Effects of feeding,starvation, and refeeding on the fatty acid composition of channel catfish,ICtalurus punctatus,tissues

• 1.1. The effects of feeding, food deprivation (14 and 28 days) and refeeding (starved 14 then fed 14 days) on the fatty acid composition of white muscle, liver and brain of pond-raised channel catfish (Ictalurus punctatus) were investigated.
• 2.2. Levels of n-3 fatty acids were significantly higher (P < 0.05) in white muscle of fish starved 28 days (10.7%) than in fish fed throughout the study (8.0%), due primarily to an increase in 22:6(n-3) docosahexaenoic acid or DHA.
• 3.3. Significantly higher levels of 20:5(n-3) (eicosapentaenoic acid or EPA) were found in livers offish starved 28 days (P < 0.05) compared to fish fed throughout the study.
• 4.4. Results suggest that the fatty acid compositions of channel catfish white muscle and liver are subject to only limited perturbation during periods of starvation and refeeding and that the brain is extremely well protected.

     

Mode of Association,Enzyme Producing Ability and Identification of Autochthonous Bacteria in the Gastrointestinal Tract of Two Indian Air-Breathing Fish,Murrel (<Emphasis Type="Italic">Channa punctatus</Emphasis>) and Stinging Catfish (<Emphasis Type="Italic">Heteropneustes fossilis</Emphasis>)

The mode of association of epithelium-associated bacteria in the gastrointestinal (GI) tract of two Indian air-breathing fish species, the murrel, Channa punctatus and the stinging catfish, Heteropneustes fossilis was demonstrated through scanning and transmission electron microscopy (SEM and TEM). The SEM examination revealed substantial numbers of rod shaped bacterial cells associated with the microvillus brush borders of enterocytes in proximal (PI) and distal regions (DI) of the GI tract of both the fish species. The TEM investigation indicated endocytosis and translocation of bacteria in the microvilli. The isolated bacterial strains (two each from the PI and DI of murrel and stinging catfish) were quantitatively evaluated for their extracellular amylase, cellulase and protease production. All the bacterial strains exhibited high cellulolytic activity than that of amylolytic and proteolytic enzymes. Only two strains, CPF1 and CPF2, isolated from the PI of murrel exhibited high proteolytic activity. Maximum amylase activity was exhibited by the strain, HFH5, isolated from the DI of stinging catfish. Totally six most promising enzyme-producing autochthonous bacterial strains were identified based on partial 16S rRNA gene sequence analytical results. All the strains showed close (92–99 %) similarity to Bacillus licheniformis.     

Fillet yield and chemical composition of farm-raised sunshine bass (Morone chrysops♀ × Morone saxatilis♂) fed high-energy diets

Fillet yield and chemical composition were evaluated in small, medium and large (about 150, 350 and 700 g, respectively) size classes of sunshine bass ( Morone chrysops ♀ ×  Morone saxatilis ♂) fed high-energy diets. Fish size had a major effect on the fillet yield, which was significantly higher (38.8 vs. 33.4–34.6%) in fish belonging to the medium-size class. The lipid contents of both whole and skinned fillets were higher than those reported in the literature, ranging from 7.8 to 10.6% in whole fillets and from 5.9 to 7.6% in skinned fillets, thereby allowing classification of sunshine bass fed high-energy diets as a medium-fat fish. A significant effect of fish size on lipid content was found only for whole fillets, therefore indicating that dietary energy excess in larger fish is mainly stored as subdermal fat. The dietary fatty acid profile was found to be the major determinant of edible muscle lipid composition in all size-classes considered. The cholesterol content of muscle tissue ranged from 23 to 29 mg/100 g tissue.     

