Isolation and characterization of polymorphic microsatellites in Labeo rohita and their cross‐species amplification in related species

The major Indian carps namely rohu (Labeo rohita), catla (Catla catla), mrigal (Cirrhinus mrigala) and calbasu (Labeo calbasu) are important freshwater species of the Indian subcontinent constituting over 65% of the fish produce. In the present study, isolation of 12 microsatellite loci from rohu has been reported. Cross‐species amplification in related carps and their implication in population genetic studies as well as selective breeding program were discussed.     

Length–weight relationships of four Cyprinid species in India

Length–weight relationships are reported for four Indian carps. The length–weight relationship was W = 0.01123L^3.355 for Cirrhinus mrigala, W = 0.07354L^3.299 for Catla catla, W = 0.04066L^3.313 for Labeo rohita and W = 0.11489L^2.732 for Labeo fimbriatus. LWR estimates for C. mrigala and L. fimbriatus had not been previously reported in FishBase.     

Characterization of IgM of Indian major carps and their cross-reactivity with anti-fish IgM antibodies

Indian major carps (IMC), rohu (Labeo rohita), catla (Catla catla) and mrigal (Cirrhinus mrigala) were immunized with bovine serum albumin and the serum immunoglobulin M (IgM) was purified by affinity chromatography. The heavy and light chain of IgM of all the three species of IMC were about 88 and 26 kDa, respectively. Anti-fish IgM antibody against all the three species were raised in mice and the reaction of anti-fish IgM antibodies with IgM of all the three species of IMC were studied by Western blot. The anti-fish IgM antibodies reacted strongly with the heavy chain of the same species against which it was raised while the reactions with the heavy chain of other species were milder indicating some degree of epitope sharing among the heavy chains of IgM of IMCs. However, there was no cross-reaction with the light chain of any of the IgM.     

Study on impact of habitat degradation on proximate composition and amino acid profile of Indian major carps from different habitats

This investigation is aimed to study an impact of habitat degradation on proximate composition and amino acid (AAs) profile of Catla catla, Labeo rohita and Cirrhinus mrigala collected from polluted, non-polluted area (upstream) and a commercial fish farm. The amino acid profile was estimated by the amino acid analyzer. C. catla collected from the polluted environment had highest lipid, protein and ash contents (12.04?±?0.01, 13.45?±?0.01 and 0.93?±?0.03%, respectively). The high protein content (14.73?±?0.01 and 14.12?±?0. 01%) was recorded in C. catla procured from non-polluted (upstream) wild habitat of River Chenab and controlled commercial fish farm. Farmed fish species showed comparatively higher moisture contents followed by upstream and polluted area fishes. C. mrigala showed significant differences in amino acid and proximate composition collected from a polluted site of the river Chenab. C. catla collected from non-polluted site of the river showed an excellent nutrient profile, followed by L. rohita (wild and farmed) and C. mrigala (polluted area), respectively. All fishes from the polluted areas of the River Chenab indicated a significant decrease in the concentration of some AAs when compared to farmed and wild (upstream) major carps. Omitting of some important AAs was also observed in the meat of fish harvested from polluted habitat of this river. C. mrigala and L. rohita exhibited a significant increase in the concentration of some of non-essential amino acids such as cysteine in their meat. The results indicated that wild fish (upstream) and farmed fish species had highest protein contents and amino acid profile and hence appeared to be the best for human consumption. The proximate composition and AAs profiles of fish harvested from the polluted area of the river clearly indicated that efforts shall be made for the restoration of habitat to continue the requirement of high quality fish meat at a low cost to the human population.     

Complete mitochondrial genome sequence of Catla catla and its phylogenetic consideration

Complete nucleotide sequence of mitochondrial genome (mitogenome) of the Catla catla (Ostariophysi: Cypriniformes: Cyprinidae) was determined in the present study. Its length is 16,594 bp and contains 13 protein coding genes, 22 transfer RNAs, two ribosomal RNAs and one non-coding control region. Most of the genes were encoded on the H-strand, while the ND6 and eight tRNA (Gln, Ala, Asn, Cys, Tyr, Ser (UCN), Glu and Pro) genes were encoded on the L-strand. The reading frames of two pair of genes overlapped: ATPase 8 with 6 and ND4L with ND4 by seven nucleotides each. The main non-coding region was 929 bp, with three conserved sequence blocks (CSB-I, CSB-II, and CSB-III) and an unusual simple sequence repeat, (TA)₇. Phylogenetic analyses based on complete mitochondrial genome sequences were in favor of the traditional taxonomy of family Cyprinidae. In conclusion present mitogenome of Catla catla adds more information to our understanding of diversity and evolution of mitogenome in fishes.     

