Evidence for independent origin of two spring‐spawning ciscoes (Salmoniformes: Coregonidae) in Germany

Combined analyses of mitochondrial DNA (mtDNA) and microsatellite loci were performed to assess the genetic differentiation of two spring‐spawning ciscoes from each other and from sympatric Coregonus albula in two German lakes. Polymorphism was screened at six microsatellite loci and mtDNA for a total of 247 and 94 ciscoes, respectively. Microsatellite data showed a weak differentiation between spring‐spawning Coregonus fontanae and sympatric C. albula in Lake Stechlin ( F _ST = 0–0·008), whereas a significant differentiation was observed between spring‐spawning Coregonus lucinensis and sympatric C. albula in Lake Breiter Luzin ( F _ST = 0·013–0·039). A more pronounced genetic difference was observed between both spring‐spawning species ( F _ST = 0·05–0·128). Shared mtDNA haplotypes among sympatric species within both Lake Stechlin and Lake Breiter Luzin were observed, whereas no haplotype was shared between C. fontanae and C. lucinensis . These results suggest an independent origin for spring‐spawning ciscoes in each lake. Evidence is also provided for mtDNA introgression of Coregonus sardinella into C. lucinensis and C. albula in Lake Breiter Luzin. Postglacially, this species or at least a population which showed mtDNA introgression has colonized the Baltic Sea basin up to the glacial margin that was located between Lakes Stechlin and Breiter Luzin.     

Modelling the growth of Weissella cibaria as a function of fermentation conditions

Aims:  To investigate the effect of pH, water activity ( a _w) and temperature on the growth of Weissella cibaria DBPZ1006, a lactic acid bacterium isolated from sourdoughs.
Methods and Results:  The kinetics of growth of W. cibaria DBPZ1006 was investigated during batch fermentations as a function of pH (4·0–8·0), a _w (0·935–0·994) and temperature (10–45°C) in a rich medium. The growth curve parameters (lag time, growth rate and asymptote) were estimated using the dynamic model of Baranyi and Roberts (1994. A dynamic approach to predicting bacterial growth in food. Int J Food Microbiol 23, 277–294). The effect of pH, a _w and temperature on maximum specific growth rate (μ_max) were estimated by fitting a cardinal model. μ_max under optimal conditions (pH = 6·6, a _w = 0·994, T  = 36·3°C) was estimated to be 0·93 h⁻¹. Minimum and maximum estimated pH and temperature for growth were 3·6 and 8·15, and 9·0°C and 47·8°C, respectively, while minimum a _w was 0·918 (equivalent to 12·2% w/v NaCl).
Conclusions:  Weissella cibaria DBPZ1006 is a fast-growing heterofermentative strain, which could be used in a mixed starter culture for making bread.
Significance and Impact of the Study:  This is the first study reporting the modelling of the growth of W. cibaria , a species that is increasingly being used as a starter in sourdough and vegetable fermentations.     

A molecular assessment of homing in the tidepool sculpin

Molecular analyses of the mitochondrial genome using restriction fragment length polymorphism (RFLP) were utilized to determine the existence of discrete populations of the tidepool sculpin Oligocottus maculosus from the Barkley Sound region of south-western Vancouver Island, British Columbia. Fifty-five haplotypes were found with three haplotypes making up 38, 16 and 8% of the total sample, respectively. Within individual tidepools, haplotype diversity ranged from 0·30 to 0·92, but haplotypes were typically closely related to one another (nucleotide diversity ranged from 0·0004 to 0·005). The majority of haplotype variation (96–97%) was located within tidepools, but a significant amount of variation ( c . 3–4%) was detected among tidepools within single islands. By contrast, there was no significant differentiation among islands or between samples from the Barkley Sound area and a single sample from north-eastern Vancouver Island. The results suggest a low level of population structure within the sample area and that the distribution of haplotypes and haplotype divergences represents a demographic expansion from a single ancestral lineage beginning between 300 000 and 1 000 000 years ago. The low level of population subdivision resolved suggests that despite their ecological specialization in tidepool habitats, tidepool sculpins are influenced by demographic and genetic processes that operate on spatial scales much larger than single tidepools or islands.     

