Molecular Evidence for Association of Chlamydiales Bacteria with Epitheliocystis in Leafy Seadragon (Phycodurus eques), Silver Perch (Bidyanus bidyanus), and Barramundi (Lates calcarifer)

Epitheliocystis in leafy seadragon (Phycodurus eques), silver perch (Bidyanus bidyanus), and barramundi (Lates calcarifer), previously associated with chlamydial bacterial infection using ultrastructural analysis, was further investigated by using molecular and immunocytochemical methods. Morphologically, all three species showed epitheliocystis cysts in the gills, and barramundi also showed lymphocystis cysts in the skin. From gill cysts of all three species and from skin cysts of barramundi 16S rRNA gene fragments were amplified by PCR and sequenced, which clustered by phylogenetic analysis together with other chlamydia-like organisms in the order Chlamydiales in a lineage separate from the family Chlamydiaceae. By using in situ RNA hybridization, 16S rRNA Chlamydiales-specific sequences were detected in gill cysts of silver perch and in gill and skin cysts of barramundi. By applying immunocytochemistry, chlamydial antigens (lipopolysaccharide and/or membrane protein) were detected in gill cysts of leafy seadragon and in gill and skin cysts of barramundi, but not in gill cysts of silver perch. In conclusion, this is the first time epitheliocystis agents of leafy seadragon, silver perch and barramundi have been undoubtedly identified as belonging to bacteria of the order Chlamydiales by molecular methods. In addition, the results suggested that lymphocystis cysts, known to be caused by iridovirus infection, could be coinfected with the epitheliocystis agent.     

Microhabitats of gill parasites (Monogenea and copepoda) of teleosts (Scomberoides spp.)

Ramasamy P., Ramalingam K., Hanna R. E. B. and Halton D. W. 1985. Microhabitat of gill parasites (Monogenea and Copepoda) of teleosts (Scomberoides spp.). International Journal for Parasitology15: 385–397. The parasites Vallisia indica, Allodiscocotyla chorinemi, Heterapta chorinemi, and Dionchus remorae exhibited site specificity on the gills of Scomberoides commersonianus, S. tol, S. lysan and S. tala, whereas the copepod Caligus sp. did not. Observations on site preference revealed microhabitat differences along the length of gill filaments, along the anterio-posterior axis of the gill, between external and internal gill filaments and on different gill arches. Site preference varied with parasite density on each gill and host. An interspecific association test between pairs of species revealed, in some cases, a positive association, and in other cases a negative association. There were apparently no association? between certain pairs of species. A comparison of intensity within pairs of parasite species infecting the same host revealed either an inverse or a direct correlation. The numerical dominance and prevalence of parasites differed on each host species. This study indicates that intra- and interspecific competition may occur among the gill parasites. The direction and speed of ventilation water-currents and certain intrinsic factors of the parasite themselves may determine their microhabitat restriction on the gills.     

Arecanut white grubs Leucopholis species (Melolonthinae: Scarabaeidae: Coleoptera) morphological,molecular identification and phylogenetic analysis

Arecanut (Areca catechu) is one of the most important commercial crops in India, which grows extensively in the malnad and coastal areas. White grubs are the most important pests on this crop and cause considerable yield loss. Surveys were conducted to record species that are infested with white grubs of Karnataka and Kerala states. Three species of white grubs were collected and described using morphological and molecular characters. A detailed study of male genitalial structures such as aedeagus and endophallus in three species of Leucopholis Dejean, 1833 (Coleoptera: Scarabaeidae: Melolonthinae) viz., Leucopholis lepidophora Blanchard, 1850, Leucopholis burmeisteri Brenske, 1894 and Leucopholis coneophora Burmeister, 1855 showed substantial differences. The mitochondrial cytochrome oxidase subunit I gene was sequenced for molecular identification and phylogenetic analysis in this respect. L. lepidophora, L. burmeisteri and L. coneophora species were successfully identified by employing COXI gene. Phylogenetic analyses revealed that the COXI DNA barcode fragment of Leucopholis species and out-group taxa, available in genetic databases, confirms the species identity.     