MtDNA diversity of the critically endangered Mekong giant catfish (Pangasianodon gigas Chevey, 1913) and closely related species: implications for conservation

Catfishes of the family Pangasiidae are an important group that contributes significantly to the fisheries of the Mekong River basin. In recent times the populations of several catfish species have declined, thought to be due to overfishing and habitat changes brought about by anthropogenic influences. The Mekong giant catfish Pangasianodon gigas Chevey, 1913 is listed as Critically Endangered on the IUCN Red List. In the present study, we assessed the level of genetic diversity of nine catfish species using sequences of the large subunit of mitochondrial DNA (16S rRNA). Approximately 570 base pairs (bp) were sequenced from 672 individuals of nine species. In all species studied, haplotype diversity and nucleotide diversity ranged from 0.118±0.101 to 0.667±0.141 and from 0.0002±0.0003 to 0.0016±0.0013, respectively. Four haplotypes were detected among 16 samples from natural populations of the critically endangered Mekong giant catfish. The results, in spite of the limited sample size for some species investigated, indicated that the level of genetic variation observed in wild populations of the Mekong giant catfish (haplotype diversity=0.350±0.148, nucleotide diversity=0.0009±0.0008) is commensurate with that of some other related species. This finding indicates that (1) wild populations of the Mekong giant catfish might be more robust than currently thought or (2) present wild populations of this species carry a genetic signature of the historically larger population(s). Findings from this study also have important implications for conservation of the Mekong giant catfish, especially in designing and implementing artificial breeding programme for restocking purposes.     

Correlation between body color and behavior in the upside-down catfish,Synodontis nigriventris

• 1.1. Unlike common fishes and as its Latin name implies, the upside-down catfish, Synodontis nigriventris, possesses dark ventral skin. Microscopic observation reveals that melanophores are present on both the ventral and the dorsal skin but differ in size and density of distribution.
• 2.2. The darkness of both sides of the fish changes in accordance with that of the background.
• 3.3. At night, the fish are very active and the body becomes pale. The change in color is more noticeable in the dorsal than the ventral skin.
• 4.4. When melatonin was added to the bathing water, the fish became pale and swam restlessly even when they were exposed to the black background.
• 5.5. It was found that the catfish preferred the black background to the white one.

     

氧化油脂对斑点叉尾鮰生长、体色和抗氧化能力的影响

为考察正常及氧化鱼油、豆油、混合油脂(鱼油﹕豆油=1﹕1)对斑点叉尾鮰(Ictalurus punctatus)生长性能、体色和肝脏抗氧化能力的影响, 设计6组等氮等脂饲料, 分别添加6%鱼油、6%豆油、3%鱼油+3%豆油、6%氧化鱼油、6%氧化豆油、3%氧化鱼油+3%氧化豆油, 饲喂6组初始体重(150.5±4.2 g)的斑点叉尾鮰8周, 每组3个重复, 每个重复14尾鱼。结果表明, 摄食6%鱼油、6%豆油、3%鱼油+3%豆油饲料的斑点叉尾鮰在增重率、饲料系数、肌肉组成、体色和肝脏抗氧化指标上均无显著差异(P>0.05); 摄食3组氧化油脂饲料后, 鱼体增重率和肌肉粗脂肪含量降低(P<0.05), 饲料系数和背部、尾部皮肤b*(黄度值)增加(P<0.05), 肝脏丙二醛(MDA)和血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(TBIL)和直接胆红素升高(DBIL)(P<0.05), 肝脏超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、还原型谷胱甘肽降低(GSH)(P<0.05), 而肌肉色度值(L*、a* 、b*)和肌肉水分、粗蛋白质、灰分含量无显著差异(P>0.05)。以上结果表明, 6%氧化油脂(鱼油、豆油或混合油脂)导致斑点叉尾鮰生长性能下降、皮肤黄度增加、肝脏抗氧化能力受损; 豆油可替代斑点叉尾鮰饲料中鱼油的使用, 而不会对生长产生负面影响。     

A dietary dairy/yeast prebiotic and flaxseed oil enhance growth,hematological and immunological parameters in channel catfish at a suboptimal temperature (15°C)

