In situ growth and photosynthetic activity of Cyanobacteria and phytoplankton dynamics after passage through the gut of silver carp (Hypophthalmichthys molitrix), bighead carp (Aristichthys nobilis), and Nile tilapia (Oreochromis niloticus)

The growth and photosynthetic activities of Cyanobacteria passed through the gut of silver carp (Hypophthalmichthys molitrix), bighead carp (Aristichthys nobilis), and tilapia (Oreochromis niloticus) were compared with those of phytoplankton taken directly from Lake Taihu during a 13-day in situ dialysis culture. After the first 3–5 days of reduced activity after excretion by silver carp and bighead carp, the photosynthetic activity of Cyanobacteria recovered and rose significantly higher (P < 0.01) than levels in the control population, whereas there was a notable reduction of photosynthetic activity after passage through tilapia gut. The phytoplankton biomass showed a 2- to 3-fold increase of growth, and extracellular polysaccharide production was also stimulated after passage through silver carp and bighead carp gut. Chlorophyta fluorescence was detected at much higher levels than that of Cyanobacteria and Bacillariophyta after passage through tilapia gut. Scenedesmus obliqnus and Chlamydomonas sp. contributed much to the growth of the Chlorophyta during the in situ cultivation. However, the total phytoplankton biomass showed a distinct reduction in the tilapia treatment during the culture. The study indicated that Nile tilapia feeding and defecation may help remove Cyanobacteria from the water column and favor a community shift to Chlorophyta.     

Comparative analysis of gastrin-immunoreactive and argyrophil cells in bony fish larvae

A comparative study was made of enteroendocrine cells in the larvae of bony fishes (Teleostei): carp (Cyprinus carpio L.), big head (Aristichyts nobilis Richardson), silver carp (Hypophthalmichthys molitrix Valenciennes), and atlantic salmon (salmo salar L.). Argyrophil cells demonstrated by the method after Grimelius were found in all examined fish species. They are located in the pyloric appendices, stomach and intestinal mucosa. Gastrin-immunoreactive cells demonstrated by the immunocytochemical method were located in the pyloric part of the stomach in all examined fishes, except for the atlantic salmon in which they were not identified. The first argyrophil cells were identified at the age of about 10 days, while gastrin-immunoreactive cells proved to appear 5-7 days later.     

Relationship of length of uterus in prepubertal pigs and number of corpora lutea and fetuses at 30 days of gestation

Relationships of the length of the uterus at one reproductive stage to the length at other stages and the effect on potential litter size were determined. In Experiment 1, the length of the uterus was measured in situ at 20, 60, or 100 days of age at laparotomy with 20 gilts in each of the three age groups. Forty days after the initial measurement, the uterus was again measured in situ, gilts were ovariectomized and hysterectomized, and associations among uterine measurements at the two different stages were determined. Correlations between uterine length in situ and between uterine length and weight 40 days later were all greater than 0.75 (P < 0.001). The length of one uterine horn increased from 13.9 cm at 20 days of age to 36.7 cm at 140 days of age (P < 0.01). In Experiment 2, 66 gilts were unilaterally hysterectomized and ovariectomized (UHOX) at 150 days of age and the ovary was weighed. Length of the one horn was measured in 36 gilts. At 10 days after first estrus, the length of the remaining uterine horn was measured at laparotomy and corpora lutea were counted in 53 gilts. In 15 of the 53 gilts the remaining uterine horn was removed to obtain uterine weights. At the second or third estrus, 38 gilts were mated and at Day 30 of gestation, 31 gilts were pregnant. The gilts were killed, and the length of the uterus measured and corpora lutea (CL) and fetuses were counted. The length of one uterine horn at 150 days of age was 70 cm with a range of 47–110 cm. At 10 days after first estrus, length had increased to a mean of 141 cm with a range of 86–194 cm and at 30 days of gestation the mean was 244 cm with a range of 186–311 cm. There were 12.2 CL at first estrus, which was not different from 12.4 CL at the second or third estrus. The mean number of fetuses in one horn at 30 days of gestation was 9.5 with 77% prenatal survival. Length of uterine horn at 150 days of age was correlated with uterine horn length (r = 0.56, P < 0.001) at 10 days after first estrus and number of live fetuses (r=0.39, P<0.05) at 30 days of gestation. At Day 10 after first estrus, uterine length was not correlated with the number of CL, whereas at Day 30 of gestation, the number of CL and uterine length were correlated with the number of live fetuses in those gilts with below the mean number of live fetuses, but not in those gilts with above the mean of live fetuses. The number of live fetuses (r=0.66, P < 0.001) and fetal survival (r=0.63; P < 0.001) were correlated with uterine horn length in pregnant UHOX gilts. Length of the prepubertal uterus gives an indication of postpubertal length and the potential litter size in pigs.     

