Characterization of Pseudomonas aeruginosa associated with diseased postlarval abalone in Shenzhen, China

Outbreaks of mass mortality of postlarval abalone, Haliotis diversicolor supertexta, have occurred in south China since 2002 and have forced many abalone farms to close. About 30 representative bacterial strains were isolated from a sample of five diseased postlarval abalone, taken 25 days post-fertilization during an outbreak of postlarval disease in Shenzhen, China, in October 2006. Bacterial challenge tests showed that the predominant strain, designated as strain 22, was highly virulent to postlarvae with an LD₅₀ value of 7.8 × 10⁴ colony forming units (CFU) ml⁻¹, while four of the other isolates were weakly virulent with LD₅₀ values ranging from 1 × 10⁶ to 1 × 10⁷ CFU ml⁻¹, and the remaining 25 isolates were classified as avirulent with LD₅₀ values greater than 1 × 10⁸ CFU ml⁻¹. By means of API 20NE and 16S rDNA and ITS sequencing analyses, strain 22 was identified as Pseudomonas aeruginosa. Antibiotic susceptibility tests showed that strain 22 exhibited around 75% of susceptibility to 16 various antibiotics tested. The results of this study show P. aeruginosa as one of the bacteria involved in the mortality of abalone postlarvae in Shenzhen, China.     

Isolation and characterization of pathogenic Vibrio alginolyticus from diseased postlarval abalone, Haliotis diversicolor supertexta (Lischke)

Outbreaks of serious mortality among cultured abalone postlarvae have occurred across Southern China since July 2002. Five motile bacterial strains were isolated from diseased abalone postlarvae on tryptic soy agar supplemented with 1% NaCl (TSA1) and/or thiosulphate citrate bile salt (TCBS) sucrose agar plates during an outbreak in August 2003 in Shanwei, Guangdong province. All isolates were characterized and identified as Vibrio alginolyticus on the basis of biochemical characteristics and comparisons with those of the reference strain V. alginolyticus ATCC 17749. Strain 19 (a representative of five similar isolates) was virulent to abalone postlarvae with an LD₅₀ value of1.00 × 10⁴ colony‐forming units mL^?1. All abalone postlarvae exhibited the same signs as in natural outbreaks. The same bacterium could be re‐isolated from abalone postlarvae after bacterial challenge using TSA1 and TCBS plates. The results reveal that V. alginolyticus is an infectious agent of abalone postlarvae.     

Isolation and identification of Vibrio campbellii as a bacterial pathogen for luminous vibriosis of Litopenaeus vannamei

Outbreak of luminescent disease was reported from Litopenaeus vannamei shrimp farms in Zhangpu County, Southern China during May–July 2011. The clinical signs included fluorescent, less food consumption and high mortality. Bacteria were isolated from the infected shrimps. The pathogen, a luminescent bacterium named VH1 was identified as Vibrio campbellii based on MLSA analysis (16S rDNA, rpoD and toxR). The haemolysin (hly) gene specific in V. campbellii was detected in strain VH1. Pathogenicity test using immersion infection confirmed that strain VH1 was virulent to L. vannamei postlarvae and juveniles, and the LC₅₀ value was 1.55 × 10⁶ CFU mL^?1 and 1.7 × 10⁶ CFU mL^?1 respectively.     

Skin‐injured Zebrafish,Danio rerio,are more Susceptible to Vibrio anguillarum Infection

Vibrio anguillarum, which is part of normal microflora on fish, is the causative agent of vibriosis in aquaculture. It is speculated that V. anguillarum does not affect the host in most situations, but can cause a severe disease once the host is compromised. In the study reported herein, skin‐injured and intestine‐injured zebrafish, Danio rerio, were established as a model to mimic the natural infection caused by V. anguillarum when fish suffered an injury to a mucosal surface. Our results showed the lethal dose to 50% of the population (LD₅₀) of skin‐injured zebrafish was 6.8 × 10³ colony‐forming unit (CFU)/mL, which was much lower than intestine‐injured zebrafish (1.9 × 10⁶ CFU/mL) or non‐injured zebrafish (5.5 × 10⁶ CFU/mL). With the quantitative polymerase chain reaction and immunohistochemical analysis, we found that V. anguillarum proliferated rapidly in the skin and muscle after the bacteria entered into the host via the skin injury. The bacteria were subsequently transported to the immune organs and then caused a systemic infection in the fish. However, mortality of skin‐injured zebrafish significantly decreased if the fish were allowed to heal. These results indicate that minimizing injury to the mucosal surfaces of fish, especially the skin, will reduce infections caused by V. anguillarum.     