Locomotory,ventilatory and metabolic responses of the subterranean Stenasellus virei (Crustacea,Isopoda) to severe hypoxia and subsequent recovery

The locomotory and ventilatory activities and the intermediary and energy metabolism modifications of the hypogean aquatic isopod crustacean Stenasellus virei were investigated in severe hypoxia (Po₂ < 0.03 kPa) and subsequent recovery. The aims of this study were i) to determine why the subterranean species displayed a greater tolerance of hypoxia than numerous other epigean crustaceans, ii) to confirm previous results obtained with four hypogean and epigean crustaceans, iii) to compare the responses to severe hypoxia in hypogean amphipods and isopods, and iv) to better understand the ecological problems of the hypogean organisms survival in subterranean habitats. S. virei responded to experimental long-term, severe hypoxia with classical anaerobic metabolism mainly characterized by a decrease in adenosine triphosphate (ATP) and phosphagen, utilization of glycogen and glutamate, and accumulation of lactate and alanine. Lactate was also largely excreted by this organism, which is unusual for crustaceans in general. Compared to most other epigean crustaceans, the isopod S. virei showed high amounts of stored glycogen and arginine phosphate. These differences in glycogen and phosphagen stores, and the ability to reduce energetic expenditures linked to locomotion and ventilation, extended the survival of S. virei under experimental anaerobiosis. During recovery, the isopod S. virei showed a higher capacity for glyconeogenesis from lactate and a faster and total replenishment of ATP and arginine phosphate levels than epigean crustaceans. Data concerning responses to hypoxia and subsequent recovery in S. virei are similar to those previously obtained with two other hypogean amphipods, except that this isopod did not synthesize succinate in anaerobiosis.     

Molecular evidence for association of chlamydiales bacteria with epitheliocystis in leafy seadragon (Phycodurus eques), silver perch (Bidyanus bidyanus), and barramundi (Lates calcarifer)

Epitheliocystis in leafy seadragon (Phycodurus eques), silver perch (Bidyanus bidyanus), and barramundi (Lates calcarifer), previously associated with chlamydial bacterial infection using ultrastructural analysis, was further investigated by using molecular and immunocytochemical methods. Morphologically, all three species showed epitheliocystis cysts in the gills, and barramundi also showed lymphocystis cysts in the skin. From gill cysts of all three species and from skin cysts of barramundi 16S rRNA gene fragments were amplified by PCR and sequenced, which clustered by phylogenetic analysis together with other chlamydia-like organisms in the order Chlamydiales in a lineage separate from the family Chlamydiaceae. By using in situ RNA hybridization, 16S rRNA Chlamydiales-specific sequences were detected in gill cysts of silver perch and in gill and skin cysts of barramundi. By applying immunocytochemistry, chlamydial antigens (lipopolysaccharide and/or membrane protein) were detected in gill cysts of leafy seadragon and in gill and skin cysts of barramundi, but not in gill cysts of silver perch. In conclusion, this is the first time epitheliocystis agents of leafy seadragon, silver perch and barramundi have been undoubtedly identified as belonging to bacteria of the order Chlamydiales by molecular methods. In addition, the results suggested that lymphocystis cysts, known to be caused by iridovirus infection, could be coinfected with the epitheliocystis agent.     

Structural and functional features of gill epithelium and the brood pouch of pipefish <Emphasis Type="Italic">Sygnathus acusimilis</Emphasis> (Syngnathidae,Gasterosteiformes) during adaptation to dilute sea water

Study of the dilate sea-water tolerance of the pipefish Syngnathus acusimilis indicated that it successfully acclimatized to an abrupt change in water salinity from 5 to 32‰. Larvae in the brood pouch also successfully acclimatized to salinity change, and, after several days, juveniles left the brood pouch and continued to live. In freshwater, adult and fry perished during several minutes. In the gill epithelium of S. acusimilis, chloride cells of only a marine type were found. A comparison of the ultrastructure of chloride cells of gill epithelium and mitochondrion-rich cells of the brood pouch of S. acusimilis demonstrated their high similarity. The ultrastructure of cells of the brood pouch indisputably attests to their involvement in maintaining an osmotic and ionic homeostasis in the cavity of the brood pouch.     