Performance of different fish species for controlling weeds in rainfed lowland rice field

A field experiment was conducted during wet season to evaluate the performance of different fish species for biocontrol of weeds in rainfed waterlogged rice fields with fingerlings of three exotic carps – grass carp, Ctenopharyngodon idella (Val.) (Cyprinidae, Cypriniformes), silver barb, Puntius gonionotus (Bleeker) (Cyprinidae, Cypriniformes) and common carp, Cyprinus carpio (L.) (Cyprinidae, Cypriniformes) – and three Indian major carps – rohu, Labeo rohita (Ham.) (Cyprinidae, Cypriniformes), catla, Catla catla (Ham.) (Cyprinidae, Cypriniformes), mrigal, Cirrhinus mrigala (Ham.) (Cyprinidae, Cypriniformes). A total of 13 major weeds under the categories of grassy, sedges, broadleaf and aquatic weeds were observed in the rice fields. Grass carp reduced maximum weed biomass (weed control efficiency [WCE] 63% at 60 days after transplanting [DAT] and 62% at 100 DAT) followed by silver barb and common carp. Among the Indian carps, only rohu was effective in control of weeds (WCE, 23% at 60 DAT). The grain yield of rice (variety Varshadhan) slightly increased (4.2–4.5 t/ha), but straw yield was significantly higher (10.2–10.7 t/ha) under rice-fish farming. Fish yield was significantly higher in exotic carps (270–288 kg/ha/90days) due to higher specific growth rates (1.8–2% body weight/day). The study indicated that exotic carps (grass carp, silver barb and common carp in order) were more effective than Indian carps for control of weed in rainfed lowland rice fields and among the Indian carps, rohu showed potential for weed control.     

Seasonal variations in protein,RNA and DNA concentrations in major carps,Catla catla,Labeo rohita andCirrhina mrigala

Seasonal variations in the concentrations of protein, RNA and DNA, and the RNA/DNA ratio were studied in the liver of major carps,Catla catla, Labeo rohita andCirrhina mrigala during their prematurity phase. These biochemical parameters maintained strikingly similar patterns of change in the three species. Alterations in nucleic acids were reciprocal to that of protein. Magnitude of such interrelation, however, varied with the season. Protein level was generally low during winter and high in the summer months, attaining peak value in March and June. Higher concentrations of RNA and DNA were registered in December and June and a sharp decline was evident in January as well as July. During the remaining months, protein, RNA and DNA varied intermittently, but not profoundly.     

Ingestion of bacteria in suspension Indian major carps (Catla catla,Labeo rohita) and Chinese carps (Hypophthalmichthys molitrix,Aristichthys nobilis)

Single-species groups of fry of the planktivorous, filter-feeding carps (silver carp Hypophthalmichthys molitrix, bighead carp Aristichthys nobilis, rohu Labeo rohita and catla Catla catla) were exposed to the bacterium Chromobacterium violaceum to assess if they could detect or ingest unattached bacteria. The results indicate that planktivorous carp fry are able to ingest unattached bacteria and that the numbers ingested increase with time and with bacterial concentration in the media, although probably by passive means. It is also suggested that unattached bacteria are of little nutritional significance to carps.     

A comparative study of the epidermis of the common carp and the three Indian major carp

A comparative study has been made of the mucogenic epidermis of the common carp, Cyprinus carpio var. communis, and the three Indian major carps, Catla catla, Labeo rohita and Cirrhina mrigala: on the basis of epidermis structural organization, these species are easily differentiated. The epithelial cells in the superficial layer, as in most fishes, show secretory activity, evidenced by positive histochemical reactions, which is high in C. carpio var. communis, moderate in C. catla and low in L. rohita and C. mrigala. The epithelial cells in the underlying two or three layers also give positive reactions, though their intensity is relatively weak. The mucous cells in C. carpio var. communis are distributed in large numbers arranged in several superimposed layers in the outer regions of the epidermis, whereas in C. catla they are fewer in number and are widely separated in the surface layers as well as in the deeper layers of the epidermis; in both species the mucous cells appear rounded, large, and open on the surface by wide pores. In contrast, in L. rohita and C. mrigala the mucous cells are smaller, restricted mainly to the superficial layer, close together in a single row, and open on the surface by narrow pores. The overall density of mucous cells in L. rohita and C. mrigala, as in C. catla, is much lower than in C. carpio var. communis. In the epidermis of C. carpio var. communis there are a large number of mucous cells, and the few club cells are restricted to the deeper layers. In contrast, in the epidermis of the three Indian major carp the overall density of the mucous cells is much lower and the club cells are very numerous. It is suggested that the high density of club cells compensates an overall low density of mucous cells as an adaptation for an effective defence mechanism. Increased mucus production in the epidermis of C. carpio var. communis, as evidenced by a large number of mucous cells in outer regions and high secretory activity of superficial layer epithelial cells, is associated with increased precipitation of mud held in suspension, needed as an adaptation to the species’peculiar bottom-scooping habits. The varied density of the taste buds in the epidermis of the four carp is associated with their feeding habits.     

