Emerging Chytrid Fungal Pathogen, Batrachochytrium Dendrobatidis, in Zoo Amphibians in Thailand

Abstract Chytridiomycosis, a disease in amphibians caused by Batrachochytrium dendrobatidis (Bd), has led to a population decline and extinction of frog species since 1996. The objective of this study was to determine the prevalence of and the need for establishing a surveillance system for monitoring chytridiomycosis in five national zoos and five free ranging protected areas across Thailand. A total of 492 skin swab samples were collected from live and dead animals and tested by polymerase chain reaction (PCR) for the presence of Bd. The positive specimens were confi rmed by amplicon sequencing and examined by histopathology and immunohistochemistry. From July 2009 to August 2012, the prevalence of Bd from frog skin samples was low (4.27%), monitored by PCR. All samples from live amphibians were negative. The positive cases were only from dead specimens (21/168, 12.5% dead samples) of two non-native captive species, poison dart frog (Dendrobates tinctorius) and tomato frog (Dyscophus antongilii) in one zoo. Immunohistochemistry and histopathology revealed the typical feature of fl ask-shaped zoosporangia and septate thalli, supporting the PCR-based evidence of chytridiomycosis in captive amphibians in Thailand, but detected Bd in only 7/21 of the PCR-positive samples. Although the introduction of a pathogenic strain of Bd from imported carriers might have a serious impact on the native amphibian populations in Thailand, chytridiomycosis has not currently been detected in native Thai amphibians. An active surveillance system is needed for close monitoring of the fungus crossing into Thai amphibian populations</jats:p

Duplex PCR for Simultaneous and Unambiguous Detection of Streptococcus iniae and Streptococcus agalactiae associated with Streptococcosis of Cultured Tilapia in Thailand บทคั ดย่ อ ปฏิ กิ ริ ยาลู กโซ่ โพลิ เมอเรสแบบดู เพล็ กซ์ สํ าหรั บตรวจหาเชื ้ อสเตรปโ

Abstract Warm-water streptococcosis outbreaks in Thai cultured tilapia are caused by Streptococcus iniae and S. agalactiae. However, distinguishing between these two species of streptococcus are very difficult due to their similar microbiological appearance and clinical signs of infected fish. In this study, we proposed a new duplex-PCR based method for simultaneous detection of these pathogens. The duplex-PCR amplified partial lctO and 16s rRNA gene of S. iniae and S. agalactiae at 870 bp and 220 bp, respectively. This technique gave 100% specificity while sensitivity of reaction was 100 fg of each bacterial genomic DNA. Detection limit of duplex-PCR applicable to clinical specimens is also evaluated as 10 6 bacterial cells per gram of fish tissue. This study suggested that this duplex-PCR based method might be a good candidate for easy, sensitive, specific and rapid detection of S. iniae and S. agalactiae associated with warm-water streptococcosis of cultured tilapia in Thailand

Development of pelleted feed containing probiotic GG and Jerusalem artichoke for Nile Tilapia and its biocompatibility studies

Growth performance and immune systems of tilapias could be improved by GG (LGG) and Jerusalem artichoke. This research aimed to determine the effects of Jerusalem artichoke on LGG viability after drying and pelleting and their subsequent exposure to simulated gastrointestinal conditions. Fresh LGG cells were added into wall material solutions, including alginate (AL), alginate + milk powder (AM), and alginate + milk powder + Jerusalem artichoke at different concentrations (AMJ). The solutions were then spray dried to obtain LGG powders. The powder with the highest cell number was then selected to mix with tilapia feed mash and pelleted using a nonthermal feed extruder to obtain pelleted feed containing LGG and Jerusalem artichoke. The LGG viability spray dried powders and pelleted feed were analyzed for their cell counts after drying and after exposure to simulated gastrointestinal conditions. The result showed that the number of viable cells in AMJ was significantly higher than AM and AL after drying. The number of viable cells under both simulated gastric and bile salt fluids was improved with the increasing of Jerusalem artichoke concentrations. The number of viable cells after pelleting process could be maintained. LGG in the pelleted feed could also survive under the simulated gastric and bile salt conditions. The study indicates that JA enhanced LGG viability after drying and exposure to simulated gastrointestinal conditions. The pelleted feed containing LGG and Jerusalem artichoke could be applied in tilapia farming, providing convenience to the farmers, and valuable effects to the fish

Use of Lactobacillus plantarum (strains 22F and 25F) and Pediococcus acidilactici (strain 72N) as replacements for antibiotic-growth promotants in pigs

