Revision of the genus Anguillicola Yamaguti, 1935 (Nematoda: Anguillicolidae) of the swimbladder of eels, including descriptions of two new species, A. novaezelandiae sp. n. and A. papernai sp. n

The genus Anguillicola is reviewed on the basis of the examination of some type specimens and other material. Two new species are described: A. novaezelandiae sp. n. from Anguilla australis (type host) from New Zealand and A. anguilla from Italy (conspecific nematodes were originally reported from Italy as A. australiensis) and A. papernai sp. n. from Anguilla mossambica from South Africa. At present the genus Anguillicola comprises the following five valid species: A. globiceps Yamaguti, 1935, A. australiensis Johnston et Mawson, 1940, A. crassus Kuwahara, Niimi et Itagaki, 1974, A. novaezelandiae sp. n., and A. papernai sp. n. Two Anguillicola species, A. crassus and A. novaezelandiae, were apparently introduced in Europe from other continents a few years ago. The genus Anguillicola is divided into two subgenera: Anguillicola (type species A. (A.) globiceps) and Anguillicoloides subgen. n. (type species A. (A.) crassus). All Anguillicola species are briefly described and illustrated. The paper is supplemented by a key to the identification of Anguillicola species.     

Granulinema gen. n. a new dracunculoid genus with two new species (G. carcharhini sp. n. and G. simile sp. n.) from the bull shark, Carcharhinus leucas (Valenciennes), from Louisiana, USA

Two new species of dracunculoid nematodes, G. carcharhini sp. n. and G. simile sp. n., representing a new genus Granulinema gen. n. (Dracunculoidea: Micropleuridae) are described from the bull shark, Carcharhinus leucas, from Louisiana (Lake Borgne), USA; the site of their localization in the host is unknown (probably abdominal cavity). The nematodes of both species were found in tissue juice and only males, juvenile females and body fragments of more advanced but nongravid females were obtained. Granulinema gen. n. differs from Micropleura, the only other genus in the family Micropleuridae, mainly by the presence of marked, dark excretory corpuscles in lateral excretory canals, pointed tail in females, greater number (6) of postanal pairs of caudal papillae in males, and by the presence of conspicuous transverse cuticular ornamentations on the body surface of mature females. The two new species can be easily distinguished from each other by the length of their spicules (0.78-0.90 mm in G. carcharhini sp. n. and 0.20-0.36 mm in G. simile sp. n.); moreover, there are two pairs of preanal papillae in the male of G. carcharhini sp. n., while there are three pairs in G. simile sp. n.     

Three new species of Gyrodactylus (Monogenoidea) from Varicorhinus capoeta gracilis (Keyserling) from the River Lenkoranka.

Three new species of Gyrodactylus Nordmann, 1832 are described from Varicorhinus capoeta gracilis (Keyserling) from the River Lenkoranka (Azebaijan SSR) as follows: G. varicorhini sp. n., G. capoetai sp.n. and G. mikailovi sp.n.     

Gyrodactylus bohemicus sp. n. (Monogenea: Gyrodactylidae) from Oncorhynchus mykiss (Walbaum) and Salvelinus fontinalis (Mitchill) (Clupeiformes: Salmonidae) in Czechoslovakia.

Gyrodactylus bohemicus sp. n (Gyrodactylidae: Monogenea) is described from the fins, skin and gill filaments of Oncorhynchus mykiss (Walbaum) and Salvelinus fontinalis (Mitchill) (Clupeiformes: Salmonidae) from a trout farm in Czechoslovakia. G. bohemicus sp. n. is most closely related to G. thymalli Zitnan, 1960 and G. magnus Konovalov, 1967 in the shape of the anchors, the ventral and the dorsal bar, but can be distinguished from both these species by the shape of the marginal hook proper.     