Parasitic copepods of the common sole, Solea solea (L.), from the Eastern Mediterranean coast of Turkey

There is increasing interest in the common sole, Solea solea (Linnaeus), as an alternative fish species in aquaculture in the Mediterranean region, and parasitic copepods are a potential hazard for farmed finfish. This paper provides taxonomic information on two species of sea lice (family Caligidae) collected from S. solea in eastern Mediterranean waters off the Turkish coast. Caligus brevicaudatus A. Scott, 1901 and Caligus apodus (Brian, 1924) were both found and this is the first report of C. brevicaudatus in Turkish waters. The discovery of C. apodus on S. solea is a new host record. Key diagnostic characters of both species are reported, supported by light and scanning electron microscopy observations. During a 12-month survey a prevalence of 28% was recorded for C. brevicaudatus, whereas for C. apodus peak prevalence was much lower (3%).     

Occurrence and assemblage composition of millipedes (Myriapoda, Diplopoda) and terrestrial isopods (Crustacea, Isopoda, Oniscidea) in urban areas of Switzerland

Terrestrial isopods and millipedes, members of the invertebrate macro-decomposer guild, were collected through pitfall traps in three Swiss cities (Zurich, Lucerne, Lugano). A total of 7,198 individuals of 17 isopod species (7093 ind.), and 10 millipede species (105 ind.) were captured. Besides the Alpine endemic isopod (Trichoniscus alemannicus) and millipede (Cylindroiulus verhoeffi), urban assemblages were mainly composed of widespread, native European and even cosmopolitan species, which are frequent in anthropogenic areas. Overall species richness (isopods and millipedes combined) was similar in Zurich (17 species) and Lucerne (16), while only 13 species were sampled in Lugano. According to the Sørensen index of similarity, species composition of Zurich and Lucerne were more alike, while the one of Lugano was more distinct from the other two cities. This result can be explained by the spatial proximity of Zurich and Lucerne in the north of the Alps compared to Lugano, which is located more distantly and in the south of the Alps. Dominant isopods and millipedes in Zurich and Lucerne were found to be widespread synanthropic species in temperate Europe(Porcellio scaber, Trachelipus rathkii and Ophyiulus pilosus) while the dominant isopod in Lugano (Trachelipus razzautii) is a species with a north-eastern Mediterranean distribution. Our study reveals that the urban millipede and isopod fauna in Swiss cities mainly consists of widespread species, but species of narrower distribution (e.g. Trichoniscus alemannicus, Cylindroiulus verhoeffi) may also find suitable habitats in cities. Despite some signs of biotic homogenization, our study also found compositional differences of millipede and isopod assemblages between northern and southern cities that suggest geographical effects of the regional species pool.     

“Candidatus Similichlamydia laticola”, a Novel Chlamydia-like Agent of epitheliocystis in Seven Consecutive Cohorts of Farmed Australian Barramundi,Lates calcarifer (Bloch)

Six consecutively hatched cohorts and one cohort of pre-hatch eggs of farmed barramundi (Lates calcarifer) from south Australia were examined for Chlamydia-like organisms associated with epitheliocystis. To identify and characterise the bacteria, 59 gill samples and three pre-hatch egg samples were processed for histology, in situ hybridisation and 16S rRNA amplification, sequencing and comprehensive phylogenetic analysis. Cases of epitheliocystis were observed microscopically and characterised by membrane-enclosed basophilic cysts filled with a granular material that caused hypertrophy of the epithelial cells. In situ hybridisation with a Chlamydiales-specific probe lead to specific labelling of the epitheliocystis inclusions within the gill epithelium. Two distinct but closely related 16S rRNA chlamydial sequences were amplified from gill DNA across the seven cohorts, including from pre-hatch eggs. These genotype sequences were found to be novel, sharing 97.1 - 97.5% similarity to the next closest 16S rRNA sequence, Ca. Similichlamydia latridicola, from Australian striped trumpeter. Comprehensive phylogenetic analysis of these genotype sequences against representative members of the Chlamydiales order and against other epitheliocystis agents revealed these Chlamydia-like organisms to be novel and taxonomically placed them within the recently proposed genus Ca. Similichlamydia. Following Fredricks and Relman’s molecular postulates and based on these observations, we propose the epitheliocystis agents of barramundi to be known as “Candidatus Similichlamydia laticola” (sp. nov.).     