Abstract The lactic acid bacteria (LAB) Lactobacillus plantarum (strains 22F and 25F) and Pediococcus acidilactici (strain 72N) have appeared promising as replacements for antibiotics in in vitro studies. Microencapsulation, especially by the spray-drying method, has been used to preserve their numbers and characteristics during storage and digestion. This study compared the efficacy of these strains and their microencapsulated form with antibiotic usage on growth performance, faecal microbial counts, and intestinal morphology in nursing-finishing pigs. A total of 240 healthy neonatal pigs were treated on days 0, 3, 6, 9, and 12 after cross-fostering. Sterile peptone water was delivered orally to the control and antibiotic groups. Spray-dried Lactobacillus plantarum strain 22F stored for 6-months was administered to piglets in the spraydry group. Three ml of each the three fresh strains (109 CFU/mL) were orally administered to piglets in each group. All pigs received the basal diets, but these were supplemented with routine antibiotic for the antibiotic group. Pigs in all the probiotic supplemented groups exhibited a better average daily gain and feed conversion ratio than those of the controls in the nursery and grower phases. Probiotic supplementation increased viable lactobacilli and decreased enterobacterial counts. Antibiotic additives reduced both enterobacterial and lactobacilli counts. Villous height and villous height:crypt depth ratio were greater in probiotic and antibiotic supplemented pigs comparing to the controls, especially in the jejunum. The results demonstrated the feasibility of using these strains as a substitute for antibiotics and the practicality of the microencapsulation protocol for use in swine farms

Effects of Jerusalem Artichoke (<i>Helianthus tuberosus</i>) as a Prebiotic Supplement in the Diet of Red Tilapia (<i>Oreochromis</i> spp.)

The aim of the present study was to evaluate the effects of a Jerusalem artichoke-supplemented diet on the blood chemistry, growth performance, intestinal morphology, expression of antioxidant-related genes, and disease resistance against Aeromonas veronii challenge in juvenile red tilapia. A completely randomized design (CRD) was followed to feed red tilapias with three experimental diets: control, 5.0 g/kg JA-supplemented (JA5), or 10.0 g/kg JA-supplemented (JA10) diets in triplicates for 4 weeks. The results revealed that the growth performance, weight gain (WG), specific growth rate (SGR), and average daily gain (ADG) of fish fed diets JA5 and JA10 were significantly higher (p p gpx1 and gst antioxidant-related genes and disease resistance against A. veronii in juvenile red tilapia. Therefore, JA5 and JA10 can be employed as promising prebiotics for sustainable red tilapia farming

Addressing Nanovaccine Strategies for Tilapia

Tilapia is the world’s most extensively farmed species after carp. It is an attractive species for aquaculture as it grows quickly, reaching harvest size within six to seven months of production, and provides an important source of food and revenue for many low-income families, especially in low- to middle-income countries. The expansion of tilapia aquaculture has resulted in an intensification of farming systems, and this has been associated with increased disease outbreaks caused by various pathogens, mostly bacterial and viral agents. Vaccination is routinely used to control disease in higher-value finfish species, such as Atlantic salmon. At the same time, many tilapia farmers are often unwilling to vaccinate their fish by injection once the fish have been moved to their grow-out site. Alternative vaccination strategies are needed to help tilapia farmers accept and use vaccines. There is increasing interest in nanoparticle-based vaccines as alternative methods for delivering vaccines to fish, especially for oral and immersion administration. They can potentially improve vaccine efficacy through the controlled release of antigens, protecting antigens from premature proteolytic degradation in the gastric tract, and facilitating antigen uptake and processing by antigen-presenting cells. They can also allow targeted delivery of the vaccine at mucosal sites. This review provides a brief overview of the bacterial and viral diseases affecting tilapia aquaculture and vaccine strategies for farmed tilapia. It focuses on the use of nanovaccines to improve the acceptance and uptake of vaccines by tilapia farmers

Immersion Vaccination by a Biomimetic-Mucoadhesive Nanovaccine Induces Humoral Immune Response of Red Tilapia (<i>Oreochromis</i> sp.) against <i>Flavobacterium columnare</i> Challenge

