, green overall pigmentation with transverse brown bands, and the absence of conducting tissue. Austrobdella coliumicus is distinguished from other species of Austrobdella by the presence of 2 pairs of ocelli on the trachelosome and a more cylindrical body. It is unusual that this leech was collected from inside the mantle cavity of the razor clam, Ensis macha. Austrobdella losmoliniensis n. sp. is described from the electric ray, Discopyge tschudii, collected at Los Molinos, Chile. It is characterized by a continuous, external coelomic canal; 5 pairs of testisacs; accessory gland cells; a body distinctly divided into trachelosome and urosome; 1 pair of eyes on the oral sucker; overall black pigmentation with unpigmented areas; and the absence of conducting tissue. Austrobdella losmoliniensis can be distinguished from other species of Austrobdella by the combination of 1 pair of eyes on the oral sucker and black pigmentation.     

Novel assay for determining the metabolic fate of juvenile hormone III: A study with Drosophila melanogaster

A thin-layer chromatographic assay was developed for the resolution of hydrolytic and conjugative catabolites of juvenile hormone (JH). A single-dimension, dual-development thin-layer system allowed complete resolution of the catabolites. Thus, this system provided a means for the rapid and economic analysis of JH hydrolysis even when different hydrolytic activities were present concurrently. Purified hydrolytic enzymes were found to be superior to chemical methods for the generation of small amounts of standards of JH catabolites. The relative levels of activities of an epoxide hydrolase and an esterase toward JH III were found to be similar in microsomal preparations from three lines of adult Drosophila melanogaster isolated from a field population. However, selection of flies by exposure to cut orange resulted in the elevation of levels of epoxide hydrolase activities, whereas esterase levels were not affected to the same extent. The formation of the JH acid-diol was not detected under the conditions of this study, suggesting that the JH acid and diol were not good substrates for epoxide hydrolase and juvenile hormone esterase, respectively.     

Regulation of JH titers: The relevance of degradative enzymes and binding proteins

Juvenile hormones play a crucial role in development, metamorphosis, and reproduction of insects. This mini-review discusses the nature of the juvenile hormones identified in insects and their changes in concentration in the hemolymph during development and reproduction. The hemolymph titer is largely determined by the rate at which juvenile hormones are synthesized and released by the corpora allata, but other factors are also involved in titer regulation, such as the affinity and concentration of juvenile hormone binding proteins in the hemolymph and the rate of juvenile hormone degradation in hemolymph and tissues. Juvenile hormone specific esterases occur in hemolymph and tissues, whereas epoxide hydrolases, which may degrade the hormone, are exclusively tissue bound. The activities of these degradative enzymes and the concentration of binding proteins change during the insect life cycle and these changes are related to fluctuations in hormone titer. However, we are still a long way from understanding the subtle interactions between these components in regulation of juvenile hormone titers. In particular, our knowledge is hampered by lack of information about the types, concentrations, and affinities of intracellular juvenile hormone receptors. © 1996 Wiley-Liss, Inc.     

Activity of hydrolytic enzymes of Candida albicans strains isolated from patients with periodontal and membrane mucosae of oral cavity diseases

Fungi are elements of the ontocenosis of the oral cavity and causal factors of inflammatory lesions in its mucous membrane. The objective of the study was to find differences in the activity of hydrolytic enzymes of Candida albicans isolated from patients with diseases of the periodontium and mucous membrane of the oral cavity. Of 235 patients examined, 31 were diagnosed with gingivitis, 38 with glossitis, 28 with leucoplakia, 37 with adult periodontitis, 25 with juvenile periodontitis, 36 with stomatitis prothetica and 40 with stomatitis atrophica. In 196 patients (83.4 ± 2.4%), fungi belonging to Candida species were detected. In the evaluation of Candida albicans strains (146) properties, bioMerieux API ZYM tests containing substrates for the detection of 19 hydrolases were used. All the investigated strains were characterized by the activity of 14 enzymes, i.e. phosphatase alcaline, esterase (C4), esterase lipase (C8), leucine and valine arylamidase, phosphatase acid, naphthol-AS-BI-phosphohydrolase, α galactosidase, β galactosidase, α glucosidase, β glucosidase, N-acetyl-β-glucosaminidase, α mannosidase and α fucosidase. Strains isolated from the oral cavity of patients with diseases of periodontium and mucous membrane are characterised by the highest phosphatase acid activity. The greatest enzymatic activity is characteristic of Candida albicans isolated from patients with stomatitis atrophica or stomatitis prothetica, and the lowest in strains from gingivitis or juvenile periodontitis cases. Differences in the activity of hydrolases are statistically significant (p<0.01) for: esterase (C4), leucine and valine arylamidase, phosphatase acid, naphthol-AS-BI-phosphohydrolase, β glucosidase, N-acetyl-β-glucosaminidase, of fungi isolated from patients with particular clinical diagnoses. This revised version was published online in June 2006 with corrections to the Cover Date.     