Studies on bacterial pathogens isolated from diseased torafugu (Takifugu rubripes) cultured in marine industrial recirculation aquaculture system in Shandong Province,China

In spring of 2011, an epidemic outbreak of torafugu with high mortality occurred in an aquafarm with marine industrial recirculation aquaculture system (MIRAS) in Yantai, Shandong Province, China. The diseased fish showed anorexia, haemorrhaging and festering fin and skin and swelling internal organs. Forty‐five dominant bacterial isolates were obtained from the diseased fish, and were found to belong to 12 species according to 16S rRNA gene sequences. One strain from each species was selected to test the pathogenicity, and five strains were showed to be virulent to zebrafish. Whereas Enterovibrio nigricans Fr42 was highly virulent with the LD₅₀ of 7.8 × 10⁴ CFU g^?1, Photobacterium swingsii Fr23, Vibrio owensii Fr40, V. harveyi Fr51 and V. rotiferianus Fr71 were moderately virulent with the LD₅₀ of 1.7 × 10⁶ to 8.4 × 10⁶ CFU g^?1. Both the bacteria and their extracellular products of the five strains were found to show phospholipase, caseinase, gelatinase, amylase and/or lipase activities. The production of N‐acyl homoserine lactones (AHLs) of the five strains was detected by three different AHLs biosensors, and three of them were found to produce AHLs by at least one kind of biosensor. This is the first study describing various opportunistic bacterial pathogens of fish cultured in MIRAS in China.     

Isolation and identification of fish pathogen Edwardsiella tarda from mariculture in China

The causative agent was isolated from diseased turbots (Scophthalmus maximus) stricken by a high‐mortality outbreak of bacterial septicaemia occurring in a mariculture farm in Yantai, a northern coastal city of China. Seven pure isolates, namely EH‐15, EH‐103, EH‐107, EH‐202, EH‐203, EH‐305 and EH‐306, belonged to Edwardsiella tarda. The phenotypic features of the cultures were analysed extensively. Three of the isolates showed high 16S rDNA sequence similarities with E. tarda sequence (GenBank accession no. EF467289). However, unlike the E. tarda ATCC 15947, all the isolates, except EH‐15, contained a novel large plasmid sized about 23.7 kb. Furthermore, pathogenicity of the isolates was addressed by experimental challenges with fish models. The isolates exhibited strong virulence to swordtail fish with LD₅₀ ranging between 3.8 × 10³ and 3.8 × 10⁵ CFU g^?1, and EH‐202 displaying the lowest LD₅₀ value among them. Antibiotic susceptibilities of E. tarda isolates were assayed. Compared with E. tarda ATCC 15947, the isolates displayed strong resistance to chloramphenicol, and the probable dominant chloramphenicol resistance determinant was cat III. Depicting the main biological properties of turbot‐borne E. tarda strains in China, the study provided useful information for further unveiling their pathogenic mechanisms.     

Isolation and characterization of bacteria associated with a syndrome disease of sea urchin Strongylocentrotus intermedius in North China

