Treatment with an aromatase inhibitor induces complete sex change in the protogynous honeycomb grouper (Epinephelus merra)

To study the mechanism of protogynous sex change in honeycomb grouper, the wild fish of both sexes and transitionals were collected for one year, and changes in gonadal structures and serum levels of sex hormones in each individual were examined. The onset of sex change was associated with low serum estradiol - 17β (E2) levels. In order to clarify whether E2 deprivation causes sex change, maturing females were implanted with the non-steroidal aromatase inhibitor (Fadrozole, AI). AI caused complete sex change from female to functional males in two and half months. The present results suggest that E2 is an important endogenous factor which regulates protogynous sex change in the honeycomb grouper.     

Influence of high dietary α-tocopherol intakes on specific immune response, nonspecific resistance factors and disease resistance of healthy and aflatoxin B1-induced immunocompromised Indian major carp, Labeo rohita (Hamilton)

The effect of aflatoxin treatment and/or feeding of a high level of α‐tocopherol on immune response and disease resistance was investigated in Indian major carp, Labeo rohita. Group A served as a healthy control, group B was treated with aflatoxin, group C was fed a high level of α‐tocopherol whereas group D was exposed both to aflatoxin and a high level of dietary α‐tocopherol for 60 days. Aflatoxin B₁ (AFB₁) was injected once intraperitoneally into fish on the first day of the experiment (groups B & D). High levels of DL‐α‐tocopherol (1000 mg kg^–1 feed) were provided to healthy as well as AFB₁‐treated immunocompromised fish for 60 days (groups C & D). At the end of the experiment blood samples were assayed for changes in nonspecific immunity and humoral protein levels. Disease resistance against two common bacterial pathogens viz., Aeromonas hydrophila and Edwardsiella tarda were evaluated in all groups. Significant (P < 0.05) suppression of specific immunity as measured through haemagglutination (HA) titre against sheep red blood cells (SRBCs) as well as bacterial (formalin‐killed E. tarda) agglutination titre; nonspecific resistance factors viz., globulin level, serum bactericidal and lysozyme activities, neutrophil activities, and disease resistance against two bacterial pathogens only in aflatoxin‐treated fish with respect to the control group, clearly indicated the immunosuppressive nature of aflatoxin. Feeding of a high level of α‐tocopherol to AFB₁‐treated immunocompromised fish significantly (P < 0.05) raised specific immunity, nonspecific resistance factors and disease resistance capacity when compared with aflatoxin‐exposed fish. Disease resistance and enhancement of immune status through feeding of high levels of α‐tocopherol to healthy as well as AFB₁‐treated immunocompromised fish confirmed the potential of α‐tocopherol in carp feed for prevention of disease and for combating natural/environmental immunosuppressants.     

Digestibility, faeces recovery, and related carbon, nitrogen and phosphorus balances of five feed ingredients evaluated as fishmeal alternatives in Nile tilapia, Oreochromis niloticus L.

This study shows that alternatives for fishmeal in a fish diet affect not only fish growth but also faeces stability and nitrogen (N) and phosphorus (P) waste production. Wheat gluten diet (WGD), soybean meal extract diet (SBE), soybean meal diet (SBM), duckweed diet (DWD) and single‐cell protein diet (SCP) were evaluated as a fishmeal replacement on a 15% weight weight^?1 basis in tilapia diets. Fishmeal replacement affected dry matter (dm), protein, ash and P digestibility significantly. Faeces recovery (6.8–11.2%) was not significantly affected, although the amount of non‐recovered faeces and total faeces showed significant differences. Duckweed diet and SCP resulted in the largest amounts of non‐recovered and total faeces (199–210, 224–225 g dm kg^?1 feed dm). Compared with fishmeal diet (FMD), the WGD and SBE resulted in similar growth, but higher non‐faecal N losses (471–495 vs. 416 g N kg^?1 N). Soybean meal diet, DWD and SCP resulted in lower growth but less non‐faecal loss (409–450 g N kg^?1 N). The DWD and FMD had the highest N retention (480 g N kg^?1 N) compared with the other diets (431–451 g N kg^?1 N). Carbon retention, faecal and non‐faecal losses and P retention were similar for all diets (302–358, 142–176 and 489–523g C kg^?1 C, 606–704 g P kg^?1 P). Phosphorus faecal loss was lower for all diets (329–381 g P kg^?1 P) than for the FMD (401 g P kg^?1 P).     

