Steroid glucuronides as male pheromones in the reproduction of the African catfish Clarias gariepinus--a brief review

After ovulation, female African catfish are strongly attracted by the odor of male conspecifics. This attraction depends on the presence of the seminal vesicle, a part of the male reproductive organs. Removal of the seminal vesicle illustrates this fact. A low dose of seminal vesicle fluid, added to the water, appears to be highly attractive for catfish which have ovulated. Fractionation of the fluid and testing of the different fractions shows that steroid glucuronides could be responsible for the attraction. These steroid glucuronides can be identified with gas chromatographic-mass spectrometric analysis. A mixture of glucuronides, prepared to resemble the composition of the seminal vesicle fluid, evokes a dose-dependent attraction. The most potent odorant, observed by measuring electrical responses from the olfactory epithelium and from the olfactory tract appears to be 3,17-dihydroxy-5β-pregnan-20-one-3-glucuronide.     

Immunohistochemically detected ontogeny of prolactin and growth hormone cells in the African catfish Clarias gariepinus

The chronological appearance of selected endocrine cells in the pituitary of African catfish Clarias gariepinus (Burchell, 1822) was studied morphologically, histologically and immunohistochemically by using antisera raised against catfish growth hormone (cgGH) and recombinant tilapia prolactin I (tPRL). cgGH- and tPRL-like immunoreactive cells were visible from day 1 post fertilisation (hatching) throughout the juvenile and the adult stage. From 1 to 90 days after hatching, the larval pituitary is oval in shape with a distinctly shaped rostral pars distalis, proximal pars distalis and pars intermedia. From day 120 onwards allometric growth of the rostral and proximal pars distalis extended the prolactin and growth hormone cells anteriorily and posteriorily, respectively. Size and activity of the prolactin and growth hormone cells, measured by the ratio of cell surface to nuclear surface remained constant until day 40 and showed a growth spurt thereafter. Growth hormone content, measured with a catfish-specific radio-immunoassay from hatching until 60 h post hatching, increased exponentially between 30 and 60 h.     

A note on alarm cues in juvenile African catfish, Clarias gariepinus Burchell: Indications for opposing behavioural strategies

Following exposure of African catfish to alarm cues, a relationship (r_p = 0.60, N = 10, P = 0.07) between feed efficiency (measured as residual feed intake) and the change in percentage of time spent swimming in response to damage-released alarm cues was observed. Feed-efficient animals responded with a decrease in percentage of time spent swimming (P < 0.05) whereas feed-inefficient animals responded with an increase in percentage of time spent swimming (P < 0.05). This study shows opposing behavioural strategies in African catfish in response to damage-released alarm cues being related to feeding efficiency.     

One-step immunoaffinity purification and partial characterization of hypophyseal growth hormone from the African catfish, Clarias gariepinus (Burchell)

Growth hormone (GH) was purified from African catfish (Clarias gariepinus) pituitary extracts in a single step by use of immunoaffinity chromatography. A monoclonal antibody to chicken GH, which labels the catfish hypophyseal somatotropes in immunocytochemistry, was coupled to CNBr-activated Sepharose, and crude alkaline pituitary extracts were run over the immunoadsorbent. Reversed-phase high-performance liquid chromatography analysis of the eluted material suggested heterogeneity, whereas silver staining upon SDS-polyacrylamide gel electrophoresis showed one single band with an estimated molecular weight between 22,000 and 23,000 Da. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis of the same preparation revealed the presence of several components with molecular weights ranging from 20,170 to 20,900 Da. The amino terminus of the protein was homogeneous, and the first 50 residues matched the proposed sequence of GH from two other siluran species (Ictalurus punctatus and Pangasius pangasius), except for one substitution at position 3. These data unequivocally confirm the identity of the purified molecule as suggested by immunochemical evidence. The bioactivity of the GH preparation was demonstrated by the short-term effect of GH on T₃ plasma levels in juvenile catfish.     

