Scanning electron microscopy of the scales of Cyprinus carpio, inhabiting two government-sponsored fish farms, with different sources of pollutants, in the North-East state of Meghalaya, India revealed extensive damage to them. Lepidonts, which are teeth-like structures present on the edge of circulii, were discovered to be either partly or completely damaged. The dislodgment of complete rows of lepidonts along with broken lepidonts was also observed. Loss of symmetry and torn areas of circulii are some of the abnormalities also observed as compared to control samples. The alterations observed could be because of the high sensitivity of the scales to environmental stress and their capacity to react to low concentrations of pollutants, which makes them a sensitive bioindicator of water pollution. The study hence not only represents an advance in pollution research, by employing a highly sophisticated analytical approach to reveal the above-mentioned microstructural scale abnormalities, but it can also have scientific impact on the society at large and the fish culturist in particular in understanding the destructive impact of pollution caused by mismanagement of city garbage and agricultural effluents.

The present study was conducted to investigate the effects of dietary fructo-oligosaccharide (FOS) (0, 1, 2 and 3 %) supplementation on the growth performance, haemato-immunological parameters, cultivable autochthonous (non-adherent) intestinal microbiota and stress resistance of common carp (Cyprinus carpio) fry (3·23 (sem 0·14) g). These parameters were measured after feeding the carp fry with the experimental diets for 7 weeks. Dietary FOS supplementation had no significant effects on the growth performance and food intake of carp fry compared with the control treatment. It also had no significant effects on the following haematological parameters: erythrocyte count; leucocyte counts (WBC); haematocrit; Hb; mean corpuscular volume; mean corpuscular Hb content; mean corpuscular Hb concentration. However, WBC and respiratory burst activity were significantly affected by dietary FOS supplementation. Evaluation of the cultivable autochthonous intestinal microbiota revealed a significant increase in the levels of total viable heterotrophic aerobic bacteria and lactic acid bacteria in fish fed diets supplemented with 2 and 3 % FOS. Furthermore, dietary FOS supplementation significantly increased the survival rate and stress resistance of carp fry compared with the control treatment. These results encourage conducting further research on the administration of FOS and other prebiotics in carp fry studies.

The aim of the present study was to investigate the effects of different dietary sustained-release microencapsulated sodium butyrate (MSB) products (0 (non-supplement), 1·5 and 3·0 h) for a control or oxidised soyabean oil (SBO) diet on fish production, intestinal mucosal condition, immunity and intestinal bacteria in juvenile common carp (Cyprinus carpio). Dietary MSB increased weight gain and reduced the feed conversion ratio within the control and oxidised SBO groups. Gut mucosa was damaged in the oxidised SBO group fed without MSB, in contrast to a normal appearance found in fish fed the MSB1·5 and MSB3·0 diets in the oxidised SBO group. Microvillus density increased in fish fed the MSB1·5 and MSB3·0 diets in the oxidised SBO group (P< 0·001); however, microvillus density was affected by the different pre-fed diets in the midgut (P< 0·001) and by the different sustained-release times of MSB in the distal gut (DG) (P= 0·003). The interaction between the pre-fed diets and the sustained-release times of dietary MSB was significant for the relative gene expression levels of gut heat shock protein-70 (HSP70), pro-inflammatory cytokines (IL-1β and TNF-α) and anti-inflammatory cytokines (transforming growth factor-β) within each gut segment, except for HSP70 in the DG and IL-1β in the foregut. Modulation of adherent bacterial communities within each gut segment investigated was not obvious when the common carp were fed the diets with MSB, as similarity coefficients of >0·79 were observed. These results indicated that MSB can be used as a dietary supplement to repair or prevent intestinal damage in carp fed oxidised SBO.

Methodology is described for the laboratory maintenance of the life-cycle of Sanguinicola inermis, using common carp (Cyprinus carpio) as the definitive host and Lymnaea peregra as the intermediate host. Lymnaea auricularia was also infected but is not considered to be a suitable laboratory host. Tench (Tinca tinca) were susceptible to infection at high cercarial doses but subsequent infection rates were low. Goldfish (Carassius auratus) and Lymnaea stagnalis were refractory to laboratory infection.

