Ozone Disinfection of Eggs from Gilthead Seabream Sparus aurata,Sea Bass Dicentrarchus labrax,Red Porgy,and Common Dentex Dentex dentex

Abstract

The risk of fish pathogen transmission via eggs can be reduced by disinfection in ozonated seawater. The aim of this study was to determine the suitable conditions for ozone disinfection of the eggs of gilthead seabream Sparus aurata, sea bass Dicentrarchus labrax, red porgy Pagrus pagrus, and common dentex Dentex dentex. The eggs were disinfected with a concentration (C) of 0.5 mg of ozone/L of water at four different exposure times (T = 2, 4, 8, and 16 min). The hatching rate was determined in triplicate for each treatment. Bacterial colonies were counted on tryptic soy agar and thiosulfate–citrate–bile salts–sucrose agar. At the end of the experiment, bacterial load and hatching rate were assessed together to determine the optimal ozone treatment values, which were estimated in CT units (i.e., C [= 0.5 mg/L] × T [min]). Optimal values were CT 2–4 (T = 4–8 min; 18°C) for gilthead seabream and red porgy, CT 2 (T = 4 min; 18°C) for common dentex, and CT 4 (T = 8 min; 15°C) for sea bass.

Received April 12, 2011; accepted February 2, 2012     

Virulence and enterobacterial repetitive intergenic consensus PCR of Vibrio alginolyticus strains isolated from Tunisian cultured gilthead sea bream and sea bass outbreaks

Vibrio alginolyticus was isolated from the internal organs of diseased gilthead sea bream (Sparus aurata) and sea bass (Dicentrarchus labrax) cultured in two fish farms located on the Tunisian Mediterranean coast, from 2003 to 2005. After phenotypic characterisation, a selection of 34 isolates from gilthead sea bream and sea bass were molecularly typed by repetitive intergenic consensus PCR (ERIC-PCR) showing a high polymorphism among the isolated strains (19 genotypes). Most of the isolates were resistant to atleast two antimicrobial agents. All the tested strains were resistant to ampicillin. However, 91.17% were resistant to nitrofurantoin and 35.29% to tetracycline. Several strains isolated from diseased gilthead sea bream and sea bass were tested for virulence in both fish species by intraperitoneal injection. The selected isolates (n=7) were pathogenic for gilthead sea bream and sea bass. LD(50) values ranged from 5.01 x 10(4) to 6.20 x 10(7)CFU/fish. This is the first report on characterisation and virulence of V. alginolyticus for sea bass and sea bream in Tunisia.     

Polymerase chain reaction amplification of repetitive intergenic consensus and repetitive extragenic palindromic sequences for molecular typing of Pseudomonas anguilliseptica and Aeromonas salmonicida

The aim of this study was to evaluate the use of two molecular techniques, repetitive extragenic palindromic polymerase chain reaction (REP-PCR) and repetitive intergenic consensus PCR (ERIC-PCR), as epidemiological tools with which to discriminate among genetically distinct strains within two bacterial fish pathogens, Pseudomonas anguilliseptica and Aeromonas salmonicida. A total of 30 A. salmonicida and 52 P. anguilliseptica were analyzed. For P. anguilliseptica, three different major fingerprints were obtained with both techniques, which defined three genomic groups: one was composed of strains isolated from eels Anguilla spp., the second of strains from turbot Scophthalmus maximus and blackspot seabream (also known as red seabream) Pagellus bogaraveo, and the third of strains from other fish species, such as gilthead seabream (also known as gilthead bream) Sparus auratus, sea bass Dicentrarchus labrax (also known as European bass Morone labrax), and salmonids. In the case ofA. salmonicida, promising results were obtained with both techniques for subspecies differentiation. Thus, two genomic profiles were obtained by ERIC-PCR. The first profile consisted of A. salmonicida subsp. salmonicida strains isolated from the different hosts. The second profile was composed of two A. salmonicida subsp. masoucida and one A. salmonicida subsp. achromogenes. Using REP-PCR, three genotypes were obtained within this pathogen that were related to the diverse subspecies analyzed. In summary, both methodologies are useful for typing distinct strains associated with different host species and therefore are helpful in epidemiological studies of P. anguilliseptica. In contrast, in the case of A. salmonicida, more studies are needed to determine their utility in discriminating the subspecies salmonicida from the other two subspecies.     

