Effect of Six Bacterial Strains on the Activity of Second Instar Larvae and Egg Hatching of Meloidogyne incognita

[Objectives]The paper was to screen effective biocontrol strains against Meloidogyne incognita.[Methods]The effect of six bacterial strains sourced from the research group s strain library on the activity of second instar larvae of M.incognita,as well as on egg hatching,was evaluated.[Results]The treatment of fermentation supernatant derived from the X-2 strain exhibited a pronounced lethal effect on M.incognita,with a corrected mortality rate reaching 97%within 72 h.Additionally,this treatment significantly inhibited egg hatching,achieving an inhibition rate of 94.69%at a 20-fold dilution.The strain was identified as Bacillus velezensis,belonging to the genus Bacillus,and was designated as RKN1111.[Conclusions]This study presents alternative strains and a theoretical framework for the biological control of M.incognita.

Study on the Quantity Dynamic Changes of Heterobacteria and Vibrios in Larvae Industrialized Culture System

[ Objective ] The dynamic change of heterobacteria and vibrios in larvae industrialized culture system was studied to provide scientific reference for healthy cultivation of shrimp. [ Method ] The heterobacteria, vibrios and pathogenic vibrio parahaemolyticus were monitored in larvae industrialized culture system. [ Result] The heterobacteria, vibrios and pathogenic vibrio parahaemolyticus were the most in fertilized eggs of shrimp but the least in nauplius, then their number would increase with growth. During whole rearing period, both boterobacteria in larvae, vibrios in water would increase by one order of magnitude, while both vibrios in larvae and heterobacteria in water would increase by two orders of magnitude. There were many heterobacteria and vibrios but few vibrio parahaemolyticus in living bait. The correlation coefficients between larvae and heterobacteria and vibrios in water were 0. 704 and 0. 840 in culture system respectively, while the correlation among heterobacteria, vibrios in living bait and larvae, water were weak or negative. [ Conclusion ] There was a dynamic relation between water and larvae in rearing period, and restrictly control of culture condition would restrain the occurrence of disease caused by vibrio parahaemolyticus, besides that bacteria number in bait was not obviously correlated with bacteria nubmer in culture system.

Research of Holotrichia diomphalia Larvae Cultivation Technology with Agaricus bisporus Residues

[Objective] The aim was to explore key technology to cultivate Holotdchia diomphalia larvae with Agaricus bisporus residues and to provide, technologicar sup- ports for processing of Agaricus bisporus residues. [Method] In the research, fodder thickness, population density and residues under different treatments were set to re- search effects on Holotrichia diomphalia larvae. [Result] The optimal thickness of fodder was 25 cm and the optimal feeding density was 44-56 larvae per hectare. The dry residues were more suitable, compared with decomposed residues and corn bran powders, for cultivation of Holotrichia diomphalia larvae. [Conclusion] Cultivation of Holotrichia diomphalia larvae with Agaricus bisporus residues is a new method to make use of Agaricus bisporus residues and of significance for extension of agricul- tural circulation chain, increase of economic benefits and ecological benefits.

Analysis on Survival Rate and Economic Benefit of Apostichopus japonicus Larvae Reared by Raising Water Temperature

In order to control the spawning quality and improve the economic benefit of Apostichopus japonicus Selenka, the indoor temperature and light control methods were employed for artificial maturation, spawning and hatching of A. japonicus, and the larval survival rate, disease incidence rate and economic benefit were analyzed. The results showed that compared with those of conventional temperature-reared larvae, the survival rate of raised temperature-reared larvae was increased by 15.8%, the disease incidence rate was reduced by 10.2%, and the economic benefit was improved by 21.6%. These results will provide theoretical basis and technical support for the artificial breeding of A. japonicus.

Study on Chironomid Larvae Recognition Based on DWT and Improved KNN

A chironomid larvae images recognition method based on wavelet energy feature and improved KNN is developed. Wavelet decomposition and color information entropy are selected to construct vectors for KNN that is used to classify of the images. The distance function is modified according to the weight determined by the correlation degree between feature and class, which effectively improves classification accuracy. The result shows the mean accuracy of classification rate is up to 95.41% for freshwater plankton images, such as chironomid larvae, cyclops and harpacticoida.