Fish Scales as Potential Substrate for Production of Alkaline Protease and Amino Acid Rich Aqua Hydrolyzate by <Emphasis Type="Italic">Bacillus altitudinis</Emphasis> GVC11

Fish processing industries generate large quantities of fish scales as processing waste, if not treated leading to environmental pollution. Fish scales are hard to degrade, hence cause difficulty in waste management. In this context present study was made to utilize fish scales as substrate for the production of alkaline protease by Bacillus altitudinis GVC11 and subsequently amino acid rich aqua hydrolyzate. B. altitudinis GVC11 efficiently utilized five types of fish scales as substrates and produced maximum alkaline protease using Labeo rohita (28,150 U/mL) followed by Catla catla (23,320 U/mL) at 48 h and Cyprinus carpio (17,146 U/mL) Mugil cephalus (18,917 U/mL), Cirrhinus mrigala (12,430 U/mL) at 72 h. The HPLC analysis of protein hydrolyzate obtained after fermentation was enriched in essential amino acids, leucine, isoleucine, phenylalanine, lysine and non-essential amino acids, tyrosine, arginine and cysteine which can be used as animal feed supplement and organic fertilizer.     

Non-specific immune responses in juveniles of Indian major carps

The non‐specific immune parameters are useful to determine the health status of fish and to evaluate the immunomodulatory substances for fish farming as markers of pollution and disease resistance. Some of the important parameters, viz. superoxide production by neutrophils through nitroblue tetrazolium (NBT) assay; haemagglutination (HA), haemolysin (HLY) and bacterial agglutination titres; myeloperoxidase (MPO), and lysozyme activities, and alternative complement levels in serum of the juveniles of three Indian major carp species (Cirrhinus mrigala, Catla catla and Labeo rohita) were measured to establish their physiological normal range. A wide variation among the individuals within a species in the ranges of the most of the immune parameters was recorded. Significantly higher levels in the mean values of HA, HLY, bacterial agglutination titres; superoxide production by neutrophils in nitroblue tetrazolium assay; serum MPO and lysozyme activities, i.e. 371.20, 4.60, 18.80, 0.40, 0.62 and 6.55 μg ml^?1, respectively, were obtained in L. rohita except a much lower alternative haemolytic complement activity (29.06 units ml^?1) compared with the other two species. In most of the parameters, L. rohita showed the highest value, possibly indicating its more natural resistance compared with the other two species.     

Ontogeny and tissue-specific expression of immune-relevant genes in Catla catla (Hamilton)

India is the second largest fish producing country in the world with production of 12.6 million tonnes (mt) in 2017–18, and Indian major carps (IMCs) contribute bulk of this fish production. Catla, Catla catla is the fastest growing species among IMCs. However, the survival rate of catla during larval rearing is normally lower than the other IMCs i.e rohu Labeo rohita, and mrigal Cirrhinus mrigala. Continuous efforts are devoted for the identification of nutritional and environmental requirements of fish larvae in order to reduce hatchery mortalities. However, very little information is available regarding physiology of the immune system, especially during the late larval and juvenile stages. Hence, understanding the ontogenetic development of immune-relevant genes in the larval stages of catla will serve as the markers for the development of immune competence and thereby, will be beneficial in developing effective immune intervention strategies. In the present study, expression profiles of some of the important innate (IL-1β, IL-10, iNOS and C3) and adaptive immune (RAG-1, Ikaros, IgM and IgZ) genes during ontogenetic developmental stages and in different tissues of catla were investigated using quantitative real-time PCR. The results revealed that immune genes IL-1β, C3, IgM and IgZ were detected in the unfertilized eggs indicating their maternal inheritance. Immune genes, IL-1β, IL-10 and iNOS were expressed significantly during initial larval developmental stages whereas C3, RAG-1, Ikaros, IgM and IgZ showed significant expression during advanced stages of larval development in catla i.e. from 23 days post hatch (dph). Study of tissue distribution pattern of the genes indicated that innate immune genes were ubiquitously expressed in different tissues with varying degree of expression, whereas adaptive immune genes were predominantly expressed in lymphoid organs of catla. The information thus generated will improve knowledge on the maturation of the immune system in catla and will aid in deciding the appropriate age for vaccination in this teleost species.     