Cercopithecine Herpesvirus 9 (Simian Varicella Virus) Infection after Total-Body Irradiation in a Rhesus Macaque (Macaca mulatta)

This case report describes a rhesus macaque (Macaca mulatta; male; age, 5 y; weight, 6.7 kg) with anorexia, dehydration, lethargy, ataxia, and generalized skin rashes that occurred 30 d after total-body irradiation at 6.5 Gy (⁶⁰Co γ-rays). Physical examination revealed pale mucus membranes, a capillary refill time of 4 s, heart rate of 180 bpm. and respirations at 50 breaths per minute. Diffuse multifocal maculopapulovesicular rashes were present on the body, including mucocutaneous junctions. The CBC analysis revealed a Hct of 48%, RBC count of 6.2 × 10⁶/µL, platelet count of 44 × 10³/µL, and WBC count of 25 × 10³/µL of WBC. The macaque was euthanized in light of a grave prognosis. Gross examination revealed white foci on the liver, multifocal generalized petechiation on serosal and mucosal surfaces of the gastrointestinal tract, hemorrhagic lymph nodes, and hemorrhagic fluid in the thoracic cavity. Microscopic examination revealed cutaneous vesicular lesions with intranuclear eosinophilic viral inclusions within the epithelial cells, consistent with herpesvirus. Immunohistochemistry was positive for herpesvirus. The serum sample was negative for antibodies against Macacine herpesvirus 1 and Cercopithecine herpesvirus 9 (simian varicella virus, SVV). Samples submitted for PCR-based identification of the etiologic agent confirmed the presence of SVV DNA. PCR analysis, immunohistochemistry, and histology confirmed that lesions were attributed to an active SVV infection in this macaque. This case illustrates the importance of screening for SVV in rhesus macaques, especially those used in studies that involve immunosuppressive procedures.Abbreviations: SVV, simian varicella virus; TBI, total-body irradiation     

Commensal Nematodes in the Glands,Genitalia, and Brood Cells of Bees (Apoidea)

Seven species of bees from the eastern United States, representing four families in the Apoidea, were dissected and examined for nematode associates. Dufour''s glands in females of Halictus ligatus, Augochlora pura mosieri, and Augochlorella gratiosa (Halictidae) from Florida were infested with dauer juveniles of Aduncospiculum halicti (Diplogasteridae). The Dufour''s glands of Colletes thoracicus (Colletidae) females from Maryland were infested with dauer juveniles of a new species of Koerneria sp. (Diplogasteridae), and abdominal glands of females of Andrena alleghaniensis (Andrenidae) from New York were infested with dauer juveniles of another new species of Koerneria. The lateral and median oviducts, Dufour''s glands, and poison sacs in females of Anthophora abrupta (Anthophoridae) from Maryland and Alabama were infested with dauer juveniles of a new species of Bursaphelenchus sp. (Aphelenchoididae). Cross sections of the nematode-infested poison sacs of A. abrupta revealed two types of humoral host defense reactions.     