Resistance of common carp (Cyprinus carpio L.) to Cyprinid herpesvirus-3 is influenced by major histocompatibility (MH) class II B gene polymorphism

The role of MH class II B (Cyca-DAB1-like) genes in resistance of common carp (Cyprinus carpio L.) to Cyprinid herpesvirus-3 (CyHV-3), also known as koi herpesvirus (KHV) was analysed. The material consisted of 934 fish from six carp crosses. Fish were challenged with CyHV-3 at an age of 7 and 10 months. During challenge experiments the peak of mortality caused by CyHV-3 was observed at days 8–12 p.i. and the overall cumulative mortality reached 79.9%. Among six Cyca-DAB1-like genotypes, revealed by PCR-RF-SSCP analysis, one genotype (E) was found associated with higher resistance to CyHV-3. Three other genotypes (B, H and J) could be linked to higher susceptibility to CyHV-3. Analysis of the alleles that compose the Cyca-DAB1-like genotypes linked one particular allele (Cyca-DAB1*05) to significantly increased, and two alleles (Cyca-DAB1*02 and Cyca-DAB1*06) to significantly decreased resistance to CyHV-3. Our data indicate that MH class II B genes could be used as potential genetic markers in breeding of common carp for resistance to this virus.     

Hormonal and ion regulatory response in three freshwater fish species following waterborne copper exposure

We evaluated effects of sublethal copper exposure in 3 different freshwater fish: rainbow trout (Oncorhynchus mykiss), common carp (Cyprinus carpio) and gibel carp (Carassius auratus gibelio). In a first experiment we exposed these fishes to an equally toxic Cu dose, a Cu level 10 times lower than their 96 h LC₅₀ value: 20, 65, and 150 µg/L Cu. In a second series we exposed them to the same Cu concentration (50 µg/L). Na⁺/K⁺-ATPase activity in gill tissue was disturbed differently in rainbow trout then in common and gibel carp. Rainbow trout showed a thorough disruption of plasma ion levels at the beginning of both exposures, whereas common carp and gibel carp displayed effects only after 3 days. Rainbow trout and common carp thyroid hormones experienced adverse effects in the beginning of the exposure. The involvement of prolactin in handling metal stress was reflected in changes of mRNA prolactin receptor concentrations in gill tissue, with an up regulation of this mRNA in rainbow trout and a down regulation in gibel carp, which was more pronounced in the latter. Overall, rainbow trout appeared more sensitive in the beginning of the exposure, however, when it overcame this first challenge, it handled copper exposure in a better manner then common and gibel carp as they showed more long term impacts of Cu exposure.     

Antigenic analysis of grass carp reovirus using single-chain variable fragment antibody against IgM from Ctenopharyngodon idella

Grass carp (Ctenopharyngodon idella) is an important species of freshwater aquaculture fish in China. However, grass carp reovirus (GCRV) can cause fatal hemorrhagic disease in yearling populations. Until now, a strategy to define the antigenic capacity of the virus’s structural proteins for preparing an effective vaccine has not been available. In this study, some single-chain variable fragment antibodies (scFv), which could specifically recognize grass carp IgM, were selected from a constructed mouse naïve antibody phage display cDNA library. The identified scFv C1B3 clone was shown to possess relatively higher specific binding activity to grass carp IgM. Furthermore, ELISA analysis indicated that the IgM level in serum from virus-infected grass carp was more than two times higher than that of the control group at 5–7 days post infection. Moreover, Western blot analysis demonstrated that the outer capsid protein VP7 has a specific immuno-binding-reaction with the serum IgM from virus-infected grass carp. Our results suggest that VP7 can induce a stronger immune response in grass carp than the other GCRV structural proteins, which implies that VP7 protein could be used as a preferred immunogen for vaccine design.     