Suitability of Hermetia illucens larvae meal and fat in broiler diets:effects on animal performance,apparent ileal digestibility,gut histology,and microbial metabolites

Background:The possibility of partially replacing soybean meal(SBM)with Hermetia illucens(HI)defatted larvae meal in broiler nutrition has frequently been suggested.For sustainability reasons,however,the larvae fat produced during defatting should also be used and could be particularly beneficial regarding gut health due to its fatty acid composition.To evaluate the suitability of HI larvae as protein and fat source,a 2×3 factorial arrangement with two types of protein,i.e.SBM(S)or SBM and 15%of its crude protein replaced by HI larvae meal(L),and three levels of fat sources,namely 0(0 L),50%(50 L)or 100%HI larvae fat(100 L)at the expense of soybean oil was applied.Results:In the starter phase,an interaction showed higher body weight(BW),average daily gain(ADG)and improved feed conversion ratio(FCR)if 50%or 100%HI larvae fat was fed with HI larvae meal.Moreover,BW,ADG and FCR improved when feeding HI larvae meal as protein source.Additionally,we observed an increased average daily feed intake in the grower,finisher,and overall phase in the L groups and an improved FCR in 0 L compared to 50 L groups during the overall period.Regarding apparent ileal digestibility,HI larvae meal feeding increased dry matter,organic matter,and fat digestibility.Feeding HI larvae meal as protein source decreased the concentrations of agmatine,spermidine,spermine and ammonia in the caecal digesta,whereas fat source affected agmatine with higher concentrations in 50 L compared to 0 L in the colonic digesta.In contrast,caecal ethanolamine concentrations increased in HI larvae meal groups compared to SBM.Caecal butyric acid concentrations decreased with HI larvae meal feeding.An interaction was found for the jejunal villus area,being higher in L+100 L compared to S+100 L.Furthermore,L groups had greater villus width.Conclusions:A partial replacement of SBM with HI larvae meal and soybean oil with HI larvae fat in broiler diets without impairing animal performance or gut health seems possible.Feeding HI larvae meal affected broiler performance positively in the starter phase and improved apparent ileal digestibility.

Hermetia illucens larvae as a potential dietary protein source altered the microbiota and modulated mucosal immune status in the colon of finishing pigs

Background: Insects, such as Hermetia illucens larvae, are rich in chitin and proteins, and represent a suitable feed ingredient replacement for animals. However, little is known about the effect of administering H. illucens larvae on intestinal microbiota, bacterial metabolite profiles, and mucosal immune status in animals. This study aimed to investigate the effects of administering H. illucens larvae on colonic microbiota and bacterial metabolites production in finishing pigs. Seventy-two crossbred(Duroc × Landrace × Large White) female pigs(initial body weight, 76.0 ± 0.52 kg) were randomly allocated to three different dietary treatments: a control diet(Control group) and two diets corresponding to 4%(H1 group) and 8%(H2 group) H. illucens larvae inclusion levels, respectively. Each treatment consisted of eight pens(replicates), with three pigs per pen. After 46 days of feeding, eight pigs per treatment(n =8) were slaughtered, and the colonic digesta and mucosa were collected for microbial composition and microbial fermentation products, and genes expression analyses.Results: The results showed that the H1 diet significantly increased the abundance of Lactobacillus,Pseudobutyrivibrio, Roseburia, and Faecalibacterium compared with those in the control group(P < 0.05), with a decrease in the abundance of Streptococcus. The numbers of Lactobacillus, Roseburia, and Clostridium cluster XIVa were significantly greater in the H1 group than in the control group(P < 0.05). Meanwhile, H2 diet increased the number of Clostridium cluster XIVa compared with the control group(P < 0.05). For colonic metabolites, total short chain fatty acids, butyrate, and isobutyrate concentrations were significantly higher in the H1 group than those in the control group(P < 0.05);the H1 treatment caused a striking decrease in protein fermentation compared with the control group, as the concentrations of total amines, cadaverine, tryptamine, phenol, p-cresol, and skatole were significantly lower(P < 0.05). Additionally, H2 diet also increased butyrate concentration compared with control group(P < 0.05), while decreased the concentrations of phenol, p-cresol, and skatole(P < 0.05). Pigs in the H1 group down-regulated the expression of TLR-4 and pro-inflammatory cytokines(IFN-γ) compared with pigs in the control group(P < 0.05), and up-regulated anti-inflammatory cytokine(IL-10) and intestinal barrier genes(ZO-1, occludin, and mucin-1). H2 diet up-regulated the expression of ZO-1 compared with control group(P < 0.05). Furthermore, the changes in the colonic mucosal gene expression were associated with changes in the bacterial composition and their metabolites.Conclusions: Collectively, dietary inclusion of Hermetia illucens larvae may enhance mucosal immune homeostasis of pigs via altering bacterial composition and their metabolites. These findings provide a new perspective on insect meal as a sustainable protein source rich in nutrient ingredients for swine.

