Study of Drug Targets Associated With Oncogenesis and Cancer Cell Survival and the Therapeutic Activity of Engineered Ashwagandha Extract Having Differential Withanolide Constitutions.

Ashwagandha (Withania somnifera) has gained worldwide popularity for a multitude of health benefits inclusive of cancer-preventive and curative effects. Despite numerous research data supporting the benefits of this wonder herb, the actual use of ashwagandha for cancer treatment in clinics is limited. The primary reason for this is the inconsistent therapeutic outcome due to highly variable composition and constitution of active ingredients in the plant extract impacting ashwagandha's pharmacology. We investigate here an engineered yield: an ashwagandha extract (Oncowithanib) that has a unique and fixed portion of active ingredients to achieve consistent and effective therapeutic activity. Using the MCF7 cell line, Oncowithanib was studied for its anti-neoplastic efficacy and drug targets associated with cell cycle regulation, translation machinery, and cell survival and apoptosis. Results demonstrate a dose-dependent decline in Oncowithanib-treated MCF7 cell viability and reduced colony-forming ability. Treated cells showed increased cell death as evidenced by enhancement of Caspase 3 enzyme activity and decreased expressions of cell proliferation markers such as Ki67 and Aurora Kinase A. Oncowithanib treatment was also found to be associated with expressional suppression of key cellular kinases such as RSK1, Akt1, and mTOR in MCF7 cells. Our findings indicate that Oncowithanib decreases MCF7 cell survival and propagation, and sheds light on common drug targets that might be good candidates for the development of cancer therapeutics. Further in-depth investigations are required to fully explore the potency and pharmacology of this novel extract. This study also highlights the importance of the standardization of herbal extracts to get consistent therapeutic activity for the disease indication.

Study of Drug Target Identification and Associated Molecular Mechanisms for the Therapeutic Activity and Hair Follicle Induction of Two Ashwagandha Extracts Having Differential Withanolide Constitutions.

Background: Ashwagandha extracts play a significant role in traditional Indian medicine to help treat a wide range of disorders from amnesia, erectile dysfunction, neurodegenerative and cardiovascular diseases, cancer, stress, anxiety, and many more. Ashwagandha root is enriched with bioactive plant metabolites of which withanolides are the most important ones. The concentration and constitution of withanolides primarily determine ashwagandha's potency and pharmacology. Various factors modulate the withanolide constitution in the plant-derived extracts, rendering inconsistent therapeutic efficacy. Standardisation of the extraction protocol and a better understanding of the pharmacology mechanism of different extracts with varied withanolide constitutions is therefore critical for developing reliable, repeatable, and effective ashwagandha-based treatment. Objectives: Here, we work toward defining indication mechanisms for two varieties of ashwagandha extract-ASHWITH (ASH-Ext1) and Regenolide (ASH-Ext2)-with different proprietary withanolide proportions. Methods: ASH-Ext1 was studied for antioxidant signaling modulation using HEK293, HeLa, and A549 cells, and ASH-Ext2 was studied for subcellular drug targets associated with the reactivation and longevity of human hair follicles, using primary human hair follicle dermal papilla cells (HFDPCs). Results: Study findings support the antioxidant activity and Nrf2 signaling modulation by ASH-Ext1 in various cell models. Of note, ASH-Ext2 was found to increase ß-catenin and telomerase reverse transcriptase (TERT) protein expression levels in HFDPCs. Conclusion: The results of drug target modulation show us that the withanolide constitution associated with different extraction protocols influences the pharmacological potential of the extract significantly and points to the value of standardisation not only of total withanolide content but also of internal withanolide proportions.

Anthelmintic efficacy of Holarrhena pubescens against Raillietina spp. of domestic fowl through ultrastructural, histochemical, biochemical and GLCM analysis.