Frequent infections of mountain stream fish with the amphibian acanthocephalan,Pseudoacanthocephalus toshimai (Acanthocephala: Echinorhynchidae)

Pseudoacanthocephalus toshimai is an intestinal acanthocephalan parasite of amphibians in Hokkaido, the northernmost island of Japan. In this study, common freshwater fish of the families Salmonidae and Cottidae in mountain streams around the Kamikawa basin of Hokkaido were examined for acanthocephalan infections with P. toshimai. A total of 160 salmonids and 14 cottids were caught in 4 streams by bait fishing during summer and autumn seasons of 2019. Adult acanthocephalans were found only from the salmonids, namely, Salvelinus leucomaenis leucomaenis, Salvelinus malma krascheninnikovi, Oncorhynchus masou, and Oncorhynchus mykiss. The maximum prevalence reached 58.1% in S. leucomaenis, but the mean worm burden was at low levels (e.g., 3.1 in S. leucomaenis and 2.2 in S. malma). All of the acanthocephalans were identified to P. toshimai by morphological observation and DNA barcoding. Although the male acanthocephalans became sexually mature, the females never reached the gravid adult stage, suggesting that salmonids are unsuitable or aberrant hosts for P. toshimai. The infected fish were found exclusively from a small stream with bush, in which a large habitat of amphibians is included. Ligidium japonicum, a terrestrial isopod, collected from the habitat was highly infected with cystacanth larvae of P. toshimai. The observation of fish stomach contents directly demonstrated that small salmonids eat L. japonicum. The terrestrial isopods, which are washed away by rain into a stream, seem to be a source of salmonid infections with P. toshimai. The habitat of intermediate hosts should be emphasized in the taxonomy of the closely related genera Acanthocephalus and Pseudoacanthocephalus.     

Histological assessment of early maturation in sea trout (Salmo trutta morpha trutta L.) males in western Pomerania (Poland)

The study was performed to assess the maturation extent of sea trout (Salmo trutta morpha trutta L.) age 0+ males from north‐western Poland on the basis of histological examination. 617 male trout were collected from seven streams that had been stocked with swimming sea trout fry and classified into groups according to maturity stages: 64% were immature (Class I) in the first year of life; 17% remained in Class II with unfinished spermatogenesis. Mature male sea trout made up about 19% of the population; 72.6% thereof underwent incomplete maturation and 47.8% did not spermiate.     

A novel betaproteobacterial agent of gill epitheliocystis in seawater farmed Atlantic salmon (Salmo salar)

Epitheliocystis, a disease characterised by cytoplasmic bacterial inclusions (cysts) in the gill and less commonly skin epithelial cells, has been reported in many marine and freshwater fish species and may be associated with mortality. Previously, molecular and ultrastructural analyses have exclusively associated members of the Chlamydiae with such inclusions. Here we investigated a population of farmed Atlantic salmon from the west coast of Norway displaying gill epitheliocystis. Although 'Candidatus Piscichlamydia salmonis', previously reported to be present in such cysts, was detected by PCR in most of the gill samples analysed, this bacterium was found to be a rare member of the gill microbiota, and not associated with the observed cysts as demonstrated by fluorescence in situ hybridization assays. The application of a broad range 16 S rRNA targeted PCR assay instead identified a novel betaproteobacterium as an abundant member of the gill microbiota. Fluorescence in situ hybridization demonstrated that this bacterium, tentatively classified as 'Candidatus Branchiomonas cysticola', was the cyst-forming agent in these samples. While histology and ultrastructure of 'Ca. B. cysticola' cysts revealed forms similar to the reticulate and intermediate bodies described in earlier reports from salmon in seawater, no elementary bodies typical of the chlamydial developmental cycle were observed. In conclusion, this study identified a novel agent of epitheliocystis in sea-farmed Atlantic salmon and demonstrated that these cysts can be caused by bacteria phylogenetically distinct from the Chlamydiae.     