Comparison of growth and survival of larvae among different shell color stocks of bay scallop Argopecten irradians irradians(Lamarck 1819)

In order to improve production and breed new broods of bay scallop Argopecten irradians irradians, different-colored orange, purple and white lines were established by two mating methods of self-fertilization and mass spawning at a commercial scallop hatchery in spring, 2002. And then larval growth and survival of different lines was compared to test whether there is a relationship between shell color and growth and survival at early developmental stage. Both growth and survival have no significant differences among different experimental larvae of self-fertilization or mass spawning. Results are as following in the order of orange, purple and white shell stock. For the self-fertilization, growth rates of larvae were 6.174, 6.412, and 6.599 μm/d, respectively. Survival rates of larvae at Day 3 were 74.41%, 76.86%, and 82.05%; Day 6 were 49.14%, 65.63%, and 52.79%; and Day 9 were 25.06%, 20.80%, and 26.47%, respectively. For the mass spawning, the growth rates were 7.836, 7.941, and 7.878 μm/d, respectively. Survival rates at Day 3 were 93.05%, 91.95%, and 92.50%; Day 6 were 79.17%, 78.05%, and 82.50%; and Day 9 were 34.72%, 36.67%, and 38.33%, respectively. The absence of any relationship between shell color and growth and survival at the larval stage may be resulted from their common genetic basis.

DISTRIBUTION AND ABUNDANCE OF EGGS AND LARVAE OF ANTARCTIC KRILL(ENPHAUSUA SUPERBADANA)IN ANTARCTIC PENINSULA AREA

During the First Chinese Antarctic Expedition zooplankton samples were collected at each of the oceanography stations in the waters around South Sheltland Islands (S. S. Is.) and in the northern part of the Bellinshausen Sea from 20 January to 12 February, 1985 by vertical tows of an 80 cm diameter conical net from 200 m to surface. Two deep tows with a closing net. were performed in slope water north of S.S. Is, and in the basin of Bransfield Strait. Eggs and larvae of Euphatisia superba Dana in zooplankton samples were identified and counted.There were four records of eggs from 27 stations. Most catches were of small number and restricted to within the nearshore area south of S.S Is.The largest number (1500/1000 cubic meters )was found in the outer side of Smith Island.No Nauplius was found . The earliest stage we found 【was Metanauplius , in two deep tows. The distribution of Calyptopis MI and Furcili I-III were quite similar. They were found mainly in three areas: 1 ) nearshore water along the

Lead levels of Culex mosquito larvae inhabiting lead utilizing factory

Objective:To determine lead level primarily in Culex quinquefasciatus(Cx.quinquefasciatus), and Culex gelidus(Cx.gelidus) larvae inhabiting lead consuming factories,and to putatively estimate eco-toxicological impact of effluents from the firms.Methods:Third instars larvae were sampled by standard dipping method and lead concentrations in the larvae and their respective surrounding factory aquatic environments were determined through standard atomic absorption spectrophotometry(AAS).Results:Cx.quinquefasciatus was the most abundant species followed by Cx.gelidus.The levels of lead were higher in the Cx.quinquefasciatus(1.08-47.47μg/g),than in the wastewaters surface(0.01-0.78 μg/mL) from the factories or closer areas around factories. Other species were not reaching the.criteria for lead determination.Conclusions:The Cx. quinquefasciatus larvae can bio-accumulate the metal and can potentially serve as a biomarker of lead contamination,to complemente conventional techniques.

