Mutualistic association of rotifer Philodina roseola with the branchiuran fish ectoparasite Argulus bengalensis at its embryonic stage.

Several rotifers including Philodina spp. are well known to make commensal and parasitic associations with different animals. The present investigation was carried out to decipher the relationship of Philodina roseola with a piscine ectoparasite Argulus bengalensis in its embryonic stage. Mechanical removal of the symbiont P. roseola from the argulid egg strips resulted in the complete (100%) failure in hatching. Several P. roseola individuals were found to feed on the solidified jelly coat of the eggs enabling the larvae to emerge under both laboratory and field conditions. Under the laboratory condition, the experimental removal of P. roseola did not affect the embryonic development, but it rendered the jelly coat intact; therefore, the larvae were unable to make hatching furrow and subsequently died. The results of our experiments thus prove this service-resource relationship to be a mutualism. Although the association is facultative for P. roseola, it is obligatory for Argulus spp. An act of intervention in this relationship thus offers a promising control of argulosis.

Histological and ultrastructural investigation of the female reproductive system of Argulus bengalensis Ramakrishna, 1951 (Crustacea: Branchiura).

In order to understand branchiuran reproductive biology, it is imperative to know the sites of oogenesis and oocyte maturation, locate the accessory reproductive glands, and identify the fertilization site with the present knowledge of the sperm transfer mechanism of the genus Argulus. With these objectives, we attempted to describe the female reproductive system of Argulus bengalensis using serial histological sections through the ovaries and associated ducts in the transverse, longitudinal, and sagittal planes. The reproductive organs include a median ovary, one pair of ovarian lumina, a median oviduct, and a pair of collateral accessory glands. A duct from each of the collateral accessory glands leads into the proximal part of the median oviduct, which opens to the exterior through a genital opening at the distal end. The glandular secretion presumably contributes to the jelly coat of the egg. The ovary is bound with a tunica propria which extends further diametrically inside the ovary forming the paired lumina. The lumina are confluent into the median oviduct. Two distinct areas, the germarium and differentiating zones, are clearly distinguishable within the ovary. The tunica propria itself houses the oogonia within a matrix, serving as the germarium. Transmission electron micrograph reveals that the matrix is made of collagen. The collagen matrix confers elasticity to the tunica propria to accommodate the postvitellogenic oocytes within the ovarian lumen. The differentiating zone is situated in between the germarium: dorsally it is covered with a chromatophore layer. The ovary is ensheathed by a circum ovarian striated muscle. The presence of spermatophores in the ovarian lumen indicates the fertilization site. J. Morphol. 277:707-716, 2016. © 2016 Wiley Periodicals, Inc.

A histological evaluation of development and axis formation in freshwater fish ectoparasite Argulus bengalensis Ramakrishna, 1951 (Crustacea: Branchiura).

The present investigation was carried out to underscore the developmental events of a crustacean ectoparasite of fish, Argulus bengalensis. Serial histological sections of the embryo were made at lateral, sagittal and longitudinal planes to explain its cleavage, gastrulation and axis specification. The centrolecithal egg of A. bengalensis underwent meroblastic superficial cleavage. The cleavage initiated at the future dorsal side of the egg within 5 h to 5 h and 30 min of incubation. Consequently, a small mass of energids appeared superficially at the future dorsal side within 6 h. Later, energids were found at the future ventral and lateral sides. A syncytial blastoderm was formed around the centrally placed yolk material which was transformed into a cellular blastoderm within 30 h of incubation. In the blastoderm, two cell masses were formed at the dorsal and ventral part which initially extended towards each other and later spread out though future anterior-posterior direction. The pressure exerted by the cell flow displaced the entire yolk material at the future postero-ventral side. At the time of egg laying, a prototype of the embryonic axes is determined. The substratum side of the egg formed the dorsal part, whilst the side facing water turned to the ventral part. The broader end of the egg formed the anterior side and the narrow end formed the posterior side of the embryo. The anterior-posterior axis formation was initiated within 72 to 96 h of incubation when the blastodermal cells displaced the yolk material at the future posterior end. Within 120 h of incubation, the germ layers of the embryo were determined. The study reveals that the cleavage pattern of A. bengalensis shows close similarities with that of the Malacostraca amongst the crustaceans and dipteran and hymenopteran amongst the insects.

Male reproductive system of the fish ectoparasite Argulus bengalensis (Crustacea: Branchiura).