The migration and development of Sanguinicola inermis in carp fingerlings were investigated by quantitative analysis of serial sections of fish and examination of flukes from hosts up to 90 days p.i. Cercariae penetrated through the epidermis and migrated through connective tissue and muscle to establish in the blood system and commence egg production within 28 days p.i. Most flukes established in the ventral blood system and released eggs which accumulated in gill epithelial tissue. A minority of flukes inhabited dorsal blood vessels and shed eggs which lodged in connective tissue and visceral sites. Maximum life-span of the parasite is between 56 and 70 days p.i. and egg production peaked at day 42 p.i.

Macrophages from the head kidney (HK) and peripheral blood leucocytes were isolated fromCyprinus carpio by Percoll gradient density centrifugation, cultured in vitro and exposed to different concentrations of the immunomodulator lentinan. To evaluate the immunostimulating effects of lentinan, proliferation of the peripheral blood leucocytes, respiratory burst of macrophages and interleukin-1β (IL-1β) gene mRNA expression of macrophages were investigated by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide], NBT (nitroblue tetrazolium) reduction, Griess reaction and real-time polymerase chain reaction (PCR). Results showed that proliferation of peripheral blood leucocytes after 24 h incubation by the induction of lentinan at 100 and 1000 μg/ml was markedly stimulated. Lentinan at 1, 100 and 1000 μg/ml could significantly induce superoxide anions in macrophages. Production of nitric oxide by HK macrophages after 96 h incubation by lentinan showed that it had no conspicuous effect on nitrogen burst activity of macrophages. Moreover, it inhibited the nitrogen burst activity at higher doses. The expression of IL-1β in the HK macrophages after 24 h of polysaccharide stimulation showed that lentinan stimulated IL-1β expression in the head kidney in carp. Lentinan can modulate the immune response of C. carpio.

The effect of Sanguinicola inermis on serum antibody and complement activity in Cyprinus carpio was assessed using an ELISA and haemolytic assays. Possible immune evasion strategies were assessed using immunodetection of host proteins on the surface of the parasite. Carp acclimatized to 20 or 25 °C were infected by exposure to 500 cercariae or injected intraperitoneally with 150 cercariae, and serum monitored over a 63-day period. In cercariae-injected carp, irrespective of time and temperature, a significant increase occurred in complement activity being greatest at 25 °C. In addition, fish exposed to the cercariae of S. inermis and maintained at 20 °C the level of complement activity was significantly higher after 5 weeks compared to controls. At 20 °C intraperitoneal injections of parasites increased serum antibody levels which peaked after 7 days. In contrast, at 25 °C, antibody levels were maintained over 63 days. Exposure of fish to infection did not appear to stimulate antibody production. Immunofluorescence studies revealed ‘host-like’ molecules on the surface of the cercarial body exposed to carp serum and adult flukes obtained directly from the fish or cultured for 24 h in L15 medium. The possible role of ‘host-like’ molecules in immune evasion is discussed and the response at different temperatures is related to infection dynamics.

Although previous studies have highlighted the inflammatory responses of fish infected with parasites and exposed to pollutants, very little is known about how these two stressors interact within the fish. In this review, which also contains original data, the effect of these two parameters on the fish inflammatory response is assessed and, in particular, the role of apoptosis and the acute phase protein, C reactive protein, is evaluated. In Cyprinus carpio exposed to 0·5 mg NH4+ l−1 or 0·1 mg Cd2+ l−1 and experimentally infected with the blood fluke, Sanguinicola inermis, the pollutant type and the order in which the fish experiences the parasite and toxicant, significantly affects the ultrastructural appearance and cellular content of the pronephros and thymus. This is reflected in the intensity of infection where the pollutant appears to have less effect on an established infection. Both stressors, pollutant and infection, may mediate their effects via the endocrine system. Studies have revealed that cortisol at 100 ng ml−1 is able to induce apoptosis in pronephric cells of carp and that an increase in apoptosis is associated with an increase in phagocytosis in this immune organ. In addition, C reactive protein, which is used as a biomarker of the inflammatory response in humans and other mammals, is evaluated as a possible indicator of physiological states in fish exposed to pathogens and pollutants.

