Toxicity Evaluation of Selected Plant Water Extracts on a Honey Bee (Apis mellifera L.) Larvae Model

So far, larval rearing in vitro has been an important method in the assessment of bee toxicology, particularly in pesticide risk assessment. However, natural products are increasingly used to control honey bee pathogens or to enhance bee immunity, but their effects on honey bee larvae are mostly unknown. In this study, laboratory studies were conducted to determine the effects of including selected aqueous plant infusions in the diet of honey bee (Apis mellifera L.) larvae in vitro. The toxicity of infusions from three different plant species considered to be medicinal plants was evaluated: tansy (Tanacetum vulgare L.), greater celandine (Chelidonium majus L.), and coriander (Coriandrum sativum L.). The impact of each on the survival of the larvae of honey bees was also evaluated. One-day-old larvae were fed a basal diet consisting of distilled water, sugars (glucose and fructose), yeast extract, and freeze-dried royal jelly or test diets in which distilled water was replaced by plant infusions. The proportion of the diet components was adjusted to the age of the larvae. The larvae were fed twice a day. The experiment lasted seven days. Significant statistical differences in survival rates were found between groups of larvae (exposed or not to the infusions of tansy, greater celandine, and coriander). A significant decrease (p < 0.05) in the survival rate was observed in the group with the addition of a coriander herb infusion compared to the control. These results indicate that plant extracts intended to be used in beekeeping should be tested on all development stages of honey bees.

DETERMINING THE OPTIMUM TEMPERATURE FOR GROWTH, FEED EFFICIENCY AND SURVIVAL OF DOMESTICATED INDONESIAN MAHSEER, Tor soro LARVAE

Temperature plays a pivotal role in the success of larvae production. Evaluation of the effects of different temperatures on growth, feed efficiency, and survival of domesticated Indonesian mahseer (Tor soro) larvae was carried out to determine an optimum rearing temperature. Five different temperature settings of 22°C, 24°C, 26°C, 28°C, and 30°C were used as the treatments arranged in triplicates, in indoor closed recirculating systems. The larvae were stocked in a rectangular glass tank with a stocking density of 150 larvae per tank and fed with a commercial feed (30% protein content) three times a day, about 10% per body weight over 61 days of observation. Measured parameters included the specific growth rate of total weight (SGRTW) and length (SGRTL), feed efficiency (FE), and survival rate (SR). The result showed that the treatments of different temperatures significantly affected the growth performance and feed efficiency of the larvae. The growth performance and feed efficiency of mahseer larva were much better at rearing temperature from 24°C to 28°C compared with those at 22°C and 30°C. There were no significant differences in SR in different temperatures tested. The SR was found to range from 86.44% to 100% in all treatments. Maintaining temperature ranging from 24°C to 28°C is the best rearing condition to achieve the optimum growth and feed efficiency of mahseer during the larval rearing period. The findings from this research could be set as a standard technique in larval rearing of Indonesian mahseer (Tor soro).

In Vitro Rearing Changes Social Task Performance and Physiology in Honeybees

In vitro rearing of honeybee larvae is an established method that enables exact control and monitoring of developmental factors and allows controlled application of pesticides or pathogens. However, only a few studies have investigated how the rearing method itself affects the behavior of the resulting adult honeybees. We raised honeybees in vitro according to a standardized protocol: marking the emerging honeybees individually and inserting them into established colonies. Subsequently, we investigated the behavioral performance of nurse bees and foragers and quantified the physiological factors underlying the social organization. Adult honeybees raised in vitro differed from naturally reared honeybees in their probability of performing social tasks. Further, in vitro-reared bees foraged for a shorter duration in their life and performed fewer foraging trips. Nursing behavior appeared to be unaffected by rearing condition. Weight was also unaffected by rearing condition. Interestingly, juvenile hormone titers, which normally increase strongly around the time when a honeybee becomes a forager, were significantly lower in three- and four-week-old in vitro bees. The effects of the rearing environment on individual sucrose responsiveness and lipid levels were rather minor. These data suggest that larval rearing conditions can affect the task performance and physiology of adult bees despite equal weight, pointing to an important role of the colony environment for these factors. Our observations of behavior and metabolic pathways offer important novel insight into how the rearing environment affects adult honeybees.