Globally, traditional knowledge systems are a powerhouse of information which can revolutionise the world, if decoded accurately and logically. Plant-based ethno-traditional and folklore curatives/medicines has a firm basis in the psyche of the common masses of West Bengal and Holarrhena pubescens is a representative example of it. This article communication on depicting the anthelmintic efficacy of ethanolic extract and Ethyl acetate fraction of the stem bark of Holarrhena pubescens against the cestode Raillietina spp. through efficacy studies, ultra-structural observations, histochemical and biochemical analysis on some tegumental enzymes i.e., Acid Phosphatase (AcPase), Alkaline Phosphatase (AlkPase), Adenosine Triphosphatase (ATPase) and 5'-Nucleotidase (5'-Nu) along with Gray Level Co-occurrence Matrix (GLCM) analysis of histochemical study. Praziquantel was used as the reference drug. Investigations revealed 10mg/ml dosage of crude extract was the most efficacious dose and amongst the fractions the ethyl acetate fraction showed the most anthelmintic property. Ultrastructural studies through Scanning Electron Microscope (SEM) and Transmission Electron Microscope (TEM) clearly depicted the damage in head, sucker, proglottids, proximal and distal cytoplasm (DC), microtriches (MT), basal lamina (BL), nuclear membrane (NM), and, nucleolus (NL) in the treated worms. Histochemical studies revealed decrease in staining intensity for all the tegumental enzymes in the treated worms compared to control. The GLCM analysis strongly supported the result of histochemical studies. Biochemical studies revealed marked reduction in enzyme activity in the treated worms with maximum reduction in the activity of 5'- Nu (77.8%) followed by ATPase (63.17%).

Biosynthesis, characterization and anthelmintic activity of silver nanoparticles of Clerodendrum infortunatum isolate.

Over the past few decades, the green synthesis of nanoparticles has gained importance for their therapeutic efficacy and eco-friendly nature. Integrating green chemistry principles into multidisciplinary nanoscience research has paved the way for developing environmentally benign and sustainable methods for synthesizing gold and silver nanoparticles. In the present study, the flowers obtained from Clerodendrum infortunatum (L.), belonging to the family Verbenaceae, have been used for biosynthesizing silver nanoparticles (AgNPs) to evaluate the anthelmintic potential. UV-Vis spectroscopy, XRD, FTIR, SEM and TEM analyses were performed to ascertain the formation of AgNPs. Clerodendrum-derived AgNP (CLE-AgNP) has significantly affected the normal physiological functions of the poultry parasite Raillietina spp., a menace to the livestock industry. Our study manifests that CLE-AgNPs cause considerable distortion of the surface tegument of this cestode parasite leading to changes in the host-parasite interface. The histochemical localization studies of the tegument-associated enzymes viz. AcPase, AlkPase, ATPase and 5'-Nu, exposed to the drug, showed a substantial activity decline, thus establishing the anthelmintic potential of the CLE-AgNPs.

Novelty and phylogenetic affinities of a new family of tapeworms (Cestoda: Rhinebothriidea) from endangered sawfish and guitarfish.

The parasites of hosts of conservation concern are often poorly known. This is the case with the iconic group of elasmobranchs known as the sawfish of the genus Pristis, all four species of which are considered as Endangered or Critically Endangered by the International Union for Conservation of Nature (IUCN, Switzerland). Examination of cestodes from three species of sawfish (Pristis pristis, Pristis clavata, and Pristis zijsron) in Australia and one of their close relatives, the also critically endangered widenose guitarfish, Glaucostegus obtusus, in India, collected over the past 25 years, yielded four new species of tapeworms which are described herein. All four belong to the previously monotypic Mixobothrium; the diagnosis of the genus is revised to accommodate the new species. Among the new taxa is a species that had been included in previous molecular phylogenies but whose identity and affinities within the order Rhinebothriidea, and thus also its familial placement, were unclear. This species exhibits the morphological features of Mixobothrium and thus its identity is, at long last, revealed. Sequence data generated for the 28S rDNA gene for three of the new species, as well as an additional new but yet undescribed species from Pristis pectinata from Florida (USA), confirms the uniqueness of this group among the rhinebothriideans. The new family Mixobothriidae is established to house these taxa. The members of this family differ from all but one of the five other families of rhinebothriideans in lacking apical suckers on their bothridia. They are also distinctive in that their bothridia are divided into three regions. The anterior and posterior regions have similar locular configurations to one another and differ from the locular configuration of the middle region. As a consequence, the bothridia are symmetrical along both their vertical and horizontal axes. We predict that a focus on species of guitarfish in the genus Glaucostegus will be the most productive approach for discovering additional diversity in this family of cestodes.