Restriction fragment length polymorphisms in Fusarium oxysporum f.sp. dianthi and other fusaria from Dianthus species

DNA restriction fragment length polymorphisms (RFLPs) among 46 isolates of Fusarium oxysporum from Dianthus spp., representing the known range of pathogenicity in carnation, were determined using total DNA digested with the restriction enzyme Hind III and a previously described probe, D4. Distinct multiple band RFLP patterns were found, which delineated RFLP groups as follows: (i) F. oxysporum f.sp. dianthi races I and 8; (ii) F. oxysporum f.sp. dianthi races 2, 5 and 6; (iii) F. oxysporum f.sp. dianthi race 4; (iv) a recently described race of F. oxysporum f.sp. dianthi (wilt-causing isolates from D. caryophyllus formerly classified as F. redolens); (v) wilt-causing isolates from D. barbatus formerly classified as F. redolens and (vi), (vii) and (viii), three further recently described races of F. oxysporum f.sp. dianthi. Isolate groups derived from analysis of RFLPs were consistent with existing and recently described vegetative compatibility groups (VCGs) in F. oxysporum f.sp. dianthi , but not in all cases with races. Isolates of F. oxysporum and F. proliferatum not associated with wilt disease had simpler RFLP patterns (with one exception) that were not associated with VCGs.     

Characterization of Fusarium oxysporum f. sp. radicis-cucumerinum attacking melon under natural conditions in Greece

A severe root and stem rot disease of melon was observed during the 2001 growing season on four glasshouse crops in Heraklio, Greece. A total of 43 isolates of F. oxysporum , obtained in Crete from glasshouse-grown melon and showing fusarium wilt or root and stem rot symptoms, were characterized by pathogenicity and vegetative compatibility. The majority of these isolates was also fingerprinted via amplified fragment length polymorphic (AFLP) analysis. Of the total number of isolates, 22 were identified by pathogenicity tests as F. oxysporum f. sp. melonis , 20 as F. oxysporum f. sp. radicis-cucumerinum , while one isolate was nonpathogenic on cucumber, melon, sponge gourd and pumpkin. All 22 isolates of F. oxysporum f. sp. melonis were assigned to vegetative compatibility group (VCG) 0134, and all 20 isolates of F. oxysporum f. sp. radicis-cucumerinum to VCG 0260. Isolates of F. oxysporum f. sp. radicis-cucumerinum were incompatible with isolates of F. oxysporum f. sp. melonis. AFLP fingerprinting allowed for the clustering of the isolates of the two formae speciales of F. oxysporum along two separate phenetic groups: f. sp. melonis to AFLP major haplotype I, and f. sp. radicis-cucumerinum to AFLP major haplotype II. Overall, pathogenicity, vegetative compatibility grouping and AFLP analysis were correlated and effectively distinguished isolates of F. oxysporum from melon. This appears to be the first report of natural infection of melon by F. oxysporum f. sp. radicis-cucumerinum worldwide.     

Single Cystosorus Isolate Production and Restriction Fragment Length Polymorphism Characterization of the Obligate Biotroph Spongospora subterranea f. sp. subterranea

ABSTRACT Spongospora subterranea f. sp. subterranea causes powdery scab in potatoes and is distributed worldwide. Genetic studies of this pathogen have been hampered due, in part, to its obligate parasitism and the lack of molecular markers for this pathogen. In this investigation, a single cystosorus inoculation technique was developed to produce large amounts of S. subterranea f. sp. subterranea plasmodia or zoosporangia in eastern black nightshade (Solanum ptycanthum) roots from which DNA was extracted. Cryopreservation of zoosporangia was used for long-term storage of the isolates. S. subterranea f. sp. subterranea-specific restriction fragment length polymorphism (RFLP) markers were developed from randomly amplified polymorphic DNA (RAPD) fragments. Cystosori of S. subterranea f. sp. subterranea were used for RAPD assays and putative pathogen-specific RAPD fragments were cloned and sequenced. The fragments were screened for specificity by Southern hybridization and subsequent DNA sequence BLAST search. Four polymorphic S. subterranea f. sp. subterranea-specific probes containing repetitive elements, and one containing single copy DNA were identified. These RFLP probes were then used to analyze 24 single cystosorus isolates derived from eight geographic locations in the United States and Canada. Genetic variation was recorded among, but not within, geographic locations. Cluster analysis separated the isolates into two major groups: group I included isolates originating from western North America, with the exception of those from Colorado, and group II included isolates originating from eastern North America and from Colorado. The techniques developed in this study, i.e., production of single cystosorus isolates of S. subterranea f. sp. subterranea and development of RFLP markers for this pathogen, provide methods to further study the genetic structure of S. subterranea f. sp. subterranea.     