Sea urchin, Strongylocentrotus intermedius, transplanted from Japan in 1989, has been widely cultured along the coasts of Liaoning and Shandong Provinces and has become the dominant and most economically important maricultured species in North China. However, a lesion syndrome symptom of S. intermedius broke out frequently these years, showing lethargy in activities, blackish peristomial membrane and body well lesions, and brought about high mortality eventually. Six representative prominent bacterial strains were isolated from diseased sea urchin from September 2009 to January 2010. By means of API 20NE and 16S rRNA gene sequences analysis, isolates were identified as Shewanella aquimarina, Pseudoalteromonas tetraodonis, Vibrio shilonii, V. harveyi, V. fortis and V. splendidus. Bacterial challenge tests showed that their representative isolates were virulent to S. intermedius with LD₅₀ values ranging from 9.2 × 10⁴ to 3.4 × 10⁶ CFU/g body weight, among which S. aquimarina , V. fortis and P. tetraodonis were highly virulent, and the other three isolates showed moderate virulence. The results indicated that a variety of bacteria including Shewanella, Pseudoalteromonas and Vibrio were involved in the mortality of S. intermedius, and the six isolates were opportunistic pathogens of sea urchins. All isolates reported herein were sensitive to ampicillin, enrofloxacin, ofloxacin, doxycycline and florfenicol.     

Identification and Detection of the Pathogenic Bacteria Responsible for Swollen Abdomen Disease in Cultured Turbot,Scophthalmus maximus,and Flounder,Paralichthys olivaceus

Swollen abdomen disease (SAD) seriously threatens the aquaculture of turbots and flounders. Two dominant bacterial strains, FS1 and FS2, were isolated from the livers and kidneys of fish with diagnosed SAD. Applications of biochemical analyses, sequence analyses of 16S ribosomal RNA gene and heat shock protein 60 gene revealed two distinct pathogenic bacterial species, Edwardsiella tarda and Vibrio alginolyticus. These two hypothesized SAD‐causing pathogens were validated by challenge trials on flounder, Paralichthys olivaceus. Challenges with E. tarda and V. alginolyticus demonstrated lethal dose 50 (LD₅₀) values at 1.51 × 10⁵ colony‐forming units (CFU) and 1.05 × 10⁵ CFU, respectively. To develop a rapid SAD diagnosis method in flounders and turbots, a multiplex polymerase chain reaction (PCR) assay method was developed to simultaneously detect E. tarda and V. alginolyticus. Our multiplex PCR assay successfully detected as low as 10⁵ CFU/mL E. tarda and V. alginolyticus in flounders and turbots. No other common fish pathogens were detected with the multiplex PCR, suggesting a high specificity of this assay. The multiplex PCR assay developed in this study showed a great reliability in detecting SAD‐causing bacterial pathogens. Further optimization of this assay may contribute to the development of a novel SAD diagnosis tool in aquaculture.     

Delivery of HUFA, probionts and biomedicine through bioencapsulated Artemia as a means to enhance the growth and survival and reduce the pathogenesity in shrimp Penaeus monodon postlarvae

One of the major problems in the shrimp culture industry is the difficulty in producing high-quality shrimp larvae. In larviculture, quality feeds containing a high content of highly unsaturated fatty acids (HUFA) and ingredients that stimulate stress and disease resistance are essential to produce healthy shrimp larvae. In the present study, Penaeus monodon postlarvae (PL15) were fed for 25 days on an unenriched Artemia diet (control; A) or on a diet of Artemia enriched with either HUFA-rich liver oil of the trash fish Odonus niger (B), probionts [Lactobacillus acidophilus (C1) or yeast-Saccharomyces cerevisiae (C2)] or biomedicinal herbal products (D) that have anti-stress, growth-promoting and anti-microbial characteristics. P. monodon postlarvae fed unenriched Artemia exhibited the lowest weight gain (227.9 ± 8.30 mg) and specific growth rate (9.95 ± 0.05%), while those fed the HUFA-enriched Artemia (B) exhibited the highest weight gain and specific growth rate (362.34 ± 12.56 mg and 11.77 ± 0.08%, respectively). At the end of the 25-day rearing experiment, the shrimp postlarvae (PL40) were subjected to a salinity stress study. At both low and high (0 and 50‰) salinities, the group fed the control diet (A) experienced the highest cumulative mortality indices (CMI) 935.7 ± 2.1 and 1270.7 ± 3.1, respectively. Those fed diet D showed the lowest stress-induced mortality, and CMI were reduced by 31.1 and 32.3% under conditions of low and high salinity stress, respectively. A 10-day disease challenge test was conducted with the P. monodon postlarvae (PL40–PL50) by inoculating the shrimp with the pathogen Vibrio harveyi at the rate of 10⁵–10⁷ CFU/ml in all rearing tanks. P. monodon postlarvae fed probiont-encapsulated Artemia diets (C1 and C2) exhibited the highest survival (94.3 and 82.3%, respectively) and lowest pathogen load (V. harveyi) in hepatopancreas (5.2 × 10² ± 9.0 × 10 and 4.6 × 10² ± 9.0 × 10 CFU g⁻¹, respectively) and muscle (2.0 × 10² ± 6 × 10 and 1.7 × 10² ± 8.6 × 10 CFU g⁻¹, respectively) tissues. The shrimp that were fed the unenriched Artemia (Control; A) showed the lowest survival (26.33%) and highest bacterial load in the hepatopancreas (1.0 × 10⁵ ± 5 × 10³ CFU g⁻¹) and muscle (3.6 × 10⁴ ± 6 × 10² CFU g⁻¹). The shrimp fed the herbal product (D)-enriched Artemia also exhibited enhanced survival and reduced V. harveyi load in the tissues tested compared to the control diet (A) group. The results are discussed in terms of developing a quality larval feed to produce healthy shrimp larvae.     

