Spatial distribution, host specificity, and affinity of Branchiura for surface and macrohabitat types in fish species from the Brazilian Amazon.

This study investigated the spatial distribution, parasite interactions, and affinities of Branchiura parasites according to macrohabitat and surface type on fish from the Araguari River, Amapá State, in the Brazilian Amazon. This study presents novel information on the interactions of branchiuran species with host fish, and how the morphology of their attachment structures is associated with different host surface morphologies, with and without scales. A total of 558 fish specimens of 26 species were collected, of which 109 fish (10 species) were parasitized by at least one branchiuran parasite; these were selected for further study. The parasites were collected from the buccal cavity, gills, fins, and integument of the host fish. Parasites with suction discs (Argulus) were mainly associated to hosts with small scales and no scales, while parasites with hooks (Dolops) were mainly associated to hosts with large scales. Overall, parasites with suction discs were more specialized per site than those with hooks.

Alteration of gut microbiota composition and function of Indian major carp, rohu (Labeo rohita) infected with Argulus siamensis.

Gut microbiome homeostasis is critical in preventing diseases. However, the effect of disease on gut microbiota assembly remains unclear. At present, there are no reports on the composition and functional analysis of intestinal microbiota of Indian major carp, rohu (L. rohita) infected with ectoparasite, Argulus. In this study, we analysed and compared the intestinal microbiota of healthy and Argulus-infected rohu by 16S rRNA amplicon sequencing. Argulus infection could significantly influence the diversity and richness of the gut microbiota. However, abundance of Actinobacteria and Patescibacteria were enriched significantly in Argulus-infected fish. Venn diagram revealed that there were many more unique genera in the infected group as compared to control fish. The genera, Stenotrophomonas and Pirellula were significantly increased in infected fish while the abundance of Reyranella was decreased. LEfSe analysis showed a significant enrichment in abundances of 11 taxa in healthy group and 17 taxa in infected group. Furthermore, genera Rubellimicrobium, Dielma, Hyphomicrobium, Reyranella, Streptomyces and Cloacibacterium performed the best in differentiating between both the groups. Predicted microbiota function by PICRUSt revealed that the gut microbiota of infected fish was mainly associated with enriched synthesis of chitinases, chitin binding proteins, osmoprotectant proteins and sulfatases enzymes. There was a positive association between the structural and functional composition of the gut microbiota. The results indicated that the Argulus infection could affect the intestinal microbiota composition and function of rohu.

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.

Immunoproteomic analysis of fish ectoparasite, Argulus siamensis antigens.

AIM: An immunoproteomic approach was followed to identify immunoreactive antigens of fish ectoparasite, Argulus siamensis with rohu (Labeo rohita) immune sera for screening of potential vaccine candidates. MATERIALS AND RESULTS: The whole adult Argulus antigen was run in 2D electrophoresis with IEF in 7 cm IPG strips of pH 4-7 and SDS-PAGE with 12% acrylamide concentration. Two parallel gels were run; one was stained with silver stain, and the other was Western blotted to nitrocellulose paper (NCP) and reacted with rohu anti-A siamensis sera. Fourteen protein spots corresponding to the spots developed in NCP were picked from the silver-stained gel and subjected to mass spectrometry in MALDI-TOF/TOF. The MS/MS spectra were analysed in MASCOT software with taxonomy 'other metazoa' and the proteins identified based on similarity with the proteins from heterologous species. The gene ontology analysis revealed a majority of proteins being involved in binding activity in 'molecular function' and belonging to metabolic processes in 'biologic process' categories. The possibility of these proteins as vaccine candidates against A siamensis is discussed in the paper. CONCLUSION: Three of the identified proteins namely, bromodomain-containing protein, anaphase-promoting complex subunit 5 and elongation factor-2 could possibly serve as vaccine candidates against argulosis in carps.

Dose-dependent co-infection of Argulus sp. and Aeromonas hydrophila in goldfish (Carassius auratus) modulates innate immune response and antioxidative stress enzymes.