The male reproductive system of the fish ectoparasite Argulus bengalensis was reconstructed with serial histological sections through longitudinal and transverse planes and the description was re-evaluated after the discovery of spermatophore. The study revealed that the testis of A. bengalensis consists of two lobes, each comprised the two enclaves. The outer enclaves from both lobes are connected proximally with an isthmus. The inner enclave is covered with two layers of chromatophores which are presumably involved in androgenic hormone production. At the proximal end of the inner enclaves a pair of vasa efferentia originates and ascends into thorax where it leads into vesicula seminalis. Like Dolops ranarum, a pair of spermatophoric glands is found in the thorax just behind the second maxilla. Glandular cells of the spermatophoric glands are columnar with a large nucleus at their basal side. A pair of spermatophoric canals carries the secretion of the glands into the vesicula seminalis through vasa efferentia. Spermatophores are delivered to the female through a median vas deference originating from the vesicula seminalis. In addition to the spermatophoric glands, a pair of accessory glands is located in the coxa of the fourth thoracic legs. At the region dorsal to the isthmus both the spermatophoric canal and accessory gland ducts unite with the vas efferens of the respective side. The secretion of the accessory gland presumably contributes to semen composition of the parasite.

Effect of aqueous extract of Azadirachta indica A. Juss (neem) leaf on oocyte maturation, oviposition, reproductive potentials and embryonic development of a freshwater fish ectoparasite Argulus bengalensis Ramakrishna, 1951 (Crustacea: Branchiura).

In present study, a microcosm experiment is carried out to investigate the efficacy of 120 and 250-ppm crude aqueous extract of Azadirachta leaf on oocyte maturation, oviposition, embryonic development and hatching of the eggs of a fish ectoparasite Argulus bengalensis. Relative abundance of different maturing oocyte stages in the ovary of the parasite from different age groups was enumerated, and marked variations were obtained. Significant depletion in the abundance of pre-vitollogenic, vitellogenic and post-vitellogenic oocytes was recorded, which indicates impairment in maturation. Chromatin condensation of the oocytes of treated parasite indicates apoptosis of oogenic cells. Strong oviposition deterrence was evident by the elevated oviposition deterrence index of 0.18 and 0.52 at respective toxin levels. The treated parasites invested less number of eggs per oviposition, and hatching percentage of the eggs reduced markedly. In vitro treatment of eggs within 70 min of incubation exhibited coagulation of yolk material and subsequent reduction in hatching percentage. However, treatment applied after this critical period, hatching was not significantly altered. In vitro treatment of eggs at 80 min of incubation resulted in normal development. It signifies that azadirachtin affects the early developmental events but not the later. Presumably, azadirachtin either affects early gene expression of the embryo or antagonizes any of the substances of the zygote required for sustaining early developmental process. The result of the present experiment suggests that azadirachtin could be a promising agent to control argulosis through inhibition of the reproductive maturity of the parasite as well as through interference of its embryonic development.

Morphological characterization of testicular cells, spermatogenesis and formation of spermatophores in a fish ectoparasite Argulus bengalensis Ramakrishna, 1951 (Crustacea: Branchiura).

The present study has been carried out to describe the cell morphology of the developing male gametes in a fish ectoparasite, Argulus bengalensis Ramakrishna, 1951. With respect to cell volume and nucleoplasmic index, spermatogonia are the smallest and primary spermatocytes are the largest in this lineage. The spermatogonia and the differentiating spermatogenic cells are in separate niches and confined to different enclaves within each testicular lobe. Spermiogenesis occurs within the inner enclave of each testicular lobe. During this process the nucleus becomes streamlined; an acrosome is formed, axoneme is originated, and residual cytoplasm is discarded through the flagellum. The sperm cell morphology displays a general pattern comprising head, mid-piece, and a full length flagellum. In the axoneme 9+2 arrangement of the microtubule is conserved. In addition to the axoneme, some more singlet microtubules are found surrounding a fiber sheath and around one of the mitochondria adjacent to the axoneme. This arrangement indicates a close phylogenetic relationship with pentastomida. In the present study, structure and formation of spermatophore are described in this branchiuran parasite.

Correlation between body size and fecundity in fish louse Argulus bengalensis Ramakrishna, 1951 (Crustacea: Branchiura).

The life history traits like fecundity and body size are useful predictors of life history strategies of organisms. The information on these aspects provided necessary input for control measures for ectoparasites. In view of this, the variations in the life history traits of the fish louse Argulus bengalensis Ramakrishna (1951) were assessed using age as an explanatory factor. The analyses revealed that the body weight (BW) is related to age in males as: y (BW) = 0.03 × (Age)-2.58: and in females as: y (BW) = 0.89 + 0.13x (Age). The body length and age relationship in males is observed as: y (BL) = 2.94 + 0.01x (Age) and in females as: y (BL) = 2.89 + 0.06x (Age). The degree of sexual dimorphism (DD) for BL is positively correlated (r = -0.358; df = 43; P < 0.001) with age while DD for BW is negatively correlated (r = -0.525; df = 43; P < 0.001) with age. The eggs/clutch remained between 02 and 43 for the 21st and 38th day old females. The fecundity as a function of age could be represented as: y (Eggs) = 1.62x (Age)-27.92. The increase in BW with age in female A. bengalensis favoured greater resource allocation for egg production, while in males it likely favours dispersal ability. Since body size and fecundity varied with age, the relative abundance and extent of infestation in fish host would vary with age composition of the population and recruitment of juveniles. The impacts of host specific variations on these features need to be tested further.