Fish photobacteriosis—The importance of rapid and accurate identification of Photobacterium damselae subsp. piscicida

MALDI‐TOF MS was tested for the identification of Photobacterium damselae subsp. piscicida on isolates grown on two media, cultured at three incubation times and applied on the target plate by the direct sample spotting (DS), by the on‐target extraction (OTE) and by the full extraction (FE) method, in triplicates. The identification of samples grown on blood agar (BA) outperformed identification on tryptic soya agar (TSA) by 0.64% for DS and OTE. The OTE gave the highest scores in both culture media, all incubation times and replicates. Reliable 24‐hr species identification was 61.54%, 84.61% and 53.85% for samples grown on TSA and identified by DS, OTE and FE, respectively. For isolates grown on BA, they were 76.92%, 96.15% and 30.77%, respectively. When identified by OTE, the 48‐hr identification was 93.58%, but for 72 hr declined to 71.79%. The reliable identification with the highest score from the first measurement was 100% only for OTE from BA (24 hr), whereas OTE from TSA gave 84.61% (24 hr), 76.92% (48 hr) and 84.61% (72 hr). The reliable MALDI‐TOF MS identification of Ph. damselae subsp. piscicida is incubation time, media, target plate preparation and replicate‐dependent.     

Evidence for the facultative intracellular behaviour of the fish pathogen Vibrio ordalii

Vibrio ordalii is an extracellular, Gram‐negative bacterium that produces vibriosis in salmonids. While pathogenesis is not fully understood, this bacterium has numerous likely genes for adhesion, colonization, invasion factors and, as recently suggested, intracellular behaviour. Therefore, this study aimed to clarify possible intracellular behaviour for V. ordalii Vo‐LM‐18 and ATCC 33509^T in the fish‐cell lines SHK‐1 and CHSE‐214. Confocal microscopy revealed Vo‐LM‐18 and ATCC 33509^T inside cytoplasm in both fish‐cell lines at 4 hr post‐inoculation (hpi). At 8 and 16 hpi, the proportion of fish cells invaded by both strains increased. Moreover, intracellular V. ordalii were observed after 8 hpi inside mouse embryonic fibroblasts (MEF), demonstrating that entry was not due to a cellular phagocytosis process. Flow cytometry confirmed immunocytochemistry results, with both V. ordalii evidencing statistically significant differences in the number of infected cells between 8 and 16 hpi. Interestingly, V. ordalii infection did not significantly damage fish cells, as determined by LDH liberation. Viable counts at 8 hpi detected, on average for both lines, 176 ± 47 CFU/ml of culturable intracellular Vo‐LM‐18 and ATCC 33509^T cells. These in vitro findings support the facultative intracellular behaviour of V. ordalii and may be of importance for understanding pathogenicity and survival in aquatic environments.     

Dietary protein requirement of juvenile marbled spinefoot rabbitfish Siganus rivulatus

Rabbitfish are an Indo‐Pacific herbivorous marine fish that have good market demand and are suitable for aquaculture. The present work was performed to determine dietary protein inclusion necessary for optimal growth of juvenile rabbitfish Siganus rivulatus. Six diets with increasing levels of protein (10, 20, 30, 40, 50 and 60 g crude protein 100 g^?1 feed) and similar levels of gross energy (20 MJ kg^?1) were prepared and offered to S. rivulatus juveniles maintained in triplicate cages placed in two large water tanks for 49 days. Growth progressively improved with dietary protein for fish offered diets from 10% to 40% protein inclusion. Diets with greater protein levels did not improve fish growth beyond that observed in the 40% group. Daily feed intake, apparent protein utilization and feed conversion ratio decreased as dietary protein increased. Protein efficiency (PE) was greatest (1.47) in fish offered the 10% protein diet and least in fish offered the 60% protein diet (0.80). No differences in PE were observed among all other treatments (20–50%). Results of the present work suggest that minimum dietary requirement for suitable growth of S. rivulatus juveniles is 40% protein when digestible energy of the diet is 16–18 MJ kg^?1.     

长江天鹅洲故道银鮈年龄、生长和死亡率的研究

根据2011年3月至2012年1月在长江天鹅洲故道采集的银鮈(Squalidus argentatus)样本873尾,用鳞片为年龄鉴定和生长退算材料,研究银鮈的年龄、生长特性及死亡率.结果表明,雌雄群体均由1～6龄组成,雌鱼87.35％为1～3龄,雄鱼87.01％为1～3龄.雌雄鱼体长(L)和体重(W)关系表现为异速生长,雌鱼w=0.009L3.2285,雄鱼w=0.009L3.2254.以退算体长拟合von Bertalanffy方程,得到生长参数为:雌鱼L∞=17.1797cm,k=0.1820,t0=-1.2673和雄鱼L∞=11.5069cm,k=0.4074,to=-0.5764.雌雄鱼的自然死亡率分别为M=0.6048/a和M=1.1459/a,远高于一般鱼群自然死亡率(M=0.2～0.3/a);开发率分别为E=0.5711和E =0.4183.长江天鹅洲故道银鮈雌鱼处于过度开发的状态,应加强管理.     