Chemical Composition of the Low-Polar Fraction of the <Emphasis Type="Italic">Copitarsia uncilata</Emphasis> Burgos &amp; Leiva (Lepidoptera: Noctuidae) Eversible Pheromone Gland

In Colombia, Copitarsia uncilata Burgos & Leiva has been reported as a pest of ornamental flowers. The leafworm causes serious economic losses to farmers because the shipments are intercepted at the detection sites. The identification and subsequent synthesis of the sex pheromone compounds can lead to the use of semiochemicals as an important strategy to complement the pest management. This paper identifies and characterizes the chemical components of the female sexual gland of C. uncilata at three different age stages in order to relate the effect of age on the gland extract composition. Eggs and larvae were collected from commercial crops. The specimens were reared and sexed. Pheromone glands of intact females were excised at (a) 1–2, (b) 3–4, and (c) 5–6 days old. The extracts were analyzed using GC-MS after two extraction time points: 20 min and 8 days. The extracts from the third age stage showed an increase in concentration for 35% of the compounds after 8 days of storage. The relative abundance as a function of the female age showed two trends: an upward trend, where the relative abundances of the compounds increased through the age stages, while some compounds shown a downward trend. Within the first trend, mostly of the compounds were found to be related with infochemicals type structures, where tetradec-9-en-1-yl acetate (within several C-14 long-chain alcohols) is the main one. The 9–14:Ac quadrupled concentration through the ages is possibly related to the increased length of calling period as female moth age.     

Development and ageing of the salivary glands of adult male Aedes aegypti (L.) and Aedes togoi (theobald) mosquitoes (Dipteria : Culcidae)

The salivary glands of adult male Aedes aegypti and Aedes togoi (Diptera : Culicidae), varying in age from less than 1 day after emergence to 42 days or 33 days respectively, were examined by light microscopy. Following emergence, the trilobed glands rapidly accumulate secretory product and attain their full size within about 48 hr. The amount of secretion in fully developed glands shows marked individual variation, but it is predominantly located in the posterior parts of the glands. Ageing changes begin to appear after about 8 days (A. aegypti) or 10–11 days (A. togoi), and although there is wide individual variation in their extent, in general, they become progressively more severe until at the age of 4–5 weeks all mosquitoes are substantially affected. Since other tissues, including ganglia, also regress with age, it is unlikely that salivary gland changes are the prime cause of death.     

Organization of the olfactory system of the Indian major carp <Emphasis Type="Italic">Labeo rohita</Emphasis> (Ham.): A scanning and transmission electron microscopy study

Catla catla, Labeo rohita, and Cirrhinus mrigala represent important alimentary fish in India. Their reproduction/breeding depends on seasons. Fish perceive external factors-stimuli and chemical signals through the olfactory system that plays the key role in central regulation of reproduction. However, no electron microscopy data are available on organization of olfactory components of these fish. We studied organization of the olfactory organ in male L. rohita using scanning (SEM) and transmission electron microscopy (TEM). This organ consists of olfactory epithelium, a short nerve, and olfactory bulb. The olfactory organ is ovoid in shape and consists of about 47–52 lamellae in adults and about 14–20 lamellae in fingerlings. These lamellae originate from the midline raphe. By SEM, microvillar sensory and ciliated non-sensory cells were observed in the lamellae. TEM revealed microvillar receptor cell with rough endoplasmic reticulum and Golgi apparatus towards apical end. Basal cells were present at the base of receptor cell, supporting cells were located adjacent to the olfactory receptor neurons, while epithelial cells—in the nonsensory part of olfactory epithelium. Mast, blastema, and macrophage cells were also found at the basement membrane. This work is the first publication on ultrastructural organization of the olfactory system of the Indian major carp, which provides information about morphological and ultrastructural organization of the olfactory system and opens new avenues for further investigation of chemical neuroanatomy, sensory signal processing, and neural regulation of reproduction in the Indian major carp.     