Solubilization of low-rank coal by Trichoderma atroviride: Evidence for the involvement of hydrolytic and oxidative enzymes by using 14C-labelled lignite

The deuteromycete Trichoderma atroviride is able to solubilize lignite in dependence on a given carbon source for growth. When cultivated on media containing glutamate, this mold excreted a set of different enzymes with hydrolytic activity. Addition of lignite to the growth media induced the synthesis of extracellular lignite-specific esterase activity but no evidence has been provided for its direct involvement in the process of lignite solubilization. Hence, the basic capability of T. atroviride enzymes to degrade a variety of ester and ether bonds at the surface or within the bulky lignite structure was tested using coal following its direct labelling with ¹⁴C-alkyl iodide. The participation of hydrolytic and oxidative enzymes in lignite degradation was assessed by measuring the release of ¹⁴C radioactivity from selectively alkylated carboxylic and phenolic OH groups. T. atroviride cleaved both carboxylic esters using esterases and the phenolic ether bonds by using oxidative enzymes, most likely laccases. Journal of Industrial Microbiology & Biotechnology (2002) 28, 207–212 DOI: 10.1038/sj/jim/7000232 Received 05 July 2001/ Accepted in revised form 08 December 2001     

A new species of Mysidobdella (Hirudinida: Piscicolidae) from mysids along the California coast

Mysidobdella californiensis n. sp. is described from the mysid Holmesimysis sculpta from Bodega Bay on the central California coast and from Holmesimysis costata var. from San Pedro on the southern California coast. The internal anatomy of M. californiensis is similar to that of the only other species in the genus, Mysidobdella borealis from the north Atlantic Ocean, except that M. californiensis lacks the medial, unpaired seminal receptacle present in M. borealis. Externally, M. californiensis is slightly larger and more robust than M. borealis, with a much larger caudal sucker. The most striking difference between the species is the unusually large, trumpet-shaped, fluted oral sucker in M. californiensis. At Bodega Bay, the prevalence of M. californiensis on its host was 17% with an average intensity of 1.46 (range 1-3) leeches per host.     

东亚飞蝗山西两地理种群酯酶特性的比较研究

对东亚飞蝗山西临猗和永济2个地理种群的酯酶特性进行了比较研究.非变性聚丙烯酰胺凝胶电泳图谱显示:以a-乙酸萘酯为底物染色,2个东亚飞蝗种群谱带差别不明显.但是,酯酶动力学研究结果表明:以a-乙酸萘酯和α-丁酸萘酯为底物时,永济种群的酯酶活性分别是临猗种群的1.81倍和1.20倍.永济种群酯酶活性的增高可能与非变性聚丙烯酰胺凝胶电泳图谱显示出较临猗种群多出的酶带有关.体外酯酶抑制动力学研究表明:永济和临猗2种群所含酯酶大都为B型酯酶,其含量分别为84.94%和91.47%.永济种群对对氧磷的耐受性要高于临猗种群,我们推测可能与2种群马拉硫磷使用背景不同有关.     

Observations on three species of branchiobdellid (Annelida: Clitellata) worms from eastern Asia

Three jars from the Hamburg Museum contained specimens of Branchiobdella minuta, presumably from the original Pierantoni (1912) material, and Cirrodrilus cirratus and C. uchidai from Yamaguchi's (1932a) collection. The head of a syntype of B. minuta was described and B. cheni becomes its junior subjective synonym. One specimen each of C. cirratus and C. uchidai was sectioned and described, particularly the male genitalia. The diet of all three branchiobdellids consists of microorganisms. In Cirrodrilus sp. basic protein granules were demonstrated in the gland cells of the intestine and its chloragogen cells. The peristomial and lateral epidermal gland cell secretions were characterized histochemically.