Immersion vaccination with a biomimetic mucoadhesive nanovaccine has been shown to induce a strong mucosal immune response against columnaris disease, a serious bacterial disease in farmed red tilapia caused by Flavobacterium columnare. However, the induction of a systemic immune response by the vaccine is yet to be investigated. Here, we examine if a specific humoral immune response is stimulated in tilapia by a biomimetic-mucoadhesive nanovaccine against Flavobacterium columnare using an indirect-enzyme-linked immunosorbent assay (ELISA), serum bactericidal activity (SBA) and the expression of immune-related genes within the head-kidney and spleen, together with assessing the relative percent survival of vaccinated fish after experimentally infecting them with F. columnare. The anti-IgM antibody titer of fish at 14 and 21 days post-vaccination was significantly higher in chitosan complex nanoemulsion (CS-NE) vaccinated fish compared to fish vaccinated with the formalin-killed vaccine or control fish, supporting the serum bactericidal activity results at these time points. The cumulative mortality of the unvaccinated control fish was 87% after challenging fish with the pathogen, while the cumulative mortality of the CS-NE vaccinated group was 24%, which was significantly lower than the formalin-killed vaccinated and control fish. There was a significant upregulation of IgM, IgT, TNF α, and IL1-β genes in the spleen and kidney of vaccinated fish. Significant upregulation of IgM and IgT genes was observed in the spleen of CS-NE vaccinated fish. The study confirmed the charged-chitosan-based mucoadhesive nanovaccine to be an effective platform for immersion vaccination of tilapia, with fish generating a humoral systemic immune response against columnaris disease in vaccinated fish

Immersion Vaccination by a Biomimetic-Mucoadhesive Nanovaccine Induces Humoral Immune Response of Red Tilapia (Oreochromis sp.) against Flavobacterium columnare Challenge

Immersion vaccination with a biomimetic mucoadhesive nanovaccine has been shown to induce a strong mucosal immune response against columnaris disease, a serious bacterial disease in farmed red tilapia caused by Flavobacterium columnare. However, the induction of a systemic immune response by the vaccine is yet to be investigated. Here, we examine if a specific humoral immune response is stimulated in tilapia by a biomimetic-mucoadhesive nanovaccine against Flavobacterium columnare using an indirect-enzyme-linked immunosorbent assay (ELISA), serum bactericidal activity (SBA) and the expression of immune-related genes within the head-kidney and spleen, together with assessing the relative percent survival of vaccinated fish after experimentally infecting them with F. columnare. The anti-IgM antibody titer of fish at 14 and 21 days post-vaccination was significantly higher in chitosan complex nanoemulsion (CS-NE) vaccinated fish compared to fish vaccinated with the formalin-killed vaccine or control fish, supporting the serum bactericidal activity results at these time points. The cumulative mortality of the unvaccinated control fish was 87% after challenging fish with the pathogen, while the cumulative mortality of the CS-NE vaccinated group was 24%, which was significantly lower than the formalin-killed vaccinated and control fish. There was a significant upregulation of IgM, IgT, TNF α, and IL1-β genes in the spleen and kidney of vaccinated fish. Significant upregulation of IgM and IgT genes was observed in the spleen of CS-NE vaccinated fish. The study confirmed the charged-chitosan-based mucoadhesive nanovaccine to be an effective platform for immersion vaccination of tilapia, with fish generating a humoral systemic immune response against columnaris disease in vaccinated fish

Effect of <i>Moringa oleifera</i> Leaf Extract on the Growth Performance, Hematology, Innate Immunity, and Disease Resistance of Nile Tilapia (<i>Oreochromis niloticus</i>) against <i>Streptococcus agalactiae</i> Biotype 2

The present study aimed to investigate the effects of Moringa oleifera leaf (MLE) extract-supplemented diets on the growth, feed utilization, hematology, innate immune response, and disease resistance of Nile tilapia against Streptococcus agalactiae Biotype 2. Four hundred and fifty Nile tilapia (32.61 ± 0.2 g/fish) were randomly allocated into fifteen tanks (30 fish/tank). Different concentrations of MLE at 0%, 0.5%, 1%, 1.5%, and 2% were fed to the Nile tilapia for 30 days, and the growth, feed utilization, hematology, and innate immune response of the Nile tilapia were determined. After the feeding trial, the Nile tilapia were challenged with a S. agalactiae Biotype 2 infection, and the relative percentage of survival (RPS) was determined. Results revealed the presence of quercetin, kaempferol, and p-coumaric acid in the MLE extract, exhibiting stronger antimicrobial activity against S. agalactiae Biotype 2. The diets supplemented with the MLE-0.5 group showed a significantly higher growth, feed utilization, hematology, and innate immune response in the Nile tilapia compared to the control and other MLE groups. Additionally, the MLE-0.5 group exhibited a significantly higher RPS of the Nile tilapia against S. agalactiae Biotype 2. Therefore, MLE-0.5 can be employed as an alternative feed supplement in sustainable Nile tilapia farming to protect against S. agalactiae Biotype 2