Effects of dietary carbohydrate/lipid ratios on growth performance,body composition and glucose metabolism of fingerling blunt snout bream Megalobrama amblycephala

This study aimed to investigate the optimum dietary carbohydrate/lipid (CHO/L) ratio for fingerling blunt snout bream Megalobrama amblycephala (average initial weight: 6.61 ± 0.03 g). Six isonitrogenous and isoenergetic diets were formulated to contain various CHO/L ratios ranging from 1.62 to 24.20. Each diet was tested in four replicates for 10 weeks. Specific growth rate (SGR), feed conversion ratio and nitrogen and energy retention all improved significantly (P < 0.05) as dietary CHO/L ratios decreased from 24.20 to 5.64 but showed little difference (P > 0.05) as dietary CHO/L ratios ranged from 2.45 to 5.64. Hepatosomatic index increased significantly (P < 0.05) as dietary CHO/L ratios increased, whereas intraperitoneal fat ratio showed an opposite trend (P < 0.05). Opposite to moisture content, lipid content of whole body and carcass all increased significantly (P < 0.05) as dietary CHO/L ratios decreased. Liver lipid content showed little difference (P > 0.05) among all the treatments, while liver glycogen content increased significantly (P < 0.05) with increasing CHO/L ratios. High dietary carbohydrate enhanced the activities of liver hexokinase, pyruvate kinase and glucose‐6‐phosphate dehydrogenase but did not induce hyperglycaemia. Based on the second‐order polynomial regression analysis of SGR, the optimal dietary carbohydrate and lipid contents for fingerling blunt snout bream were 291.7 and 81.4 g kg⁻¹, respectively, with a corresponding dietary CHO/L ratio of 3.58.     

Protection of Fenneropenaeus chinensis (Osbeck, 1765) against the white spot syndrome virus using specific chicken egg yolk immunoglobulins by oral delivery

Two kinds of specific chicken egg yolk immunoglobulins (IgYs), IgY‐WSSV and IgY‐VP28, were, respectively, raised against the 2 mM binary ethylenimine (BEI)‐inactivated white spot syndrome virus (WSSV) and a principal envelope protein VP28. The activity of purified specific IgYs was stable under the conditions of 20–70 °C, pH 3.0–10.0 and 0–700 g L^?1 sucrose solution. In the neutralization assay, these high‐affinity IgY antibodies can specifically bind with the virus particles to protect shrimp (Fenneropenaeus chinensis) against WSSV infection. After oral delivery for 20 days, the IgY‐WSSV exerted a higher protection effect (RPS: 71.5%) than IgY‐VP28 (RPS: 63.7%). Moreover, an increase in RPS (79.2%) was found on addition of IgY‐WSSV:VP28 (0.1% IgY‐VP28 plus 0.2% IgY‐WSSV). This may indicate that neutralization of WSSV refers to the multiple‐hit model. By time‐course study of the levels of the specific IgYs in vivo, the data showed that the titre was enhanced to a relatively high level (P/N=8.35±0.45) at 3 days post administration, declined slightly (P/N=7.13±1.01) at 7 days post administration and then remained stable for further investigation. The stable antibody level potentially contributes towards blocking a large number of WSSV particles from entering and infecting on the major tissues at the early and late stages after challenge in shrimp.     

Effect of Luteolin on Blood Indexes,Liver and Kidney in Mice with Acute Mercury Poisoning

To study the effect of luteolin on blood indexes,liver and kidney in mice with acute mercury poisoning,28 mice were randomly divided into four groups:Control group (intraperitoneal injection 0.9% saline),luteolin group (lavage 100 mg/kg luteolin),mercuric chloride group (intraperitoneal injection 4 mg/kg mercury chloride) and mercury chloride+luteolin group (intraperitoneal injection 4 mg/kg mercuric chloride,lavage 100 mg/kg luteolin).The activities of ALT in serum,AST,CREA and BUN contents,blood WBC,RBC,HGB content and GSH and MDA contents of liver tissue were detected.Morphological changes of liver and kidney tissues were observed.The results showed that compared with the control group,the activities of ALT and AST of mercuric chloride group were extremely significantly increased (P < 0.01),serum CREA,BUN,blood WBC and liver tissue MDA contents were significantly increased (P < 0.05) while blood RBC,HGB and liver tissue GSH contents were significantly decreased (P < 0.05).Liver,kidney pathological changes were obviously.Compared with mercuric chloride group,the activities of ALT,AST in serum,CREA,BUN,blood WBC and liver tissue MDA contents of mercuric chloride+luteolin group were significantly decreased (P < 0.05),while blood RBC,HGB and liver tissue GSH contents were significantly elevated (P < 0.05).Liver,kidney pathological changes were attenuated obviously.The poisoning were characterized by inflammation and the occurrence of anemia when acute mercury poisoning occurred,liver and kidney showed different degrees of injury in mice.Luteolin could reduce the toxic effects of acute mercury poisoning on blood,liver and kidney.