Evolution of Fusarium oxysporum f. sp. vasinfectum Races Inferred from Multigene Genealogies

ABSTRACT Fusarium wilt of cotton is a serious fungal disease responsible for significant yield losses throughout the world. Evolution of the causal organism Fusarium oxysporum f. sp. vasinfectum, including the eight races described for this specialized form, was studied using multigene genealogies. Partial sequences of translation elongation factor (EF-1alpha), nitrate reductase (NIR), phosphate permase (PHO), and the mitochondrial small subunit (mtSSU) rDNA were sequenced in 28 isolates of F. oxysporum f. sp. vasinfectum selected to represent the global genetic diversity of this forma specialis. Results of a Wilcoxon Signed-Ranks Templeton test indicated that sequences of the four genes could be combined. In addition, using combined data from EF-1alpha and mtSSU rDNA, the phylogenetic origin of F. oxysporum f. sp. vasinfectum within the F. oxysporum complex was evaluated by the Kishino-Hasegawa likelihood test. Results of this test indicated the eight races of F. oxysporum f. sp. vasinfectum appeared to be nonmonophyletic, having at least two independent, or polyphyletic, evolutionary origins. Races 3 and 5 formed a strongly supported clade separate from the other six races. The combined EF-1alpha, NIR, PHO, and mtSSU rDNA sequence data from the 28 isolates of F. oxysporum f. sp. vasinfectum recovered four lineages that correlated with differences in virulence and geographic origin: lineage I contained race 3, mostly from Egypt, and race 5 from Sudan; lineage II contained races 1, 2, and 6 from North and South America and Africa; lineage III contained race 8 from China; and lineage IV contained isolates of races 4 and 7 from India and China, respectively.     

Genetic diversity in Fusarium oxysporum f.sp. dianthi and Fusarium redolens f.sp. dianthi

Pathogenic isolates were selected representing all known vegetative compatibility groups (VCGs) and races of Fusarium oxysporum sensu lato from Dianthus spp. On basis of differences in the internal transcribed spacer region of the ribosomal DNA, six VCGs were classified as F. oxysporum f.sp. dianthi and four as F. redolens f.sp. dianthi. All VCGs of F. oxysporum f.sp. dianthi were characterized by unique restriction fragment length polymorphisms (RFLPs), unique overall esterase profiles, and unique virulence spectra, supporting a clonal lineage concept. Two VCGs of F. oxysporum f.sp. dianthi nevertheless comprised more than one race, but races within the same VCG shared the same distinct overall virulence spectrum. VCGs belonging to F. redolens f.sp. dianthi also had unique RFLPs and unique virulence spectra, but had grossly identical esterase profiles. Three new races (9, 10 and 11) are described for F. oxysporum f.sp. dianthi, and four for F. redolens f.sp. dianthi. Two races previously considered lost were recovered; race 7 was identified as a member of VCG 0021 of F. oxysporum f.sp. dianthi while race 3 was identified as a distinct VCG and race of F. redolens f.sp. dianthi. A summary of races and VCGs in F. oxysporum f.sp. dianthi and F. redolens f.sp. dianthi is presented.     

Two new nematodes,Paraseuratoides ophisterni gen. et sp. n. (Quimperiidae) and Philometra ophisterni sp. n. (Philometridae), from the swamp-eel Ophisternon aenigmaticum in Mexico

Two new nematodes, Paraseuratoides ophisterni gen. et sp. n. (Seuratoidea: Quimperiidae) and Philometra ophisterni sp. n. (Dracunculoidea: Philometridae) are described based on specimens recovered from the intestine and mesentery, respectively, of the swamp-eel Ophisternon aenigmaticum Rosen et Greenwood (Synbranchiformes: Synbranchidae) from a canal of the Papaloapan River in Tlacotalpan, State of Veracruz, Mexico. The genus Paraseuratoides is mot similar to Paraseuratum Johnston et Mawson, 1940, differing from it mainly in the absence of a bulbous inflation on the anterior end of the oesophagus and in the structure of the mouth (presence of 6 spines in addition to 6 oesophageal teeth). Neoquimperia Wang, Zhao, Wang et Zhang, 1979 and Wuinema Yu et Wang, 1992 are synonymised with Ezonema Boyce, 1971 and Paragendria Baylis, 1939, respectively, and Haplonema hamulatum Moulton, 1931 is considered a junior synonym of Ichthyobronema conoura Gnedina et Savina, 1930. Philometra ophisterni (only females) is mainly characterised by minute cephalic papillae, a greatly developed anterior oesophageal bulb separated from the cylindrical part of the oesophagus, anterior extension of the oesophageal gland anterior to the nerve ring, and by the character of large caudal projections. This is the first Philometra species recorded from inland fishes in Mexico.     