Effects of rearing temperature and density on growth,survival and development of sea cucumber larvae,Apostichopus japonicus(Selenka)

In laboratory conditions,effects of rearing temperature and stocking density were examined on hatching of fertilized egg and growth of auricularia larvae of Apostichopus japonicus respectively.Data series like larval length and density,metamorphic time,and survival rate of the larvae were recorded.Statistics showed that for A.japonicus,survival rate(from fertilized egg to late auricularia) decreased significantly with the increasing rearing temperature(P<0.05).At different temperatures SGR was statistically significant as well(P<0.05) from day 1,and maximal SGR was found on day 9 at 24℃(159.26±3.28).This study clearly indicated that at low temperature(<24℃),metamorphic rate was remarkably higher than at higher temperature(>26℃).Hatching rate was significantly different between 0.2-5 ind./ml groups and 20-50 ind./ml groups.Rearing larvae at the higher density had the smaller maximal-length,whereas needed longer time to complete metamorphosis.This study suggested that 21℃ and 0.4 ind./ml can be used as the most suitable rearing temperature and stocking density for large-scale artificial breeding of A.japonicus's larvae.

Toxic effects of several phthalate esters on the embryos and larvae of abalone Haliotis diversicolor supertexta

As the most widely used plasticizers in the world, phthalate esters （PAEs） are potential endocrine disruption compounds （EDCs）. In the present study, the toxicity of dimethyl phthalate （DMP）, diethyl phthalate （DEP）, dibutyl phthalate （DBP）, di （2-ethylhexyl） phthalate （DEHP） on embryogenesis and larvae development of the marine univalve Haliotis diversicolor supertexta was examined in laboratory. The results show that the malformation of embryos appeared during the experiment, such as embryos died or lysed, small transparent flocculent rings studded on the periphery of the embryo, and the larvae could failed to hatch. In embryo toxic test, embryos incubated at the highest concentration of DMR DEP and DBP solutions showed significantly high abnormal rate compared with the control, while DEHP solutions displayed no significant difference. In larval toxic test, in all concentrations of DMP, DEP and DBP solutions, larval settlement rates were low significantly than that of the control. Similarly, DEHP solutions show nearly no effect on the larval settlement. The order of toxicity on embryos and larvae is DBP〉DEP〉DMP〉DEHE Being a simple and easy stimulation to indoor spawn, sensitive to environmental factors, and short culture time, the embryos of H. diversicolor supertexta can be used to indicate toxicity of the PAEs.

Effects of temperature and salinity on the development of embryos and larvae of the veined rapa whelk Rapana venosa(Valenciennes, 1846)

The major population of the veined rapa whelk Rapana venosa （Valenciennes, 1846）, which is an important fishery resource, is facing a large decline in China. We studied the effects of incubation temperature （16-34℃ at salinity 30） and salinity （5-45 at 25℃） on the incubation period and subsequent larval development. In the temperature experiment, the shortest incubation period was 12 days at 34℃, the lower temperature limit was 16~C, the longest mean shell length （1 193＋17 pan） occurred at 25℃ and the highest survival rate 72.28%＋5.62% was observed at 28℃. In the salinity experiment, the shortest incubation period was 15 days at 25. The salinity tolerance range was 15-40, the longest mean shell length （855-4-9 μm） and the highest survival rate 72.93%~4.85% were both observed at 35. This study demonstrated that, during the egg-mass stage, temperature and salinity regimes influence later growth and survival of larvae. These observations deepen our understanding of the ecology and conservation of natural populations of Rapana venosa.

Development Study on Some Digestive Enzymes of Takifugu rubripes Larvae and Juvenile

The activities of some digestive enzymes are studied for Takifugu rubripes larvae and juvenile from the first feeding to 27 d after hatching at selected stages of development. The homogenate of whole larvae body is used for enzymatic determination. Activity of acid protease decreases notably during the beginning days after the commencement of completely exogenous feeding and the days before the beginning of the juvenile stage. Alkaline protease specific activity also decreases at metamorphosis. The activities are associated with the morphology of the developing digestive tract. Amylase activity increases before the first feeding, followed by a decreasing and then a rather constant level. Lipase activity remains low during the larvae and juvenile periods. Alkaline phosphatase activity increases gradually. This reflects the development of brush border membranes of enterocytes.