Regular observation of survival of the carp breeding lines constituting a living gene bank at the Institute of Ichthyobiology and Aquaculture in Golysz (Poland) over a period of at least 15 years showed different survival rates for various lines. In this study, we have examined the polymorphism of the major histocompatibility complex (MHC) gene class II B in nine carp lines. The class II B gene encodes for the part of the MHC class II molecule which presents peptides from pathogens and protein antigens that are present in the extracellular milieu and have been taken up into the endocytic vesicles of antigen-presenting cells. Polymerase chain reaction was used to amplify Cyca-DAB gene fragments comprising part of exon 1, complete intron 1 and almost complete exon 2. Exon 2 encodes for the β1 domain which is the most polymorphic fragment of MHC class II molecules. Single-strand conformational polymorphism (SSCP) was applied to detect different MHC class II B haplotypes. The analysis revealed the presence of seven different haplotypes occurring with various frequencies.

This study summarizes optimization of techniques for common carp artificial propagation including improvements of activation solution (AS), the process of insemination, and elimination of egg stickiness. The optimum gamete ratio for good fertilization and hatching rate ranged from 8490 to 23672 spermatozoa per egg, when dechlorinated tap water was used. Optimal ratio between eggs (weight in g) and AS (in ml) was defined as 1:1 to 1:2. Different concentrations of AS such as NaCl from 0 to 34 mM (0–68 mOsmol kg–1) did not change fertilization and hatching rates. An AS adopted for carp spermatozoa (45 mM NaCl, 5 mM KCl, 30 mM Tris–HCl, pH 8) was compared with other saline AS; only the 51 mM (102 mOsmol kg–1) NaCl solution decreased fertilization and hatching rate. The AS containing 20 mM Tris–HCl at pH 9 increased fertilization and hatching rates compared to dechlorinated tap water of pH 7 or to AS of pH 6 and 7. Adhesiveness from the eggs was successfully removed by incubation in Alcalase DX (PLN 04715) using two successive steps with different enzyme concentrations. The first step with an enzyme concentration of 2 ml l–1 was applied from 8 to 20 min after fertilization. Later in a second step, the best time for application of alcalase enzyme at a concentration of 20 ml l–1 was for 45 and 60 s at 20 min after fertilization leading to fertilization and hatching rates of 80–87%. The α-Chymotrypsin (EC 3.4.21.1. Merck) was also found effective for elimination of stickiness. Results withα-Chymotrypsin enzyme indicate that the response to success in elimination of stickiness is highly variable mainly due to differences in the environment, quality of water and carp strains.

Domesticated/captive stocks and wild/feral populations of common carp from Europe, Central Asia and East/South-East Asia were examined for allozyme (23 populations), microsatellite (11 populations) and mitochondrial DNA (21 populations) variation. Allozyme variability (1.06–1.81 alleles per locus, expected heterozygosity 0.006–0.136 at 16 loci) was much lower than microsatellite variability (2.5–14.0 alleles per locus, expected heterozygosity 0.426–0.887 at four loci). Differences in variability between domesticated/captive stocks and wild-caught ones were more pronounced at microsatellite loci than at allozyme loci, suggesting that microsatellites are better suited to detect population bottlenecks and loss of variation due to inbreeding. All but one European population were fixed for a single composite mtDNA haplotype, which also dominated in Central Asia but was completely missing in East/South-East Asia, indicating a single origin of European carp in Central Asia. All three classes of genetic markers clustered populations into two highly divergent groups: Europe/Central Asia and East/South-East Asia. Hierarchical partition of genetic diversity showed that for microsatellite loci most of variation was due to the within-population component while the highest proportion of mtDNA variation and substantial proportion of allozyme variation was accounted for by differences between geographical regions. Genetic data support the subspecies status of C. c. carpio assigned to the European carp and C. c. haematopterus assigned to the East/South-East Asian carp but do not justify a separate subspecies status (C. c. aralensis) for the Central Asian carp. As demonstrated for a wild/feral carp population from R. Danube, Germany, the genetic markers used in our study may be effectively applied to detect mixing and introgression of intra-species units in the presence of sufficient genetic differentiation.

The common carp is one of the main aquaculture species in the world. Despite this, most of the production is carried out using unselected strains. Selective breeding for fast growth has not proven to be effective in this species, but other traits (disease resistance, shape) could be successfully selected for. Most heritability estimates in the literature are unreliable due to environmental biases, but complementary results from population genetics and comparison of strains seem to indicate that there should be a potential for selective breeding in this species, including selection for growth rate, provided the base populations are variable enough (e.g. synthetic strains). New techniques such as parentage assignment with microsatellites and use of doubled haploid progenies may help describe much more accurately, without environmental bias, the genetic determination of traits of interest in the carp. This could be a new opportunity to design efficient breeding programs in this important species.

