Shining a light on parasite behaviour: daily patterns of Argulus fish lice.

Parasites display a wide range of behaviours that are frequently overlooked in favour of host responses. Understanding these behaviours can improve parasite control through a more precise application or development of new behaviour-based strategies. In aquaculture fish lice are an ongoing problem, infections reduce fishery production and control options are limited. Fish lice are distinct in their ability to survive and swim off hosts, allowing the transmission to multiple fish hosts across their lifespan. Here we assessed the off-host behaviour of Argulus foliaceus (a freshwater fish louse) and observed a diurnal rhythmical pattern in their behaviour. This pattern was lost when lice were exposed to constant darkness, indicating that the behaviour is not endogenously driven. Males were consistently active in light with reduced activity in darkness. In contrast, females were active during light and dark phases with peak activity at the start of dark periods. A. foliaceus was also strongly attracted to a light stimulus, preferring white- and blue-coloured lights over green- or red-coloured lights. Light is a strong driver of fish louse activity and could be used to trap parasites. Aquaculture light regimes could also be altered to reduce parasite attraction and activity.

Bet hedging and cold-temperature termination of diapause in the life history of the Atlantic salmon ectoparasite Argulus canadensis.

Argulus canadensis is a crustacean ectoparasite observed increasingly on wild migrating adult Atlantic salmon. We investigated temperature and salinity tolerance regarding development, survival and hatch of A. canadensis eggs to help understand spatiotemporal features of transmission. Argulus canadensis eggs differentiate to pharate embryos by 35 days buttheir hatch is protracted to ~7 months. Cold treatment ⩾75 days mimics overwintering and terminates egg diapause, with 84.6% (72.1-100%) metanauplius hatch induced ⩾13 °C and synchronized to 3-4 weeks. Inter- and intra-clutch variability and protracted hatch in the absence of cold-temperature termination of diapause is compatible with bet hedging. Whereas diapause likely promotes phenological synchrony for host colocalization, bet hedging could afford temporal plasticity to promote host encounter during environmental change. Our egg storage and hatch induction/synchronization methodologies can be exploited for empirical investigations. Salinity tolerance reveals both significantly higher embryonic development (94.4 ± 3.5% vs 61.7 ± 24.6%) and metanauplius hatch (53.3 ± 7.5% vs 10.1 ± 8.2%) for eggs in freshwater than at 17 ppt. Unhatched embryos were alive in freshwater by the end of the trial (213 days) but were dead/dying at 17 ppt. Eggs did not develop at 34 ppt. Salinity tolerance of A. canadensis eggs supports riverine transmission to adult Atlantic salmon during return to freshwater for mating each year.

Life in the fast lane: Temperature, density and host species impact survival and growth of the fish ectoparasite Argulus foliaceus.

With expanding human populations, the food sector has faced constant pressure to sustainably expand and meet global production demands. In aquaculture this frequently manifests in an animal welfare crisis, with fish increasingly farmed under high production, high stress conditions. These intense environments can result in fish stocks having a high susceptibility to infection, with parasites and associated disease one of the main factors limiting industry growth. Prediction of infection dynamics is key to preventative treatment and mitigation. Considering the climatic and technology driven changes facing aquaculture, an understanding of how parasites react across a spectrum of conditions is required. Here we assessed the impact of temperature, infection density and host species on the life history traits of Argulus foliaceus, a common palearctic fish louse, representative of a parasite group problematic in freshwater aquaculture and fisheries worldwide. Temperature significantly affected development, growth and survival; parasites hatched and developed faster at higher temperatures, but also experienced shorter lifespans when maintained off the host. At high temperatures, these parasites will likely experience a short generation time as their life history traits are completed more rapidly. A. foliaceus additionally grew faster on natural hosts and at lower infection densities. Ultimately such results contribute to prediction of population dynamics, aiding development of effective control to improve animal welfare and reduce industry loss.

Stable isotopes reveal contrasting trophic dynamics between host-parasite relationships: A case study of Atlantic salmon (Salmo salar) and parasitic lice (Lepeophtheirus salmonis and Argulus foliaceus).