Effect of three bacterial isolates from a commercial hatchery on early red abalone (Haliotis rufescens) postlarvae

Three bacterial strains, GHrC11, GHrC13 and GHrC15, were isolated from an abalone postlarval culture system in a commercial farm at Baja California, México. The strains were phenotypically characterized and sequenced (16S rDNA). Strain GHrC11 was a Gram-positive coccobacillum, while strains GHrC13 and GHrC15 were Gram-negative bacilli. Strain GHrC11 was identified as Exiguobacterium sp. The strains GHrC13 and GHrC15 were identified as Vibrio splendidus. The effects of these strains for the development of early abalone postlarvae (2 days old) were evaluated following a completely randomized design with three replicates using 5-mL-volume Petri dishes as experimental units. The experiment considered two different bacterial concentrations of each strain (10³ and 10⁵?cells?ml^?1) and two controls (with and without the benthic diatom Navicula incerta). After 10?days of experimentation, the highest mortality (90?±?5.8?%) and the lowest growth rate (4.1?±?0.1?μm?day^?1) were recorded for the strain GHrC11. In contrast, the lower mortality (16.7?±?3.3?%) and the highest growth rate (11.2?±?0.9?μm?day^?1) corresponded to the control fed N. incerta. Our results suggest that pathogenic effects of these bacterial strains were stronger than any potential benefits derived from the ingestion of bacteria by early abalone postlarvae. In conclusion, the most pathogenic strain was GHrC11, and the intensity of pathogenicity could be ordered as Exiguobacterium sp.?>?V. splendidus (C13)?>?V. splendidus (C15).     

ELECTRICITY AND PROPANE CONSUMPTION OF A SHRIMP HATCHERY IN THE STATE OF BAJA CALIFORNIA SUR,MEXICO

The consumption of electricity and propane for producing larvae and postlarvae of the whiteleg shrimp (Litopenaeus vannamei) was examined in a commercial shrimp hatchery on the Baja California Peninsula of Mexico. Between January and August 2005, 6 × 10⁶ postlarvae (average age PL16) were produced from 1.43 × 10⁹ nauplii. During that production period, the hatchery used 2.48 × 10⁹ kcal of fossil fuel energy (30% for electricity, 70% for liquid propane), which was equivalent to 16% of the operating costs. Electricity was used mainly for larval and postlarval rearing (36% and 10%) and microalgae culture (27%). During this production period, 1.47 × 10⁹ kcal of propane were consumed for heating daily more than 600 m³ seawater daily. Of that total, 33% was used for broodstock maintenance, 65% for larvae culturing, and 2% for postlarvae rearing. With increasing costs for shipping postlarvae to the mainland Mexican coastal areas (15% of operating costs) and the need to remain competitive with hatcheries in the coastal areas of Sonora and Sinaloa, alternative energy sources to reduce overall operational costs in hatcheries of Baja California Sur are discussed.     