Comparative Study of Malathion Toxicity and General Esterases in Larvae and Adults from a Field Population of Oxya chinensis (Thunberg) (Orthoptera: Acridoidea)

The susceptibility of Oxya chinensis to malathion was compared in larvae and adults from a field population, collected from Jinyuan outskirt, Shanxi Province. The results showed that Oxya chinensis was more susceptible to malathion in the adult stage than in the larval stage. The LD50 values for malathion susceptibility of Oxya chinensis were 4.94 and 2.44 mg g-1 body weight in the larvae and adults respectively. The results indicated that the larvae were 2.02-fold less susceptible to malathion than the adults. The general esterases and the kinetics were characterized and compared between the two life stages and between females and males. Larval preparations of Oxya chinensis were more active than adult preparations in females and males. The larvae showed 1.18-, 1.49-, and 1.17- fold higher specific activities than the adults in females with α-NA, α-NB and β-NA respectively. In males, the ratios were 1.34-, 1.70-, and 1.06-fold. Female preparations were more active than those of males in the adults. The reverse results were observed in the larvae where male preparations were more active than female preparations. Kinetic studies showed that Km values of general esterases hydrolyzing α-NA, α-NB, and β-NA in the adult stage were 1.36-, 1.32- and 1.39-fold respectively, higher than those in the larval stage in females. In males, the ratios were 1.24-, 2.14-, and 1.20-fold. The esterase from male insects had a higher affinity (lower Km value) to the substrate than those from females. The results also showed that the Vmax values of general esterase hydrolyzing α-NA, α-NB, and β-NA in the two stages were similar. From the results of bioassays and biochemical analyses, it has been inferred that a higher level of resistance to malathion in larvae than in adults would appear to result from differences in the expression of resistance mechanisms in these two life stages. Enhanced esterase activities appeared to play a major role in resistance to malathion in both larvae and adults. From the analysis of inhibition in vitro, the esterases in the two life stages were B-type, and carboxylesterases were predominant enzymes in the composition of the esterases in the two stages.

Settlement and metamorphosis of Styela canopus Savigny larvae in response to some neurotransmitters and thyroxin

The larvae of ascidian Styela canopus Savigny were treated with epinephrine, norepinephrine, L-DOPA, GABA and thyroxin to test the ability of these compounds to induce or inhibit larval settlement and metamorphosis. The results showed that epinephrine, norepinephrine and L-DOPA at the concentration of 1 μmol/dm^3 induced larval settlement and metamorphosis in S. canopus, with short exposure （ 1 h） to 1 μmol/dm^3 of L-DOPA inducing rapid settlement. In contrast, GABA at the concentrations of 0.1 ～1130.0 μmol/dm^3 significantly inhibited the settlement and metamorphosis of S. canopus larvae. In addition, thyroxin at 1 -50 μg/dm^3 had no effect on larval settlement and metamorphosis in S. canopus. These results suggest the importance of neurotransmitters in the settlement and metamorphosis of S. canopus larvae.

Pilot-scale study of removal effect on Chironomid larvae with chlorine dioxide

Chironomid larvae propagated excessively in eutrophic water body and could not be effectively inactivated by the conventional disinfection process like chlorination due to its stronger resistance to oxidation. In this paper, a pilot-scale study of chlorine dioxide preoxidation cooperating with routine clarification process for Chironomid larvae removal was conducted in Shenzhen Waterworks in Guangdong Province, China. The experimental results were compared with that of the existing prechlorination process in several aspects, including the Chironomid larvae removal efficiencies of water samples taken from the outlets of sedimentation tank, sand filter, the security of drinking water and so on. The results showed that chlorine dioxide might be more effective to inactivate Chironomid larvae than chlorine and Chironomid larvae could be thoroughly removed from water by pre-dosing chlorine dioxide process. The GC-MS examination and Ames test further showed that the sort and amount of organic substance in the treated water by chlorine dioxide preoxidation were evidently less than that ofprechlorination and the mutagenicity of drinking water treated by pre-dosing chlorine dioxide was substantially reduced compared with prechlorination.