Channel catfish raised in the southern United States require two growing seasons to reach market size. Growing seasons are separated by a cool period of about 3 months when feed intake and growth are greatly reduced. A cool-weather feeding strategy to improve feed intake, growth or health of catfish might improve survival and reduce the time needed to achieve market size. We conducted a feeding trial with channel catfish at a suboptimal temperature (15°C) to determine the effects of supplementing diets with either a dairy/yeast prebiotic or flaxseed oil (high in 18:3n-3) compared with a control with soybean oil (high in 18:2n-6). The trial was conducted in recirculating systems with 1140-l tanks containing 100 fish each (mean initial weight 61.4 g±0.43 s.e.m.). A 28%-protein basal diet was supplemented with 20 g/kg cellulose and 20 g/kg soybean oil (SBO, control), 20 g/kg cellulose and 20 g/kg flaxseed oil (FLAX) or 20 g/kg of a dairy/yeast prebiotic and 20 g/kg soybean oil (PREB). Fish were fed once daily to satiation and weighed every 3 weeks to track growth. Hematology, non-specific immune responses, proximate and fatty acid composition of muscle were determined to assess diet effects. Catfish-fed FLAX or PREB had higher weight gain, feed consumption and lysozyme activity than fish fed SBO. Total n-3 fatty acids in muscle were higher in fish fed SBO or FLAX than those fed PREB. Total n-6 long-chain polyunsaturated acids were higher in muscle of fish fed PREB than those fed SBO. Fatty acids in the PREB and SBO diets were similar, so the PREB appeared to increase elongation and desaturation of n-6 fatty acids in muscle. Flaxseed oil and the dairy/yeast prebiotic both have potential to increase catfish performance at a low temperature.     

Improving of lipid productivity of the biodiesel promising green microalga <Emphasis Type="Italic">Chlorella pyrenoidosa</Emphasis> via low-energy ion implantation

High lipid content in microalgae is an essential parameter for adopting of microalgal biomass as a feedstock for biodiesel. Mutation is one approach to obtain desired algal strain with high lipid production. In this study, a mutant strain of Chlorella pyrenoidosa was isolated using 1.5?×?10¹⁵ ions cm^?2 s^?1 of N⁺ ion beam implantation technique, which has been widely used in mutagenesis of agricultural crops. N⁺ implantation slightly improved the growth of the mutant over the corresponding wild strain with significant increase in lipid content (32.4 % higher than the wild strain), which resulted in significant increase in lipid productivity by 35 %. In addition, ion implantation mutagenesis of C. pyrenoidosa resulted in 21.4 % decrease in total saturated fatty acids (SFAs) compared to the wild type, with a noticeable increase in polyunsaturated fatty acids (PUFAs). The increase in PUFAs was due mainly to stimulation of hexadecadienoic acid (C16:2) and octadecadienoic acid (C18:2) production. However, the SFA content of wild and mutant strains was 31.7 and 24.9 % of total fatty acids, respectively, highlighting the oxidative stability of biodiesel produced by both strains according to the European standards. Cultivation of C. pyrenoidosa mutant in selenite enrichment medium for five successive cultivation experiments showed insignificant changes in biomass productivity, lipid content, and lipid productivity alongside the study period, which confirms the genetic stability of the produced mutant. The present study confirmed the feasibility of generation of microalgae mutants with significant high lipid production using ion beam implantation.     

A laboratory study of host use by the cuckoo catfish <Emphasis Type="Italic">Synodontis multipunctatus</Emphasis>

The only known non-avian vertebrate obligate brood parasite is the cuckoo catfish (Synodontis multipunctatus), a Lake Tanganyikan endemic. The cuckoo catfish parasitizes Tanganyikan mouthbrooding cichlids, and under captive conditions, will also parasitize cichlids from other Rift Valley lakes. Here we examine the frequency of parasitism by the cuckoo catfish of Ctenochromis horei from Lake Tanganyika and three species from Lake Malawi and the greater Lake Victorian system in a laboratory setting. C. horei was parasitized significantly less (17%) than the allopatric species Haplochromis latifasciatus, Haplochromis nubilus, and Metriaclima estherae (combined parasitism rate of 28%). The lower rates of parasitism in C. horei may be due to differences in the mating ritual, oviposition (e.g., long periods of pseudo-spawning before actual oviposition), and behavioral adaptations (e.g., increased aggression towards the cuckoo catfish). The number of catfish eggs per parasitized brood was similar between C. horei, H. latifasciatus, H. nubilus, and M. estherae. Our results are comparable to findings from the field for C. horei parasitism frequency and number of cuckoo catfish per brood. We also analyzed the parasitism rate of the albino morph of Metriaclima zebra, a domestic strain. Parasitism rates and number of catfish per brood were the highest in the albino morphotype suggesting that the higher levels of parasitism may be related to lower aggressive behavior, lower visual acuity, or captive influence. Cuckoo catfish and mouthbrooding cichlids provide a model system for testing brood parasitism in a laboratory setting.     