Biochemical and Serological Characterization of Flavobacterium columnare from Freshwater Fishes of Eastern India

Flavobacterium columnare is an important pathogen of freshwater fishes often causing high mortality. Seven strains of F. columnare have been isolated from gill necrosis, skin lesions and internal organs of Catla catla, Labeo rohita, Cirrhinus mrigala, Carassius auratus, Anabas testudineus and Clarias batrachus and characterized by biochemical reactions and serological tests viz indirect enzyme‐linked immunosorbent assay (ELISA), Dot‐ELISA, agglutination test. All the strains showed binding to Congo red dye as well as hemolysis. All the seven strains were susceptible to amikacin, gentamycin and ofloxacin. In vivo LD₅₀ dose of virulent strain MS2 was found to be 6 × 10⁴ CFU/mL after experimental infection to L. rohita. Serologically all the seven strains showed a positive result to agglutination, Dot‐ELISA and indirect ELISA. The agglutination titer was found to be in the range of 256‐131,072. The lysozyme activity of hyperimmune sera was found to be 39.37 ± 0.80 units/mL. This is the first extensive study that report about the various strains of F. columnare from freshwater fishes of Eastern India and their differentiation by biochemical and serological methods.     

Hafnia alvei as an opportunistic pathogen causing mortality in brown trout, Salmo trutta L.

Strains of Hafnia alvei caused mortalities in brown trout, Salmo trutta L., following intraperitoneal injection with LD₅₀ values ranging between 1.3 × 10⁴ and 2.5 × 10⁷ bacteria fish^??1. These values are considered to represent a high to moderate degree of virulence. Virulent strains were isolated from non-fish sources. Fish surviving the LD₅₀ values continued to harbour the organism in the kidney, suggesting the establishment of a carrier state.     

Mortality and pathology of hybrid catfish,Clarias macrocephalus (Günther) × Clarias gariepinus (Burchell), associated with Edwardsiella ictaluri infection in southern Thailand

Enteric septicaemia of catfish (ESC) caused by Edwardsiella ictaluri is becoming an increasing problem in aquaculture and has been reported worldwide in a variety of fish species. This study reports ESC in hybrid catfish, Clarias macrocephalus (Günther) × Clarias gariepinus (Burchell), cultured in southern Thailand. The bacteria were identified as E. ictaluri by conventional and rapid identification systems, as well as by genetic and phylogenetic characterization. Analysis of 16S rRNA indicated 100% homology to the 16S rRNA sequence of several E. ictaluri strains in GenBank. Plasmid profiles demonstrated 4.0‐ and 5.6‐kb plasmids, compared with the 4.8‐ and 5.6‐kb plasmids in the US isolates, and representative genes of three of the four known pathogenicity islands of US isolates were present. Serologically, lipopolysaccharide (LPS) purified from the Thai isolates was not recognized by a monoclonal antibody against the LPS of US isolates. Fish experimentally infected with E. ictaluri showed 23–100% mortality within 14 days with a 168‐h LD₅₀ of 6.92 × 10⁷ CFU mL^?1 by immersion and a 96‐h LD₅₀ of 1.58 × 10⁶ CFU fish^?1 by intraperitoneal injection. Examination of tissue sections obtained from both naturally and experimentally infected fish indicated that infection of hybrid catfish with E. ictaluri produced lesions in several organs including liver, kidney, spleen, heart and brain. Histopathology findings included cellular necrosis, focal haemorrhage, infiltration of lymphocytes and multifocal granulomatous inflammation in the infected organs.     