The maturation of the salmon GnRH system and its regulation by gonadal steroids in masu salmon

Pituitary gonadotropin (GTH) secreting cells and brain gonadotropin-releasing hormone (GnRH) secreting neurons are known to be subjected to feedback control by gonadal steroid in teleosts. In masu salmon, Oncorhynchus masou, salmon GnRH (sGnRH) neurons in the ventral telencephalon (VT) and the preoptic area (POA) are involved in the control of GTH cells because sGnRH synthesis in these areas is activated with gonadal maturation. In this study, we attempted to clarify mechanisms of feedback control of sGnRH neurons by gonadal steroids. We examined the effects of 17-methyltestosterone (MT) on sGnRH synthesis in yearling and 2-year-old female fish (which were immature during experimentation in May), and the effects of castration on sGnRH synthesis in underyearling precocious male fish in August. sGnRH synthesis in the POA, but not in the VT, was increased by MT administration in 2-year-old females only, indicating higher sensitivity to MT in the preoptic sGnRH neurons. Castration increased sGnRH synthesis in the VT but not in the POA. These results suggest that sGnRH neurons in the VT and those in the POA are differentially regulated by gonadal steroids.     

Light and electron microscopic evidence of white spot disease in the giant tiger shrimp, Penaeus monodon (Fabricius), and the kuruma shrimp, Penaeus japonicus (Bate), cultured in Taiwan

Since 1992, mass mortalities among cultured giant tiger shrimp, Penaeus monodon (Fabricius), and kuruma shrimp, Penaeus japonicus (Bate), have been observed in Taiwan. The condition is known as 'white spot disease' (WSD), based on the characteristic white spots on the cuticle of diseased shrimp. With the scanning electron microscope, two sizes of white spots were observed. Each spot represented a protrusion on the inside surface of the carapace. The composition of white spots was similar to that of the cuticule, most calcium, as determined with an energy dispersive spectrometer. Histological studies of moribund, infected specimens revealed degenerated cells, characterized by hypertrophied nuclei, in various meso- and ectodermal tissues. Infected tissues included cuticular epidermis, connective tissue, lymphoid organ, antennal gland, and haematopoietic, gill and nervous tissue. Nuclei were Feulgen-positive and no occlusion body was found in the necrotic tissue. Transmission electron microscopy revealed the presence of rod-shaped and enveloped virions in the hypertrophied nuclei. The virions measured 298 ± 21 × 107 ± 8 nm in the giant tiger shrimp and 248 ± 12 × 104 ± 8 nm in the kuruma shrimp. In an experimental infection trial, cumulative mortality was 40% within 14 days under stress conditions. No mortality was observed in controls or in non-stressed infected shrimp. Experimental infections show that environmental stressors such as ammonia may enhance the severity of WSD virus infections in cultured shrimp.     

Establishment of a new cell line susceptible to Cyprinid herpesvirus 3 (CyHV‐3) and possible latency of CyHV‐3 by temperature shift in the cells

A new cell line named CCF‐K104 predominantly consisting of fibroblastic cells showed optimal growth at temperatures from 25 °C to 30 °C. Serial morphological changes in the cells induced by Cyprinid herpesvirus 3 (CyHV‐3) included cytoplasmic vacuolar formation, cell rounding and detachment. Mature virions were purified from CyHV‐3‐infected CCF‐K104 cells by sucrose gradient ultracentrifugation and had a typical herpesvirus structure on electron microscopy. Infectious CyHV‐3 was produced stably in CCF‐K104 cells over 30 viral passages. Our findings showed that CCF‐K104 is a useful cell line for isolation and productive replication of CyHV‐3. A temperature shift from 25 °C to 15 °C or 35 °C did not allow serial morphological changes as observed at 25 °C for 14 days. Under the same conditions, real‐time PCR showed that CyHV‐3 was present with low viral DNA loads, suggesting that CyHV‐3 may establish latent infection in CCF‐K104 cells. Amplification of the left and right terminal repeat sequences of the CyHV‐3 genome arranged in a head‐to‐tail manner was detected by nested PCR following an upshift in temperature from 25 °C to 35 °C. The PCR results suggested that the circular genome may represent a latent form of CyHV‐3.     