Activation and aponeurosis morphology affect in vivo muscle tissue strains near the myotendinous junction

Hamstring strain injury is one of the most common injuries in athletes, particularly for sports that involve high speed running. The aims of this study were to determine whether muscle activation and internal morphology influence in vivo muscle behavior and strain injury susceptibility. We measured tissue displacement and strains in the hamstring muscle injured most often, the biceps femoris long head muscle (BFLH), using cine DENSE dynamic magnetic resonance imaging. Strain measurements were used to test whether strain magnitudes are (i) larger during active lengthening than during passive lengthening and (ii) larger for subjects with a relatively narrow proximal aponeurosis than a wide proximal aponeurosis. Displacement color maps showed higher tissue displacement with increasing lateral distance from the proximal aponeurosis for both active lengthening and passive lengthening, and higher tissue displacement for active lengthening than passive lengthening. First principal strain magnitudes were averaged in a 1cm region near the myotendinous junction, where injury is most frequently observed. It was found that strains are significantly larger during active lengthening (0.19 SD 0.09) than passive lengthening (0.13 SD 0.06) (p<0.05), which suggests that elevated localized strains may be a mechanism for increased injury risk during active as opposed to passive lengthening. First principal strains were higher for subjects with a relatively narrow aponeurosis width (0.26 SD 0.15) than wide (0.14 SD 0.04) (p<0.05). This result suggests that athletes who have BFLH muscles with narrow proximal aponeuroses may have an increased risk for BFLH strain injuries.     

Effect of feeding and lack of food on the growth, gross biochemical and fatty acid composition of juvenile catfish

The effect of feeding and starvation on growth, gross body composition and fatty acid composition of body muscle and liver were investigated in juvenile African catfish ( Clarias gariepinus ). The relative composition of fatty acids was influenced by starvation. In both liver and muscle there were marked relative decreases of 14: 0, 16: 1n9 and 18: 1n9 fatty acids and relative increases of 20: 5n3 and 22: 6n3 when these were compared to the controls. Preference of utilization of fatty acids from liver and body muscle during starvation was highest for 14: 0>16: 1n9>18: 1n9 and conservation highest for 22: 6n3>20: 5n3. The changes of these fatty acids were smaller in muscle than in the liver.     

Some morphometric and biochemical features of ready-to-migrate silver and pre-migratory yellow stages of the shortfin eel of south-eastern Australian waters

The mean total length ( L _T), mass and age of ready to migrate female silver shortfin eels Anguilla australis from the Hopkins River estuary and the mouth of the Merri River in south-eastern Australia, were 83·2 ± 1·2 cm, 1051 ± 51 g, and 17·2 ± 1·79 years, respectively. The eye index ( I _E) of the silver shortfin eels was < 5·2 (mean 7·64 ± 0·29) and differed significantly from that of the yellow shortfin eels collected from two other sites. The I _E increased with L _T (mm) and was related by log I _E= 2·656 log L _T6·925. The per cent moisture, protein and ash content of the liver of silver shortfin eels was significantly lower than in yellow shortfin eels, but lipid content was significantly higher in the former (35·5 ± 2·0%). The mean mass μg mg lipid ⁻) of saturates (230·4 ± 2·6 v . 181·7 ±2·6), monoenes (367·4 ± 6·3 v . 290·8 ± 8·9) and PUFA (177·3 ± 5·3 v . 159·7 ± 4·6) in muscle was significantly higher, and the great majority of individual fatty acids was found also in higher quantities in silver shortfin eels. In the liver, the PUFA found in the highest quantity was 22:6n-3, except in shortfin eels from Hopkins River estuary, and the amount of 18:2n-6 in the liver of silver shortfin eels was significantly higher than that in yellow shortfin eels but the reverse was true of 20:4n-6. In both muscle and liver tissues the saturate 16:0 and the monoene 18:ln-9 collectively accounted for >50% of all the fatty acids in the lipid.     