Impact of temperature on food intake and growth in juvenile burbot

The effect of temperature on food consumption, food conversion and somatic growth was investigated with juvenile burbot Lota lota (age 0 years). Juvenile burbot showed a significant dome shaped relationship between relative daily food consumption ( C _R) and temperature ( T ) with C _R = − 0·00044 T ² + 0·01583 T  − 0·06010; ( n  = 90, r ² = 0·61). Maximum C _R was at 17·9° C (95% CL 17·2–18·6° C). The temperature related instantaneous growth rate ( G ) also followed a dome shaped function with G  = − 0·000063 T ² + 0·002010 T  − 0·007462; ( n  = 95, r ² = 0·57), with maximum growth rate at 16·0° C (95% CL 15·3–16·6° C). A significant linear relationship was found between the water temperature and the conversion coefficient ( C _C) with C _C = − 1·63 T  + 59·04; ( n  = 80, r ² = 0·74). The results indicate that juvenile burbot in large lakes benefit from higher water temperatures in the littoral zone, by increased food uptake and growth, especially during the warm summer months. Because profundal water temperatures do not reflect the optimal temperature for food consumption in large burbot, temperature is unlikely to be the main proximate factor for the obligate littoral‐profundal migration of juvenile burbot observed in many lake populations.     

Mitochondrial cytochrome b DNA sequence variation of Atlantic cod from Iceland and Greenland

Sequence variation of a 250–bp (base pair) fragment of the mitochondrial cytochrome b gene has been studied using polymerase chain reaction and direct sequencing of 519 Atlantic cod Gadus morhua from Iceland and 78 cod from Greenland. Twenty-four variable nucleotide sites, mostly silent, define 34 haplotypes. The amount of variation is high ( ĥC =0·73, π=0·52 per 100 bp) with five haplotypes at polymorphic frequencies in Iceland and a number of widely dispersed rather rare haplotypes. A tree of genetic relationships among haplotypes has considerable homoplasy yet it is relatively shallow implying a high turnover of variants of the polymorphism. Net nucleotide genetic divergences among localities are nil. Geographic locality overall area, and inshore/offshore comparison explain none of the variation in an AMOVA, all the variation is among individuals and a null hypothesis of non-differentiation of haplotype frequencies among localities or overall areas cannot be rejected. A temporal year-class effect is found. The evolutionary difference between Greenland and Iceland cod is not significant and the percentage of variation accounted for by the Greenland/Iceland difference is half of what a temporal effect within Iceland explains. There is no evidence for considering the cod at Greenland and Iceland to consist of separate evolutionary units and the question of separate management units must address the lack of diagnostic genotypes and evidence for gene flow from clinal variation.     

Changes in movement, range and habitat preferences of adult grayling from late summer to early winter

Radio‐tagged adult grayling Thymallus thymallus ( n  = 22), monitored from mid August to mid December 1999 in the River Kuusinkijoki, Finland, shifted by the end of September (water temperature 10·0–14·5° C) from riffle sites to deeper and slower pool sites 0·7–1·6 km up‐ or downstream. In early winter ( c . 0° C water temperature), eight of 13 fish still under study made a further shift into new pool sites, possibly triggered by ice formation. The summer range of grayling in the riffles was smaller (mean ±  s . d . length: 75 ± 146 m) than the autumn range (99 ± 46 m) in the pools, but gross daily movements were equally short in both the seasons (18 ± 34 m and 15 ± 7 m, respectively). In late summer, adult grayling preferred water depths 80–120 cm and mean velocities >40 cm s⁻¹. In autumn, the preferred ranges were 100–240 cm and <30 cm s⁻¹, respectively. Substratum was mainly boulders in the summer sites, and gravel or pebbles in the autumn sites.     

Genetic variation in skipjack tuna Katsuwonus pelamis(L.) using PCR‐RFLP analysis of the mitochondrial DNA D‐loop region

A high level of genetic diversity was observed in Katsuwonus pelamis populations from India ( h  = 0·952, ne  = 14·3) and Japan ( h  = 0·897, ne  = 8·9). The log‐likelihood ( G )‐based exact test revealed significant heterogeneity in the distribution of haplotypes between the two populations ( P  < 0·01, s . e . = 0·001). This result suggests that the two populations should now be treated as demographically independent and managed separately.     

Auxin production by plant associated bacteria: impact on endogenous IAA content and growth of Triticum aestivum L.