Primary stress response and biochemical profile of Labeo rohita (Hamilton, 1822) experimentally parasitized with Argulus bengalensis (Ramakrishna, 1951).

Argulosis is a major problem that causes huge economic loss in aquaculture. In a microcosm, an infested condition was developed upon Labeo rohita with 100 ± 10 adult morphs of Argulus bengalensis per fish. Primary stress response and biochemical profiles of the host were evaluated to underscore the pathogenicity of the parasites. Significant alterations in biochemical parameters were monitored at four different post-infestation time points: days 1, 3, 6 and 9. The overall increasing trends of both plasma cortisol and plasma epinephrine indicate parasite-induced primary stress response among experimental fish. The study revealed a hyperglycaemic trend throughout the infestation period, which has been correlated with hypoxia-associated glycogenolysis. Decreasing level of plasma cholesterol has also been correlated with the development of anaemia and subsequent hypoxia among the infested fish. Plasma protein of the experimental fish initially increases as an outcome of the immediate innate immune response against Argulus infestation, whereas the decrease in plasma protein at the later period of infestation results from less-dietary protein intake due to loss of appetite, reduced digestibility and metabolic proteolysis. Plasma Na+ concentration showed an overall decreasing trend throughout the infestation period, which may be due to excess production of catecholamine under stress. Nonetheless, plasma K+ concentration showed an increasing trend up to day 6 of infestation, and thereafter the value declined to the control level. Plasma ionic imbalances reflect changes in cell permeability under tissue hypoxia and the wounds produced on the skin for intensive feeding activity of the parasites. Both serum glutamate oxaloacetate transaminase and serum glutamate pyruvate transaminase activities were significantly elevated throughout the infestation period, which indicates excess synthesis or release of those enzymes from the damaged cells of the tissues. Activities of some renal, hepatic and branchial antioxidant enzymes, viz., superoxide dismutase, catalase and glutathione-S-transferase, increased in the infested fish. The fact is explained as an effort of the fish for gaining adaptive adjustment to neutralize the oxidative stress generated under the parasitic stress. The overall experimental result points towards the generation of potential stress upon host fish by this branchiuran parasite. The biochemical alterations of the fish under argulosis are centred around the two stress-sensitive hormones, cortisol and epinephrine. The outcome of the study will be the important physiological determinants in adopting a suitable control measure as well as assessing the nutritional value of the fish under diseased condition.

New Additions to the CRISPR Toolbox: CRISPR-CLONInG and CRISPR-CLIP for Donor Construction in Genome Editing.

CRISPR-Cas has proven to be the most versatile genetic tinkering system of our time, predominantly as a precision genome editing tool. Here, we demonstrate two additions to the repertoire of CRISPR's application for constructing donor DNA templates: CRISPR-CLONInG and CRISPR-CLIP. CRISPR-CLONInG (CRISPR-Cutting and Ligation Of Nucleic acid In vitro via Gibson) was devised to enable efficient cut-and-paste of multiple complex DNA fragments by using CRISPR-Cas9 as a digestion alternative with precision and exclusivity features, followed by joining the digested products via Gibson Assembly, to construct double-stranded DNA and adeno-associated virus (AAV) donor vectors rapidly without cloning scars. CRISPR-CLIP (CRISPR-Clipped Long ssDNA via Incising Plasmid) was devised as a DNA clipping tool to retrieve long single-stranded DNA (lssDNA) efficiently from plasmid, up to 3.5 kbase, which can be supplied as the donor template for creating genetically engineered mice via Easi-CRISPR. We utilized two different Cas types (Cpf1 and Cas9n) to induce two distinct incisions at the respective ends of the lssDNA cassette junctions on the plasmid, yielding three independent single-stranded DNA units of unique sizes eligible for strand separation, followed by target strand clip-out through gel extraction. The retrieval of the lssDNA donor circumvents involvements of restriction enzymes and DNA polymerase-based steps. Hence, it not only retains sequence fidelity but also carries virtually no restriction on sequence composition, further mitigating limitations on the current Easi-CRISPR method. With the add-on feature of universal DNA-tag sequences of Cpf1-Cas9 duo protospacer adjacent motif, CRISPR-CLIP can be facile and applicable to generate lssDNA templates for any genomic target of choice. Additionally, we demonstrate robust gene editing efficiencies in the neuroblastoma cell line, as well as in mice attained with the AAV and lssDNA donors constructed herein.