Comparison between superficial and solid-state cultures of <Emphasis Type="Italic">Isaria fumosorosea</Emphasis>: conidial yields,quality and sensitivity to oxidant conditions

Conidia production and quality from mycoinsecticides in solid-state cultures (SSC) are frequently inferred from superficial culture (SC) results. Both parameters were evaluated for two Isaria fumosorosea strains (ARSEF 3302 and CNRCB1), in SC and SSC, using culture media with the same chemical composition. For both strains, conidia production was higher in SC than SSC in terms of conidia per gram of dry substrate. Germination in both strains did not show significant differences between SC and SSC (>90 %). Similarly, conidia viability in ARSEF 3302 strain did not show differences at early stages between SC and SSC, but was higher in SC compared to SSC in the late stage of culture; in contrast, conidia from CNRCB1 strain did not differ between both culture systems. Some infectivity parameters improved in conidia from SSC, compared to SC at the early stages, but these differences disappeared at the final stage, independently of the strain. Both strains showed decreased conidia production when 26 % O₂ pulses were applied; nevertheless, conidiation in SSC was two orders of magnitude more sensitive to oxidant pulses. In SC with 26 % O₂ pulses, conidia viability for both strains at early stages, was higher than in normal atmospheric conditions. Infectivity towards Galleria mellonella larvae was similar between conidia from normal atmosphere and oxidant conditions; notably, for the strain ARSEF 3302 infectivity decreased at the final stage. This study shows the intrinsic differences between SC and SSC, which should be considered when using SC as a model to design production processes in SSC.     

Dependence of the Phenotypic Characteristics of <Emphasis Type="Italic">Acidithiobacillus ferrooxidans</Emphasis> on the Physical,Chemical, and Electrophysical Properties of Pyrites

Comparison of Acidithiobacillus ferrooxidans strains TFV-1 and TFBk with respect to their capacity to oxidize pyrite 1, with an electron-type (n-type) conductivity, or pyrite 2, with hole-type (p-type) conductivity, showed that, at a pulp density of 1%, both before and after its adaptation to the pyrites, strain TFBk, isolated from a substrate with a more complex mineral composition, grew faster and oxidized the pyrites of both conductivity types more efficiently than strain TFV-1, which was isolated from a mineralogically simple ore. At a pulp density of 3–5%, the oxidation of pyrite 2 by strain TFV-1 and both of the pyrites by strain TFBk began only after an artificial increase in Eh to 600 mV. If the pulp density was increased gradually, strain TFBk could oxidize the pyrites at its higher values than strain TFV-1, with the rate of pyrite 2 oxidation being higher than that of pyrite 1. During chemical oxidation of both of the pyrites, an increase was observed in the absolute values of the coefficients of thermoelectromotive force (K_TEMF); during bacterial-chemical oxidation, the K_TEMF of pyrite 1 changed insignificantly, whereas the K_TEMF of pyrite 2 decreased.     

The haematology of gynogenic tench, Tinca tinca L., and of recessively homozygous colour tench strains

Two wild‐coloured strains of tench (the first meiotic gynogenic generation MeiG₁, and their control diploid half siblings) and three recessively homozygous colour strains (golden, blue and alampic) were examined for the determination of basic haematological indices. The MeiG₁ strain had higher erythrocyte counts than diploid controls or the blue and alampic strains (P < 0.001), and had a higher blood haemoglobin content than all three colour strains (P < 0.001). No differences were detected among strains for haematocrit, mean corpuscular haemoglobin, or mean corpuscular volume. Both the lowest leucocyte count (P < 0.001) and leucocrit value (P < 0.001) were found in the alampic tench, and may result from a negative pleiotropic effect of this recessive homozygous genotype (bbgg). In agreement with previous findings in tench, the differential leucocyte count revealed lymphocytes to be the dominating white blood cells; their rate was about 90% in both the wild‐coloured and blue strains, and less in the other two strains (83–84%). Neutrophil granulocytes were most abundant in the MeiG₁ strain. Eosinophil granulocytes were detected only in the golden strain, and were not common (0.2%).     