Aims:  The aim of this study was to investigate the potential of bacterial strains of Bacillus, Pseudomonas, Escherichia, Micrococcus and Staphylococcus genera associated with wild herbaceous flora to enhance endogenous indole-3-acetic acid (IAA) content and growth of Triticum aestivum var. Inqalab-91.
Methods and Results:  Gas chromatography and mass spectrometric (GC–MS) analysis revealed that bacterial strains produced 0·6–8·22 μg IAA ml⁻¹ in the presence of L-tryptophan. Plant microbe experiments showed a significant positive correlation between auxin production by bacterial strains and endogenous IAA content of T. aestivum for GC–MS ( r  = 0·618; P  = 0.05) and colorimetric analysis ( r  = 0·693; P  = 0.01). Similarly, highly significant positive correlation for shoot length ( r  = 0·627; P  = 0.01) and shoot fresh weight ( r  = 0·626; P  = 0.01) was observed with auxin production under axenic conditions. Bacterial inoculations also enhanced shoot length (up to 29·16%), number of tillers (up to 97·35%), spike length (up to 25·20%) and seed weight (up to 13·70%) at final harvest.
Conclusions:  Bacterial strains have the ability to increase the endogenous IAA content and growth of T. aestivum var. Inqalab-91.
Significance and Impact of the Study:  Microbial strains of wild herbaceous flora can be effectively used to enhance the growth and yield of agronomically important crops.     

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.     

High genetic diversity, large inter-oceanic divergence and historical demography of the striped mullet

The striped mullet Mugil cephalus , L. is a circumtropical species whose extreme conservative morphology stands in contrast with the degree of genetic differentiation at a global scale. One hundred and fourteen mitochondrial control region DNA sequences were analysed from four localities in the Gulf of Mexico (Texas, Louisiana, Mississippi, Florida), one on the U.S.A. east coast (N. Carolina), and one in Hawaii, giving very high levels of molecular diversity ( h =1·0 every haplotype was unique in all samples, π=1·1–2·0% in Gulf–Atlantic and π=3·1% in Hawaii). With no genetic evidence of dramatic population expansions, mismatch distributions were still very different in each ocean. Stable population levels in Hawaii have fostered the generation and persistence of very high molecular diversity, but Gulf–Atlantic samples suggest a shorter span of population stability. Ninety-five per cent of the molecular variance was allocated between ocean basins and virtually none among Gulf–Atlantic samples. A neighbourjoining reconstruction revealed Atlantic- and Pacific-specific lineages separated by more than 24% uncorrected sequence divergence ( d =0·49 Tamura-Nei Gamma-corrected). The lack of phylogeographic structure among Gulf–Atlantic samples corroborated the AMOVA results and supported the existence of a single population with high levels of gene flow along the Gulf of Mexico and north-west Atlantic coasts. The genetic differentiation between oceans points to the absence of gene flow and an accelerated rate of mitochondrial evolution in the genus Mugil .     

Migration of hatchery‐reared Atlantic salmon and wild anadromous brown trout post‐smolts in a Norwegian fjord system

Hatchery‐reared Atlantic salmon Salmo salar ( n  = 25) and wild anadromous brown trout (sea trout) Salmo trutta ( n  = 15) smolts were tagged with coded acoustic transmitters and released at the mouth of the River Eira on the west coast of Norway. Data logging receivers recorded the fish during their outward migration at 9, 32, 48 and 77 km from the release site. Seventeen Atlantic salmon (68%) and eight sea trout (53%) were recorded after release. Mean migratory speeds between different receiver sites ranged from 0·49 to 1·82 body lengths (total length) per second (bl s⁻¹) for Atlantic salmon and 0·11–2·60 bl s⁻¹ for sea trout. Atlantic salmon were recorded 9, 48 and 77 km from the river mouth on average 28, 65 and 83 h after release, respectively. Sea trout were recorded 9 km from the release site 438 h after release. Only four (23%) sea trout were detected in the outer part of the fjord system, while the rest of the fish seemed to stay in the inner fjord system. The Atlantic salmon stayed for a longer time in the inner part than in the outer parts of the fjord system, but distinct from sea trout, migrated through the whole fjord system into the ocean.     