Die Nachzucht des Roten Felsenhahns,Rupicola peruviana (Latham, 1790) und des Schildschmuckvogels,Querula purpurata (Müller, 1776) im Zoologischen Garten Wuppertal

Incubation, handrearing and motherrearing of 5 male Andean Cocks of the Rock (Rupicola peruviana) in Wuppertal Zoo is described, in particular the development of the juvenile plumage to adulthood, and the change of colour of the beak and the iris of the eye. Incubation lasted 26–28 days. The mother-reared nestling fledged after 28 days. The young male Cock of the Rock attained full adult plumage at the age of 18 months, not at 3 years as Del Hoyo et al. (2004) describe from free-living Cocks of the Rock. A report on the supposed world's first breeding of the Purple-throated Fruitcrow (Querula purpurata) is given. The incubation period was 25 days.     

Temporal changes in body mass, body composition and metabolism of gibel carp Carassius auratus gibelio during food deprivation

To investigate the response to starvation, gibel carp Carassius auratus gibelio [12.5 ± 0.03 g (mean ± SE, n = 24)] were deprived of food at 25.8 ± 0.2°C (mean ± SE, n = 56) for 56 days. Body mass, proximate composition in whole body and muscle, and respiration were measured at 7‐day intervals. Body mass decreased with prolongation of deprivation, with a significant decline recorded after 7 days deprivation. Fish lost 22% of their fresh mass and 34% of dry mass after 56 days. Fish lost 38% of the body lipid over the first 7 days, and lost body lipid at a rate of 0–11% per week over the remaining 49 days. Body protein was lost at 1–5% per week throughout deprivation. Compared with the initial composition, body lipid concentration was lower and ash concentration higher on day 7. Water as a percentage of body mass was higher after 28 days, and protein concentration lower after 42 days, than at the start of deprivation. Muscle lipid and protein concentration was lower, and % water higher, after 7 days than at the start of deprivation, whereas muscle ash concentration was relatively constant during deprivation. After 56 days, fish lost body water by 18%, body lipid by 84%, body protein by 30%, and body energy by 45%. Oxygen consumption rate dropped from day 1 to day 3, increased from day 4 to day 14, gradually decreased from day 15 to day 35, and maintained a relatively constant level from day 36 to day 56. Results of the present experiment reveal that gibel carp utilize body lipid as a major energy source in the first 7 days of food deprivation, then turn to body protein as an energy fuel when lipid reserves are heavily depleted. Oxygen consumption is maintained at a relatively low and constant level when most lipid reserves are exhausted.     

A Set of Polymorphic Trinucleotide and Tetranucleotide Microsatellite Markers for Silver Crucian Carp (Carassius auratus gibelio) and Cross-Amplification in Crucian Carp

Silver crucian carp (Carassius auratus gibelio Bloch), as a gynogenetic fish, is a promising model for the study of the evolutionary genetics of vertebrates. We have developed 59 polymorphic trinucleotide and tetranucleotide markers for the silver crucian carp through the biotin capture method and radioactive-labeling hybridization. The number of alleles ranged from 2 to 12 in the population, and the average proportion of heterozygotes (including tri- and diallelic) at polymorphic loci was 76.8%. In addition, these loci were successfully applied to a close relative, the crucian carp (Carassius auratus), by cross-amplification, as shown by the range of alleles (2–19), observed heterozygosity (0.1765–0.9706), expected heterozygosity (0.2392–0.9421), and polymorphism information content (0.2186–0.9236).     

Contrasting Patterns of Larval Mortality in Two Sympatric Riverine Fish Species: A Test of the Critical Period Hypothesis

Understanding the causal mechanisms that determine recruitment success is critical to the effective conservation of wild fish populations. Although recruitment strength is likely determined during early life when mortality is greatest, few studies have documented age-specific mortality rates for fish during this period. We investigated age-specific mortality of individual cohorts of two species of riverine fish from yolksac larvae to juveniles, assaying for the presence of a “critical period”: A time when mortality is unusually high. Early life stages of carp gudgeons (Hypseleotris spp.) and unspecked hardyhead (Craterocephalus stercusmuscarum fulvus)—two fishes that differ in fecundity, egg size and overlap between endogenous and exogenous feeding—were collected every second day for four months. We fitted survivorship curves to 22 carp gudgeon and 15 unspecked hardyhead four-day cohorts and tested several mortality functions. Mortality rates declined with age for carp gudgeon, with mean instantaneous mortality rates (-Z) ranging from 1.40–0.03. In contrast, mortality rates for unspecked hardyhead were constant across the larval period, with a mean -Z of 0.15. There was strong evidence of a critical period for carp gudgeon larvae from hatch until 6 days old, and no evidence of a critical period for unspecked hardyhead. Total larval mortality for carp gudgeon and unspecked hardyhead up to 24 days of age was estimated to be 97.8 and 94.3%, respectively. We hypothesise that life history strategy may play an important role in shaping overall mortality and the pattern of mortality during early life in these two fishes.     