Parasitic castration of a vertebrate: Effect of the cymothoid isopod, Anilocra apogonae, on the five-lined cardinalfish, Cheilodipterus quinquelineatus

Parasitic castration, the specific blocking of host reproductive output by an individual parasite, is a host-parasite interaction common to many invertebrates, particularly crustaceans, echinoderms and molluscs. It can reduce host density, alter host population dynamics and the evolution of host life history traits. Here we show that parasitisation by a single female cymothoid isopod, Anilocra apogonae, castrates its vertebrate host, the five-lined cardinalfish, Cheilodipterus quinquelineatus. Parasitised male fish fail to mouthbrood their young. The gonads of parasitised fish are smaller and parasitised female fish have substantially fewer and smaller ova than do the gonads of unparasitised fish. As for parasitic castrators of invertebrate hosts, A. apogonae on C. quinquelineatus are uniformly dispersed amongst infested hosts (one adult female isopod per host), are site specific, and their body size is highly correlated with that of their host. These isopods are large relative to the body size of their hosts, averaging 3.8% of the weight of the host. Parasitised fish also weigh less and are shorter than unparasitised fish of the same age. Despite the presence of other potential hosts, A. apogonae only infests C. quinquelineatus. The consistency of the ecological correlates amongst known parasitic castrators suggests that the parasitic castrator host-parasite relationship will be recognised for other parasites of vertebrates.     

External Microflora of a Marine Wood-Boring Isopod

Bacteria associated with the marine wood-boring isopod Limnoria lignorum were enumerated by acridine orange epifluorescence microscopy and by plate counts on several media; the plate-viable bacteria were isolated and identified. Similar procedures were followed to enumerate and identify bacteria associated with the wood substrate from which the isopods were collected and with the surrounding water from the isopod habitat. Approximately 1.4 × 10⁷ bacterial cells were associated with each individual L. lignorum. Aeromonas hydrophila, Pseudomonas, and Vibrio were the most common genera in the isopod microflora. Wood from L. lignorum burrows had an associated bacterial flora of 1.6 × 10⁷ cells per mg (damp weight). A. hydrophila also predominated in the wood microflora. The water from which the isopod population was collected contained 2.3 × 10⁶ bacteria per ml. Pseudomonas and Vibrio species were very common in the water microflora, but A. hydrophila was not detected. Interactions between the isopod, its associated microorganisms, and the microorganisms within the wood substrate are discussed in the light of the known absence of a resident digestive tract microflora in these animals.     

Variation of specific proteins, mitochondria and fatty acid composition in gill of Scylla serrata (Crustacea, Decapoda) under low temperature adaptation

The mud crab Scylla serrata is an important commercial crustacean inhabiting estuarine water along the coast of southeast China. Metabolism in the gill is affected continuously by fluctuating water temperature and, therefore, the ability to cope with temperature change is essential to maintain physiological function. This experiment was conducted to help understand the mechanism of low temperature adaptation in S. serrata gill. In this study, 40 healthy juvenile male S. serrata from the same broodstock were grouped randomly into four groups, which were kept at 5 °C, 10 °C, 15 °C and 27 °C, with the same feeding regime during a 3-week adaptation period. Two-dimensional electrophoresis of the proteome was conducted to separate the specific proteins responsible for low temperature adaptation. Variations in the mitochondria were observed using transmission electron microscopy, and fatty acid composition was determined using gas chromatography. The results showed that different numbers of specific proteins were expressed under different low temperature adaptation, with more expressed at 5 °C and 10 °C than at 15 °C. Mitochondrial morphology also varied under different low temperature adaptation, but there was no linear relationship between microbial density and adaptation temperature. The composition of different fatty acids in the gill varied considerably with adaptation temperature, but elongation of the carbon chain and transition from fatty acids occurred at lower temperatures. Thus, changes in the specific proteins, mitochondria and fatty acid composition of the gill were the positive effects of low temperature on metabolism, leading to improved adaptation ability in S. serrata.     

Habitat specific growth rates and condition indices for the sympatric soles Solea solea (Linnaeus, 1758) and Solea senegalensis Kaup 1858, in the Tagus estuary, Portugal, based on otolith daily increments and RNA-DNA ratio