Across existing fish host-parasite literature, endoparasites were depleted in δ15 N compared to their hosts, while ectoparasitic values demonstrated enrichment, depletion and equivalence relative to their hosts. δ13 C enrichment varied extensively for both endo- and ectoparasites across taxa and host tissues. In our case study, sea lice (Lepeophtheirus salmonis) were enriched in δ15 N relative to their farmed Atlantic salmon (Salmo salar) hosts, although the value contradicted the average that is currently assumed across the animal kingdom. Common fish lice (Argulus foliaceus) did not show a consistent trend in δ15 N compared to their wild S. salar hosts. Both parasitic species had a range of δ13 C enrichment patterns relative to their hosts. Farmed and wild S. salar had contrasting δ13 C and δ15 N, and signals varied across muscle, fin and skin within both groups. L. salmonis and A. foliaceus subsequently had unique δ13 C and δ15 N, and L. salmonis from opposite US coasts differed in δ15 N. Given the range of enrichment patterns that were exhibited across the literature and in our study system, trophic dynamics from host to parasite do not conform to traditional prey to predator standards. Furthermore, there does not appear to be a universal enrichment pathway for δ13 C nor δ15 N in parasitic relationships, which emphasizes the need to investigate host-parasite linkages across species.

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.

Apolipoprotein A-I in Labeo rohita: Cloning and functional characterisation reveal its broad spectrum antimicrobial property, and indicate significant role during ectoparasitic infection.

Apolipoprotein A-I (ApoA-I) is the most abundant and multifunctional high-density lipoprotein (HDL) having a major role in lipid transport and potent antimicrobial activity against a wide range of microbes. In this study, a complete CDS of 771 bp of Labeo rohita (rohu) ApoA-I (LrApoA-I) encoding a protein of 256 amino acids was amplified, cloned and sequenced. Tissue specific transcription analysis of LrApoA-I revealed its expression in a wide range of tissues, with a very high level of expression in liver and spleen. Ontogenic study of LrApoA-I expression showed presence of transcripts in milt and 3 h post-fertilization onwards in the larvae. The expression kinetics of LrApoA-I was studied upon infection with three different types of pathogens to elucidate its functional significance. Its expression was found to be up-regulated in the anterior kidney of L. rohita post-infection with Aeromonas hydrophila. Similarly following poly I:C (poly inosinic:cytidylic) stimulation, the transcript levels increased in both the anterior kidney and liver tissues. Significant up-regulation of LrApoA-I expression was observed in skin, mucous, liver and anterior kidney of the fish challenged with the ectoparasite Argulus siamensis. Immunomodulatory effect of recombinant LrApoA-I (rApoA-I) produced in Escherichia coli was demonstrated against A. hydrophila challenge in vivo. L. rohita administered with rApoA-I at a dose of 100 µg exhibited significantly higher protection (∼55%) upon challenge with A. hydrophila 12 h post-administration of the protein, in comparison to that observed in control group, along with higher level of expression of immune-related genes. The heightened expression of ApoA-I observed post-infection reflected its involvement in immune responses against a wide range of infections including bacterial, viral as well as parasitic pathogens. Our results also suggest the possibility of using rApoA-I as an immunostimulant, particularly rendering protection against A. hydrophila.

Transcriptional changes in three immunoglobulin isotypes of rohu, Labeo rohita in response to Argulus siamensis infection.

Immunoglobulin heavy chains of three isotypes viz., IgM, IgD and IgT/IgZ are described in teleosts. In this study, a challenge experiment with an ectoparasite Argulus siamensis was conducted to evaluate the changes in adaptive immune response by quantitation of expression of Ig heavy chains in skin, head kidney and mucus of infected rohu, Labeo rohita. Rohu were challenged with 100 metanauplii of A. siamensis/fish. Head kidney, skin and mucus samples were collected at 0 h, 12 h, 24 h, 3 d, 7 d, 15 d and 30 d by sacrificing four fish each from infected and control groups at each time point. The expression of IgM, IgD and IgZ in these tissues were measured by reverse transcription real time quantitative PCR. IgM level was found to reach its peak significantly 30 d post-infection in head kidney tissue, while IgM transcripts were below detectable range in skin and mucus at all time points. IgZ and IgD levels were significantly up-regulated post-infection in all the three tissue samples. Early up-regulation of IgD was observed in skin and mucus, compared to head kidney. This study showed that parasitic invasion can trigger varied expressions of immunoglobulin types to provide systemic as well as local protection in the host. In particular, the appearance of high level of expression of IgZ and IgD in skin and mucus will pave the way for vaccine development against A. siamensis which feeds on those tissues.