Ammonia-Assisted Growth of CoSn(OH)6 Nanostructures and Their Electrochemical Performances for Supercapacitor.

In this work, ammonia-assisted one step growth of SnO2 and mixed metal hydroxide CoSn(OH)6 on Ni foam is illustrated. The nanostructured films grown on Ni foam are highly porous in nature. The CoSn(OH)6 nanostructures were deposited using three different molar ratios of Sn/Co precursors i.e., 1:1, 1:2 and 2:1 and their performances as supercapacitor have been investigated and compared with pristine SnO2. Interestingly, the CoSn(OH)6 thin films prepared with 1:2 molar ratio of Sn/Co showed remarkably high areal capacitance and good cyclic stability. It is believed that the presence of different metal cations and their valence states along with the porous nanostructure makes the material outstanding as supercapacitor. The highest areal capacitance of 4189 mF cm-2 was obtained at a current density of 2 mA cm-2 for CoSn(OH)6 thin films offering an excellent cyclic stability with a capacity retention of 90.3% after 2000 cycles.

Facile synthesis of flower-like morphology Cu0.27Co2.73O4 for a high-performance supercapattery with extraordinary cycling stability.

The partial replacement of Co by Cu in cobaltite to give Cu0.27Co2.73O4 with unique flower-like morphology is found to be very beneficial for supercapacitor-battery hybrid applications. The 3D architecture of the material on a conductive substrate resulted in outstanding supercapattery performance. Asymmetric assembly of the material with activated carbon in a two-electrode system delivered high energy and power densities as well as a high specific capacity. The device also showed excellent cycling stability over 20 000 cycles, with a capacity retention value of 86.9%.

Deposition of Tin Oxide Thin Films by Successive Ionic Layer Adsorption Reaction Method and Its Characterization.

Tin oxide thin films were uniformly deposited by successive ionic layer adsorption reaction (SILAR) method on glass substrates using ethylene diamine as a complexing agent. The proper annealing treatment in air converts as-deposited amorphous films into crystalline and removes defects, reducing strain in the crystal lattice. The films were characterized by X-ray diffraction (XRD), scanning electron microscope (SEM), Atomic Force Microscopy (AFM), Fourier Transform Infrared (FTIR) spectroscopy. The film shows good optical transparency in the range of 200-1000 nm wavelength and electrical resistivity decreases upon annealing.

Ultrastructural changes in Raillietina (Platyhelminthes: cestoda), exposed to sulfonoquinovosyldiacylglyceride (SQDG), isolated from Neem (Azadirachta indica).

Neem (Azadirachta indica), has been known to be a curative for various ailments and diseases in the traditional Indian medicinal system from times immemorial. A glycolipid sulfonoquinovosyldiacylglyceride (SQDG) isolated from the leaves of neem has been found to be a proactive antibacterial and antiviral agent in previous studies. The current communication pertains to the anthelmintic activity of SQDG in vitro against a model cestode Raillietina spp. The results of efficacy tests showed a paralysis time of 1.0 ± 0.1 and 0.7 ± 0.01 h, whereas death time of 1.6 ± 0.3 and 0.9 ± 0.02 h, following treatments with dosages of 0.5 and 1.0 mg/mL, respectively. The scanning electron microscopic studies showed significant and unique changes in the ultrastructure of the worms with prominent breakages and furrows on the surface.

Anthelmintic efficacy of gold nanoparticles derived from a phytopathogenic fungus, Nigrospora oryzae.