The Effect of Freezing Rate on the Survival of Cryopreserved African Sharptooth Catfish (Clarias gariepinus) Spermatozoa

Cryoprotective agents were evaluated to find the optimal concentration of the cryoprotectant and most suitable combination of solution and cryoprotectant. A cryoprotective agent composed of 4% glucose and 9% glycerol yielded the best results. It was established that the optimal freezing rate is dependent on the composition of the cryoprotective agent. Maximal survival of catfish spermatozoa (60%) occurs at 5°C min^-1 and faster and slower freezing rates result in poor survival or no survival at all. Incorporation of an isothermal holding period into the freezing rate led to remarkable increase (20-30%) in sperm survival when Me₂SO was present in the cryoprotective agent. Cryoprotective agents containing glucose also showed improved survival when a three phase freezing rate was used. These results lead to the conclusion that the presence of an isothermal holding period in the freezing rate is beneficial for the cryoprotective action of Me₂SO and glucose.     

Cyanolipids in Cordia verbenacea—a correction

Seeds of materials previously identified as Cordia verbenacea were re-examined and found to be from a sapindaceous plant, probably of the genus Allophyllus. The anomalous occurence of cyanolipids in this member of the Boraginaceae is thus resolved.     

Control of reproduction in female cockroaches with special reference to Nauphoeta cinerea—I. First pre-oviposition period

Prior to the first oviposition, a receptivity centre, perhaps neurosecretory cells in the brain, controls the female's acceptance of courting males. In L. maderae this centre is affected by starvation. A brief exposure to food can induce mating but is inadequate for oöcyte development. Before the first ovulation starvation has no effect on receptivity in N. cinerea.

In N. cinerea mechanical stimulation caused by the firm insertion of the spermatophore in the bursa copulatrix releases stimuli via the nerve cord to the brain which render the female unreceptive and, at the same time, increases the activity of the corpora allata resulting in rapid development of the oöcytes.

The mechanical presence of the oötheca in the uterus also has two principal effects. Like spermatophore insertion, it inhibits mating. But its effect on the corpora allata is inhibitory, rather than stimulatory, and, consequently, the oöcytes remain underveloped for almost the entire gestation period. The effectiveness of inhibitory stimulation from the stretched uterus depends upon the period in the reproductive cycle in which it occurs-i.e. on the physiological state of the female. In N. cinerea uterine stretching inhibits mating and oöcyte development after oviposition or during gestation but is not effective when exerted during the first pre-oviposition period. In P. surinamensis, uterine stretching does not inhibit the corpora allata prior to the first ovulation but does prevent oöcyte development during gestation.

In fed L. maderae and N. cinerea there appears to be a synergistic action of nutrition and mating in controlling the rate of oöcyte development. Mating (mechanical) and feeding (chemical) stimuli are both usually required for activating the corpora allata to their fullest extent so that the oöcytes mature at their maximum rate. There is some indication that mating stimuli in N. cinerea and L. maderae are effective in further stimulating the corpora allata only if the corpora allata have reached a certain level of activity, if activating stimuli have begun to occur in the brain, or if the mating stimulus occurs in combination with nutritional factors. Thus, the corpora allata in starved virgin females of N. cinerea become sufficiently active so that some yolk is deposited in the oöcytes but these oöcytes do not mature; mating is effective in further stimulating the endocrine glands in these starved females and oviposition occurs in about the normal period. In starved virgins of L. maderae the corpora allata are virtually inactive and yolk is not deposited in the oöcytes; mating has no effect on oöcyte development in starved females. D. punctata differs from both the above species in that the corpora allata in the virgin female usually remain inactive whether she feeds or starves. Mating stimuli alone can activate the corpora allata, in fed or starved females, and consequently the oöcytes mature.     

Mutagenic and physiological responses in the juveniles of African catfish, Clarias gariepinus (Burchell 1822) following short term exposure to praziquantel

