The dorso-lateral pit organs of the Port Jackson shark contribute sensory information for rheotaxis

Resting Port Jackson sharks Heterodontus portusjacksoni with dorso-lateral pit organs ablated oriented with a mean angle of 263° to the current direction in a flume. This was significantly different ( P <0·01) to controls (normal and sham operated) who had a pooled mean angle of 44° to the current. Thus the dorso-lateral pit organs of H. portusjacksoni , like the free neuromasts of some teleosts, provide sensory information for rheotaxis.     

Influence of dietary protein level on growth performance and body composition of juvenile blackspot seabream, Pagellus bogaraveo(Brunnich, 1768)

Blackspot seabream Pagellus bogaraveo is a potential candidate species for southern European aquaculture. A 12‐week feeding trial was then conducted to evaluate the effect of dietary protein level on growth performance, body composition and feed use efficiency of blackspot seabream juveniles (22·7 g each). Five isolipidic diets (12·5%) with graded levels of protein (20, 30, 40, 50 and 60%) were hand‐distributed, twice a day, to duplicate groups of fish (120 fish per tank), until satiation. Growth (DGC), survival and feed utilization (FCR) were recorded through monthly sampling. Growth performance was significantly affected by dietary protein level ( P  < 0·05), and fish fed at the 50% protein level exhibited the highest final body mass. Feed conversion ratio decreased with increasing levels of dietary protein, fish fed 60% protein being the most efficient. No significant differences were found in whole body composition among treatments ( P > 0·05). Financed by FCT (Project POCTI/CVT/39239/2001, PhD Grant SFRH‐BD‐14068–2003)     

Muscle fine structure reflects ecotype in two nototheniids

The fine structure of swimming (pectoral) and myotomal (axial) skeletal muscle and myocardium of two species of Antarctic nototheniid fishes were studied by electron microscopy, comparing the cryopelagic Pagothenia borchgrevinki and the benthic Trematomus bernacchii . Mean fibre size varied by a factor of four among muscles within each species and may have reflected the locomotory power available, being larger in pectoral oxidative (red) and axial glycolytic (white) muscle of P. borchgrevinki . Both species use labriform locomotion, and the more active P. borchgrevinki had a greater capillary supply, expressed as a capillary to fibre ratio, than T. bernacchii to both red (3·48 ± 0·36 v . 1·63 ± 0·14, mean ±  s . e .; P  < 0·01) and white (2·70 ± 0·20 v . 1·53 ± 0·18, mean ±  s . e .; P  < 0·01) regions of the pectoral musculature. The greater aerobic scope of P. borchgrevinki was strikingly demonstrated in the higher mitochondrial content of all skeletal muscle types sampled, and the ventricular myocardium (0·269 ± 0·011 v . 0·255 ± 0·012 mean ±  s . e .; P  < 0·05). Minor differences were found in other elements of fibre composition, with the exception of a five‐fold greater lipid content in pectoral red fibres of P. borchgrevinki (0·074 ± 0·014 mean ±  s . e .) v . T. bernacchii (0·010 ± 0·003; P  < 0·05). Differences in muscle fine structure among species clearly reflected differences in their ecotype.     

Free radical scavenging activity of Lactobacillus fermentum in vitro and its antioxidative effect on growing–finishing pigs

Aims:  To evaluate the free radical-scavenging capacity of Lactobacillus fermentum and its effects on antioxidant enzyme levels in finishing pigs.
Methods and Results:  The free radical-scavenging activity of Lact. fermentum was analysed in vitro . The tested Lactobacillus showed a high scavenging ability against DPPH (1,1-diphenyl-2-picrylhydrazyl), superoxide and hydroxyl radicals which was dose dependent. Subsequently, 108 crossbred pigs weighing 20·67 BW, were allotted to dietary treatments including a basal diet or the basal diet supplemented with either aureomycin or 10·2 × 10⁷  Lact. fermentum CFU g⁻¹ diet. Supplementation of Lact. fermentum increased total antioxidant capacity ( P  < 0·01) in serum from 50 kg pigs, while serum superoxide dismutase ( P  = 0·01) and glutathione peroxidase ( P  < 0·01) increased, and malondialdehyde levels decreased ( P  < 0·01) in 90 kg pigs. Hepatic catalase ( P  = 0·04), muscle superoxide dismutase ( P  < 0·01) and copper–zinc-superoxide dismutase were enhanced ( P  = 0·01), whereas malondialdehyde levels were reduced ( P  = 0·05) by Lact. fermentum .
Conclusions:  The free radical-scavenging capacity of Lact. fermentum was dose dependent and its supplementation improved the antioxidant status of pigs.
Significance and Impact of the Study:  Lactobacillus fermentum could be used to alleviate oxidative stress and increase pig performance and improve pork quality.     