Molecular characterization of interleukin 15 mRNA from rohu, Labeo rohita (Hamilton): Its prominent role during parasitic infection as indicated from infection studies.

Interleukin 15 (IL-15) is an important cytokine of fish immune system. Sequence characterization of IL-15 from rohu, Labeo rohita revealed a mRNA sequence of 1064 bp with coding sequence of 567 bp and signal peptide of 16 amino acids. There are four characteristic sequence features viz., presence of four out-of-frame AUG initiation codons, four highly conserved cysteine residues, constitutive expression in all tissues and evolutionary similarity. The ontogeny study revealed maternal transfer of this molecule and higher expression up to 3 h post-fertilization in fertilized embryos. Its expression was down-regulated in anterior and posterior kidneys, intestine and liver tissues of rohu infected with Aeromonas hydrophila. Mild up-regulation in liver and higher expression in spleen was noticed in rohu stimulated with poly I:C (poly ionosinic:cytidylic), whereas down-regulation was observed in intestine and kidney tissues. However, a consistent higher expression was noticed in kidney and skin tissues during Argulus siamensis infection. Therefore, rohu IL-15 might possess more defensive role during early development and parasitic infection.

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.

Additional morphological information on Dipteropeltis hirundo Calman, 1912, and a description of Dipteropeltis campanaformis n. sp. (Crustacea: Branchiura) from two characiform benthopelagic fish hosts from two Northern rivers of the Brazilian Amazon.

Dipteropeltis is a monotypic genus of the Branchiura and endemic to South America. Twelve specimens of Dipteropeltis sp. and micrographs of a thirteenth specimen were obtained from four institutions. Measurements and light micrographs were taken of all the specimens. The specimens were compared to all known descriptions of D. hirundo and sorted into two groups, those that conformed to the description of D. hirundo; and those that differed morphologically. Two specimens, one from each group¸ were stained with lignin pink and subsequently prepared for Scanning Electron Microscopy. The differences observed in the specimens indicated that a new species exists. Thus, Dipteropeltis campanaformis n. sp. is described from seven specimens collected from two characiform benthopelagic fish species and compared to D. hirundo. Dipteropeltis campanaformis n. sp. differs from D. hirundo in the head shape, the length and shape of the carapace lobes, the membrane composition of the maxillules, the shape of the maxillae, the shape of the mouth, and the shape of the natatory lobes.

De novo whole transcriptome analysis of the fish louse, Argulus siamensis: first molecular insights into characterization of Toll downstream signalling molecules of crustaceans.

Argulus siamensis is a major ectoparasitic pathogen of freshwater fish capable of causing substantial economic loss. None of the available control measures have been able to address the problem of argulosis resourcefully. To combat this pathogen effectively, it is necessary to have a comprehensive understanding of its life processes with information on various genes involved. The transcriptome studies can generate introductory information about genes participating in physiological processes of the parasite which could be targeted for their control. In this study, the transcriptome sequencing of A. siamensis was performed on Illumina HiSeq 2000 platform which generated 75,126,957 high quality reads. A total of 46,352 transcript contigs were assembled with average length of 1211bp and N50 length of 2302bp. In total, 19,290 CDS including 184 novel CDS and 59,019 open reading frames (ORFs) were identified from the assembled contigs. Gene ontology and Kyoto Encylopedia of Genes and Genomes pathway analysis were performed to classify contigs into their functional categories and regulation pathways. Additionally, 1171 simple sequence repeats were identified from the assembled contigs. Further, twelve contigs with high similarity with downstream molecules of the mammalian toll like receptor (TLR) pathway were validated by their inductive expressions in response to lipopolysaccharide (LPS) of Gram negative bacteria, Escherichia coli and Gram positive bacteria, Staphylococcus aureus. The transcriptome of an ectoparasite A. siamensis was sequenced, assembled, annotated, and the downstream signalling molecules of Toll pathway characterized. The transcriptome data generated will facilitate studies on functional genomics that will subsequently be applied for vaccine development and other control strategies against the parasite.