Effect of light limitation on the water quality,bacterial counts and performance of Litopenaeus vannamei postlarvae reared with biofloc at low salinity

The aim of this study was to evaluate the effect of light limitation on the water quality, bacterial counts and performance of Litopenaeus vannamei postlarvae reared with biofloc at low salinity (≈9 g L^?1). Two treatments were designed: T₁ = culture with natural sunlight and T₂ = culture in darkness. After 28 days, in both treatments, the final weight of shrimp was over 0.6 g with a specific growth rate over 7.4% d^?1, and a survival rate over 70%. In both treatments, Vibrio sp. concentration presented low values (culture with natural sunlight = 0.1 to 9.9 × 10² CFU mL^?1, culture in darkness = 0.4 to 11.7 × 10² CFU mL^?1) and Bacillus sp. had high values (culture with natural sunlight = 0.7 to 66.0 × 10⁴ CFU mL^?1, culture in darkness = 0.7 to 65.8 × 10⁴ CFU mL^?1). All water quality parameters remained within the ranges suitable for shrimp culture, except for alkalinity during the first stage of the study. Although in some sampling periods some significant differences were found in bacterial counts and water quality parameters, shrimp productive performance under culture with biofloc at low salinity was not affected significantly by light limitation.     

BALB/c小鼠用于评价尼罗罗非鱼无乳链球菌毒力的研究

实验采用BALB/c小鼠作为实验动物,旨在建立尼罗罗非鱼无乳链球菌毒力测定的BALB/c小鼠模型。BALB/c小鼠经腹腔注射尼罗罗非鱼源无乳链球菌建立感染模型,比较了尼罗罗非鱼源无乳链球菌分别感染尼罗罗非鱼和小鼠的LD_(50)差异,分别测定了不同毒力尼罗罗非鱼无乳链球菌对尼罗罗非鱼和小鼠的毒力。结果显示,小鼠经腹腔注射无乳链球菌,在24 h内出现死亡现象,且对小鼠脑、肝脏、脾脏、肾脏等组织造成损伤。3次测定尼罗罗非鱼无乳链球菌TFJ0901对尼罗罗非鱼和小鼠LD_(50)分别为7.7×10~7、2.2×10~8、3.5×10~9 CFU/mL和405、361、419 CFU/只。将无乳链球菌TFJ0901和THN0901感染尼罗罗非鱼(1.0×10~7 CFU/mL)和小鼠(100 CFU/只),尼罗罗非鱼和小鼠存活率分别为100%、6.7%±5.8%和100%、0,其存活率都具有显著性差异。将无乳链球菌TFJ0901和TFJ-F感染尼罗罗非鱼(3.0×10~8 CFU/mL)和小鼠(2 500 CFU/只),尼罗罗非鱼的存活率分别为73.3%±11.5%和80.0%±10.0%,存活率差异不显著,小鼠存活率分别为13.3%±11.5%和100.0%,存活率具有显著性差异。研究表明,本实验成功建立了BALB/c小鼠作为尼罗罗非鱼源无乳链球菌毒力测定的稳定模型,测定不同毒力的尼罗罗非鱼源无乳链球菌对小鼠毒力与对尼罗罗非鱼毒力一致,且该模型能够区分尼罗罗非鱼模型难以区分的毒力相近的无乳链球菌。     

患腹水病牙鲆病原菌分离、鉴定及病原菌的特性

为确定引起河北北戴河地区养殖牙鲆患腹水病死亡的病原,本实验从患腹水病牙鲆体内分离到3株优势菌,对分离株进行生理生化鉴定和16S rRNA序列比对来确定分离株的生物学地位,进一步通过毒力基因(toxR、vhhA、vhhB)鉴定及组织病理学分析其病理特征,并通过人工回感实验分析分离株毒性。结果显示,通过生理生化实验及16S rRNA序列比对,确定此次从患病牙鲆中分离到的3株菌株均为哈维氏弧菌;经鉴定,3株分离株毒力基因(toxR、vhhA、vhhB)结果均为阳性,病理组织切片结果显示,该分离株对牙鲆多器官(肠、肾脏、脾脏和肝脏)均可造成不同程度的病理损伤,呈全身性感染;人工回感实验显示,菌株BDHYPFS-Y1G对牙鲆的半致死浓度为LD50=5.88×106 CFU/mL,低于自然状态毒性;药敏实验表明,3株分离株均对呋喃妥因高度敏感。本实验确认了此次牙鲆腹水病的病原菌,并初步研究了病原菌致病性以及药物敏感性,以期为牙鲆工厂化养殖疫病防控提供科学依据。     