Scarabaeid Larvae- and Herbicide-Resistant Transgenic Perennial Ryegrass (Lolium perenne L.) Obtained by Agrobacterium tumefaciens-Mediated Transformation of cry8Ca2, cry8Ga and bar Genes

Insect pest and weeds are two major problems for forage and turf grasses. In this study, scarab larvae- and herbicide-resistant transgenic perennial ryegrass （Lolium perenne L.） was obtained by transforming it with cry and bar genes simultaneously via the Agrobacterium-mediated method. To optimize the callus induction and plant regeneration conditions, various concentrations of 2,4-dichlorophenoxyacetic acid and 6-benzylaminopurine were assayed. The transformation efficiencies of different Agrobacterium suspension media, used during Agrobacterium-mediated transformation, were compared. Then, plasmids of pCAMBIA3301 containing cry gene （cry8Ca2 or cry8Ga） and bar gene, driven by ubiquitin promoter, were transformed into perennial ryegrass. The transformants were generated and confirmed by both Southern hybridization analysis and Western hybridization analysis. Further, the resistance of transgenic perennial ryegrass plants to scarab larvae and herbicide were analyzed. After 30 d of co-cultivation with scarab larvae, the damage to the root system of transgenic plants was less than that of non-transgenic control plants. Additionally, the leaves of transgenic plants were resistant to Basta, while leaves of the wild plants wilted after Basta spraying. These results show that cry gene and bar gene were successfully transferred into perennial ryegrass by the Agrobactgerium-mediated method, and convey resistance to scarab larvae and herbicide in transgenic perennial ryegrass plants.

Advances in Studies of the Effects of Starvation on Growth and Development of Fish Larvae

Starvation has important effects on early development of fish. It determines the survival and growth of fish larvae,and plays an important role in the dynamics of fish population and fisheries recruitment. In this review,we discuss the current studies about the effects of starvation on growth and development of fish larval stage. The goals of this review are to understand some adaptive mechanisms and ecological countermeasures of starved fish larvae and to provide the scientific guideline for exploring early life history processes,evaluating the nutrition condition and growth of larval fish,protecting fish resource and breeding fish larvae.

Effects of florfenicol exposure on growth,development and antioxidant capacity of flounder Paralichthys olivaceus larvae at different developmental stages

Antibiotics are widespread in various environmental media,and may pose a potential threat to aquatic ecosystems and non-target aquatic organisms.Florfenicol(FLO)is one of the most commonly used antibiotics in aquaculture,and extensively used to substitute chloramphenicol with its strong sterilization and low adverse ef fect.In this study,flounder Paralichthys olivaceus,an important economic fish species in seawater was used as an experimental subject.Five exposure concentrations of FLO(including environmentrelated concentrations)were set at 0,0.01,0.1,1,and 10 mg/L.Ef fects of FLO exposure for 168 h on growth and development,motor behavior,antioxidant enzyme activity,malondialdehyde(MDA)content,and thyroid hormone level of P.olivaceus larvae were studied in pre-larvae(1 dpf)and post-larvae(20 dpf).The results show that the short-term FLO exposure could promote the larvae growth to some degrees,but inhibit them as the exposure time prolonged.For pre-larvae,FLO at 0.01 mg/L could stimulate the motor nerve system and increase the swimming ability,but inhibited it at 1 mg/L.With the increasing dosage of FLO,the superoxide dismutase(SOD)and MDA contents were elevated,reaching the maximum in the 1 mg/L FLO group.The pre-larvae were more sensitive than the post-larvae to FLO in the environment,and the growth and immune resistance could be damaged with long exposure.Post-larvae were more tolerant to external pollutants,FLO at 1 mg/L could promote the motor behavior and reduce SOD and MDA contents.Therefore,FLO can be used as an antibiotic at a proper concentration but as a drug to prevent disease in a long-term way.