Detection of Penaeus monodon-type baculovirus (MBV) infection in Penaeus monodon Fabricius by in situ hybridization

Abstract. A digoxigenin-labelled DNA probe was used for in situ detection of the Penaeus monodon -type baculovirus (MBV) derived from cloned MBV polyhedrin genome in cultured Penaeus monodon Fabricius. First, the specificity of the probe against MBV DNA with dot blot hybridization analysis was verified. This probe indicated that cloned MBV polyhedrin fragment can be used as an MBV-specific probe. This was then used to microscopically examine sections of MBV-infected tissues for a blue-purple precipitate indicative of a positive reaction for MBV. MBV-positive cells were located only in the epithelium of the hepatopancreatic tubules and of the midgut. Furthermore, comparison of the susceptibility to MBV infection among several life-stages of the shrimp showed that the MBV genome was found in the zoea, mysis, post-larva, and adult stages, whereas MBV DNA was not detected in either eggs or nauplii. The results were quantified from in situ hybridization with an image analyser to compare the degree of cell infection among groups of cultured P. monodon collected from various farms in Taiwan.     

A structured approach to using biological and environmental parameters to forecast anchovy recruitment

Simulation studies have indicated that a valuable increase in mean annual yield could be achieved in the South African anchovy, Engraulis capensis , fishery if below-average recruitment could be forecast at the start of the fishing season, 6 months before recruitment can he reliably measured. This paper reports on initial investigations into biological and environmental indicator- which could be used to make such forecasts. Valid estimates of recruitment strength in the species are available from 1985 to 1992 and the environmental and biological indicators of interest have been monitored for periods starting from between 1984 and 1988, to the present. Most of the data were obtained from monitoring on anchovy spawner biomass surveys which have been undertaken each November since 1984.
A conceptual model of the factors influencing the recruitment process was constructed and the available data were examined for empirical evidence of the importance of each event included in the model. Fluctuations in copepod biomass and production on the spawning grounds, the incidence of oocyte atresia in adult females, the incidence of southerly winds at Cape Point and the distance offshore of the 16°C isotherm at Cape Columbine were associated with fluctuations in anchovy recruitment. It is suggested that particular attention should be paid to monitoring of these variables to investigate further these associations. If persistent relationships are found, it may be possible to construct an expert system to forecast recruitment from observed values of these variables at the start of a fishing season. A preliminary set of three decision rules is presented.     

Ein Schnelltest zur Ermittlung der individuellen SO2-Empfindlichkeit bei Kiefern

Zusammenfassung Es wird eine Schnellmethode zur Rauchh?rteprüfung von Kiefernarten beschrieben, die vom Terminaltrieb abgetrennte, in Pillengl?ser gesteckte einj?hrige Kurztriebe verwendet. Die Nadeln reagieren auf akute SO₂-Immissionen kloneinheitlich und weitgehend in übereinstimmung mit der Mutterpflanze. Bei den Kurztrieben lassen sich endogene, vom Mutterbaum abh?ngige Differenzen in der Austrocknungsgeschwindigkeit nachweisen, die aber ohne wesentlichen Einflu? auf ihre Reaktion gegenüber SO₂ bleiben, sofern die Expositionsdauer kurzgehalten wird. Die entwickelte Methode ist schneller, wirtschaftlicher und auch zuverl?ssiger als Rauchh?rteprüfungen mit eingetopften Pflanzen. Sie testet lediglich die Reaktion auf akute Begasungen.

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Summary A short-time technique of testing pines for resistance against SO₂-emissions has been developed, using one-year-old needle fascicles, put into water-filled glass vials. Against actual SO₂-exposure, fascicles of the same tree react uniformly, and their reaction is very similiar to that of the parent tree. Besides, needle fascicles show endogenous clonal variation in the rate of drying out. This, however, does not influence the reaction against SO₂, provided the time of exposure is kept short. The new technique is quicker, accurate, and more economical than SO₂-tests with potted seedlings or graftings, but it only tests the reaction against short time-high doses exposure to SO₂.

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Durchgeführt mit finanzieller Unterstützung der Landesanstalt für Immissions- und Bodennutzungsschutz des Landes Nordrhein-Westfalen in Essen.