Molecular insights into the genetic and haplotype diversity among four populations of Catla catla from Madhya Pradesh revealed through mtDNA cyto b gene sequences

In the present investigation, the genetic structure of four populations of Catla catla, sequences of mitochondrial gene, cytochrome b (cyto b) from four populations were sequenced and analyzed. The sequences of mitochondrial regions revealed high haplotype diversity and low nucleotide diversity. The lowest 249 polymorphic sites and 0.00 parsimony informative sites were detected in populations of Fish Federation Pond (CCFFB) whereas highest 330 polymorphic sites and 56 parsimony informative sites were detected in populations of Narmada River (CCNRH) in the cyto b gene sequences in Catla catla populations. The twelve different haplotypes were detected among the four populations studied, lowest population specific haplotype as 2.00 was observed in Fish Federation Pond (CCFFB) and highest was in Population of Narmada River and Tighra reservoir. Sequencing of cyto b gene revealed 12 number of haplotypes (h) with haplotype (gene) diversity (Hd) 0.8736 and nucleotide diversity (π) 0.6474. These data clearly indicated that, feral/wild population showing highest values of polymorphisms, parsimony, haplotype diversity showing good, healthy habitat is lotic water (Narmada River) and lentic water body (Tighra reservoir). The results also concluded that the partial cyto b is polymorphic and can be a potential marker to determine ecological habitat based genetic differentiation among the populations.     

Comparison of plankton intake by Indian carp fingerlings under different feeding conditions

The influence of different culture systems on the amount of plankton consumed by fingerlings of three carp species (Labeo rohita, Catla catla and Cirrhinus mrigala) was examined in outdoor culture tanks during a 90 day growing period. The fish were raised under two basically different feeding conditions: (a) fed with allochthonous live plankton; (b) fed with plankton grown autochthonously in the fish growing tanks fertilized with manures. Both feeding regimes were compared with control groups. It was found that the plankton intake for carp held in the live food system was significantly higher than with the manured and control systems. Related to maximum intake, maximum abundance of plankton in the live food system was a consequence of improved water quality expressed in terms of lower values of BOD and COD and higher values of DO and pH; this was conducive to fast reproduction of some of the zooplankton which constituted the major food items for the test carp, as well as to the regular exogenous introduction of live plankton. The ingestion of plankton was found to be related to carp body weight raised to the power b. The exponent b ranged from 0.29 to 0.93 during the 90-day growing period.     

Mitochondrial control region and protein coding genes sequence variation among phenotypic forms of brown trout Salmo trutta from northern Italy

The Pô River basin of northern Italy is the home of distinctive and endemic morphological forms of brown trout Salmo trutta. We used PCR-direct sequencing and RFLP techniques to study variation in the mitochondrial control region of 225 trout in order to assess genetic relatedness among 18 populations from that region. The distribution analysis of these genotypes among north Italian populations confirmed the phylogenetic differentiation of marbled trout Salmo trutta marmoratus populations and the postglacial origin of S. t. fario. Extensive genetic heterogeneity was observed among morphologically identical S. t. fario populations. Introgression with domestic strains of Atlantic basin origin was detected in all forms. In order to assess the phylogenetic congruence detected in coding and noncoding regions of the mitochondrial genome, we also analysed sequence variation in segments of the cytochrome b and ATPase subunit VI genes among representatives of all variants detected in the analysis of the control region. Variation in protein coding genes was only slightly less than that observed in the control region of the same individuals, both in terms of number of variants detected and of pairwise sequence divergence estimates among variants. Phylogenetic analysis based on protein coding genes sequences identified the same phylogenetic groupings defined by the control region analysis and also allowed a partial resolution of their phyletic relationships that was previously unresolved. However, coding and noncoding segments differed substantially in the transition-transversion ratio (17:0 in coding segments vs. 17:6 in control region segments).     

Molecular cloning of growth hormone encoding cDNA of Indian major carps by a modified rapid amplification of cDNA ends strategy

A modified rapid amplification of cDNA ends (RACE) strategy has been developed for cloning highly conserved cDNA sequences. Using this modified method, the growth hormone (GH) encoding cDNA sequences ofLabeo rohita, Cirrhina mrigala andCatla catla have been cloned, characterized and overexpressed inEscherichia coli. These sequences show 96–98% homology to each other and are about 85% homologous to that of common carp. Besides, an attempt has been made for the first time to describe a 3-D model of the fish GH protein.     