Microbial arsenite oxidation with oxygen,nitrate, or an electrode as the sole electron acceptor

The purpose of this study was to identify bacteria that can perform As(III) oxidation for environmental bioremediation. Two bacterial strains, named JHS3 and JHW3, which can autotrophically oxidize As(III)–As(V) with oxygen as an electron acceptor, were isolated from soil and water samples collected in the vicinity of an arsenic-contaminated site. According to 16S ribosomal RNA sequence analysis, both strains belong to the ?-Proteobacteria class and share 99% sequence identity with previously described strains. JHS3 appears to be a new strain of the Acinetobacter genus, whereas JHW3 is likely to be a novel strain of the Klebsiella genus. Both strains possess the aioA gene encoding an arsenite oxidase and are capable of chemolithoautotrophic growth in the presence of As(III) up to 10 mM as a primary electron donor. Cell growth and As(III) oxidation rate of both strains were significantly enhanced during cultivation under heterotrophic conditions. Under anaerobic conditions, only strain JHW3 oxidized As(III) using nitrate or a solid-state electrode of a bioelectrochemical system as a terminal electron acceptor. Kinetic studies of As(III) oxidation under aerobic condition demonstrated a higher V _max and K _m from strain JHW3 than strain JHS3. This study indicated the potential application of strain JHW3 for remediation of subsurface environments contaminated with arsenic.     

<Emphasis Type="Italic">Deinococcus rubellus</Emphasis> sp. nov., bacteria isolated from the muscle of antarctic fish

Two new bacterial strains designated as Ant6^T and Ant18 were isolated from the muscle of a fish which had been caught in the Antarctic Ocean. Both strains are Gram-stain-positive, catalase positive, oxidase negative, aerobic, and coccoid bacteria. Phylogenetic analysis based on the 16S rRNA gene sequences of strains Ant6^T and Ant18 revealed that the strains Ant6^T and Ant18 belong to the genus Deinococcus in the family Deinococcaceae in the class Deinococci. The highest degrees of sequence similarities of strains Ant6^T and Ant18 were found with Deinococcus alpinitundrae LMG 24283^T by 96.4% and 96.8%, respectively. Strain Ant6^T exhibited a high level of DNA- DNA hybridization values with strain Ant18 (82 ± 0.6%). Chemotaxonomic data revealed that the predominant fatty acids were C_{17: 0} cyclo, 16:0, and feature 3 (C_16:1ω6c/ω7c) for both strains. A complex polar lipid profile consisted of major amounts of unknown phosphoglycolipids (PGL) and unknown aminophospholipid (APL). Based on the phylogenetic, phenotypic, and chemotaxonomic data, strains Ant6^T (=KEMB 9004-169^T =JCM 31434^T) and Ant18 (=KEMB 9004-170) should be classified as a new species, for which the name Deinococcus rubellus sp. nov. is proposed.     

Comparative study of mannose-binding C-type lectin isolated from channel catfish (Ictalurus punctatus) and blue catfish (Ictalurus furcatus)

Mannose-binding C-type lectin (MBL) was isolated from channel catfish (Ictalurus punctatus) NWAC 102 and 103 strains, blue catfish (Ictalurus furcatus) D+B and Rio Grande strains, hybrid catfish (channel catfish female NWAC 103 x blue catfish male D+B) sera, and purified by affinity chromatography from channel catfish Norris strain serum. Reduction of purified channel catfish MBL with 2-ME yielded a single band of 62 kDa by SDS-PAGE and Western blot analysis using guinea pig anti-MBL IgG as primary antibody. Channel catfish NWAC 102 strain, channel catfish NWAC 103 strain and hybrid catfish sera had molecular masses of 63 kDa for MBL. Blue catfish (D+B strain) serum MBL had a molecular mass of 66 kDa. Rio Grande blue catfish serum MBL had a molecular mass of 65 kDa. Amino acid composition analysis (mol%) of the affinity-purified channel catfish MBL found a high content of serine present. Functional binding studies of channel catfish and blue catfish MBLs binding to Edwardsiella ictaluri were done using a dot-immunoblot ELISA method. A dot-immunoblot ELISA binding assay was done to compare nine different strains and species of channel catfish and blue catfish for their levels of serum MBL. Blue catfish had higher levels of MBL than did the various strains of channel catfish tested. MBL could be used as a genetic marker for selection of disease resistance in the different strains of catfish used in aquaculture. This study describes the presence of serum MBL in catfish and evidence for a C-type lectin complement pathway of innate immunity.     