Habitat specific growth rates and condition indices were estimated for Solea solea and Solea senegalensis, in two nursery areas within the Tagus estuary, at the end of the estuarine colonization process, in 2005. While in the uppermost nursery area the two species of sole live in sympatry, in the lower nursery only S. senegalensis is present. Daily increments of left lapillar otoliths were used to estimate age (in days) and determine growth rates (mm per day). Condition indices were assessed through RNA‐DNA ratio in muscle samples. Growth rates were higher for S. senegalensis (0.970and 1.180 mm per day in nursery A and B, respectively) than for S. solea (0.767 mm per day in nursery A). Growth rates of S. senegalensis from the uppermost nursery area were lower when compared to those obtained for the other nursery. The RNA/DNA condition index followed the general trend given by the growth rate estimates, i.e. values were higher for S. senegalensis than for S. solea. However, no significant differences were detected in S. senegalensis from the two nurseries. Larger variations in salinity (10‰ amplitude in the uppermost nursery vs 0.2‰ in the lower nursery) and highest pollution loads may be important factors lowering the habitat quality of the uppermost nursery in comparison to the lower nursery. The use of growth rate estimates based on otolith readings and the RNA/DNA index as tools for habitat quality assessment was discussed.     

Ultrastructural and molecular phylogenetic delineation of a new order,the Rhizophydiales (Chytridiomycota)

In the order Chytridiales, Rhizophydium is a morphologically defined genus based upon the production of a monocentric, inoperculate, epibiotic sporangium, an endobiotic rhizoidal axis which branches, and an epibiotic resting spore. Despite its simple morphology, over 220 species of Rhizophydium have been described. Recent phylogenetic analyses using nuLSU rRNA (28 S rRNA) gene sequences of a geographically diverse sampling of Rhizophydium cultures revealed that the classical genus Rhizophydium is genetically more variable than previously understood and actually represents multiple genera. In the present study, we use zoospore ultrastructural characters and 28 S rRNA and 5.8 S ribosomal gene sequences of 96 isolates in culture to circumscribe the monophyletic Rhizophydium clade as a new order, Rhizophydiales. Correspondingly, zoospores of members of the Rhizophydiales exhibit a unique suite of ultrastructural character states that further define the order and distinguish it from the order Chytridiales. Molecular analyses reveal several strongly supported clades within the Rhizophydiales. Three of those clades encompass a broad range of isolates and are defined as new families Rhizophydiaceae, Terramycetaceae, and Kappamycetaceae. To resolve close relationships within Terramycetaceae, combined 28 S rRNA and ITS1–5.8 S–ITS2 sequences were analysed and details of zoospore ultrastructural character states determined, with two new genera, Terramyces and Boothiomyces, described. Two species formerly classified in Rhizophydium are transferred to the new genera. This work provides a framework for additional taxonomic revisions within the new order Rhizophydiales and compares genetic variation useful in defining genera, species, and populations within this lineage of chytrids. A broader sampling of representatives is needed before taxonomic decisions can be made for remaining clades within the Rhizophydiales.     

Herbivore- and Elicitor- Induced Resistance in Groundnut to Asian armyworm,Spodoptera litura (Fab.) (Lepidoptera: Noctuidae)

Induced defense was studied in three groundnut genotypes ICGV 86699 (resistant), NCAc 343 (resistant) and TMV 2 (susceptible) in response to Spodoptera litura infestation and jasmonic acid (JA) application. The activity of the oxidative enzymes [peroxidase (POD) and polyphenol oxidase (PPO)] and the amounts other host plant defense components [total phenols, hydrogen peroxide (H2O2), malondialdehyde (MDA), and protein content] were recorded at 24, 48, 72 and 96 h in JA pretreated (one day before) plants and infested with S. litura, and JA application and simultaneous infestation with S. litura to understand the defense response of groundnut genotypes against S. litura damage. Data on plant damage, larval survival and larval weights were also recorded. There was a rapid increase in the activities of POD and PPO and in the quantities of total phenols, H2O2, MDA and protein content in the JA pretreated + S. litura infested plants. All the three genotypes showed quick response to JA application and S. litura infestation by increasing the defensive compounds. Among all the genotypes, higher induction was recorded in ICGV 86699 in most of the parameters. Reduced plant damage, low larval survival and larval weights were observed in JA pretreated plants. It suggests that pretreatment with elicitors, such as JA could provide more opportunity for plant defense against herbivores.