Effect of fasting on resting metabolic rate and postprandial metabolic response in Silurus meridionalis

Resting metabolic rate in southern catfish of 2 and 5 day fasting groups were significantly higher than that of the 15 day fasting group ( P  < 0·05). After feeding, peak metabolic rate of specific dynamic action (SDA) of the 15 day fasting group was significantly lower than that of the 2 and 5 day fasting groups ( P  < 0·05). The duration of the SDA of the 15 day fasting group was significantly longer than that of the 2 day fasting group ( P  < 0·05) and the SDA coefficient of the 15 day fasting group was significantly lower than that of the 2 day fasting group ( P  < 0·05).     

Effects of a synbiotic product on blood antioxidative activity in subjects colonized with Helicobacter pylori

Aim:  To evaluate the impact of the consumption of a synbiotic product on the antioxidative activity markers of blood in asymptomatic H. pylori -colonized persons.
Methods and Results:  Fifty-three healthy adult volunteers without gastric symptoms participated in a randomized, double-blind placebo-controlled study. The crossover consumption of the enterocoated capsules containing antioxidative Lactobacillus fermentum ME-3, Lact. paracasei 8700:2 and Bifidobacterium longum 46 with Raftilose P95 lasted for 3 weeks and did not change the H. pylori colonization. In H. pylori -positive subjects the sera values of total antioxidative status (TAS) were significantly lower compared to H. pylori -negative subjects (0·97 vs 1·05 mmol l⁻¹, P  = 0·008). After the consumption of the synbiotic, TAS values (0·97 vs 1·03 mmol l⁻¹, P  = 0·004) increased, while the ratio between oxidized and reduced glutathione (0·035 vs 0·030, P  = 0·016) decreased in H. pylori -positive subjects.
Conclusions:  The consumption of a synbiotic containing an antioxidative probiotic strain improved the reduced systemic antioxidative activity in H. pylori -colonized asymptomatic subjects.
Significance and Impact of the Study:  A synbiotic product containing an antioxidative probiotic strain may be useful in the reduction of systemic oxidative stress in H. pylori infection.     

Growth variability of juvenile soles Solea solea and Solea senegalensis, and comparison with RNA : DNA ratios in the Tagus estuary, Portugal

Growth variability and condition of juvenile soles Solea solea and Solea senegalensis , were assessed through RNA : DNA estimates and compared to absolute growth rates. Higher mean cohort RNA : DNA ratios were observed for cohort I at the beginning of estuarine occurrence for both species (4·42 and 4·87, for S. solea and S. senegalensis respectively). Despite different estuarine colonization habits, no significant differences were observed between RNA : DNA monthly variation for both sole species within the same year ( P  > 0·05 for 2003 and 2004). Juvenile S. senegalensis showed significant differences between RNA : DNA ratios obtained for the two nursery areas ( P  < 0·001). The decrease of seasonal growth rates with fish age was similar to seasonal variation of mean RNA : DNA values. Thus the RNA : DNA pattern of juvenile S. solea and S. senegalensis reflected growth and estuarine colonization patterns.     