Muscle Development in Gilthead Sea Bream (Sparus aurata, L.) and Sea Bass (Dicentrarchus labrax, L.): Further Histochemical and Ultrastructural Aspects

The histochemical profiles - mATPase and NADH-TR reactions - of the red and white muscle fibres of gilthead sea bream and sea bass were determined from the first week after hatching. Modifications of the mATPase technique by combinations of pH/time/molarity were carried out in order to compare the sensitivity of the myosin ATPase of each muscle fibre type of the lateral muscle. Results showed that the staining of muscle fibres was independent of small modifications in the technique. The intermediate ‘pink’ muscle was histochemically defined towards the end of the larval life and is considered to be implicated in the growth of the myotome. A layer of external cells was observed, by electron microscopical examination, between the connective tissue of the skin and the superficial red muscle fibres of larvae and postlarvae. It is suggested that the external cells are unlikely to be a source of red muscle fibres and implicated on the growth of the myotome, but rather a part of the dermatome. The timing, areas and mechanisms of hyperplastic growth of the myotome were defined and discussed.     

Preliminary assessment of dietary mannanoligosaccharides on growth performance and health status of gilthead seabream Sparus auratus

A feeding trial was performed to assess the potential beneficial effect of two levels of mannanoligosaccarides (MOS) on the growth performance, feed utilization, hematological parameters, and liver histopathology of gilthead seabream Sparus auratus (also known as gilthead bream). Mannanoligosaccarides were added at the rates of 2 and 4 g/kg to a fish-meal-based control diet, and each diet was given (twice daily [midmorning and midafternoon] to apparent satiation) to triplicate groups of gilthead seabream growers (mean weight = approximately 170 g) in sea cages. The trial lasted 12 weeks, and the average ambient water temperature ranged from 19.6 degrees C to 24.7 degrees C during the experimental period. At the end of the experiment, fish attained market size (350-450 g) and their health status was evaluated by blood analysis and liver histology. There were no differences in survival rates among fish fed experimental diets. However, there were significant improvements in both growth and feed utilization among fish fed diets supplemented with MOS. Hemoglobin (Hb) and hematocrit (Ht) levels and erythrocyte, leukocyte, and thrombocyte (Thr) counts were unaffected by any dietary MOS. The levels of Hb (g/dL; mean +/- SD) and Ht (%; mean +/- SD) were 11.0 +/- 2.5 and 45.6 +/- 6.7 for the control group, 11.1 +/- 1.7 and 39.3 +/- 8.0 for the 2-g/kg group, and 11.2 +/- 1.9 and 40.2 +/- 8.4 for the 4-g/kg group. The mean Thr count ranged from 47.6 to 53.8 x 10(3)/mm3. Despite the apparently higher Thr counts for fish fed diets supplemented with MOS, these differences were not significant. Moreover, no histopathological differences were observed in liver tissue cross sections between control and treatment groups. These results suggest that supplementation of diets with MOS had no significant effects on general fish health.     

Effects of temperature and relative humidity on development and survival of the free-living stages of Trichostrongylus colubriformis, T. rugatus and T. vitrinus

Temperatures of 4 and 10 degrees C reduced the rate of hatching of eggs and the development of the pre-infective stages of T. colubriformis and T. rugatus, but not T. vitrinus. Eggs of T. rugatus hatched at 4 degrees C, but did not develop to the infective stage. At 20 and 30 degrees C, no differences were detected between species in egg hatching or the development of infective larvae. Third-stage larvae of T. rugatus survived for significantly longer periods of time at 33 and 56% relative humidities at a temperature of 20 degrees C than did either T. colubriformis or T. vitrinus. At 30 degrees C, differences were less marked, but T. rugatus and T. colubriformis survived longer than T. vitrinus. The results are discussed in relationship to the known distribution of these species in eastern Australia.     

Effects of ozone and ultraviolet radiation treatments on the infectivity of Toxoplasma gondii oocysts

Clinical toxoplasmosis in humans has been epidemiologically linked to the consumption of drinking water contaminated by Toxoplasma gondii oocysts. We evaluated killing of T. gondii oocysts after ultraviolet (UV) or ozone treatments by bioassay in mice and/or cell culture. A 4-log inactivation of the oocyst/sporozoite infectivity was obtained for UV fluences >20 mJ cm(-2). In contrast, oocysts were not inactivated by ozone with an exposure (Ct) up to 9.4 mg min l (-1) in water at 20 degrees C. In conclusion, UV treatment can be an effective disinfection method to inactivate T. gondii oocysts in drinking water, but ozone did not show promise in this research.     