Immunological and structural changes in the thymus and pronephros of Cyprinus carpio infected with the blood fluke, Sanguinicola inermis for 30 days, and exposed to 0·5mg NH+4/l for 48 and 168h were investigated. Ultrastructural observations revealed cell disruption and highly vacuolated cytoplasm in the thymus. Of the cells that remained intact there was a significant increase in thrombocytes after 48h exposure to the pollutant. In addition, there was a decrease in lymphocytes following exposure to ammonia at both time-periods studied. In contrast the pronephros of fish exposed to the pollutant underwent relatively mild changes in cellular architecture although ammonia and time of exposure had significant effects on the proportions of several leucocyte types. A significant decrease in neutrophils, thrombocytes and lymphocytes occurred in fish exposed to the pollutant for 168h. Pronephric lymphocyte stimulation (cpm) by Con A and PWM increased in vitro, whereas the stimulation index was reduced in infected fish exposed to ammonia. Changes in the immune organs of S. inermis-infected carp treated with pollutant were both organ- and time-specific. The possible reasons for this are discussed and significance in relationship to parasitization assessed.

The effects of dietary medium-chain fatty acids (MCFA) on survival, growth and fatty acid composition of first-feeding carp larvae were investigated in two 21-day feeding trials. In trial 1, one diet CO was supplemented with 10% coconut oil, two diets, M1 and M2 with 1.7 or 4.2% interesterified medium-chain triacylglycerols (MCT), and two other diets T1 and T2 with 1% tricaprylin and 0.7% tricaprin, or 2.5% tricaprylin and 1.7% tricaprin. The five diets were made isolipidic (18% of dry diet) with triolein and compared to a control diet without MCFA. With a low dietary level (1% of dry diet) of caprylic acid (C8:0), final survival and mean weight of larvae fed CO, M1 and T1 were not significantly different from those of control (68 ± 6% and 62 ± 8 mg, respectively). In contrast, with a high dietary C8:0 level (2.5% of dry diet), final survival and mean weight of larvae fed M2 and T2 were markedly decreased (23 ± 2% and 40 ± 8 mg, respectively), without effect of the form of supply. In trial 2, larvae were fed six diets with graded tricaprylin contents (from 0 to 10% in diet). Survival was not significantly affected but growth was decreased by C8:0 levels higher than 2% of dry diet. Fatty acid composition of larval total lipids revealed high levels of lauric and myristic acids in larvae fed coconut oil. Deposition of C8:0 and capric acid (C10:0) was low after MCT feeding, but depended of the form of supply, being higher with pure than with interesterified MCT. Relatively high amounts of C8:0 and C10:0 were recorded in total lipids of larvae fed 10% tricaprylin. It is concluded that high levels of caprylic acid decrease growth but that low levels are well utilised by carp larvae, irrespective of the form of supply.

The humoral antibody response and the number of pronephric antibody-secreting cells were examined in naturally Bothriocephalus acheilognathi-infected carp. Cyprinus carpio, and in those injected intraperitoneally with an extract of the cestode. In the extract-injected fish, specific antibody was detected 3 weeks after a second injection given 2 weeks after the primary injection, and antibody levels persisted for more than 200 days. A third injection also enhanced the antibody level in the extract-injected carp. The numbers of antibody-secreting cells were significantly higher in carp injected 3 times with the extract than in the control. In naturally-infected fish, the serum antibody levels and the number of pronephric antibody-secreting cells were higher in infected fish than in uninfected individuals although this difference was not statistically significant. The relevance of these results to immune protection against infection is discussed.

The technique already adopted by the authors for the cryopreservation of rainbow trout blastomeres was applied to carp. Three stages were utilised for cell freezing: the morula stage, the early blastula stage and the late blastula stage. Survival rates were 89 ± 1.6 % for morula cells, 94 ± 0.6 % for early blastula cells and 96 ± 0.4 % for late blastula cells. Early blastula and late blastula cells reaggregated into morula-like bodies within 2 h in culture medium. Results suggest that the freezing technique adopted for rainbow trout may be successfully applied to carp. The morula stage blastomeres were more sensitive to freezing than blastula stage blastomeres, as already found in rainbow trout.

