斑马鱼幼鱼嗜中性粒细胞对副溶血弧菌清除的动态变化

副溶血弧菌(Vibrio parahaemolyticus)是人-兽-鱼共患的致病菌,严重威胁食品安全和人类的健康。为了研究宿主嗜中性粒细胞清除细菌感染的动态变化过程,利用红色荧光蛋白标记的副溶血弧菌Vp57~(RFP)菌株感染受精后48 h(hours post fertilization,hpf)的斑马鱼(Danio rerio)幼鱼耳泡,建立局部感染模型,并以大肠杆菌(Escherichia coli)Ec01菌株为对照,系统比较了副溶血弧菌和大肠杆菌的半数致死剂量(median lethal dose,LD_(50))、存活曲线以及和嗜中性粒细胞相互作用的动态过程,RT-qPCR验证炎症相关基因il1b和il10在感染前后的表达量变化。结果显示副溶血弧菌和大肠杆菌感染斑马鱼幼鱼的LD_(50)分别为7.90×10~8 CFU/mL和1.14×10~(11) CFU/mL,与感染大肠杆菌相比,斑马鱼感染副溶血弧菌后招募嗜中性粒细胞的速度更快,且数量更多。RT-qPCR结果发现斑马鱼感染副溶血弧菌后在相同时间点能够激活更高的il1b和il10基因的表达,引起强烈的炎症反应。基于以上研究,我们建立的副溶血弧菌-斑马鱼幼鱼耳泡局部感染模型,为进一步研究嗜中性粒细胞清除副溶血弧菌感染的动态过程,以及深入揭示天然免疫应答机制提供了依据。     

Effects of music stimulus on behavior response,cortisol level,and horizontal immunity of growing pigs

An enriched environment is widely used to improve domestic animals’ welfare and promote their natural behaviors. Music can reduce abnormal behavior in humans, nonhuman primates, and rodents. However, little is known about the effects of music on pigs. This study aims to explore the effects of repeated music stimulation on the behavior, physiology, and immunity of growing pigs. A total of 72 hybrid piglets (Large White × Duroc × Minpig) were randomly divided into three groups, including music (Mozart K.448, 60 to 70 dB), noise (recorded mechanical noise, 80 to 85 dB), and control (natural background sound, <40 dB), and 6 h sound stimulation was given per day (1000 to 1600 hours) from 40 to 100 d of age. The behavioral activities of the pigs were observed during the music stimulation, and their serum cortisol, salivary cortisol, and serum immune indices were also measured. Compared with the control group, the music group and noise group increased activity but decreased lying of pigs (P < 0.05). A significant increase in tail-wagging, playing, and exploring behaviors of pigs was found in the music group (P < 0.05), and the noise significantly increased the aggressive behavior of the pigs (P < 0.05). Tail-wagging, playing, exploring, manipulating, and aggressive behaviors decreased over time. Short-term (8 d) music stimulus had a lower cortisol level than that of the noise and control groups (P < 0.05), whereas long-term (60 d) music stimulus increased immunoglobulin G (IgG), interleukin-2 (IL-2), and interferon-gamma (IFN-γ) levels (P < 0.05) and decreased interleukin-4 (IL-4) level (P < 0.05). Long-term noise stimulus significantly reduced the level of IgG (P < 0.05) but did not affect the level of IL-2, IL-4, and IFN-γ levels (P > 0.05). In conclusion, short-term music stimulus (8 d) reduced the stress response, whereas long-term music stimulus (60 d) enhanced the immune responses. In addition, the noise increased the aggressive behavior, and long-term noise reduced the immunity of the growing pigs.     

口蹄疫疫苗对牛的应用效果调查

牛口蹄疫是传染性疾病,为研究青海省峰堆乡口蹄疫疫苗的免疫效果,于2018—2019年对该乡的7个村开展牛口蹄疫疫苗免疫效果调查,共调查牛634头,7个村总体的口蹄疫合格率为100.00%和97.56%,最低合格率为81.63%和88.63%。为更好地控制该类疾病发生流行,提升免疫效果,该文探讨口蹄疫疫苗免疫失败原因,并制定了相应的预防措施。     

Dietary profile of n‐3 series LC‐PUFAs in rainbow trout under regular stripping condition: Semen production and quality,hepato‐somatic index,haemato‐immunologic values,oxidative stress and fatty acid composition of liver,muscle and semen