Effect of pre-incubation storage conditions on hatchability,chick weight at hatch and hatching time in broiler breeders

1. Eggs were stored for two different times at varying temperatures. The effects on hatchability, chick weight at hatch and hatching time were examined in two broiler breeder lines from 33 to 58 weeks of age. 2. Short storage (1 to 3 d). Storage at 20 degrees C compared with 16.5 degrees C reduced hatchability of all eggs set. No effect was observed on hatchability of fertile eggs, hatching time or chick weight. 3. Long storage (9 to 11 d). Storage at 16.5 degrees C compared with 10 degrees C decreased both hatchability of fertile eggs and chick weight at hatch. Incidence of early embryonic death increased and incubation time decreased at 16.5 degrees C compared with 10 degrees C. 4. Chicks from morning eggs were heavier than those from afternoon eggs irrespective of storage conditions. 5. Hatchability (all eggs set and fertile eggs) and chick weight varied with hen age irrespective of storage conditions. During long storage, hatching time varied with hen age independently of breeder line, storage temperature or egg laying time. 6. Hatchability (all eggs set and fertile eggs) was higher in line A than in line B. Line B eggs hatched later and produced heavier chicks than line A eggs irrespective of storage time.     

Effective,appropriate and simple culture,egg hatching and cryopreserving of the nematode Cheilospirura hamulosa

1. Successful invasion by nematode parasites is associated with several factors including egg hatching at the right time in their hosts. To determine a simple and appropriate medium for culture and egg hatching of the highly pathogenic species of the Acuariidae family, Cheilospirura hamulosa were cultured in three different media. In addition the viability of C. hamulosa eggs was determined after storage in frozen infected gizzards.

2. Eggs removed from the uteri of the female worms in infected gizzards were pooled and washed in distilled water and screened under a stereo dissecting microscope. Eggs were counted and cultured in three different media, nutrient agar, normal saline 0.9% and Bearman, at room temperature. Additionally, 10 infected gizzards were kept at ?20°C for 2 and 8 months.

3. After 4–5 d there had been no growth in the nutrient agar medium, whereas 11% of the cultured eggs in the Bearman medium contained larvae 2–3 d after culturing. In 0.9% normal saline medium the two polar knobs appeared on the two poles of the eggs at 2 d post cultivation, and 74% of the eggs contained a larva on the third day. Mature larvae gradually exited from the eggs.

4. Eggs collected from female worms in gizzards frozen at ?20°C were cultured in the same three culture media at room temperature. Larvae were visible in the eggs after 2–3 d in the Bearman and 0.9% normal saline media and hatched thereafter.

5. The 0.9% normal saline medium is recommended for egg hatching and cultivation of C. hamulosa due for simplicity, efficacy and cost effectiveness. Moreover, freezing of the infected gizzards at ?20°C is proposed for long-term storage of the eggs.     

The Post-larval Development of Lateral Musculature in Gilthead Sea Bream Sparus aurata (L.) and Sea Bass Dicentrarchus labrax (L.)

Fibre-type differentiation of lateral musculature has been studied in gilthead sea bream Sparus aurata (L.) and sea bass Dicentrarchus labrax (L.) during post-larval development using ultrastructural, histochemical and morphometric techniques. The study showed three muscle layers: red, intermediate (or pink) and white. Initially, most of the red muscle showed low myosin ATPase (m-ATPase) activity fibres, whereas near the transverse septum some small high m-ATPase activity fibres appeared and later acquired a rosette aspect. Afterwards, during adult growth the red muscle showed a histochemical mosaic appearance. The pink muscle in sea bass was observed at the beginning of juvenile development by the oxidative technique (NADH-RT reaction) whereas in gilthead sea bream it was also observed at the end of larval development. The pink layer consists of high m-ATPase activity fibres. However, along the muscle development other low and moderate m-ATPase activity fibres were observed close to the red and white muscles, respectively. The white muscle of juvenile fish showed a histochemical mosaic appearance near the pink muscle. In adult specimens the mosaic white muscle spread out occupying the whole of the myotome. Morphometric analysis shows a significant increase in mean fibre diameter during post-larval development, as shown by the Student's t-test (hypertrophic growth). Skewness and kurtosis values of fibre diameters point to the generation of a new fibres from the myosatellite cells (hyperplastic growth).     