Exploring a green chemistry approach, this study brings to the fore, the anthelmintic efficacy of gold nanoparticles, highlighting the plausible usage of myconanotechnology. Gold nanoparticles of ∼6 to ∼18 nm diameter were synthesized by treating the mycelia-free culture filtrate of the phytopathogenic fungus with gold chloride. Their size and morphology were confirmed by UV-Vis spectroscopy, DLS data, AFM and TEM images. The XRD studies reveal a crystalline nature of the nanoparticles, which are in cubic phase. The FTIR spectroscopic studies before and after the formation of nanoparticles show the presence of possible functional groups responsible for the bio-reduction and capping of the synthesized gold nanoparticles. The latter were tested as vermifugal agents against a model cestode Raillietina sp., an intestinal parasite of domestic fowl. Further, ultrastructural and biochemical parameters were used to corroborate the efficacy study.

Revision of Gangesia (Cestoda: Proteocephalidea) in the Indomalayan region: morphology, molecules and surface ultrastructure.

Tapeworms of Gangesia Woodland, 1924 (Cestoda: Proteocephalidea) parasitic in freshwater fishes in the Indomalayan Region were critically reviewed. Evaluation of type specimens and newly collected materials from Bangladesh, Cambodia and India, as well as critical examination of extensive literature have shown that only the following four species, instead of 48 nominal species of Gangesia and Silurotaenia Nybelin, 1942 reported from this region (36 new synonymies proposed), are valid: Gangesia bengalensis (Southwell, 1913), type-species of the genus and most common parasite of Wallago attu (Siluridae), G. macrones Woodland, 1924 typical of Sperata seenghala (Bagridae), both species characterized by the possession of two circles of hooks on the rostellum-like organ and several rows of hooklets on the anterior margins of suckers; G. agraensis Verma, 1928 from W. attu (typical host), which has the scolex with only one circle of hooks and 1-3 incomplete rows of tiny hooklets on the suckers; and G. vachai (Gupta and Parmar, 1988) n. comb. from several catfishes, which possesses 4-6 circles of hooks and 5-11 rows of hooklets on the anterior half of suckers. Scolex morphology, including surface ultrastructure (microtriches), of all but one species (G. vachai) is described for the first time using scanning electron microscopy. A phylogenetic analysis based on the partial sequences encoding the large nuclear ribosomal subunit RNA gene has shown that three Indomalayan species, namely G. bengalensis, G. macrones and G. vachai, form a monophyletic group within Gangesia, whereas G. agraensis tends to form a clade with the Palaearctic species of the genus. A table with differential characters of all species from the Indomalayan Region is also provided together with a key to identification of genera of the subfamily Gangesiinae. The present study demonstrates that species of Silurotaenia do not occur in the Indomalayan region.

A new monozoic tapeworm, Lobulovarium longiovatum n. g., n. sp. (Cestoda: Caryophyllidea), from barbs Puntius spp. (Teleostei: Cyprinidae) in the Indomalayan region.

A new caryophyllidean cestode is described from barbs Puntius spp. (Cypriniformes: Cyprinidae), with P. sophore (Hamilton) as its type-host, in the Ganges and Brahmaputra river basins in India and Bangladesh, and a new genus, Lobulovarium n. g., is proposed to accommodate it. The genus belongs to the Lytocestidae because its vitelline follicles are situated in the cortex. It is typified by: (i) a peculiar ovary, which is roughly H-shaped, but with asymmetrical, irregular lobes on its ventral and dorsal sides; (ii) an extensive vitellarium formed by numerous vitelline follicles scattered throughout the cortex; (iii) a long, conical postovarian part of the body with numerous vitelline follicles; (iv) a broadly digitate scolex with a slightly protrusible central cone; (v) a single gonopore (male and female genital ducts open via a single pore and a common genital atrium is absent); and (vi) a small number of testes (< 60). Molecular data (partial sequences of the lsrDNA) indicate that Lobulovarium longiovatum n. sp. belongs among the most basal caryophyllidean cestodes, being unrelated to species from siluriform catfishes in the Indomalayan region. Paracaryophyllaeus osteobramensis (Gupta & Sinha, 1984) Hafeezullah, 1993 (syn. Pliovitellaria osteobramensis Gupta & Sinha, 1984) from another cyprinid fish, Osteobrama cotio (Hamilton), in Uttar Pradesh, India, is tentatively transferred to Lobulovarium as L. osteobramense (Gupta & Sinha, 1984) n. comb. It differs from L. longiovatum by having much smaller eggs (length <50 µm versus >90 µm in L. longiovatum), which are spherical (length/width ratio 1:1 versus 2.5-3:1 in the new species), and the presence of vitelline follicles alongside the ovarian lobes (almost completely absent in L. longiovatum).