Co-infection with parasites and bacteria is of frequent occurrence in aquaculture, leads to growth impedance otherwise mortality in fish depending on the varying degree of a load of primary pathogen either parasite or bacteria. The mechanistic regulation of immune response during co-infection in fish has merely documented. The aim of this study was to determine the impact of co-infection with Aeromonas hydrophila at three exposure doses of Argulus sp. on the innate immune responses and antioxidative stress enzymes of goldfish (Carassius auratus). The experimental fish were randomly distributed into eight treatment groups viz. T1 (control group without Argulus and A. hydrophila infection), T2 (fish exposed to a sub-lethal dose of A. hydrophila), T3 (low Argulus-infested fish), T4 (T3 + sub-lethal dose of A. hydrophila), T5 (moderate Argulus-infested fish), T6 (T5 + sub-lethal dose of A. hydrophila), T7 (high Argulus-infested fish) and T8 (T7+ sub-lethal dose of A. hydrophila) in duplicates. After distributing experimental fish into their respective treatment group, A. hydrophila was injected to T2, T4, T6 and T8. After the bacterial challenge, four fish from each experimental group were randomly sampled on 24, 72, and 168 h and subjected to the hematological, innate immune parameters and enzymatic analysis. In the co-infection group T8, a high degree of enhanced pathogenicity of A. hydrophila was noticed with increased mortalities (84.2%) in comparison to other groups. The current study shows a declining pattern in RBC, PCV and Hb values with the degree of parasite infestation without co-infection groups. Moreover, in the T8 group, exposure of a sub-lethal dose of bacteria resulted in a drastic reduction of the recorded parameters. Furthermore, a decreased value for WBC, monocyte and neutrophil was found in higher parasite group co-infected with a sub-lethal dose of bacteria relative to other co-infected groups during the experimental period. Also, a decrease in innate immune parameters and antioxidative stress enzymes were observed in the T8 group compared to T7 and T2 groups throughout the trial period. These findings indicate that a rise in the dose of Argulus infection improves A. hydrophila colonization in goldfish and contributes to suppression of the innate immune system and increased mortality.

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.

Estimating cage-level mortality distributions following different delousing treatments of Atlantic salmon (salmo salar) in Norway.

This retrospective descriptive study estimates cage-level mortality distributions after six immediate delousing methods: thermal, mechanical, hydrogen peroxide, medicinal, freshwater and combination of medicinal treatments. We investigated mortality patterns associated with 4 644 delousing treatment of 1 837 cohorts of farmed Atlantic salmon (Salmo salar) stocked in sea along the Norwegian coast between 2014 and 2017. The mortality is expressed as mortality rates. We found distributions of delta mortality rate within 1, 7 and 14 days after all six delousing treatments, using mortality rate within 7 days before treatments as baseline. The results show that we can expect increased mortality rates after all six delousing methods. The median delta mortality rates after thermal and mechanical delousing are 5.4 and 6.3 times higher than medicinal treatment, respectively, for the 2017 year-class. There is a reduction in the delta median mortality for thermal and freshwater delousing from 2015 to 2019. There is a wide variability in the mortality rates, in particular for thermal delousing. Our results suggest that the variability in delta mortality for thermal delousing has been reduced from the 2014 to 2017 year-class, indicating an improvement of the technique. However, a significant increase in the number of thermal treatments from 14 in 2015 to 738 in 2018 probably contributes to the overall increased mortality in Norwegian salmon farming.

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.

Lethal concentration of Cymbopogon citratus (Poaceae) essential oil for Dolops discoidalis and Argulus sp. (Crustacea: Argulidae).

This study investigated the acute toxicity (LC50-24 hr ) effects of the essential oil of Cymbopogon citratus for adult Argulus sp. and Dolops discoidalis, before and during oviposition. In vitro acute toxicity (LC50-24 hr ) was tested using 20, 40, 60, 80, 100, 120, 140 and 160 µg/L of C. citratus essential oil, and two control groups (one with cultivation tank water and one with cultivation tank water + alcohol) were used. Specimens of Argulus sp. and D. discoidalis submitted to acute toxicity were evaluated using histological procedures. The major chemical compounds of C. citratus essential oil were geranial (47.5%), neral (35.6%) and myrcene (6.7%). The LC50-24 hr for Argulus sp. adults was 67.97 µg/L, while for D. discoidalis it was 59.55 µg/L. In the oviposition of both species of argulids, maximum mortality began with treatments of 140 µg/L, while the LC50-24 hr for Argulus sp. and D. discoidalis was 83.98 µg/L and 82.48 µg/L, respectively. In both argulid species exposed to C. citratus essential oil, morphological alterations were observed only in the eyes, and they occurred in the ommatidium and rhabdomeres and were dependent on the concentration of C. citratus essential oil and the parasite species.

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.

Under stress conditions, pacu Piaractus mesopotamicus modulates the metabolic allostatic load even after Dolops carvalhoi challenge to maintain self-protection mechanisms.