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 water activity and nutrients on growth and production of squalestatin S1 by a Phoma sp

D. ALDRED, N. MAGAN and B.S. LANE.1999.This study investigated the effects of temperature, nutrient status and water activity (a_W) on the production of squalestatin S1 by a Phoma sp. The fungus was grown on malt extract (MEA), wheat extract (WEA), oat extract (OEA) and oil seed rape extract (OSREA) agars at 15, 20 and 25 °C and 0·998, 0·995, 0·990, 0·980 and 0·960 a_W levels. The growth rate and secondary metabolite formation were followed over a total of 30 d. The maximum growth rate was observed at 25 °C and 0·998–0·990 a_W for all media types, which was significantly reduced ( P = 0·05) for most media at 0·96 a_w. The growth rate was greatest for WEA and OEA but the growth form was an effuse exploitative type compared with the dense assimilative type on the richer MEA. The lipid-based OSREA appeared to be a poor growth substrate for this fungus. In contrast to the growth rate data, squalestatin S1 production was maximal for all media types at slightly reduced a_w in the range 0·990–0·980. There was greater production of the secondary metabolite under significant water stress (0·960 a_W) compared with that with freely available water (0·998 a_W). Maximum production was observed in WEA. Production began earlier in WEA and OEA compared with MEA. Squalestatin S1 production was not significantly affected by incubation temperature ( P = 0·05). This study has shown that nutritionally depleted substrates may be usefully employed in the production of squalestatin S1 and perhaps also for other secondary metabolites.     

Evaluation of PCR assays for quantifying seed-borne infection by Fusarium and Microdochium seedling blight pathogens

Aims:  To evaluate competitive PCR assays for quantifying seed-borne Microdochium and Fusarium seedling blight pathogen DNA and to determine test and year repeatability and sources of variability.
Methods and Results:  Relationships between DNA and plate counts were significant for Fusarium and Microdochium seedling blight pathogens in 152 seed batches from 3 years. Coefficient of determinations, however, differed greatly ( Fusarium ; R ² = 0·25, P  =   0·029, Microdochium ; R ² = 0·73, P  <   0·001). Significant differences between years were observed in the regression slopes for Microdochium . Pathogen DNA quantified in 16 extractions after sampling was highly correlated to results following storage for 1–2 years ( R ² > 0·90). Residual maximum likelihood analysis showed that the least and greatest variance components of the testing procedure were DNA extraction subsampling and PCR assay respectively.
Conclusions:  Amount of pathogen DNA is a useful estimator of seed batch contamination for Microdochium but not Fusarium seedling blight pathogens. Although reproducible over time, improvements to the testing procedure should focus on repeated PCR amplifications to reduce assay variability.
Significance and Impact of the Study:  Replacing plate counts with competitive PCR for determining the severity of seed batch contamination is feasible in areas where Microdochium seedling blight pathogens predominate.     

Ontogenetic changes in temperature preference of Atlantic cod

Final thermal preferendum ( T ) experiments were conducted in a horizontal thermal gradient tank from the beginning of August 2001 to mid‐November 2001 using Atlantic cod Gadus morhua from 6·5 to 79·0 cm fork length ( L _F). The value of T varied significantly ( P  < 0·005) with L _F( T  = 7·23–0·054 L _F), with smaller (younger) fish choosing higher temperatures than larger (older) fish. The preferendum varied from 6·9° C for fish of 6·5 cm to 3·0° C for those of 79·0 cm. Experiments comparing fish positions in the gradient tank between thermal gradients of 0·5–11·0 and 4·5–14·5° C demonstrated that fish positions were determined by temperature selection instead of undesirable tank effects. This study is the first to demonstrate the effect of ontogeny on temperature preferences of a marine fish species.     

Big and beautiful? Quantitative genetic parameters for appearance of large rainbow trout

A quantitative genetic analysis of body condition, body shape, skin colour and spottiness of large, farmed rainbow trout Oncorhynchus mykiss showed that the traits can be modified through selective breeding. This was indicated by their high heritabilities ( h ² = 0·46–0·61 on the underlying liability scale and 0·29–0·50 on the observed scale). Correlations calculated using linear models showed that skin colour and the amount of spots displayed positive phenotypic ( r _P = 0·33) and genetic correlations ( r _A = 0·83), the relationship being advantageous for the genetic improvement of the traits. Body shape and condition factor displayed disadvantageous correlations with body mass at both ages, the genetic correlations between the traits ranging from 0·36 to 0·57. It was concluded that there are no strong genetic constraints for the genetic improvement of appearance, the only limitation being that rapidly growing fish become rotund.     