Stocking methods and parasite-induced reductions in capture: modelling Argulus foliaceus in trout fisheries.

Argulus foliaceus is a macroparasite which can have a significant impact on yield in recreational trout fisheries, partly by increasing fish mortalities but also by reducing the appetite of infected fish, making them less likely to respond to bait. The aim of this paper is to determine the impact of four commonly used fish stocking methods both on the parasite dynamics, and on fisheries' yields. The wider consequences of the resultant reduction in host feeding are also of interest. To this end four different stocking methods were incorporated into Anderson and May's macroparasite model, which comprises three differential equations representing the host, attached parasite and free-living parasite populations. To each of these a reduction in the fish capture rate, inversely linked to the mean parasite burden, is added and the effects interpreted. Results show that (1) the common practise of increasing the stocking rate as catches drop may be counterproductive; (2) in the absence of any wild population of reservoir hosts, the parasite will be unable to survive if the stocking rate does not exceed the rate of capture; (3) compensatory stocking to account for fish mortalities can have disastrous consequences on yield; and (4) the parasite can, under certain circumstances, maintain the host population by preventing their capture.

Freshwater abiotic components' impact on the viability of fish lice, Argulus sp., in Guangdong province, China.

Parasite intensity dynamics can be strongly affected by ecosystem abiotic components. We conducted this study to investigate how the intensity of Argulus sp. is influenced by physicochemical factors in three water bodies - river, fish farm, and reservoir - and to find the occurrence of Argulus sp. under different conditions. Argulus sp. specimens were collected using different methods - gill-netting 30 × 1.5 m, 40 mm mesh monofilament net and set from a small boat, beach seining and plankton net - from July 2010 until Jun 2011 at Guangdong province, China. Main water parameters taken for the assessment consisting of temperature, pH, DO, biochemical oxygen demand of 5 days (BOD5), chemical oxygen demand (COD), NH (4) (+) , total nitrogen (TN) and total phosphorus (TP). The results show that the ratio of female to male Argulus sp. during the studied period was 2.3:1. The presence and intensity of Argulus sp. were temperature-dependent and variable; pH values were lower than standers with high intensity of Argulus sp. Argulus sp. has the ability to live in low dissolved oxygen (DO) concentration. Relatively high concentrations of BOD5, COD, NH (4) (+) , TN and TP were detected with related variable intensity of Argulus sp., which is supposed to be a freshwater fish ectoparasite found in polluted ecosystems. It is concluded that the fish ectoparasite Argulus sp. have a strong potency of genetic adaptation impacted by environmental factors. However, this needs further study in order to gain insight into the question of the likelihood of adaptation to abiotic factors' variation.

A bridge between original and novel states: ontogeny and function of "suction discs" in the Branchiura (Crustacea).

The emergence of novel structures in the course of evolution faces an explanatory problem, leaving the gap from the ancestral structures difficult to bridge. This difficulty is caused by the lack of intermediate stages. Branchiurans are ectoparasitic crustaceans which use a pair of "suction discs" to attach to their host. These structures are modified first maxillae. During ontogeny, the first maxillae transform from a normal cephalic appendage to the specialized suction disc. However, supposedly ancestral branchiurans lack the suction discs in the adults and the first maxilla remains a normal appendage throughout. We describe the muscular arrangements in the developing first maxillae in Argulus coregoni. The suction discs originate as a fusion of the first and second podomeres. The sucker muscles of the suction discs are homologous to the muscles that insert in the second podomere at the early larval stages. The developmental process of the suction disc can be seen as a "recapitulation" of the evolutionary process. We thus show how the first maxilla can maintain not just the biological role but also a functional continuity during the evolution of the novel structure. From this example it is obvious that the intermediate stages of the emerging novelty, if present in the ontogeny, can help solve at least some of the enigmatic appearances of novel structures.