In this study, the semen production and quality, hepato‐somatic index, haemato‐immunologic values, oxidative stress and the fatty acid contents in liver, muscle and semen of rainbow trout fed diets supplemented n‐3 series long‐chain polyunsaturated fatty acids (LC‐PUFAs) under regular stripping condition were investigated. For this aim, three diets (Control, D1 and D2) were prepared. These diets were contained n‐3 LC‐PUFAs (as a percentage of dietary total fatty acid) at 3.14%, 7.84% and 13.63% respectively. Experimental fish were fed with the control and test diets. The highest hepato‐somatic index, spermatologic (semen pH and volume, sperm motility and density), haematologic (haematocrit value, haemoglobin, erythrocyte count, corpuscular volume, haemoglobin and its concentration in corpuscular), immunologic (nitroblue tetrazolium activity, leucocyte count, phagocytic index, protein and immunoglobulin [IgM] in total plasma) and antioxidants (reduced glutathione, catalase, glutathione peroxidase and superoxide dismutase) values were found in fish fed the D2, D1 and control diets respectively (p < 0.01). Increase in the dietary n‐3 LC‐PUFAs was not significantly (p > 0.01) increased the oxidative stress (malondialdehyde) in fish. The results indicated that the n‐3 LC‐PUFAs at 13.63% level of total fatty acid in the diet could increase the semen production and quality, hepato‐somatic index, haematologic and immunologic values, and the n‐3 LC‐PUFA contents in liver, muscle and semen of rainbow trout broodstock under regular stripping condition.     

红枣提取物对虹鳟血清指标和头肾免疫相关基因表达的影响

为了探讨添加红枣提取物对促进虹鳟(Oncorhynchus mykiss)机体健康的可能性,选取体长为7.2~8.4 cm的健康虹鳟,随机分成4个试验组,分别投喂红枣提取物添加量为0%(对照组)、0.25%、0.50%和1.00%的等氮等脂饲料,饲养56 d,养殖结束后测定12项血清生化指标和头肾的6个免疫相关基因mRNA表达量。结果显示:与对照组相比较,血清中肌酐(CREA)和尿素氮(BUN)含量三个红枣提取物组均出现极显著下降;谷丙转氨酶(GPT)活性在0.50%和1.00%组极显著下降,谷草转氨酶(GOT)活性也在0.50%和1.00%组出现了一定程度下降,但差异不显著;酸性磷酸酶(ACP)和碱性磷酸酶(AKP)活性无显著变化;总蛋白(TP)和白介素6(IL-6)含量在0.25%组出现显著增加;溶菌酶(LZM)活性三个红枣提取物组均显著升高;超氧化物歧化酶(SOD)活性在0.50%组极显著增加;补体4(C4)和肿瘤坏死因子(TNF-α)含量在0.25%和0.50%组均极显著增加。在头肾中,与对照组相比,SOD、白介素1β(IL-1β)和补体3(C3)基因表达量在0.50%和1.00%组均为显著或极显著上调;Factor H基因表达量在0.50%组显著上调;过氧化氢酶(CAT)和LZM基因表达量在0.50%和1.00%组仅出现差异不显著上调。上述结果表明,在虹鳟饲料中添加一定量的红枣提取物,可以起到促进肾功能,保护肝脏和提高其非特异性免疫功能的作用。     

Marine-Derived Chitosan Nanoparticles Improved the Intestinal Histo-Morphometrical Features in Association with the Health and Immune Response of Grey Mullet (Liza ramada)