Inactivation effect of X-ray treatments on Cronobacter species (Enterobacter sakazakii) in tryptic soy broth, skim milk, low-fat milk and whole-fat milk*

Aims:  To determine the inactivation effect of X-ray treatments on Cronobacter ( E. sakazakii ) in tryptic soy broth (TSB), skim milk (0% fat), low-fat milk (1% and 2%) and whole-fat milk (3·5%).
Methods and Results:  X-rays were produced using the RS 2400 generator system (Rad Source Technologies Inc.). Cronobacter (in TSB), inoculated skim milk (0% fat), low-fat milk (1% and 2% fat) and whole-fat milk (3·5% fat) were treated with 0·0, 0·1, 0·5, 0·75, 1·0, 2·0, 3·0, 4·0, 5·0 and 6·0 kGy X-ray doses. Surviving bacteria in the TSB and inoculated milk, before and after treatment, were enumerated using plating method onto trypticase soy agar. Greater than 7·0-log CFU reduction in Cronobacter population was observed with 4·0, 5·0, 6·0, 6·0 and 6·0 kGy X-ray in the TSB, skim milk, 1% fat milk, 2% fat milk and 3·5% fat milk, respectively.
Conclusions:  Treatment with X-rays significantly ( P  <   0·05) reduced Cronobacter to less than detectable limits (<1 log CFU ml⁻¹) in skim milk at 5·0 kGy and milk with 1% fat content and greater at 6·0 kGy dose levels. The D-value for Cronobacter in TSB was significantly ( P  <   0·05) lower than those in milk samples.
Significance and Impact of the Study:  Treatment with X-rays could be an effective and safe alternative technology to control pathogenic bacteria ( Cronobacter ) in the dairy industry.     

Finlets and the steady swimming performance of Thunnus albacares

The functional significance of finlets on the steady swimming performance of yellowfin tuna Thunnus albacares was evaluated by measuring the speed and tail‐beat frequency of the fish with and without them. It was hypothesized that if finlets do improve swimming performance, fish without finlets would have to work harder to maintain the same swimming speed as fish with them and that this would be reflected in kinematic differences. Two‐way ANOVA showed significant effects between individuals on speed (d.f. = 5 and 228, P  < 0·001) and tail‐beat frequency (d.f. = 5 and 48, P  < 0·001), but no significant effects of treatment on speed (d.f. = 1 and 228, P  = 0·25) and tail‐beat frequency (d.f. = 1 and 48, P  > 0·1). No interaction effects on speed (d.f. = 5 and 228, P  > 0·1) and tail‐beat frequency (d.f. = 5 and 48, P  > 0·25) were found. This suggested that finlets were unlikely to function as significant drag reduction and thrust enhancing devices in routine steady swimming. Though not statistically significant, small percentage differences between the mean swimming speeds and tail‐beat frequency of the untreated and treated groups (fish with and without finlets respectively) of the order of 0·5% may be meaningful over the life of a fish. Also, finlets may improve performance at high sustained speeds in rapid accelerations and turns.     

Heat stress‐induced alterations of antioxidants in the freshwater fish Channa punctata Bloch

The effect of elevated temperature on the antioxidants in the freshwater fish Channa punctata was investigated. Fish stressed with an elevated temperature of 12° C, range ± 1° C over the ambient temperature for 3 h showed a significant ( P  < 0·05–0·01) reduction in the levels of antioxidants: reduced glutathione (GSH) and non‐protein thiols. Activity of glutathione reductase was also reduced in all the tissues (liver, kidney and gills) after 3 h of heat stress and 24 h recovery. Catalase (CAT) showed enhanced activity in liver in both the conditions while gills and kidney showed a decreased CAT activity. Glutathione S‐transferase (GST) activity in kidney and liver decreased significantly ( P  < 0·05–0·01) after 3 h of heat stress. At 24 h GST activity showed a tendency to normalize in all the tissues along with a concomitant increase in the GSH level in the kidney. Total and protein thiols in heat stressed fish when matched with controls, showed significant ( P  < 0·05) reduction in the kidney only with a transient increase in liver and gills. Heat shock also induced lipid peroxidation in 3 h heat‐treated and recovery groups when compared with controls. Elevated temperature therefore resulted in tissue specific and time‐dependent alterations of antioxidants in the fish. It also induced lipid peroxidation in various tissues.     