Praziquantel (PZQ) is an acylated quinoline-pyrazine originally developed for veterinary application but now one of the most used anti-helminthic drugs for treatment of certain trematodes and cestodes in both human and other animals. The present study investigated the mutagenic and physiological responses in the juveniles of African catfish, Clarias gariepinus following short term exposure to praziquantel. Based on the 53.52 mg/l 96 h LC₅₀ of PZQ obtained, two sublethal concentrations of 5.35 and 10.70 mg/l of the drug were selected and fish were exposed to these concentrations and control for 15 days. Micronuclei induction in the peripheral blood of PZQ-exposed fish was highest on day 10 but the fish morphological parameters were not affected. The packed cell volume (PCV) was significantly reduced (p < 0.05) from day 5 while red blood cells (RBC) and hemoglobin (Hb) significantly declined (p < 0.05) on day 15. Macrocytic anemia was observed on day 1 of study and thereafter microcytic anemia developed on day 5 of study. The white blood cell (WBC) was significantly (p < 0.05) elevated from day 10 of exposure while values of mean cellular volume (MCV), mean cellular hemoglobin (MCH) and mean cellular hemoglobin concentration (MCHC) were not significantly different (p > 0.05) from the control. Alanine aminotransferase (ALT), aspartate aminotransferase (AST) and glucose levels significantly increased while protein reduced (p < 0.05) throughout the exposure period but a mixed trend was observed in the leukocyte differentials. PZQ should be used with caution as sublethal exposure elicited micronucleus induction and alterations of hematological and biochemical parameters in the fish.     

Seasonal acclimatization in the migratory hamster Cricetulus migratorius—the role of photoperiod

1. 1.|The migratroy hamster Cricetulus migratorius, a small nocturnal rodent, inhabits ecosystems characterized by dramatic seasonal fluctuations of ambient temperatures. The aim of this study was to assess seasonal acclimatization of its thermoregulatory system.

2. 2.|Heat production by means of oxygen consumption and body temperature in various ambient temperatures as well as non-shivering thermogenesis were measured in C. migratorius. The hamsters were acclimated to two different photoperiod regimes (16L:8D and 8L:16D) at a constant ambient temprature of 24°C. Overall thermal conductance was calculated for such hamsters.

3. 3.|The results of this study indicate that photoperiod manipulations adjust the thermoregulatory system of the migratory hamster mainly by affecting overall thermal conductance.

Taenia hydatigena—V. Surface structure of the adult worm and evaginated scolex

Scanning electron microscopy (SEM) of the surface of the adult tapeworm and the freshly evaginated scolex from the cysticercus indicated that in Taenia hydatigena there was a variety of microthrix form. This variety was found between different areas on the same specimen and between the adult and freshly evaginated scolex. In the latter there was a noticeable absence of pointed spikes from most areas. Artifacts attributable to techniques of preparation are also discussed.     

Pullulans produced by strains of Cryphonectria parasitica—I. Production and characterisation of the exopolysaccharides

Studies published during the last four decades indicated that pullulan has mainly -(1→6) maltotriose subunits, accompanied by -(1→6) maltotetraose subunits and other anomalies in the polymer chain. The presence and the extent of these anomalies depend on the strain of Aureobasidium pullulans used for obtaining the pullulan as exocellular material, and on the culture conditions as well. Besides A. pullulan, other fungi produce this polysaccharide. In all cases, the maximum amount of -(1→6) maltotetraose subunits was believed to be 7%. This paper reports the characterization of pullulan samples, obtained by liquid cultures of a virulent strain (CP159) and two hypovirulent strains (CP263 and CP102) of Cryphonectria parasitica. It was found that in all cases pullulan was much richer in -(1→6) maltotetraose subunits than the pullulan(s) so far investigated. Chestnut blight caused by C. parasitica is a disease of worldwide importance in chestnut production. The structure of pullulan could be correlated with the pathogenesis and symptoms expressed by infected plants.     

Juvenile hormone involvement in Drosophila melanogaster male reproduction as suggested by the Methoprene-tolerant mutant phenotype

Juvenile hormone (JH) involvement in male reproduction is poorly understood. In Drosophila melanogaster adults, JH deficiency has been shown to result in lowered protein synthesis in male accessory glands. To probe additional roles, we have examined males homozygous for a null allele of Methoprene-tolerant (Met). This gene is involved in the action of JH, possibly at the JH receptor level, and Met²⁷ null mutants reflect a diminution of JH action. Met²⁷ males were found to have reduced protein accumulation in male accessory glands and to court and mate wild-type females much less avidly than do either Met⁺ or Met²⁷; Met⁺ transgenic males. Exposure of Met²⁷ males to methoprene partially rescued the courtship deficiency. However, sperm transfer as reflected by fertility of Met²⁷ fathers was found to be similar to that of Met⁺. Taken together with previous work examining the JH-deficient mutant apterous, these results corroborate JH involvement in protein synthesis in the male accessory glands and suggest a role for JH in promoting male mating behavior in these flies.     