Fish metabolic allostatic dynamics, when animal present physiological modifications that can be strategies to survive, are important for promoting changes to ensure whole body self-protection and survival in chronic states of stress. To determine the impact of sequential stressors on pacu (Piaractus mesopotamicus), fish were subjected to two trials of stressful treatments, administration of exogenous dietary cortisol, and parasite challenge. The first experiment consisted of a two-day acute stress trial and the second, an eight-day chronic stress trial, and after both experiments, fish parasite susceptibility was assessed with the ectoparasite Dolops carvalhoi challenge. Physiological changes in response to acute trial were observed in glycogen, cortisol, glucose, osmolarity, sodium, calcium, chloride, potassium, hematocrit, hemoglobin, red blood cells and mean corpuscular volume, and white blood cell (P < 0.05), whereas response to chronic trial were observed in glycogen, osmolarity, potassium, calcium, chloride, mean corpuscular volume, white blood cell, neutrophil, and lymphocyte (P < 0.05). Acute trials caused physiological changes, however those changes did not induce the consumption of hepatic glycogen. Chronic stress caused physiological changes that induced hepatic glycogen consumption. Under acute trial, stress experience was important to fish to achieve homeostasis after chronic stress. Changes were important to modulate the response to stressor, improve body health status, and overcome the extra stressor with D. carvalhoi challenge. The experiments demonstrate that pacu initiate strategic self-protective metabolic dynamics in acute states of stress that ensure the maintenance of important life processes in front of sequential stressors.

Molecular evidence linking the larval and adult stages of Mexiconema cichlasomae (Dracunculoidea: Daniconematidae) from Mexico, with notes on its phylogenetic position among Dracunculoidea.

We describe the larval developmental stages and life cycle of the dracunculid nematode Mexiconema cichlasomae in both the intermediate, Argulus yucatanus (Crustacea: Branchiura), and definitive hosts, Cichlasoma urophthalmus (Perciformes: Cichlidae), from the Celestun tropical coastal lagoon, Yucatan, Mexico. The morphological analyses showed significant differences between the total length of L1 found in M. cichlasomae gravid female and L2-L3 in A. yucatanus. This result indicates that the M. cichlasomae larval development occurs in the intermediate host. We obtained sequences from the small subunit (SSU) ribosomal marker from larval stages of M. cichlasomae in A. yucatanus and adult nematodes in C. urophthalmus. Our morphological and molecular results support conspecificity between M. cichlasomae larvae in A. yucatanus and the adult stages in C. urophthalmus. We briefly discuss the phylogenetic position of M. cichlasomae among the Daniconematidae, and provide evidence of the monophyly of the daniconematids associated with branchiurid intermediate hosts. Based on the phylogenetic results, we support the transfer of the Mexiconema genus to the family Skrjabillanidae and do not support the lowering of family Daniconematidae to subfamily.

Development of a western blot method for detection of fish ectoparasite Argulus siamensis antigens.

A study was undertaken to develop a western blot method for detection of immunogenic proteins of fish ectoparasite, Argulus siamensis for its further use as potential vaccine candidates. Argulus antigens were prepared by homogenization and injected to rohu (Labeo rohita) juveniles for development of immune serum. The serum was used to immunostain the antigens in western blot. The other reagents added in sequence were rabbit anti-rohu serum, goat anti-rabbit ALP conjugate and substrate (BCIP-NBT). However, similar banding patterns were observed with both control and immunized rohu serum. Hence, a possible cross-reaction was suspected and verified in number of western blot experiments. A typical cross-reaction observed was of rabbit serum reacting directly with Argulus antigens. Hence, the rabbit anti-rohu serum was replaced with guinea pig anti-rohu serum. Another cross-reaction of goat anti-guinea pig ALP conjugate with rohu serum was eliminated by using goat anti-rabbit ALP conjugate with guinea pig serum. Thus, the final western blot method consisting of Argulus antigens → rohu serum → guinea pig anti-rohu serum → goat anti-rabbit ALP conjugate → substrate, yielded distinguishing results between control and Argulus-immunized rohu serum samples. The developed test has tremendous downstream applications, particularly in immunoproteomic studies of Argulus antigens.

Molecular cloning, characterization and expression of immunoglobulin D on pathogen challenge and pathogen associated molecular patterns stimulation in freshwater carp, Catla catla.

The primordial immunoglobulin class, IgD, was the first non-IgM isotype discovered in teleosts. The crucial roles of IgM and IgZ in imparting systemic and mucosal immunity, respectively, in various fish species have been widely established. However, the putative function of a unique IgD isotype during pathogenic invasions has not been well explored. The present study reports the existence of an IgD ortholog in freshwater carp, Catla catla, and further evaluates its differential expression profile in response to bacterial, parasitic and viral antigenic exposure and pathogen associated molecular patterns (PAMPs) stimulation. The IgD of C. catla (CcIgD) cDNA sequence was found to encode 226 amino acids and confirmed homology with heavy chain delta region of Cyprinidae family members. Phylogenetic analysis of CcIgD exhibited greatest similarity with Ctenopharyngodon idella. qRT-PCR analysis revealed significant upregulation (P < 0.001) of IgD gene expression in kidney with respect to other tissues at 24 hr post-Aeromonas hydrophila challenge. CcIgD gene expression in skin was enhanced following Streptococcus uberis infection and in blood following Argulus infection and inactivated rhabdoviral antigen stimulation. Further, the treatment of bacterial and viral products (PAMPs) also triggered significant (P < 0.05) increases in CcIgD mRNA expression in kidney. These findings indicate the functional importance of teleost IgD in orchestrating tissue specific neutralization of antigens on stimulation with different pathogens and PAMPs.