Growth of Red Abalone,Haliotis rufescens,Fed Cold‐stored Benthic Diatoms

We measured the proximal composition of four benthic diatoms that were stored for 24 mo in the dark at low temperature (4 C by refrigeration) and examined their potential as feed for abalone, Haliotis rufescens, postlarvae. The proximal composition of the four diatoms was modified by species‐specific responses as a function of time in storage. The cultures of all stored diatoms contained low or undetectable concentrations of Vibrio‐like bacteria (<0.01 VLB/mL). As feed for abalone postlarvae, the four diatoms promoted growth under all experimental conditions. Greater shell lengths were measured on Day 14 when Navicula sp. and Navicula incerta were used as feed. Postlarvae that were fed N. incerta and Navicula sp. had higher growth rates. In contrast, lower growth rates were observed on Day 7 with fresh and stored Nitzschia thermalis as food. Survival was high in postlarvae that were fed the four stored diatoms (100%). This report demonstrates that cultures of benthic diatoms that have been stored by refrigeration for 24 mo can be used to feed abalone postlarvae and have an effect on improving growth and survival.     

Impacts of Lactobacillus plantarum in Depuration for Reducing Vibrio parahaemolyticus in Pacific Oysters (Crassostrea gigas)

This study investigated potential application of lactic acid bacteria (LAB) in depuration for reducing Vibrio parahaemolyticus in oysters. Lactobacillus plantarum ATCC 8014, which exhibited strong bactericidal effects against V. parahaemolyticus in vitro, was added to artificial seawater for depuration of Pacific oysters (Crassostrea gigas) inoculated with V. parahaemolyticus BE 98-2029 (O3:K6) to levels of about 10⁴ MPN/g at 15 ± 1 and 10 ± 1°C. Application of L. plantarum ATCC 8014 treatment (10⁷ CFU/mL) in oyster depuration did not enhance reductions of V. parahaemolyticus in oysters depurated at 15 ± 1°C but significantly decreased (p < 0.05) levels of V. parahaemolyticus in oysters depurated at 10 ± 1°C after 5 days (3.40 log reductions) when compared with controls (2.75 log reductions). It is not clear if a competitive exclusion by LABs to compete with V. parahaemolyticus binding sites in oyster tissues plays a role in the reduction of V. parahaemolyticus in the oysters. Further studies utilizing different types of LABs in oyster depuration might provide additional knowledge for application of LAB in depuration for decontaminating V. parahaemolyticus in oysters.     

The effects of dietary probiotic Bacilli (Bacillus subtilis and Bacillus licheniformis) on growth performance,feed efficiency,body composition and immune parameters of whiteleg shrimp (Litopenaeus vannamei) postlarvae

The present study investigates the effects of dietary commercial Bacilli probiotic (Bacillus subtilis and Bacillus licheniformis) on the growth performance, feed efficiency, body composition and immune parameters of Litopenaeus vannamei. The L. vannamei postlarvae were supplied and acclimated (in 500‐L tanks) to laboratory conditions for 14 days. The shrimps were fed with diets containing 1 × 10⁴ and 1 × 10⁸ CFU/g probiotic Bacilli for 60 days. At the end of the feeding trial, growth performance parameters, body composition, serum biochemical parameters and the hemocytes count were evaluated. Shrimps fed diets supplemented with 1 × 10⁴ and 1 × 10⁸ CFU/g probiotic Bacilli showed improved weight gain, total length, specific growth rate, FCR and survival compared with the control group. The body composition studies revealed higher dry matter, crude protein and ash in shrimps fed with 1 × 10⁴ and 1 × 10⁸ CFU/g probiotic Bacilli. Also, dietary administration of 1 × 10⁴ and 1 × 10⁸ CFU/g probiotic Bacilli decreased serum glucose and cortisol levels. However, significantly increased total protein, lysozyme and hemocyte cell count were noticed in shrimps fed 1 × 10⁴ and 1 × 10⁸ CFU/g probiotic Bacilli. In general, the findings of this study proved that oral administration of 1 × 10⁴ and 1 × 10⁸ CFU/g commercial probiotic Bacilli improved growth performance, feed utilization and immune parameters in whiteleg shrimp (Litopenaeus vannamei).