Morphology and molecular characterization of Acanthogyrus (Acanthosentis) bilaspurensis Chowhan,Gupta & Khera, 1987 (Acanthocephala: Gyracanthocephala: Quadrigyridae) from the common carp Cyprinus carpio Linnaeus (Cypriniformes: Cyprinidae) in Pakistan

Acanthogyrus (Acanthosentis) bilaspurensis Chowhan, Gupta & Khera, 1987 is a poorly known acanthocephalan species reported from the reba carp Cirrhinus reba (Hamilton) (Cypriniformes: Cyprinidae) in India. In the present study, the detailed morphology of A. (Acanthosentis) bilaspurensis were studied using light microscopy and, for the first time, scanning electron microscopy (SEM), based on newly collected specimens from the common carp Cyprinus carpio Linnaeus (Cypriniformes: Cyprinidae) in Pakistan. The SEM observations revealed the outer shell of eggs with numerous remarkable protuberances and the gonopore of female located at a discoid protrusion surrounded by some very small spines. This is the first time that A. (Acanthosentis) bilaspurensis was reported in Pakistan, and the common carp represents a new host for this species. In addition, the molecular characterization of the 18S and 28S rDNA of A. (Acanthosentis) bilaspurensis is provided for the first time. There was no intraspecific genetic variation detected in the 18S and 28S regions between different individuals of A. (Acanthosentis) bilaspurensis, but high level of interspecific nucleotide divergence was found in the 18S (4.15–16.7%) and 28S regions (3.60–15.4%). Phylogenetic results based on the 18S, 28S and 18S + 28S sequence data, respectively, all revealed the Quadrigyridae, Pallisentinae and the genus Neoechinorhynchus are not monophyletic groups, and the genera Acanthogyrus and Pallisentis have far relationship. The systematic status of the genus Acanthogyrus and some species of Neoechinorhynchus need further clarify based on different mtDNA and nuclear DNA data including broader representatives of the class Eoacanthocephala.     

Migration of thymus cells to the developing gut-associated lymphoid tissues of the young rabbit

Thymus cell migration to the gut-associated lymphoid tissues (GALT) as compared to other lymphoid tissues in young rabbits was determined following in vivo intrathymic inoculation of tritiated thymidine. The GALT received as many or more thymus cells than the spleen or lymph nodes during the first few postnatal days. Migration to the GALT and nonGALT decreased with age, and seeding appeared to be essentially complete by 30–40 days.     

Effects of three strains of intestinal autochthonous bacteria and their extracellular products on the immune response and disease resistance of common carp,Cyprinus carpio

The study isolated three strains of intestinal autochthonous bacteria Aeromonas veronii BA-1, Vibrio lentus BA-2, and Flavobacterium sasangense BA-3 from the intestinal tract of the common carp (Cyprinus carpio). To reveal the effects of these three strains of bacteria on the innate immunity of carp, the lysozyme, complement C3, total serum protein, albumin and globulin levels, respiratory burst activity, phagocytic activity by blood leucocytes and the expression of IL-1b, lysozyme-C, and TNF-α were examined after feeding with seven different diets for up to 28 days. Also the survival of carp against Aeromonas hydrophila was challenged for 14 days. The carp were fed seven different diets: one control, three diets supplemented with 1 × 10⁸ cell g⁻¹ of carp intestinal bacteria BA-1 (Group D-I), BA-2 (Group D-II) and BA-3 (Group D-III), and three diets supplemented with extracellular products FA-1 (Group E-I), FA-2 (Group E-II) and FA-3 (Group E-III) which were corresponding to the strains BA-1, BA-2, and BA-3, respectively, up to 28 days. For groups D-I, D-III, E-I and E-III, the innate immune parameters of carp were significantly increased, the expression of three immune-related genes in blood was significantly up-regulated examined during 7, 14, and 21 days of feeding, and the survival rate was improved. The study indicates that the two isolated intestinal autochthonous bacteria A. veronii BA-1 and F. sasangense BA-3 could positively influence immune response and enhance disease resistance of carp against A. hydrophila infection.     