Chonopeltis australis (Crustacea) male reproductive system morphology; Sperm transfer and review of reproduction in Branchiura

The morphology of the male reproductive system as well as sperm transfer in Branchiura has been described for Dolops ranarum and Argulus japonicus. In this study, the reproductive system and accessory structures are described for male Chonopeltis australis using histology, light microscopy, and scanning electron microscopy. For the first time, we describe sperm transfer by means of a spermatophore in this genus. The internal and external morphology and mechanism of sperm transfer is compared with other Branchiura, where it has been described. The morphology of the reproductive system of C. australis is similar to that of D. ranarum while the accessory structures and the spermatophore produced are similar to that of A. japonicus. A revision of the definition of Branchiura with respect to reproduction is provided. J. Morphol. 276:209–218, 2015. © 2014 Wiley Periodicals, Inc.     

Biotransformation of glucose to gluconic acid by Aspergillus niger—study of mass transfer in an airlift bioreactor

A glucose–gluconic acid biotransformation system was suggested for the experimental study of oxygen transfer in bioreactors. This biosystem was used for the investigation of the effect of the flow rate and biomass concentration on the volumetric oxygen transfer coefficient k_La in a 10 dm³ internal-loop airlift bioreactor. For this purpose, the fermentation broth of the mycelial strain Aspergillus niger was employed, representing a three-phase system, where bubbles come into contact with dense rigid pellets. The results showed that the presented biotransformation system can be successfully utilised for the determination of the oxygen transfer rate in airlift bioreactors. The experiments showed a strong positive influence of the air flow rate on the rate (r_Glu), specific rate of gluconic acid production (k_Glu/X) as well as on the volumetric oxygen transfer coefficient (k_La). This confirmed an expected limitation of production rate by the oxygen transport from the gas to the liquid phase in the whole range of air flow rates applied. Moreover, consistent curves of the production rate r_Glu and k_La values vs. biomass concentration c_X (amount of enzymes) were observed. These exhibited a local maximum for c_X equal to 6.68 g dm⁻³. On the other hand, the specific production rate monotonously decreased with increasing biomass concentration. A decline of k_La values at higher c_X values was attributed to a bubble coalescence promoting effect of mycelial pellets.     

Changes in amino acid composition in the tissues of African catfish (Clarias gariepinus) as a consequence of dietary L-carnitine supplements

A study was undertaken to examine the effect of different amounts of dietary lysine (13 and 21 g kg^?1 diet), lipid (80 and 160 g kg^?1 diet) and L ‐carnitine (0.2 and 1.0 g kg^?1 diet) on growth performance, proximate composition and amino acid metabolism of the African catfish (Clarias gariepinus). Juvenile African catfish (23 ± 1.5 g/fish) were stocked into 70‐L aquaria (16 aquaria, 28 fish/aquarium) connected to a recirculation system during a maximum period of 74 days. All groups were fed at a level of 24 g kg^?0.8 day^?1 in an experiment run at pair feeding. Animals receiving 1.0 g carnitine accumulated up to six times more carnitine in their tissues than animals receiving 0.2 g (P < 0.05). Acyl‐carnitine and free L ‐carnitine levels increased in the whole body and in tissues. Dietary L ‐carnitine supplements increased protein‐to‐fat ratios in the body, but did not affect growth rate. Protein‐to‐fat ratios were only affected when the biosynthesis capacity of L ‐carnitine was restricted due to low lysine levels and when there was a shortage of dietary fat. When lysine was offered at 21 g kg^?1 feed, dietary L ‐carnitine supplements did not affect the amino acid concentrations of body tissues. Dietary L ‐carnitine supplements raised the concentration of glutamic acid > aspartic acid > glycine > alanine > arginine > serine > threonine in skeletal muscle tissue (P < 0.05). Total amino acid concentration in muscle and liver tissues (dry‐matter basis) increased from 506 to 564 and from 138 to 166 mg g^?1, respectively, when diets were offered with high L ‐carnitine, low lysine and low fat levels. These data suggest that dietary L ‐carnitine supplementation may increase fatty acid oxidation and possibly decrease amino acid combustion for energy.     