Pathogenic, Genetic and Molecular Characterisation of Fusarium oxysporum f.sp. lilii

Isolates of Fusarium oxysporum from lily were screened for pathogenicity, vegetative compatibility and DNA restriction fragment length polymorphisms, and compared to reference isolates of F. oxysporum f.sp. gladioli and F. oxysporum f.sp. tulipae to justify the distinction of F. oxysporum f.sp. lilii. Twenty-four isolates from different locations in The Netherlands (18 isolates), Italy (4 isolates), Poland and the United States (1 isolate each) shared unique RFLP patterns with probes D4 and pFOM7, while hybridization did not occur with a third probe (F9). Except for a self-incompatible isolate, these 24 isolates all belonged to a single vegetative compatibility group (VCG 0190). Isolates belonging to VCG 0190 were highly pathogenic to lily, but not to gladiolus or tulip, except for a single nonpathogenic isolate. Six saprophytic isolates of F. oxysporum from lily were nonpathogenic or only slightly aggressive to lily, gladiolus and tulip, belonged to unique VCGs and had distinct RFLP patterns. Three pathogenic isolates previously considered to belong to F. oxysporum f.sp. lilii were identified as F. proliferatum var. minus; all three belonged to the same VCG and shared unique RFLP patterns. These three isolates were moderately pathogenic to lily and nonpathogenic to gladiolus and tulip. The reference isolates of F. oxysporum f.sp. tulipae were pathogenic to tulip, but not to lily and gladiolus; they shared a distinct RFLP pattern, different from those encountered among pathogenic and saprophytic isolates from lily, and formed a separate new VCG (VCG 0230). Reference isolates of F. oxysporum f.sp. gladioli belonging to VCG 0340 proved pathogenic to both gladiolus and lily, but not to tulip. These isolates, as well as isolates belonging to VCGs 0341, 0342 and 0343 of F. oxysporum f.sp. gladioli, had RFLP patterns different from those encountered among the isolates from lily or tulip. These findings identify F. oxysporum f.sp. lilii as a single clonal lineage, distinct from F. oxysporum f.sp. gladioli and f.sp. tulipae.     

Posterotestes unelen gen. et sp. n. (Digenea: Apocreadiidae) from Percichthys trucha (Osteichthyes: Percichthyidae) in Patagonia, Argentina

Posterotestes gen. n. (Digenea: Apocreadiidae, Apocrcadiinae) is proposed for specimens with the following features: spiny body, posterior position of gonads, extension of vitelline follicles up to level of intestinal bifurcation and absent at post-testicular space, long oesophagus and extension of caeca up to anterior testis. Posterotestes unelen sp. n. is described from the intestine of the native fish, Percichthys trucha (Cuvier et Valenciennes) (Osteichthyes: Percichthyidae) from Patagonian Andean lakes.     

小麦白粉病菌有性生殖与自然群体交配型检测

 本研究根据MAT1-1-1(GenBank登录号HQ171902)和MAT1-2-1(GenBank登录号HQ171899)的部分基因序列设计和筛选其特异性引物,使用PCR方法对2012年度采自四川、陕西、云南、贵州、河南、安徽、江苏、河北、山东和甘肃10个省30个采集地点共516个小麦白粉病菌菌株的交配型基因进行检测。结果显示30个采集地点两种交配型基因都能检测到,其中28个采集地的两种交配型菌株比例(MAT1-1-1 :MAT1-2-1)符合1:1。选择具不同交配型基因的小麦白粉病菌株配对接种,16个随机杂交组合都能产生闭囊壳。对获得的闭囊壳进行子囊孢子释放,均可获得子囊孢子。该研究结果为深入研究有性生殖对小麦白粉病菌株毒性多样性的作用奠定了基础。     

Dispersal of Formulations of Fusarium oxysporum f. sp. erythroxyli and F. oxysporum f. sp. melonis by Ants