Effect of luteinizing hormone/choriogonadotropin receptor (LHCGR) gene on chicken reproductive traits

Luteinizing hormone/choriogonadotropin receptor (LHCGR) gene, potentially related to reproductive traits in chickens, was genotyped by using the Pooled DNA Sequencing, PCR–SSCP and Directing Sequencing techniques. 306 Erlang Mountain chickens form one line (SD03, a line that has been selected for egg quality from a local chicken breed in Sichuan province, China) were genotyped in this study. The associations between LHCGR polymorphisms and six reproductive traits [body weight at first egg (BWAFE), weight of first egg, age at first egg (AFE), number of eggs at 300 days of age (EN), body weight at 300 days of age and egg weight at 300 days of age (EWTA)] were estimated using the one-way analysis of variance method. Results showed that SNP +G4058A and SNP +T4099G of the LHCGR gene were significantly associated with BWFE and AFE. Birds with the AG genotype for the +G4058A SNP exhibited shorter AFE (P < 0.05) and greater EN than those of the GG and AA genotypes, suggesting a balancing selection (overdominance); the effect of allele C in SNP +C3021T and allele C in SNP +T4490C on EN and AFE is additive and may reflect the influence of positive selection. These alleles have promise as genetic markers for future marker-assisted selection.     

Larval development,growth and age determination in the sharpnose pufferfishCanthigaster valentini (Teleostei: Tetraodontidae)

The eggs, early larvae and juveniles of the sharpnose pufferfishCanthigaster valentini are described, based on material collected in Great Barrier Reef waters. Eggs were obtained in the field by divers and reared in the laboratory. The eggs are spherical, strongly adhesive, 0.68–0.72 mm in diameter, possess a dense cluster of small oil droplets, and hatch around sunset 3 to 5 days after fertilization. Newly hatched larvae have a small yolk sac, pectoral fin folds, 17 myomeres (6 pre-anal, 11 post-anal) and measure 1.30–1.40 mm in notochord (standard) length. The eggs ofC. valentini differ from those of other tetraodontids in being much smaller and having a longer incubation time. The larvae can be distinguished from other tetraodontid larvae by pigmentation, myomere count and size at hatching. Growth is most rapid during the first day of larval life. Age determinations (based on otolith microstructure) of field collected juveniles, both pelagic and newly settled, indicate a pelagic phase of between 64 and 113 days for this species. This estimate appears consistent with the extended pelagic juvenile stages observed in other tetraodontiform fishes and could indicate thatC. valentini can delay settlement for some time after becoming competent to settle at a minimum age of 64 days.     

Food Provisioning in Red-Necked Grebes (Podiceps Grisegena) at Common Carp (Cyprinus Carpio) Ponds

Parental feeding patterns were studied in red-necked grebe (Podiceps grisegena) broods throughout the entire period of parental care in a common carp (Cyprinus carpio) fish-pond area in SE Poland in 1993–2002. Fish formed a substantial part of prey provided to the flightless young from their second week of life. Although the numbers of large invertebrates and tadpoles, the alternative prey to fish, did not decrease during the chick rearing period, grebe parents gradually shifted from delivering predominately invertebrates to delivering fish, and the average size of fish fed to chicks increased with brood age. Broods with relatively high fledging success (at least two chicks fledged) had a larger proportion of fish in their diet than broods seriously reduced because of undernourishment. The dive duration of foraging grebe parents did not differ between carp, wild fish and non-fish prey, but carp prey required significantly more time for handling. The percentage of prey rejected by chicks increased over the prefledging period from 2 to 24%. Of the prey rejected, 82% were fish apparently too large for the young to swallow. Fish prevalence in the diet of red-necked grebe chicks at carp ponds contradicts the results of other studies on the feeding habits of the nominative subspecies during breeding season. However, the red-necked grebe is a gape-limited predator and the piscivory of the chicks is limited to small-bodied fish.