Structure of parasites community in Chaetobranchopsis orbicularis (Cichlidae), a host from the Amazon River system in northern Brazil.

The aim of this paper was to investigate the component communities of parasites in Chaetobranchopsis orbicularis from a tributary of the Amazon River system, in Northern Brazil. In 32 fish examined, 902,551 parasites were collected, including Ichthyophthirius multifiliis, Piscinoodinium pillulare, Sciadicleithrum geophagi, Posthodiplostomum sp., Clinostomum marginatum, Echinorhynchus paranensis, Neoechinorhynchus pterodoridis, and Dolops longicauda. I. multifiliis was the dominant and abundant parasite species. The ectoparasites presented aggregate dispersion, but the endoparasites showed random dispersion pattern. Mean species richness was 4.0 ± 1.5 parasites, mean Brillouin diversity (HB) was 0.33 ± 0.28, mean evenness was 0.15 ± 0.13, and Berger-Parker dominance (d) was 0.85 ± 0.17. The species richness of parasites and HB were positively correlated with the length of hosts. There was positive correlation between the abundance of P. pillulare and length and weight, between the abundance of I. multifiliis and weight, as well as between the abundance of E. paranensis and N. pterodoridis and the length of hosts. Body condition of the hosts was not affected by moderate parasitism. The low diversity of endoparasites indicates that C. orbicularis is a host with low position in the food web. This is the first record of all these parasites for C. orbicularis.

Involvement of aurantiactinomyxon in the life cycle of Thelohanellus testudineus (Cnidaria: Myxosporea) from allogynogenetic gibel carp Carassius auratus gibelio, with morphological, ultrastructural, and molecular analysis.

During the investigation of actinosporean fauna diversity from commercial fish ponds in Hubei Province, China, a novel aurantiactinomyxon type was found from Branchiura sowerbyi. Spore body of the aurantiactinomyxon was ellipsoidal in side view and triangular in apical view, 15.5 ± 0.5 (14.5-16.4) µm in diameter; three leaf-like caudal processes were approximately equal, measuring 13.2 ± 0.9 (11.5-16.2) µm long and 7.4 ± 0.4 (6.7-8.0) µm wide at the base; three polar capsules were located at the apex of spore body, globular in apical view, 2.2 ± 0.1 (2.0-2.3) µm in diameter, and pyriform in side view, 2.5 ± 0.2 (2.3-2.9) µm in length and 2.0 ± 0.2 (1.8-2.4) µm in width; a total of 32 germ cells were observed within the sporoplasm. Ultrastructural analysis showed that the development was asynchronous between pansporocysts but synchronous within a pansporocyst. The formation of sporoblast and the development of sporogonic stage were also described and discussed. The 18S ribosomal DNA sequences of the current aurantiactinomyxon type corresponded to that of a previously reported Thelohanellus testudineus, suggesting that the newly identified aurantiactinomyxon type is the actinosporean stage in the life cycle of T. testudineus.

Rediscovery of Chonopeltis meridionalis Fryer, 1964 (Crustacea: Branchiura) from Labeo rosae Steindachner in the River Olifants, Mpumalanga, and the taxonomic status of C. victori Avenant-Oldewage, 1991 and C. koki Van As, 1992.

Chonopeltis Thiele, 1900 presently comprises 14 species, it is endemic to Africa and its species show a high degree of host-specificity towards fish families and in some cases, individual fish species. Chonopeltis meridionalis Fryer, 1964 was originally described from Labeo rosae Steindachner collected in the River Nuanetsi in Rhodesia, now Zimbabwe, Limpopo River System. At the time of description C. meridionalis was the most southerly species of the genus. Chonopeltis victori Avenant-Oldewage, 1991 was described from the River Olifants, Mpumalanga, South Africa, which also forms part of the Limpopo River System. The host fish was Labeo rosae and Labeobarbus marequensis (A. Smith), whilst C. koki Van As, 1992 was described from Labeo cylindricus Peters, collected in the River Zambezi, Eastern Caprivi, Namibia. During surveys conducted in 2012, additional material of a species of Chonopeltis was collected from Labeo rosae in the River Olifants. Upon closer examination, the new material was identified as C. meridionalis. Further investigation revealed that C. victori and C. koki share a number of characteristics with C. meridionalis. This paper provides the first scanning electron microscopy of C. meridionalis, includes additional information on fully-mature as well as sub-adult males and females. Finally, it was concluded that C. victori and C. koki are junior synonyms of C. meridionalis.

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.