Insulin,IGF-I,and muscle MAPK pathway responses after sustained exercise and their contribution to growth and lipid metabolism regulation in gilthead sea bream

Herein, we studied whether sustained exercise positively affects growth of gilthead sea bream by alterations in a) plasma concentrations of insulin and IGF-I, b) signaling pathways in muscle, or c) regulation of lipid metabolism. Specifically, we evaluated the effects of moderated swimming (1.5 body lengths per second; BL/s) on the circulating concentrations of insulin and IGF-I, morphometric parameters, and expression of genes related to lipid metabolism in gilthead sea bream (80–90 g BW). Exercise increased the specific growth rate (P < 0.05) and reduced the hepatosomatic index (P = 0.006). Plasma IGF-I concentrations increased in exercised fish (P = 0.037), suggesting a role for this endocrine factor in the control of muscular growth and metabolic homeostasis during swimming. The observed decrease in plasma insulin concentrations (P = 0.016) could favor the mobilization of tissue reserves in exercised fish. In this sense, the increase in liver fatty acid content (P = 0.041) and the changes in expression of peroxisome proliferator-activated receptors PPARα (P = 0.017) and PPARγ (P = 0.033) indicated a hepatic lipid mobilization. Concentration of glycogen in both white and red muscles was decreased (P = 0.021 and P = 0.017, respectively) in exercised (n = 12) relative to control (n = 12) gilthead sea bream, whereas concentrations of glucose (P = 0.016) and lactate (P = 0.0007) were decreased only in red muscle, indicating the use of these substrates. No changes in the glucose transporter and in lipoprotein lipase mRNA expression were found in any of the tissues studied. Exercised sea bream had decreased content of PPARβ mRNA in white and red muscle relative to control sea bream expression (P = 0.001 and P = 0.049, respectively). Mitogen-activated protein kinase phosphorylation was significantly down-regulated in both white and red muscles of exercised sea bream (P = 0.0374 and P = 0.0371, respectively). Tumor necrosis factor-α expression of white muscle was down-regulated in exercised gilthead sea bream (P = 0.045). Collectively, these results contribute to the knowledge base about hormonal regulation of growth and lipid metabolism in exercised gilthead sea bream.     

Effect of two thermal regimes on the muscle growth dynamics of sea bass larvae,Dicentrarchus labrax L

Muscle growth was studied in larvae of sea bass, Dicentrarchus labrax L., reared at two temperatures: real ambient temperature ( congruent with 15 degrees C during vitelline phase and increased gradually) and 19 degrees C from fertilization until the end of larval development. Muscle cellularity, body length and body weight were measured. Early temperature influenced larval development and so, pre-larval phase finished earlier at 19 degrees C than at ambient temperature (4 and 6 days, respectively). Temperature also affected muscle growth such that at hatching and at mouth opening hypertrophy of muscle fibres was greater at 19 degrees C (P < 0.05), whereas hyperplasia was similar in both groups. After 25 days, the cross-sectional area of the white muscle was greater at 19 degrees C (P < 0.05), which was mainly associated with a higher proliferation of new white muscle fibres. At this stage the body length was also higher at 19 degrees C. Metamorphosis finished earlier in fish reared at 19 degrees C (52 days) than at natural temperature (82 days). At this developmental stage body length and cross-sectional area of the myotome were similar in both groups. However, muscle cellularity differed between groups. Thus, hypertrophy of muscle fibres was higher in fish reared at ambient temperature (P < 0.05), whereas proliferation of new muscle fibres was higher at 19 degrees C (P > 0.05).     

Influence of Slaughtering Method on Some Aspects of Quality of Gilthead Seabream and Smoked Rainbow Trout

The effects of several slaughter methods on the quality of fresh and smoked trout and fresh gilthead seabream were evaluated during storage at 2^°C. Electrically stunned trout had slower ATP depletion of raw muscle and lower lipid oxidation in smoked product during storage. Gilthead seabream immersed in an ice slurry (IS group) after the harvest showed a more regular ATP depletion than in fish exposed to CO₂. Nevertheless, in the case of the IS group, self-initiated behaviour, response to handling and breathing all ceased only after 15–20 min, whereas carbon dioxide-stunned fish appeared dead after 5 min. However, gilthead seabream group having slower ATP depletion also showed lower lipid oxidation of muscle during storage. In both species this could be due to the rapid conversion of xanthine dehydrogenase to xanthine oxidase induced by the rapid consumption of ATP. Xanthine oxidase, in the presence of redox iron and reintroduced oxygen, can produce hydrogen peroxide and, consequently, hydroxyl radicals.     