Marine-derived substances are known for their beneficial influences on aquatic animals’ performances and are recommended to improve intestinal health, immunity, and anti-oxidative status. The present study investigates the role of chitosan nanoparticles on the intestinal histo-morphometrical features in association with the health and immune response of Grey Mullet (Liza ramada). Chitosan nanoparticles are included in the diets at 0, 0.5, 1, and 2 g/kg and introduced to fish in a successive feeding trial for eight weeks. The final body weight (FBW), weight gain (WG), and specific growth rate (SGR) parameters are significantly increased while feed conversion ratio (FCR) decreases by chitosan nanoparticles compared to the control (p < 0.05). The morphometric analysis of the intestines reveals a significant improvement in villus height, villus width, and the number of goblet cells in chitosan-treated groups in a dose-dependent manner. Additionally, there is a positive correlation between the thickness of the enterocyte brush border and the chitosan dose, referring to an increasing absorptive activity. Histologically, the intestinal wall of Grey Mullet consists of four layers; mucosa, sub-mucosa, tunica muscularis (muscular layers), and serosa. The histological examination of the L. ramada intestine shows a normal histo-morphology. The epithelial layer of intestinal mucosa is thrown into elongated finger-like projections, the intestinal villi. The values of hemoglobin, hematocrit, red blood cells (RBCs), total protein (TP), albumin, and globulin are significantly increased in fish fed 1, and 2 g/kg of chitosan nanoparticles compared to fish fed 0 and 0.5 g/kg (p < 0.05). The highest levels of TP and albumin are observed in fish fed 1 g/kg diet (p < 0.05). The lysozyme activity and phagocytic index are significantly enhanced by feeding chitosan nanoparticles at 0.5, 1, and 2 g/kg, whereas the phagocytic activity is improved in fish fed 1 and 2 g/kg (p < 0.05). The highest lysozyme activity and phagocytic index are observed in fish fed 1 g/kg. SOD is significantly activated by feeding chitosan nanoparticles at 1 g/kg. Simultaneously, glutathione peroxidase (GPx) and catalase (CAT) activities also are enhanced by feeding chitosan at 1 and 2 g/kg, compared to fish fed 0 and 0.5 g/kg (p < 0.05). The highest GPx and CAT activities are observed in fish fed 1 g/kg (p < 0.05). Conversely, the malondialdehyde (MDA) levels are decreased by feeding chitosan at 1 and 2 g/kg, with the lowest being in fish fed 1 g/kg (p < 0.05). To summarize, the results elucidate that L. ramada fed dietary chitosan nanoparticles have a marked growth rate, immune response, and anti-oxidative response. These improvements are attributed to the potential role of chitosan nanoparticles in enhancing intestinal histo-morphometry and intestinal health. These results soundly support the possibility of using chitosan nanoparticles at 1–2 g/kg as a feasible functional supplement for aquatic animals.     

Effects of dietary Bacillus pumilus on growth performance,innate immunity and digestive enzymes of giant freshwater prawns (Macrobrachium rosenbergii)

The present study was conducted to investigate the effects of Bacillus pumilus on the growth performance, innate immunity and digestive enzymes of the giant freshwater prawn, Macrobrachium rosenbergii. Prawns (mean weight: 1.81 ± 0.01 g) were fed different levels of B. pumilus at doses of 0, 1 × 10⁷, 1 × 10⁸ and 1 × 10⁹ CFU/g to produce one control group and three experimental groups (G1, G2 and G3), respectively. After a 60‐day experimental feeding period, the results showed that the final weight, weight gain rate (WGR) and specific growth rate (SGR) were significantly higher in experimental groups than in the control (p < 0.05), whereas these improvements did not differ significantly (p > 0.05) among experimental groups. The improvements in phagocytic activity, respiratory burst (RBs) activity and the activity of catalase (CAT), nitric oxide synthase (NOS) and acid phosphatase (ACP) were significantly higher in G2 and G3 than in the control and G1 (p < 0.05). Further, the activity of CAT, NOS and ACP significantly increased with an increase in the dietary level of B. pumilus (p < 0.05). The activity of phenoloxidase (PO) in G3 was significantly higher than that in the control (p < 0.05). The activity of amylase was significantly higher in the groups G2 and G3 than that in the control group. G3 improved the activity of protease compared with the control (p < 0.05). In conclusion, the results of this study indicated that the potential use of B. pumilusat a dose of 1 × 10⁸ CFU/g feed could improve the growth, immunity and digestive enzymes of M. rosenbergii.     

Dietary simultaneous replacement of fish meal and fish oil with blends of plant proteins and vegetable oils in yellowfin seabream (Acanthopagrus latus) fry: Growth,digestive enzymes,antioxidant status and skin mucosal immunity