Gonadal steroids and the maturation of the species-specific gonadotropin-releasing hormone system in brain and pituitary of the male African catfish (Clarias gariepinus)

The effect of testosterone (T), 11-ketotestosterone (KT) and estradiol (E(2)) on the development of the catfish gonadotropin-releasing hormone system (cfGnRH) of male African catfish (Clarias gariepinus), at the onset of puberty [between 10 and 12 weeks post hatching (ph)] was investigated. The cfGnRH neurons, located in the ventral forebrain, were visualized by immunofluorescence and their numbers were determined and the amounts of cfGnRH-associated peptide (cfGAP) in the pituitary were measured by RIA. Steroid treatments did not significantly alter the numbers of immunoreactive GnRH neurons. However, T and E(2) caused an increase in the amount of GnRH, demonstrated by the intensity of the immunostaining of GnRH neurons and fibers in the brain and the amount of cfGAP in the pituitary. Treatment with KT, the main circulating androgen in adult male catfish, neither changed the number of cfGnRH neurons, nor elevated the cfGnRH content in the pituitary. In previous experiments with younger, prepubertal fish (2-6 weeks ph), T caused an elevation of the number of cfGnRH neurons to the same level as present in pubertal fish of 12-14 weeks. We conclude that the onset of puberty in the male African catfish coincides with the completion of the steroid-dependent structural maturation of the cfGnRH system in the brain. T and/or E(2), however, are still able to exert a positive influence on the amounts of cfGnRH during the later stages of pubertal development, thus still playing a role in the control of the cfGnRH system.     

Patterns of reproduction and growth of the catfish Iheringichthys labrosus (Lütken, 1874) after a reservoir formation

This work contributes to the knowledge of the reproduction, growth parameters and mortality rates of Iheringichthys labrosus in a newly‐formed reservoir. A total of 554 males and 1227 females were collected over 12 consecutive months, 1998–1999, from sites in the Corumbá Reservoir, Brazil, using gillnets (meshes: 2.4–16 cm). Information on each individual, i.e. standard length (cm), weight (g), sex, and gonadal development phase was recorded. The pectoral spines were removed to estimate age. The number of juveniles and adults, males and females, reproductive sites and seasons were estimated. First maturation length was estimated using a likelihood function fitted by binomial distribution. Growth parameters were estimated using the von Bertalanffy equation. Total instantaneous mortality was obtained through a linearized catch curve method. Standard length varied from 6.0 to 20.5 cm. Growth showed negative allometry for both sexes. The reproduction period was August to December in all environments sampled and first maturation length was 11.5 cm. All individuals were adults with 17.0 cm standard length. Ages varied from zero to 7 years. Asymptotic length, growth coefficient and t₀ for the entire population were 27.79 cm, 0.12 and ?2.64, respectively. Instantaneous and annual mortality rates were 0.90 and 0.59, respectively.     

Study of LH response to GnRH in the young male as a criterion of genetic merit for female reproduction in sheep

A high and a low response line in sheep were selected on the basis of the mean concentration of LH in 10-week-old Finn-Dorset ram lambs after an i.v. injection of 5 micrograms GnRH. After 8 male generations the mean LH response of the high line was more than 5-fold that of the low line and the heritability of the selected trait was estimated at 0.44 +/- 0.015. Highly significant line differences in mean LH response to GnRH were also found in males at 20 weeks of age and females at 10 and 20 weeks of age and the genetic correlations between the four LH response traits appear to be close to unity. Large line differences in the mean FSH response to GnRH were also found in both males and females at 10 and 20 weeks of age. Selection had little effect on the physical characteristics of lambs. High-response line ewes entering their first breeding season at about 7 months of age showed oestrus earlier in the season and had higher ovulation rates and numbers of lambs born per ewe lambing than did low-response line ewes. In the second breeding season, at about 19 months of age, the only line difference was a higher ovulation rate early in the breeding season in high-line ewes. It is suggested that these changes may be mediated by a more rapid response in high-line ewes to increased GnRH stimulation at puberty or at the beginning of the breeding season.