Immersion booster vaccination effect on sea bass (Dicentrarchus labrax L.) juveniles

In each challenge 30 sea bass juveniles (mean weight 3.3 +/- 0.2 g SD) were used. During the whole experiment (water T: 18 +/- 1 degrees C) the fish were held in four 50l seawater independent recirculation systems (one fish group per 50l system). The protection to the pathogen Vibrio anguillarum was tested on booster vaccinated sea bass (Dicentrarchus labrax L.) juveniles. The vaccination was performed by immersion for 60 s in a commercial anti-V. anguillarum vaccine suspension. Booster vaccination took place 60 days after the initial immunization. Thirty days after the booster vaccination all the fish received intraperitoneally (IP) 3.0 x 10(6) cfu/fish (colony forming units) virulent V. anguillarum bacteria. The booster vaccination showed a strong protection effect on the challenged sea bass. In the next 20 days after the challenge the mortality was 0% among the booster vaccinated sea bass, 10% among the once vaccinated fish and 50% in the control group (unvaccinated fish). No mortality was observed among the unvaccinated sea bass injected IP with sterile normal saline by the challenge.     

Red muscle development of gilthead sea bream Sparus aurata (L.): structural and ultrastructural morphometry

The transverse red and white muscle area, the superficial red muscle fibres area and their percentages of mitochondria (%mit), myofibrils (%myof) and sarcoplasm (%sarc) were determined in the Mediterranean teleost gilthead sea bream, Sparus aurata (L.). Fish aged from hatching to 78 days were studied. The proportional growth of the red and white muscles was higher for the red muscle in the first half of the larval stage (1-35 days). Then the opposite relationship was observed. The hypertrophic growth of the superficial muscle fibres was continuous except in the first week after hatching. The percentage of mitochondria and percentage of myofibrils showed a significant change just after the half of the larval stage. Whereas the %mit of the superficial muscle fibres was higher than %myof from the first week after hatching to 35 days (average 66.64%), then the %mit decreased significantly and at 73-78 days both parameters were close to an average value of 50%. The meaning of these morphological changes is discussed in relation to the functional role of the red muscle of larvae and the onset of the gills respiration.     

Caligus sclerotinosus (Copepoda: Caligidae), a serious pest of cultured red seabream Pagrus major (Sparidae) in Korea

Caligid copepods (Crustacea) known as sea lice are pests of cultured fish, causing serious diseases and economic losses in fish aquaculture worldwide. One species, Caligus sclerotinosus Roubal, Armitage & Rohde, 1983 (Caligidae), is considered a serious pest of the highly prized red seabream Pagrus major (Temminck and Schlegel, 1843) (Sparidae) cultured in Japan. Recently, in neighboring Korea, red seabream culture has intensified and almost replaced yellow tail culture. However, until now, there have been no reports on infection of this sea louse from red seabream in Korea. We surveyed 120 (20 fish per month) P. major from a sea ranched Tongyeong Marine Research Center aquaculture facility, Gyeongsangnamdo, Korea for six months in 2011 (June to November). We recorded severe infection by the sea louse C. sclerotinosus on the skin of P. major. Prevalence was 100%, mean intensity 7.06, maximum intensity 49, and minimum intensity 2. Adult females (624), males (219) and few chalimi (5) were observed and identified by their morphology. As an average of all our collections, less than 0.6% of individuals were chalimi. We suggest, therefore, that adults of C. sclerotinosus undergo ontogenetic host switching after their final moult. No infection of C. sclerotinosus was found on wild P. major collected from Tongyeong and Yeosu fish markets on the southern coast of Korea. Severe infection by this sea louse may cause secondary infections of the host. This copepod is already reported from Australia and Japan and hence, this is the first report from Korea. We expect this pest to have an impact on Korean red seabream fisheries equally serious to that being experienced in Japan.     

Preliminary Assessment of Dietary Mannanoligosaccharides on Growth Performance and Health Status of Gilthead Seabream Sparus auratus