We propose a simple method to quantify the morphology of 2-3-year-old carps (Cyprinus carpio) which is related to their fillet yield. This method is based on the automated image analysis of the lateral projection of the fish (mask). Seven morphological measurements specifically related to fillet yield of carps were defined. The comparison between this method and the conventional quantification of fish shape, based on measurements of skeletal landmarks, increases the significance of this fitting. From the analysis of the morphometric data, we propose a profitable carp morphotype which is defined by four angles and two distances which quantify the shape of the head and the position of the caudal peduncle, respectively. The main characteristic of this morphotype is the absence of a large dorsal development, and the ventral position of the caudal peduncle.

Apparent digestibility coefficients (ADCs) of macro- and micronutrients and energy of two diets were measured with common carp Cyprinus carpio reared at two water temperatures (18 and 25 °C). Two diets containing either soy protein concentrate (SPC) or Norwegian herring meal (FM) as a major protein source were formulated. Dicalcium phosphate was incorporated at a level of 4 % of the former diet. Duplicate groups of 20 fish (body weight: 55 g) were fed by hand twice a day to apparent satiety. After a 7-day conditioning period, faecal collection was made over 10 days using an automatic faeces collector. ADC of dry matter was not different (P > 0.05) between fish fed the two diets at 18 °C, but at 25 °C, we observed a higher ADC value in fish fed the FM diet than in those fed the SPC diet. ADC of protein was higher in fish fed the SPC diet than in the FM groups at either temperature. ADC of lipids was not different between the two groups at 25 °C but was found to be higher in fish fed the SPC diet than in the FM group at 18 °C. ADCs of energy and ash of the two fish groups remained constant at both water temperatures. Availability of phosphorus was significantly higher (P < 0.05) from fish meal based diets than from SPC based diets at both temperatures. A more striking effect of dietary protein source was found in the availability of calcium, which was lower in carp fed SCP based diets.

Nicotine injected intravenously or on pituitary in vivo stimulated maturational gonadotropin (GtH2) secretion in domperidone (D-2 receptor antagonist) pretreated carps. The effective dose was 1 mg·kg−1 and 100 μg·kg−1, respectively. No stimulatory effects of nicotine were observed in normal (non-treated) carps. When nicotine was injected together with gonadotropin releasing hormone (GnRH) analogs (D-Arg6 sGnRH or D-Ala6 LHRH) a clear potentiation effect of nicotine on GnRH analogs' action was observed. This potentiation was seen even in normal fish where nicotine itself had no effect on GtH2 release. A synergism between nicotine and GnRH analogs appeared when low doses of GnRH analogs were used. In domperidone pretreated fish, this synergism occurred after injection of 1 μg·kg−1 of sGnRHa. In normal fish, the same phenomenon was observed after injection of 5 μg·kg−1 of sGnRHa or 20 μg·kg−1 of LHRHa. Higher doses of GnRH analogs mask this effect. Results confirm previous in vitro findings on the stimulatory action of nicotine on GtH2 secretion. They also indicate a possible synergy between nicotine and GnRH as well as an inhibitory influence of dopamine on nicotine's action.

Scanning electron microscopic observation of embryos and larvae of the carp (Cyprinus carpio) in the course of development was undertaken to further understanding of the relationship between larval behaviour and their chemo- and mechanoreceptors. Free neuromasts and olfactory pits equipped with mechano- and sensory-cilia were found on carp embryos. During larval development, the number of neuromasts and the olfactory mechano- and sensory cilia steadily increases and they become more morphologically differentiated. The olfactory features, presented in early larval stages, indicate that although the olfactory organ at this early life history has not yet attained its final configuration, it can perceive odour stimulation. The neuromasts found on the forehead of the newly hatched larvae seem to assist in the early behavioural selection, approach and attachment to a substratum, before the stage of filling of the gas bladder. The increasing number of neuromasts during development reflects the larvae's capability of capturing food items in the absence of visual stimulation. The rapid maturation of free neuromasts, the lateral-line system and the olfactory organ, is evidence of their post-metamorphosis position in the search for, location and successful capture of food.