Fish farms, parasites, and predators: implications for salmon population dynamics.

For some salmon populations, the individual and population effects of sea lice (Lepeophtheirus salmonis) transmission from sea cage salmon farms is probably mediated by predation, which is a primary natural source of mortality of juvenile salmon. We examined how sea lice infestation affects predation risk and mortality of juvenile pink (Oncorhynchus gorbuscha) and chum (O. keta) salmon, and developed a mathematical model to assess the implications for population dynamics and conservation. A risk-taking experiment indicated that infected juvenile pink salmon accept a higher predation risk in order to obtain foraging opportunities. In a schooling experiment with juvenile chum salmon, infected individuals had increased nearest-neighbor distances and occupied peripheral positions in the school. Prey selection experiments with cutthroat trout (O. clarkii) predators indicated that infection reduces the ability of juvenile pink salmon to evade a predatory strike. Group predation experiments with coho salmon (O. kisutch) feeding on juvenile pink or chum salmon indicated that predators selectively consume infected prey. The experimental results indicate that lice may increase the rate of prey capture but not the handling time of a predator. Based on this result, we developed a mathematical model of sea lice and salmon population dynamics in which parasitism affects the attack rate in a type II functional response. Analysis of the model indicates that: (1) the estimated mortality of wild juvenile salmon due to sea lice infestation is probably higher than previously thought; (2) predation can cause a simultaneous decline in sea louse abundance on wild fish and salmon productivity that could mislead managers and regulators; and (3) compensatory mortality occurs in the saturation region of the type II functional response where prey are abundant because predators increase mortality of parasites but not overall predation rates. These findings indicate that predation is an important component of salmon-louse dynamics and has implications for estimating mortality, reducing infection, and developing conservation policy.

[Monoxenous and heteroxenous parasites of fish manipulate behavior of their hosts in different ways].

Adaptive host manipulation hypothesis is usually supported by case studies on trophically transmitted heteroxenous endoparasites. Trematodes and cestodes are among efficient manipulators of fish, their common intermediate hosts. In this review paper, new data on modifications of host fish behavior caused by monoxenous ectoparasitic crustaceans are provided together with a review of effects caused by heteroxenous parasites. Differences in modifications of host behavior caused by heteroxenous and monoxenous parasites are discussed. Manipulation by heteroxenous parasites enhances availability of infected fish to predators--definitive hosts of the parasites. Fine-tuned synchronization of modified anti-predator behavior with a certain phase of the trematode Diplostomum spathaceum development in the eyes of fish, their second intermediate host, was shown. Modifications of behavior are habitat specific. When juvenile salmonids are in the open water, parasites impair their cooperative anti-predator behavior; in territorial bottom-dwelling salmonids, individual defense behavior such as sheltering is the main target of manipulation. It was shown that monoxenous ectoparasitic crustaceans Argulus spp. decreased motor activity, aggressiveness and increased shoal cohesiveness of infected fish. Such a behavior facilitates host and mate searching in these parasites, which often change their hosts, especially during reproduction. Reviewed experimental data suggest that heteroxenous parasites manipulate their host mainly through impaired defense behavior, e.g. impairing shoaling in fish. Alternatively, monoxenous parasites facilitate shoaling that is profitable for both parasites and hosts. Coordination of modified host behavior with the parasite life cycle, both temporal and spatial, is the most convincing criterion of the adaptive value of host manipulation.

Argulus japonicus: sperm transfer by means of a spermatophore on Carassius auratus (L).