Abstract

A feeding trial was performed to assess the potential beneficial effect of two levels of mannanoligosaccarides (MOS) on the growth performance, feed utilization, hematological parameters, and liver histopathology of gilthead seabream Sparus auratus (also known as gilthead bream). Mannanoligosaccarides were added at the rates of 2 and 4 g/kg to a fish-meal-based control diet, and each diet was given (twice daily [midmorning and midafternoon] to apparent satiation) to triplicate groups of gilthead seabream growers (mean weight = approximately 170 g) in sea cages. The trial lasted 12 weeks, and the average ambient water temperature ranged from 19.6°C to 24.7°C during the experimental period. At the end of the experiment, fish attained market size (350–450 g) and their health status was evaluated by blood analysis and liver histology. There were no differences in survival rates among fish fed experimental diets. However, there were significant improvements in both growth and feed utilization among fish fed diets supplemented with MOS. Hemoglobin (Hb) and hematocrit (Ht) levels and erythrocyte, leukocyte, and thrombocyte (Thr) counts were unaffected by any dietary MOS. The levels of Hb (g/dL; mean ± SD) and Ht (%; mean ± SD) were 11.0 ± 2.5 and 45.6 ± 6.7 for the control group, 11.1 ± 1.7 and 39.3 ± 8.0 for the 2-g/kg group, and 11.2 ± 1.9 and 40.2 ± 8.4 for the 4-g/kg group. The mean Thr count ranged from 47.6 to 53.8 × 10³/mm³. Despite the apparently higher Thr counts for fish fed diets supplemented with MOS, these differences were not significant. Moreover, no histopathological differences were observed in liver tissue cross sections between control and treatment groups. These results suggest that supplementation of diets with MOS had no significant effects on general fish health.

Received February 20, 2011; accepted September 12, 2011     

Effect of heat-stress on bovine embryo development in vitro.

Chronic elevation of uterine temperature has long been known to increase embryo mortality in dairy cattle. Short-term elevation in temperature of mouse embryos to 43 degrees C (acute) has been shown to induce intracellular production of heat-shock proteins. In this study, in vitro development of bovine embryos was assessed during short-term (60 h) coculture with oviduct epithelial cells at 38.6 degrees C (T1), 40 degrees C (T2), 38.6 degrees C after a prior pulse treatment (20 min) at 43 degrees C with 5% CO2 (T3), or 38.6 degrees C after a prior pulse treatment (20 min) at 43 degrees C with 100% CO2 (T4). During incubation, embryos cocultured at 40 degrees C had a greater (P < .05) mean embryo development score at 36 h than embryos cocultured at 38.6 degrees C. At 60 h of incubation, embryo development scores were greater (P < .05) for embryos cultured at 38.6 degrees C than for those cocultured at 40 degrees C. The number of embryos hatched at 60 h was similar after coculture at 38.6 degrees C (T1) or a prior pulse treatment with 5% CO2 and 43 degrees C (T3), but the embryo development score at 60 h was greater (P < .05) for the pulse-treated embryos. Embryos in T4 had greater (P < .05) embryo development scores than did T1 embryos from 36 through 60 h. Pulse treatment (T4) resulted in a greater (P < .05) number of hatched embryos at 60 h than T1, T2, and T3. These results indicate a detrimental effect of a chronic elevation in temperature that was evident shortly after embryo hatching.(ABSTRACT TRUNCATED AT 250 WORDS)     

Kinetics of the inactivation of Salmonella during thermal disinfection of liquid manure]

Technical processes for thermal liquid manure disinfection usually reach temperatures between 50 degrees C and 70 degrees C. The destruction of important infectious micro-organisms can be expected in this temperature range. The purpose of the investigations reported here was to study the thermal resistance of Salmonellae during heat treatment of liquid manure. Salmonella senftenberg 775W survived much longer than each of 12 other strains from 8 different Salmonella serovars. Resulting from a regression analysis kinetics of thermal death were determined for this strain and decimal reduction times were calculated in cases of an exponential die-off. D60-values ranged from 47 to 138 sec depending on the type of slurry used. Further investigations on the inactivation of Salmonella senftenberg 775W at 50 degrees C, 55 degrees C, 60 degrees C, and 65 degrees C were carried out and following D-values were obtained: D50 = 56.7 min, D55 = 11.5 min, D60 = 2.3 min, D65 = 0.47 min. The resulting ZD-value was 7.2 degrees C. Minimum requirements concerning temperature and heating time can be derived from the results of this study. The given recommendations may only be applied, if technical processes work without any functional deficiencies and thermal energy is evenly distributed in the heated slurry. Combinations of temperature and heating time should not fall below the following values: 50 degrees C/15 h, 55 degrees C/3 h, 60 degrees C/30 min, 65 degrees C/5 min.     

Early development of artificially spawned southern mullet,Liza richardsonii (Smith, 1846)

The embryonic development of artificially spawned southern mullet, Liza richardsonii, eggs and the development of the larvae to 46 days are described and illustrated using drawings and photographs. The floating eggs hatched in sea water at 18–24°C after 46–60 h. Newly stripped eggs usually had more than one oil droplet (up to 16) which merged during development. Free embryos averaged 2,36 mm at hatching and had distinct yellow pigment.