Studies on the lipopolysaccharide of a virulent and an avirulent strain of Vibrio vulnificus.

Vibrio vulnificus is a marine bacterium associated with both primary septicemias and wound infections in humans. The lipopolysaccharides of a virulent and an avirulent strain of Vibrio vulnificus were compared with respect to their chemical constituents and electrophoretic characteristics. 2-Keto-3-deoxyoctonic acid, a normal constituent of the lipopolysaccharide of typical Enterobacteriaceae, was not found in the lipopolysaccharide of either strain. Hexadecenoate (C16:1) was the predominant fatty acid of the lipid A moiety of the lipopolysaccharides and of the membrane phospholipids of both strains. Hydroxy fatty acids composed 44% of the total fatty acids of the lipid A of the avirulent and 40% of those in the virulent strain. In addition, odd-numbered fatty acids were detected in both lipopolysaccharides. The electrophoretic profile was similar for both strains, but demonstrated no "ladder-like" pattern characteristic of "smooth" lipopolysaccharides. The result of this study showed no significant differences between the lipopolysaccharides of the virulent and avirulent strains of Vibrio vulnificus. The possible role for lipopolysaccharide in pathogenesis of Vibrio vulnificus infections is discussed.     

<Emphasis Type="Italic">Cupriavidus malaysiensis</Emphasis> sp. nov., a novel poly(3-hydroxybutyrate-<Emphasis Type="Italic">co</Emphasis>-4-hydroxybutyrate) accumulating bacterium isolated from the Malaysian environment

Bacterial classification on the basis of a polyphasic approach was conducted on three poly(3 hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] accumulating bacterial strains that were isolated from samples collected from Malaysian environments; Kulim Lake, Sg. Pinang river and Sg. Manik paddy field. The Gram-negative, rod-shaped, motile, non-sporulating and non-fermenting bacteria were shown to belong to the genus Cupriavidus of the Betaproteobacteria on the basis of their 16S rRNA gene sequence analyses. The sequence similarity value with their near phylogenetic neighbour, Cupriavidus pauculus LMG3413^T, was 98.5%. However, the DNA–DNA hybridization values (8–58%) and ribotyping analysis both enabled these strains to be differentiated from related Cupriavidus species with validly published names. The RiboPrint patterns of the three strains also revealed that the strains were genetically related even though they displayed a clonal diversity. The major cellular fatty acids detected in these strains included C15:0 ISO 2OH/C16:1 ω7c, hexadecanoic (16:0) and cis-11-octadecenoic (C18:1 ω7c). Their G+C contents ranged from 68.0  to 68.6 mol%, and their major isoprenoid quinone was Ubiquinone Q-8. Of these three strains, only strain USMAHM13 (= DSM 25816 = KCTC 32390) was discovered to exhibit yellow pigmentation that is characteristic of the carotenoid family. Their assembled genomes also showed that the three strains were not identical in terms of their genome sizes that were 7.82, 7.95 and 8.70 Mb for strains USMAHM13, USMAA1020 and USMAA2-4, respectively, which are slightly larger than that of Cupriavidus necator H16 (7.42 Mb). The average nucleotide identity (ANI) results indicated that the strains were genetically related and the genome pairs belong to the same species. On the basis of the results obtained in this study, the three strains are considered to represent a novel species for which the name Cupriavidus malaysiensis sp. nov. is proposed. The type strain of the species is USMAA1020^T (= DSM 19416^T = KCTC 32390^T).