The process of sperm transfer is somewhat enigmatic in Argulus, even though copulation has been witnessed. A breeding colony of Argulus japonicus was kept under laboratory conditions in order to study reproduction in the species. Pairs in copula were removed and studied with histology and scanning electron microscopy to describe the mechanism of sperm transfer. Sections of copulating pairs revealed sperm on the accessory copulatory structures of the male's swimming legs; and scanning electron microscopy showed that sperm transfer occurs in three phases which can be differentiated to 10 different stages. Sperm transfer occurs via a spermatophore which is extruded from the genital aperture of the male and is then transferred to the socket on the third pair of legs of the male, before being transferred into the spermathecae of the female via the spermathecal spines. This is the first observation of a spermatophore in Argulus.

Crustacean parasites of Leporinus affinis, an Anostomidae fish from the Brazilian Amazon.

This study investigated the fauna of ectoparasitic crustaceans in Leporinus affinis from Reservoir Coaracy Nunes, in eastern Amazon (Brazil), as well as the parasite-host interactions. The mouth, gills and tegument of 50.9% of the fish examined were parasitized by Argulus chicomendesi, Ergasilus turucuyus and Excorallana berbicensis, and a total of 118 parasites were collected. The dominance was found for E. berbicensis and the higher infestation levels were caused by E. berbicensis on the body surface of the hosts, but E. turucuyus had the highest prevalence on the gills of this host. The cluster analysis revealed higher similarity in the infestations by E. berbicensis and A. chicomendesi in relation to infestation site in hosts. Host sex and relative condition factor (Kn) were not influenced by moderate parasitism, but the abundance of parasites presented negative correlation with weight and Kn of the fish. This is the first study on the parasites of L. affinis showing low species diversity, with moderate prevalence and low parasite abundance.

The influence of risk factors on the abundance, egg laying habits and impact of Argulus foliaceus in stillwater trout fisheries.

Fishery managers perceive the ectoparasitic crustacean, Argulus foliaceus to be a cause of significant economic loss through reduced fish capture rates. This study investigates the influence of previously identified risk factors on the abundance, egg laying habits and impact of this parasite through a longitudinal study of five trout fisheries of varying management intensity. Low water clarity, slow stock turnover and high temperatures showed a significant association with a high abundance of A. foliaceus. High infection levels, low water clarity and low temperature were also associated with reduced rates of fish capture, suggesting abundance of A. foliaceus alone may not affect the catch rates. Depth of egg laying varied in each site throughout the study, increasing in depth as temperature and water clarity increased. Eggs were found to be most abundant in natural fish holding areas and it is hypothesized that the location of egg laying is determined by the habitat usage of host fish.

Identification and functional characterization of a g-type lysozyme gene of Labeo rohita, an Indian major carp species.

Lysozyme, an important secretory innate immune component, possesses antimicrobial activity against broad spectrum of bacteria and viruses. In the present study, complete CDs (558 bps) of g-type lysozyme of rohu (Labeo rohita) was amplified and translated for a putative protein of 185 amino acids. The domain architecture and tertiary structure was also predicted for the protein. Its expression profile was studied in three infection models (bacteria: Aeromonas hydrophila, poly I:C, a dsRNA viral analogue and an ectoparasite: Argulus siamensis) in liver and kidney tissues of rohu. An up-regulation of 630-fold and 420-fold of the gene was observed at 48 h in liver and anterior kidney tissues respectively, after A. hydrophila infection. Significant increase in transcript level was noticed in both liver (0.8-fold) and kidney (480-fold) after 1 h and 12 h of poly I:C induction, respectively. Similarly, expression of lysozyme g transcripts was increased 6000-fold after 7 d of A. siamensis infection in liver tissue. The recombinant protein of g-type lysozyme of rohu (rLr-lysG) of 20.19 kDa was produced in Escherichia coli system and the lysozyme activity of rLr-lysG was found to be most active at pH 6.0 and temperature 35 °C. The potential lytic activity was found to be against A. hydrophila (UL = 0.53) followed by for E. tarda (UL = 0.45) whereas the lytic activity was the least against S. aureus (UL = 0.35) and M. lysodeikticus (UL = 0.34), at pH 6.0 and temperature 35 °C. The normal serum level of protein was estimated using indirect ELISA and was found to be very low (0.12-0.15 µg/ml). These results suggested that g-type lysozyme of rohu might be a potent immunostimulant against microbial infections, with a major role in innate immunity.