A 56‐day nutritional research was performed to examine the influence of alternative vegetal protein and lipid sources on performance of yellowfin seabream fry (Acanthopagrus latus) (0.5 ± 0.0 g). In this regard, five isoproteic (Ca. 500 g/kg) and isolipidic (Ca. 150 g/kg) diets were formulated in which fish meal (FM) and fish oil (FO) were simultaneously replaced with blends of plant proteins (PP, soybean meal and corn gluten) and vegetal oils (VO, canola and soybean oils) at 20% (SR20), 40% (SR40), 60% (SR40) and 80% (SR80) levels, respectively; meanwhile, a control diet (SR0) was formulated based on FM and FO. Growth and feed utilization were not influenced by experimental diets. The fatty acid profile of fillet drastically altered by dietary treatments. Fish fed with the SR60 and SR80 feeds had higher total protease, trypsin and α‐amylase activities than other treatments. The antioxidant enzyme activities and glutathione content in liver were enhanced in fish fed with the SR40, SR60 and SR80 diets. Skin mucosal immune parameters including total protein content, alkaline phosphatase and alternative complement pathway activities in the control group were relatively lower than the vegetal treatments. According to these results, it is recommended that 410 g/kg of FM and 45 g of FO/kg can be replaced with alternative vegetal sources in diet for A. latus fry.     

Dietary Moringa oleferia leaf meal induce growth,innate immunity and cytokine expression in grass carp,Ctenopharyngodon idella

The present research was designed to investigate the growth promoting and immunostimulating properties of Moringa oleferia leaf meal (MLM) in grass carp. Juvenile grass carp (22.03 g ± 1.164) were fed with diets supplemented with 0, 50, 100 and 150 g/kg MLM for 48 days. At the end of feeding trial, skin mucus was used for analysis of lysozyme, protease, antiprotease and peroxidase activity. Head kidney was used for expression analysis of tumour necrosis factor‐alpha, interleukin‐8 and interferon‐γ. The obtained results showed that fish fed with 100 and 150 g/kg MLM had significant increase in weight gain and specific growth rate (p < .05). However, condition factor was not altered. The MLM (50 and 100 g/kg) inclusion resulted in higher mucus lysozyme and protease activity (p < .05), while peroxidase activity increased only in fish fed with 100 g/kg MLM and antiprotease activity was not altered. Expression of tnf‐α increased in a dose‐dependent manner, and significant (p < .05) increase was recorded in fish fed with 150 g/kg MLM. The expression of il‐8 and ifn‐γ increased in fish fed with 50 and 150 g/kg MLM; however, the increase was not significant (p > .05). In conclusion, supplementing juvenile grass carp feed with MLM up to 150 g/kg has growth promoting and immunostimulating effects.     

Effects of fish origin probiotics on growth performance,immune response and intestinal health of shrimp (Litopenaeus vannamei) fed diets with fish meal partially replaced by soybean meal

This study was conducted to evaluate the application of four fish origin probiotics to relieve the side effects induced by SBM in shrimp. Shrimps were fed with high fish meal diet (C0, positive control containing 500 g/kg FM and 0 g/kg SBM), low fish meal diet (C, control containing 250 g/kg FM and 340 g/kg SBM) or the control diet (C) supplemented, respectively, with 1.0 × 10⁸ CFU/g Bacillus pumilus SE5 (T1), Psychrobacter sp. SE6 (T2), Enterococcus faecium MM4 (T3) or Bacillus claussi DE5 (T4). After 6 weeks of feeding, the growth performance, hepatopancreatic digestive enzymes, intestinal histological structure and immune parameters were determined. The results showed that compared with high FM group (C0), significantly lower WGR and SGR as well as higher FCR were observed in the low FM group (C) (p < .05). Compared with the control, significantly lower FCR was observed in treatment T1 (p < .05), but not in treatments T2–T4. Protease, amylase and lipase activities in all the treatments (T1‐T4) were significantly higher than the control (p < .05). GOT and GPT activities in the control were significantly lower than the C0 (p < .05), while higher MDA level was recorded in the control (p < .05). Meanwhile, higher GPT activities were observed in treatments T1‐T4 compared with the control (p < .05). Compared with the C0, lower SOD, ACP and AKP activities were observed in the control. Significantly improved SOD and AKP activities were observed in all probiotic feeding groups compared with the control (p < .05). The control diet led to significant reduction of intestinal wall thickness and villus height compared with the C0 (p < .05), while significantly higher intestinal wall thickness and villus height were exhibited in all the probiotic groups compared with the control (p < .05), except intestinal wall thickness in treatment T3. Thus, these results demonstrated that the four fish origin probiotic strains could relieve the side effects induced by high level of SBM in shrimp, while probiotic B. pumilus SE5 showed the best performance.