Energy density of anchovy Engraulis encrasicolus L. in the Adriatic Sea

European anchovy Engraulis encrasicolus , with total lengths ranging from 40·0 to 132·5 mm, were sampled during October 2002 and May 2003 in the northern Adriatic Sea in order to estimate their energy densities ( E _D). A highly significant ( P  < 0·001) relationship between E _D(y)(J g⁻¹wet mass) and per cent dry mass ( x ) was found: y  = 321 x  − 3316·9 ( n  = 161, r ² = 0·82).     

Effects of temperature, body size and feeding on rates of metabolism in young‐of‐the‐year haddock

The mean rate of oxygen consumption (routine respiration rate, R _R, mg O₂ fish⁻¹ h⁻¹), measured for individual or small groups of haddock Melanogrammus aeglefinus (3–12 cm standard length, L _S) maintained for 5 days within flow‐through respiratory chambers at four different temperatures, increased with increasing dry mass ( M _D). The relationship between R _R and M _D was allometric ( R _R = α  M ^b ) with b values of 0·631, 0·606, 0·655 and 0·650 at 5·0, 8·0, 12·0 and 15·0° C, respectively. The effect of temperature ( T ) and M _D on mean R _R was described by     indicating a Q ₁₀ of 2·27 between 5 and 15° C. Juvenile haddock routine metabolic scope, calculated as the ratio of the mean of highest and lowest deciles of R _R measured in each chamber, significantly decreased with temperature such that the routine scope at 15° C was half that at 5° C. The cost of feeding ( R _SDA) was c . 3% of consumed food energy, a value half that found for larger gadoid juveniles and adults.     

Post mortem shrinkage of four species of temperate and tropical marine fishes, without freezing or preservation

The present study indicated significant post mortem decreases in length for four Australian teleost species from temperate to tropical habitats: black bream Acanthopagrus butcheri , King George whiting Sillaginodes punctata , summer whiting Sillago ciliata and redthroat emperor Lethrinus miniatus . Shrinkage averaged 5·0 mm after 24 h (range 0–10 mm) for A. butcheri , 4 mm (0–8 mm) for S. punctata , 2·2 mm (0·7–3·7 mm) for S. ciliata and 5·4 mm (−2·5–15·0 mm) for L. miniatus . For A. butcheri held under three different post mortem treatments, the mean length of fish in all treatments decreased but there was no significant effect of treatment type on the extent of shrinkage. The rate of shrinkage of S. ciliata varied with treatment, but the ultimate extent did not. For A. butcheri shrinkage was most rapid (2·5 mm h⁻¹) between 1 and 2 h postcapture. The results from these studies confirm that a post mortem decrease in length is a common phenomenon, even in fishes that are not frozen or preserved. Such shrinkage has implications for the enforcement of minimum legal length legislation, and may cause bias in biological investigations, including growth estimates from tag and recapture studies.     

Haematology and leucocyte morphology of wild caught Thunnus maccoyii

The haematology of wild southern bluefin tuna Thunnus maccoyii was described using blood samples collected from fish immediately after they were caught. Cytology and cytochemistry revealed that the blood in peripheral circulation is comprised of erythrocytes, reticulocytes, ghost cells, lymphocytes, thrombocytes, eosinophilic granulocytes, neutrophilic granulocytes and monocytes. Reference ranges established were 41·09–55·50% for haematocrit, 0·62–3·00% for leucocrit, 13·25–17·92 g dl⁻¹ for haemoglobin and 2·1–2·9 million erythrocytes μl⁻¹ for erythrocyte count. Differential cell counts showed 94·58 ± 2·15% erythrocytes, 3·99 ± 1·44% leucocytes and 1·43 ± 1·03% thrombocytes (mean ±  s . d .). Normal ranges for differential leucocyte counts were 0·00–5·45% for neutrophils, 0·69–12·06% for eosinophils, 0·00–5·03% for monocytes, 46·97–74·32% for lymphocytes and 14·47–43·92% for thrombocytes. Erythrocyte indices, leucocyte types and cytochemistry were comparable to other species of scombrids. Packed cell volume was sensitive to the physiological state of the fish and to sample handling technique.