Effect of acclimated temperature on thermal tolerance,immune response and expression of HSP genes in Labeo rohita,Catla catla and their intergeneric hybrids

The ability of a species and population to respond to a decrease or an increase in temperature depends on their adaptive potential. Here, the critical thermal tolerance (CTmax and CTmin) of four populations: Labeo rohita, Catla catla, and their reciprocal hybrids L. rohita♀× C. catla♂ (RC) and C. catla♀ × L. rohita♂ (CR) being acclimatized at four acclimation temperatures (22, 26, 30 and 34 °C) were determined. All populations indicated substantial variations (P < 0.05) in CTmax and CTmin values. L. rohita displayed, comparatively the highest CTmax with largest total and intrinsic polygon zones as well as the upper and lower acquired thermal tolerance zones followed by RC and CR hybrids, while C. catla showed significantly the highest CTmin value and the smallest intrinsic and acquired thermal tolerance zones. Both hybrids illustrated low parent heterosis (≤11%). Additionally, the highest expression of Hsp70 and Hsp90 (heat shock proteins) genes, serum lysozyme level, respiratory burst activity and lowest lipid peroxidation level under lower and higher temperature shock further illustrated strong physiological mechanism of L. rohita in contrast to C. catla, to deal with acute temperature, while hybrids, especially F1 RC hybrid appeared as a good option to replace C. catla in relatively higher and lower temperature areas.     

Food selection by Labeo Rohita (Ham.) and its feeding relationship with other major carps

Selectivity of food byLabeo rohita (Ham.) was studied in a stocking pond (Moat), by calculating an Electivity index (E) for each food organism as described byIvlev (1961). It was found thatL. rohita was definitely selective in its feeding. In case of fingerlings, there was a strong selection for zooplanktonic organisms (Arcella andDifflugia among protozoans,Keratella andBrachionus among rotifer andDaphnia andCyclops among crustaceans) and smaller algae (Cosmarium andClosterium among desmisd,Euglena andVolvox among phytoflagellates and algal spores and zygotes) while most of the phytoplanktonic organisms, belonging to green algae, diatoms and blue green algae, were avoided. In case of adults, a strong negative selection was observed for all zooplanktonic organisms and a strong positive selection for most of the green algae and diatoms (Ankistrodesmus, Zygnema, Spirogyra, Selenastrum, Pediastrum, Scenedesmus, Tetraspora, Stephanodiscus, Naviculla, Diatoma, Synedra andNitzchia). However, all blue green algae were avoided.The feeding relationship ofL. rohita with other major carps,Cirrhina mrigala (Ham.) andCatla catla (Ham.) was studied in two different habitats, pond and river. It was observed that in both habitats all three species were found to feed on almost similar types of food organisms, but the quantity of any food item eaten by adults differed markedly from one species to another and the food items which were dominant in one species, were of secondary importancce for the other two species. The adults ofL. rohita were found to feed mainly on phytoplankton and macrovegetation, the main food of adultC. mrigala was decayed organic matter, sand and mud supplimented by plankton, while the food of adultC. catla was chiefly composed of zooplankton, and some phytoplankton. Hence there was no true identity of feeding habits between the adults of any two species. However, there was an indication of competition for food between the fingerlings of all three species, because all of them feed mainly on zooplankton (Crustaceans, rotifers and protozoans). However, such feeding habits lasted a very short time only and as the fishes grew, their feeding habits diverged.     

Genetic divergence in wild population of <Emphasis Type="Italic">Labeo rohita</Emphasis> (Hamilton, 1822) from nine Indian rivers,analyzed through MtDNA cytochrome b region

The present study examined partial cytochrome b gene sequence of mitochondrial DNA for polymorphism and its suitability to determine the genetic differentiation in wild Labeo rohita. The 146 samples of L. rohita were collected from nine distant rivers; Satluj, Brahmaputra, Son, Chambal Mahanadi, Rapti, Chauka, Bhagirathi and Tons were analyzed. Sequencing of 307 bp of Cyto b gene revealed 35 haplotypes with haplotype diversity 0.751 and nucleotide diversity (π) 0.005. The within population variation accounted for 84.21% of total variation and 15.79% was found to among population. The total Fst value, 0.158 (P < 0.05) was found to be significant. The results concluded that the partial cyto b is polymorphic and can be a potential marker to determining genetic stock structure of L. rohita wild population.