Determination of optimal growth parameters for the bioincising fungus Physisporinus vitreus by means of response surface methodology

Aim:  To evaluate the influence of water activity ( a _w), temperature and pH on the radial growth and lag phase of Physisporinus vitreus (E-642), a basidiomycete was used in the biotechnological process of bioincising.
Methods and Results:  Radial growth was monitored for 20 days on malt extract agar medium. Five levels of a _w (0·998, 0·982, 0·955, 0·928, 0·892) were combined with three incubation temperatures (10, 15, 20°C) and three pH values (4, 5, 6). Data analyses showed a highly significant effect of a _w and temperature ( P <  0·0001) and a significant effect of pH ( P <  0·05). The radial growth rate and lag phase of P. vitreus were very sensitive to a _w reduction. Although P. vitreus was able to grow at all the selected temperatures and pH values, the lag phase increased with decreasing a _w and growth became inhibited at a _w = 0·955. Optimal conditions for growth of P. vitreus were a _w = 0·998, 20°C and pH 5. The response surface model provided reliable estimates of these growth parameters and confirmed a greater dependence on a _w than on temperature or pH under in vitro conditions.
Conclusions:  Low levels of a _w can prevent growth of P. vitreus , so wood moisture content should be adjusted accordingly.
Significance and Impact of the Study:  Implementation of these results should contribute towards the optimization and efficiency of bioincising.     

Detection of infectious rotavirus in naturally contaminated source waters for drinking water production

Aims:  To assess chlorine susceptibility of Legionella pneumophila grown from two amoebic hosts, Acanthamoeba castellanii and Hartmannella vermiformis .
Methods and Results:  After being released from amoebae, Leg. pneumophila were chlorinated at 2 and 5 mg l⁻¹ for 5 min–24 h. Bacterial culturability and cytoplasmic membrane deterioration were quantified by culture assay on BCYEα agar and BacLight stains coupled with a fluorescent microscope, respectively. Chlorination reduced the culturability of Leg. pneumophila by 2·93–4·59 log CFU ml⁻¹ and damaged cellular membrane by 53·8–99·2%. Moreover, cells released from H. vermiformis exhibited significantly lower degrees in culturability reduction ( P  = 0·0008) and membrane deterioration ( P  < 0·0001) when compared with those from A. castellanii . The amoebic genus is the most significant parameter affecting cytoplasmic membrane integrity of chlorinated Legionella ( P  < 0·0001), followed by free chlorine concentration ( P  = 0·042).
Conclusions:  Legionella pneumophila replicated from H. vermiformis possess greater chlorine resistance than the cells from A. castellanii .
Significance and Impact of the Study:  This study shows the heterogeneity of amoebae-grown Leg. pneumophila in chlorine susceptibility, which should be considered in the control of legionellae proliferation, particularly in the systems where H. vermiformis is dominant, e.g. hot water plumbing.     

Influence of rearing temperature during the larval and nursery periods on growth and sex differentiation in two Mediterranean strains of Dicentrarchus labrax

European sea bass Dicentrarchus labrax of the north‐western (NW) and south‐eastern (SE) Mediterranean Sea strains were exposed to different temperatures (13, 17 or 21° C) during the larval rearing (11–51 days post hatching, dph) or nursery periods (55–95 dph), in order to examine the effects of temperature on sex differentiation and subsequent growth during the first year of life. Higher growth was observed during exposure to higher temperatures, but fish of the NW strain exposed to 13 or 17° C during larval rearing exhibited compensatory growth once exposure to the lower temperatures finished, and as a result their final size at 300 dph was similar or greater to the group exposed to 21° C. Fish exposed to 17° C during the nursery period also had similar size to fish exposed to 21° C after 300 days of rearing, but the fish exposed to 13° C remained significantly smaller (ANOVA, n  = 55–100, P  < 0·05). There were significant differences in the sex ratio among the fish exposed to different temperatures during the two periods of rearing, with high temperature (21° C) resulting in a significantly higher percentage of males in the population, both in the NW (ANOVA, n  = 2, P  < 0·04) and SE populations (ANOVA, n  = 2, P  < 0·01). The masculinization effect of high temperature was significantly stronger during the larval rearing stage, both in the NW (ANOVA, n  = 2, P  < 0·005) and SE populations (ANOVA, n  = 2, P  < 0·01). None of the temperature manipulations could produce 100% females, suggesting that there is a part of the genetic component in sex differentiation which is not labile to environmental influence.     