ABSTRACT A natural epidemic of Fusarium wilt on coca (Erythroxylum coca) in Peru prompted the suggestion of possibly using the pathogen Fusarium oxysporum f. sp. erythroxyli as a mycoherbicide against this narcotic plant. During field trials conducted in Kauai, HI, to test the pathogenicity of the coca wilt pathogen, ants were observed removing formulations from test plots. While removal of formulations by ants was considered detrimental with respect to conducting field tests, ant removal was considered potentially beneficial in disseminating the mycoherbicide. Thus, research was initiated to assess the ability of formulation additives to alter the preference of ants for the formulated mycoherbicide. In Hawaii, preference of indigenous ants for removing formulations was tested using three different food bases (rice, rice plus canola oil, and wheat flour [gluten]). Similar tests were conducted at Beltsville, MD, using F. oxysporum f. sp. melonis, in which the formulation based on wheat flour was replaced by a formulation based on canola meal. Formulations based on wheat were preferred by ants in both locations; up to 90% of the wheat plus rice flour granules (C-6) and the wheat gluten plus kaolin granules (pesta) were removed within 24 h, while only 20% of those containing rice without oils were taken. However, when either canola, sunflower (Maryland only), or olive oil was added to the rice formulation, up to 90% of the granules were taken. The formulation based on canola meal was less attractive to ants, as only 65% of the granules were removed within a period of 24 h. Ants showed no preference with respect to presence or absence of fungal biomass. To alter the attractiveness of the C-6 formulation to ants, C-6 was amended with three natural products. Canna and tansy leaves were added to C-6 at a ratio of 1:5 (wt/wt), while chili powder was added at 1:25 or 1:2.5 (wt/wt). Canna, tansy, and the higher rate of chili powder significantly reduced the number of C-6 granules removed by ants. Canna and tansy leaves affected neither germination nor sporulation of the mycoherbicide, while the high concentration of chili powder reduced viability of propagules in the formulation. More F. oxysporum f. sp. erythroxyli-type colonies were recovered from inside ant nests (9 cm depth) than from nest surfaces, indicating that ants may distribute the mycoherbicide in the soil profile. Ants passively carried propagules of F. oxysporum f. sp. erythroxyli outside their bodies, as well as either very closely adhering to the outside or within their bodies.     

Philometrid nematodes infecting fishes from the Everglades National Park, Florida, U.S.A

The following three species of the Philometridae (Nematoda: Dracunculoidea) are described from marine perciform fishes of the Everglades National Park (northern Gulf of Mexico), Florida, U.S.A.: Philometra brevispicula sp. n. (male and females) from the subcutaneous tissue of mouth of the gray snapper Lutjanus griseus (Linnaeus) (Lutjanidae), Philometroides grandipapillatus sp. n. (only females) from pectoral fin muscle of the crevalle jack Caranx hippos (Linnaeus) (Carangidae), and Caranginema americanum Moravec, Montoya-Mendoza et Salgado-Maldonado, 2008 (females) from the subcutaneous fascia of trunk muscle in crevalle jack C. hippos. Philometra brevispicula is mainly characterized by small cephalic papillae of the external circle, the absence of oesophageal teeth and the presence of small caudal projections in gravid female, markedly short spicules (45 microm) in male, and by its location in the host. Philometroides grandipapillatus differs from congeners mainly in the shape of the cephalic region (narrow, conspicuously protruding), large cephalic papillae of the external circle and the absence of caudal projections in female, and by the site of infection in the host. Caranginema americanum is for the first time recorded from the northern Gulf of Mexico.     

Three new species of the ectoparasitic mites of the genus Syringophiloidus Kethley, 1970 (Acari: Syringophilidae) from passeriform birds from Slovakia

Three new quill mite species of the genus Syringophiloidus Kethley, 1970 (Acari: Syringophilidae) from Slovakia are described and figured: S. bombycillae sp. n. from the waxwing Bombycilla garrulus (L.) (Passeriformes: Bombycillidae), S. schoeniclus sp. n. from the reed bunting Emberiza schoeniclus L. (Passcriformes: Emberizidae) and S. montanus sp. n. from the tree sparrow Passer monlanus (L.) (Passeriformes: Ploceidae). A key to all known species of the genus Syringophiloidus is given.     