Cestodes (Caryophyllidea) of the stinging catfish Heteropneustes fossilis (Siluriformes: Heteropneustidae) from Asia.

The stinging catfish Heteropneustes fossilis (Bloch) (Siluriformes: Heteropneustidae) has been reported to harbor as many as 19 species of caryophyllidean tapeworms (Cestoda) of 11 genera in tropical Asia (Indomalayan zoogeographical region). However, a critical review of the species composition has shown that only 1 species, Lucknowia fossilisi Gupta, 1961 (Lytocestidae), is a specific parasite of H. fossilis. Three other species, Djombangia penetrans Bovien, 1926 (syn., Djombangia caballeroi Sahay and Sahay, 1977 ), Pseudocaryophyllaeus ritai Gupta and Singh, 1983 (syn. Pseudocaryophyllaeus lucknowensis Gupta and Sinha, 1984 ), and Pseudocaryophyllaeus tenuicollis (Bovien, 1926) Ash, Scholz, Oros and Kar, 2011 (syn. P. mackiewiczi Gupta and Parmar, 1982 ), were found only once. Lucknowia fossilisi is redescribed on the basis of new material collected in West Bengal and voucher specimens from Maharashtra, India. A total of 9 species of Capingentoides, Lucknowia, Lytocestus, Pseudoadenoscolex, Pseudocaryophyllaeus, Pseudoheteroinverta, and Sukhapatae are newly synonymized with L. fossilisi and previous synonymies of 9 other species, proposed by Hafeezulah (1993), are confirmed. Generic diagnosis of Lucknowia Gupta, 1961 is amended. In addition, 1 species of Pseudobatrachus and 2 species of the monotypic genera Pseudoneckinverta and Sudhaena are invalidated as nomina nuda.

Tapeworms (Cestoda: Caryophyllidea), parasites of Clarias batrachus (Pisces: Siluriformes) in the Indomalayan region.

Revision of monozoic cestodes (Caryophyllidea) parasitic in commercially important walking catfish Clarias batrachus (L.) in tropical Asia (Indomalayan zoogeographical region) was carried out, based on the evaluation of newly collected material from India, Indonesia, and Thailand, as well as a study of type specimens. Instead of the 59 nominal taxa of 15 genera from 3 caryophyllidean families previously described, only 8 species of the Lytocestidae are considered to be valid: Bovienia indica (Niyogi, Gupta and Agarwal, 1982) n. comb.; Bovienia raipurensis (Satpute and Agarwal, 1980) Mackiewicz, 1994; Bovienia serialis (Bovien, 1926) Fuhrmann, 1931; Djombangia penetrans Bovien, 1926; Lucknowia microcephala (Bovien, 1926) n. comb.; Lytocestus indicus (Moghe, 1925) Woodland 1926; Pseudocaryophyllaeus ritai Gupta and Singh, 1983; and Pseudocaryophyllaeus tenuicollis (Bovien, 1926) n. comb. All valid species are redescribed and SEM photomicrographs of their scolices and photomicrographs of their eggs are provided for the first time. Crescentovitus Murhar, 1963, Heeradevina Srivastav and Khare, 2005, Pseudobatrachus Pathak and Srivastav, 2005, Pseudobilobulata Srivastav and Lohia, 2002, Pseudoclariasis Pathak, 2002, and Pseudoinverta Pathak, 2002 are invalidated and 50 nominal species are newly synonymized, including 4 species described from other fish hosts. Taxonomic status of the remaining caryophyllidean taxa reported from C. batrachus (at least 6 taxa) could not be clarified because of the unavailability of their original descriptions. A key to identification of caryophyllidean tapeworms parasitic in C. batrachus is provided. To avoid current inflation of descriptions of invalid taxa, researchers are strongly encouraged to work only with well-fixed material; damaged, decomposed, or strongly flattened specimens should not be used for taxonomic studies, and type specimens must always be deposited in internationally recognized collections.