Cyclic feeding and subsequent compensatory growth do not significantly impact standard metabolic rate or critical swimming speed in rainbow trout

Standard metabolic rate ( R _s) and critical swimming speed ( U _crit) were used to assess the aspects of physiological status (stamina) of rainbow trout Oncorhynchus mykiss . Fish were fed either 1·5% body mass daily, 1·5% body mass cyclically (3 weeks of food deprivation followed by 3 weeks of refeeding), a ration based on Stauffer's formula (a maximum temperature-specific ration level) daily or on Stauffer's ration cyclically for 18 weeks. It was hypothesized that if cyclic feeding had no impact on the status of the fish, R _s and U _crit would not cycle with the feeding regime. This hypothesis was supported. No significant difference was found between the mean mass and the fork length of the four groups at the end of the experiment ( P > 0·05). Feeding had no effect on changes in R _s among the four groups, which were significantly different throughout the experiment ( P ≤ 0·05). No significant difference in U _crit was found ( P > 0·05) until at week 12 between groups fed 1·5% body mass ration cyclically and Stauffer's ration daily ( P ≤ 0·05). For groups fed a 1·5% body mass ration cyclically and daily, significant differences occurred at week 15 ( P ≤ 0·05) but no significant difference was found by week 18 ( P > 0·05), suggesting that cyclic feeding does not affect the aspects of physiological status (stamina) of the fish.     

A field and experimental evaluation of the effect of data storage tags on the growth of cod

The effect of electronic data storage tags (DSTs) on the growth of cod Gadus morhua was evaluated in laboratory and field experiments. In the first laboratory experiment, large DSTs (60 × 18 mm, 3 g in water ) attached externally for 3 months did not have any effect ( anova , P  > 0·05) on the growth of adult cod (mean ±  s . d . 65 ± 4·5 cm total length) relative to untagged adult cod. In a second experiment, small DSTs (34 × 11 mm, 1·5 g in water) implanted into young cod (48·1 ± 4·4 cm) for an 8 month period did not have any effect upon the growth relative to untagged controls ( anova , P  > 0·05). Length data returned from tagging experiments conducted on adult cod (57·3 ± 7·5 cm) in the North Sea showed that the growth of fish tagged either externally or internally with large DSTs was not different ( t ‐test, P  > 0·05). Attachment wounds, however, provided evidence that external attachment of DSTs should be avoided unless sensor configuration requires access to the external environment, and that internal implantation should be preferred whenever possible.     

Effect of delayed first feeding on development and feeding ability of Paralabrax maculatofasciatus larvae

The impact of delayed feeding early in development on late feeding ability and development of spotted sand bass Paralabrax maculatofasciatus larvae was examined. Larvae were sampled from hatching until day 19 after delayed feeding for zero (control), 1, 2, or 3 days. Feeding incidence was evaluated as the percentage of larvae with food in the gut and feeding intensity was measured by direct counting of prey after dissection of the gut. Delayed feeding effects due to starvation were observed early in development. By day 5, significant differences ( P  < 0·05) were observed between controls and larvae submitted to degrees of delayed feeding, including total length ( L _T), eye diameter, and the size of the head, liver, gut, muscles and body. Differences were still apparent in L _T, and body, liver and muscle heights at the end of the studied period. Larvae under total starvation did not survive beyond day 5. Initial feeding incidence was 35, 60, 90 and 10% for larvae first fed on day 2, 3, 4 or 5, respectively. Mean ±  s . d . feeding intensity was 3·6 ± 0·8, 2·8 ± 1·3, 5·2 ± 0·3, and 10·2 ± 1·5 rotifers per larva depending on whether larvae were initially fed on day 2, 3, 4 or 5, respectively. The point of no return occurred between 4 and 5 days after hatching.     