PCR-based differentiation of Fusarium oxysporum ff. sp. lycopersici and radicis-lycopersici and races of F. oxysporum f. sp. lycopersici

The pathogenic type (form and race) of Fusarium oxysporum, which generates wilt symptoms on tomato, was rapidly identified with a polymerase chain reaction (PCR)-based technique. We compared the partial nucleotide sequences of endo polygalacturonase (pg1) and exo polygalacturonase (pgx4) genes from isolates of F. oxysporum ff. sp. lycopersici (FOL) and radicis-lycopersici (FORL) from Japan and designed specific primer sets (uni, sp13, sp23, and sprl) based on the nucleotide differences that appeared among the pathogenic types. PCR with the uni primer set amplified a 670∼672-bp fragment from all isolates of FOL and FORL. With the sp13 primer set, an amplicon of 445 bp was obtained only from isolates of FOL race 1 and 3. With the sp23 primer set, a 518-bp fragment was obtained from isolates of FOL race 2 and 3. The sprl primer set yielded a 947-bp fragment from isolates of FORL, but not from FOL. A combination of amplifications with these primer sets effectively differentiated the pathogenic types of F. oxysporum in tomato.     

Caryophyllaeides ergensi sp. n. (Cestoda: Caryophyllidea) from Leuciscus Leuciscus baicalensis from Mongolia

The tapeworm Caryophyllaeides ergensi sp. n. (Cestoda: Caryophyllidea) is described from Leuciscus leuciscus baicalensis from the River Selenga basin in Mongolia. It differs from the only hitherto recognized species of the genus C. fennica (Schneider, 1902), mainly in its extremely long uterus with coils extending anteriorly to anterior half, sometimes up to anterior third of the body of gravid specimens.     

Genetic Diversity in Australian Populations of Puccinia graminis f. sp. avenae

ABSTRACT Sequence-tagged microsatellite profiling was used to develop 110 microsatellites for Puccinia graminis f. sp. tritici (causal agent of wheat stem rust). Low microsatellite polymorphism was exhibited among 10 pathogenically diverse P. graminis f. sp. tritici isolates collected from Australian cereal growing regions over a period of at least 70 years, with two polymorphic loci detected, each revealing two alleles. Limited cross-species amplification was observed for the wheat rust pathogens, P. triticina (leaf rust) and P. striiformis f. sp. tritici (stripe rust). However, very high transferability was revealed with P. graminis f. sp. avenae (causal agent of oat stem rust) isolates. A genetic diversity study of 47 P. graminis f. sp. avenae isolates collected from an Australia-wide survey in 1999, and a historical group of 16 isolates collected from Australian cereal growing regions from 1971 to 1996, revealed six polymorphic microsatellite loci with a total of 15 alleles. Genetic analysis revealed the presence of several clonal lineages and subpopulations in the pathogen population, and wide dispersal of identical races and genotypes throughout Australian cereal-growing regions. These findings demonstrated the dynamic population structure of this pathogen in Australia and concur with the patterns of diversity observed in pathogenicity studies.     

Experimental infection of Atlantic salmon (Salmo salar) with marine Eubothrium sp. (Cestoda: Pseudophyllidea): observations on the life cycle, aspects of development and growth of the parasite.

The life cycle of marine Eubothrium sp. (Cestoda: Pseudophyllidea), from Atlantic salmon (Salmo salar L.) was experimentally completed in one year and included only one intermediate host (Acartia tonsa Dana) (Copepoda: Calanoida). Adult cestodes were collected from farmed salmon, and ripe eggs released by the cestodes were fed to Acartia tonsa. Ingested eggs hatched in the gut and the larvae developed in the haemocoel of the copepod for 15 days at 16 degrees C. A total of 170 seawater-reared salmon were exposed to infected copepods and the total prevalence of Eubothrium sp. in the salmon after infection was 95.3%, with a mean intensity of 15.0 (range 1-87). The infected salmon were kept in the laboratory where the growth of the cestodes was studied for eleven months. Mean length of the cestodes increased with time, but a large variation among the cestodes was observed. Growth and maturation of the cestodes were dependent on host size and the number of worms present in the intestine. No evidence of mortality of Eubothrium sp. was observed during the experimental period.