Ritacestus gen. n. (Cestoda: Proteocephalidea) and redescription of R. ritaii comb. n., a parasite of Rita rita (Siluriformes) in India.

A new genus, Ritacestus, is proposed to accommodate Ritacestus ritaii (Verma, 1926) comb. n. (syn. Proteocephalus ritaii), a parasite of the catfish Rita rita (Hamilton) in India. The new genus, which is placed in the Gangesiinae, is characterized by (i) a small, subspherical scolex formed by four large lobes separated from one another by longitudinal grooves, with a large, widely oval to pyriform rostellum-like apical organ, larger than suckers and possessing an apical hemispherical depression; (ii) paramuscular and cortical position of some vitelline follicles (most follicles are situated medullary); (iii) ventral and dorsal bands of vitelline follicles usually uninterrupted ventral to terminal genitalia and reaching to the posterior margin of proglottides; (iv) the vagina always anterior to the cirrus-sac; (v) a large size of the body (length up to 51 cm); and (vi) development of the uterus of type 2. In its morphology, especially shape of the scolex and apical organ, and paramuscular and cortical position of some vitelline follicles, Ritacestus resembles Postgangesia Akhmerov, 1969, but differs in the presence of a genital atrium (both genital pores of Postgangesia are separate), the anterior position of the vagina (almost always posterior in the latter genus), position of vitelline follicles in cross sections (dorsal and ventral bands in Ritacestus versus only a lateral band in the latter genus), and dorsal excretory canals indistinguishable in mature and gravid proglottides of R. ritaii (well developed in Postgangesia spp.). The type and only species of the genus, R. ritaii, is redescribed on the basis of new material from the type host from the Ganges River basin in India and its neotype is designated.

New data on the morphology and taxonomy of three species of Rhabdochona (Nematoda:Rhabdochonidae) parasitizing fishes in India.

Specimens of three little-known species of Rhabdochona (Nematoda: Rhabdochonidae) were collected during occasional examinations of some freshwater fishes in India: R. (Rhabdochona) hellichi turkestanica (Skryabin, 1917) in Schizothorax sp. (Cyprinidae, Cypriniformes) from the Lodhomakhola and Rangit Rivers, West Bengal and Sikkim, respectively; R. (R.) hospeti Thapar, 1950 in Tor sp. (Cyprinidae) from the Rangit River; and R. (Globochona) mazeedi Prasad et Sahay, 1965 in Clupisoma garua (Hamilton) (Schilbeidae, Siluriformes) from the Farakka Dam Lake, West Bengal. Their detailed light and electron microscopical studies revealed some taxonomically important, previously not observed features and made possible their redescription. Fourth-stage larvae of R. hospeti are described for the first time. Rhabdochona hellichi turkestanica (syns. R. denudata filamentosa Bykhovskaya-Pavlovskaya, 1936, R. kashmirensis Thapar, 1950, R. schizothoracis Siddiqi et Khattak, 1984) is proposed as a subspecies, differing from the nominotypical subspecies R. hellichi hellichi (Srámek, 1901) mainly in the shape of the distal end of the left spicule, molecular data and geographical distribution. Rhabdochona moraveci Katoch et Kalia, 1991 (a homonym to R. moraveci Duggal et Kaur, 1987) is renamed R. indica nom. n. The following six species are considered new junior synonyms of R. hospeti: Comephronema [sic] mackiewiczi Malhotra et Rautela, 1984, Rhabdochona moraveci Duggal et Kaur, 1987, R. bifidum Kakar et Bilqees, 2007, R. uvaginus Kakar et Bilqees, 2007, R. bolani Kakar, Bilqees et Ahmad, 2008 and R. cephalodiverticula Kakar, Bilqees et Ahmad, 2008. Rhabdochona edentati Paul et Majumdar, 1994 is considered a species incertae sedis.