Protein synthesis, nitrogen excretion and long-term growth of juvenile Pleuronectes flesus

This study aimed to measure protein synthesis using a stable isotope method, investigate protein-nitrogen flux in a flatfish Pleuronectes flesus , and use the data to test the hypothesis that individual differences in growth efficiency were related to individual differences in protein-nitrogen flux mediated through differences in protein synthesis and degradation. Three measurements of protein-nitrogen flux via consumption, protein synthesis and nitrogenous excretion were made for individual flounder during a 212-day period and fractional rates of protein-nitrogen flux were scaled for a 50–g flounder to provide mean values for protein consumption (2·11 ± 0·21% day⁻¹), protein synthesis (2·08±0·23% day⁻¹), protein growth (0·71±0·06% day⁻¹) and protein degradation (1·37±0·24% day⁻¹). Mean rates of nitrogenous excretion were 0·142 mg N g⁻¹ day⁻¹ and 0·047 mg N g⁻¹ day⁻¹ for ammonia and urea, respectively. Individual flounder had different protein growth efficiencies and this was correlated negatively and significantly with mean rates of protein synthesis ( r - 0·70; P <0·05) and degradation ( r - 0·67; P < 0·05) and correlated positively and significantly with the efficiency of retaining synthesized protein ( r +0·63, P <0·05). This supported the proposed hypothesis that flounder which grow more efficiently achieve this through adopting a low protein turnover strategy.     

Climate variables and spawning migrations of the striped mullet and white mullet in the north‐western area of the Gulf of Mexico

The 'smoothed'(4253e HT smoother) 6 year time series with a monthly periodicity of catches (where the increases of the catches reflect the spawning migration) of Mugil cephalus and Mugil curema , were correlated (Pearson correlation index) against the 'smoothed' data on rainfall, water temperature, daylight and tide monthly averages. Water temperature and M. curema spawning migrations had a negative correlation ( r  = −0·77), while tide monthly averages and M. cephalus spawning migrations were positively correlated ( r  = 0·84). The two environmental variables that showed highly significant correlations were water temperature and daylight ( r  = 0·85, P  < 0·05). Meteorological tides were used as a reflection of the effect of the north winds from the Gulf of Mexico which produce an average tide rise from September to November. The relationship of these variables with spawning migrations is discussed, and it is suggested, in particular, that meteorological tides, as a reflection of the north winds, are the trigger (zeitgebers) for M. cephalus spawning in the ocean.     

Polyaluminum chloride-enhanced concentration efficiency of poliovirus and f2 phage from sewage water

Aims:  To enhance the recovery of f₂ bacteriophage and poliovirus by an established method based on the adsorption to and elution from positively-charged Al(OH)₃-treated silica gel.
Methods and Results:  Polyaluminum Chloride (PAC) was added to water samples to neutralize the negatively charged materials, which can reduce virus recovery by providing a competing adsorption mode on the media surface. Using this improved process (PAC 30 mg l⁻¹, pH 6·5, temperature 20∼30°C), the recoveries of Poliovirus I and f₂ from small-volume sewage (100 ml) were 110·76 ± 36·0% and 92·06 ± 8·65%, respectively ( P  < 0·05 vs. traditional methods). Recovery from a 20-L volume of sewage averaged 85·65 ± 4·43% for f₂ and 88·73 ± 9·76% for poliovirus, significantly higher than the recoveries in the traditional methods ( P  < 0·05).
Conclusions:  PAC could enhance concentration efficiency of poliovirus and f₂ phage from sewage water.
Significance and Impact of the Study:  This method should significantly improve the recovery of viruses from sewage.     

Digestive enzymes of Anarhichas minor and the effect of diet composition on their performance

The effect of feeding two different diets, containing 38 and 45% protein and 35 and 28% lipid, respectively, on growth as well as performance of the digestive proteases (total protease at pH 1·5, 7·0, 9·0 and 10·0) and carbohydrases (total carbohydrase, α-amylase and α-glucosidase) of a new candidate species for aquaculture, the carnivorous spotted wolffish Anarhichas minor (Teleostei, Anarhichadidae), was investigated. No significant growth differences were observed when feeding high protein (HP) or low protein (LP) levels ( P > 0·05), although the LP group exhibited slightly higher protease activity and capacity ( P > 0·05), mostly at the anterior end of the intestine and at alkaline pH. Carbohydrase levels were generally low, apart from α-glucosidase, which was significantly increased for the HP group ( P < 0·05). Implications concerning the performance of digestive enzymes under varying dietary macronutrient levels and effects of digestive enzyme activity on the formulation of efficient artificial diets for carnivorous fishes are discussed in this study.