Predatory efficiency of the sewage drain inhabiting larvae of Toxorhynchites splendens Wiedemann on Culex quinquefasciatus Say and Armigeres subalbatus (Coquillett) larvae.

The rate of predation by stage IV instar Toxorhynchites splendens larvae on the equivalent instar stage larvae of Culex quinquefasciatus and Armigeres subalbatus, co-occurring in sewage drains, were noted for a period of three consecutive days in the laboratory using different prey densities and combinations. The rate of predation varied by age of the predator, density of prey, and prey type. The number of Ar. subalbatus larvae consumed by a single Tx. splendens larva ranged between 0.50 +/- 0.71 and 16.40 +/- 2.01; while for Cx. quinquefasciatus larvae, the number consumed ranged from 0.20 +/- 0.42 to 20.40 +/- 1.43 per day. The pupation rates of the prey species varied in respect to control, with a minimum of 0.20 +/- 0.42 pupa/day to a maximum of 12.20 +/- 2.30 pupa/day in the presence of Tx. splendens. The values for the controls were 1.00 +/- 0.87 and 14.44 +/- 2.83 pupa/day, respectively. Irrespective of prey densities and combinations, a single Tx. splendens fourth instar larvae was found to consume on average 10.07 larvae on the first day 16.57 larvae on the second day and 4.38 larvae on the third day, killing a total of 17.70 to 45.10 larvae, in three days. In the presence of Tx. splendens, the cumulative pupation, irrespective of prey, remained between 12.20 and 45.10, and differed significantly from control where the values were between 13.90 and 54.70. The results indicate that Tx. splendens can significantly reduce immature numbers and lower the rate of pupation of Cx. quinquefasciatus and Ar. subalbatus. Tx. splendens may be a potential biological resource in the control of mosquitoes inhabiting sewage drains.

Anthelmintic efficacy of genistein, the active principle of Flemingia vestita (Fabaceae): alterations in the free amino acid pool and ammonia levels in the fluke, Fasciolopsis buski.

The crude root-peel extract of Flemingia vestita, its active principle genistein and the reference flukicide oxyclozanide were tested against Fasciolopsis buski, the giant intestinal trematode. The amino acid composition of F. buski was demonstrated using HPLC and it was observed that the free amino acid (FAA) pool of the control worm consisted of aspartate, threonine, serine, glutamic acid, glutamine, proline, glycine, alanine, valine, methionine, isoleucine, leucine, tyrosine, lysine, histidine, arginine, phosphoserine, taurine, citrulline, ornithine, beta-alanine, and gamma-amino butyric acid (GABA). Of the amino acids detected valine was found to be the maximum in quantitative analysis. In qualitative analysis the FAA pool of the parasites under various treatments remained same as that of the control; however, quantitatively the level of various FAAs in the parasite was significantly affected. The treated parasites showed a marked decrease in the levels of arginine, ornithine, tyrosine, leucine, isoleucine, valine, alanine, glycine, proline, serine, threonine, and taurine following treatment with 20 mg/ml of crude peel extract, 0.5 mg/ml of genistein and 20 mg/ml of the reference drug, though an increase in the levels of glutamic acid, glutamine, phosphoserine, citrulline and GABA was noticeable. Enhanced levels of GABA and citrulline under the influence of genistein may be implicated in alterations of nitric oxide release and consequent neurological change (e.g. paralysis) in the parasite. Ammonia in the tissue homogenate as well as in the incubation medium showed a quantitative increase compared to the controls after treatment with the various test materials. The ammonia level increased by 40.7%, 66.4% and 18.16% in treatments with F. vestita, genistein and oxyclozanide, respectively, at the mentioned dosages. The changes in the levels of the amino acids and nitrogen components post treatment suggest that the amino acid metabolism in the parasite may have been altered under the influence of the test materials.