Genome editing in cultured fishes

With external fertilization, high fecundity, and established methods for propagation and larval rearing for cultured species, fish provide systems well suited to genome-editing procedures. While early experiments utilized zinc-finger nucleases and transcription activator-like effector nucleases (TALENs), most recent ones have used the CRISPR/Cas9 editor, and achieved rates of targeted genomic insertion well above those of classical transgenic methods, with lower frequencies of off-site integration. Genome-editing experiments with cultured fishes have focused on improving growth rate and disease resistance, achievement of reproductive confinement, and other valued traits. As reviewed here, advances in knowledge of key molecular pathways and, in some cases, favorable alterations of phenotype have been achieved. For example, loss-of-function of myostatin, a negative regulator of muscle growth, led to increased muscle mass, greater weight, and greater fillet yield in genome-edited lines of red sea bream, tiger puffer, and Nile tilapia than in their unedited counterparts. The red sea bream line become the first genome-edited animal to reach commercial production. As for all animals, wide adoption of genome-edited fishes will depend upon addressing issues of regulation, consumer acceptance, and breeding infrastructure.

Increased Coral Larval Supply Enhances Recruitment for Coral and Fish Habitat Restoration

Loss of foundation reef-corals is eroding the viability of reef communities and ecosystem function in many regions globally. Coral populations are naturally resilient but when breeding corals decline, larval supply becomes limiting and natural recruitment is insufficient for maintaining or restoring depleted populations. Passive management approaches are important but in some regions they are proving inadequate for protecting reefs, therefore active additional intervention and effective coral restoration techniques are needed. Coral spawning events produce trillions of embryos that can be used for mass larval rearing and settlement on degraded but recoverable reef areas. We supplied 4.6 million Acropora tenuis larvae contained in fine mesh enclosures in situ on three degraded reef plots in the northwestern Philippines during a five day settlement period to initiate restoration. Initial mean larval settlement was very high (210.2 ± 86.4 spat per tile) on natural coral skeleton settlement tiles in the larval-enhanced plots, whereas no larvae settled on tiles in control plots. High mortality occurred during early post-settlement life stages as expected, however, juvenile coral survivorship stabilised once colonies had grown into visible-sized recruits on the reef by 10 months. Most recruits survived and grew rapidly, resulting in significantly increased rates of coral recruitment and density in larval-enhanced plots. After two years growth, mean colony size reached 11.1 ± 0.61 cm mean diameter, and colonies larger than 13 cm mean diameter were gravid and spawned, the fastest growth to reproductive size recorded for broadcast spawning corals. After three years, mean colony size reached 17 ± 1.7 cm mean diameter, with a mean density of 5.7 ± 1.25 colonies per m–2, and most colonies were sexually reproductive. Coral cover increased significantly in larval plots compared with control plots, primarily from A. tenuis recruitment and growth. Total production cost for each of the 220 colonies within the restored breeding population after three years was United States $17.80 per colony. A small but significant increase in fish abundance occurred in larval plots in 2018, with higher abundance of pomacentrids and corallivore chaetodontids coinciding with growth of A. tenuis colonies. In addition, innovative techniques for capturing coral spawn slicks and larval culture in pools in situ were successfully developed that can be scaled-up for mass production of larvae on reefs in future. These results confirm that enhancing larval supply significantly increases settlement and coral recruitment on reefs, enabling rapid re-establishment of breeding coral populations and enhancing fish abundance, even on degraded reef areas.

First hatchery bred streaked spinefoot rabbitfish (Siganus javus) in Sabah, Malaysia

The present study aims to expand the market potential of the streaked spinefoot rabbitfish (Siganus javus) in Malaysia and provide consumers more choices of affordable protein source from aquaculture. Spawning hehavior of S.javus was recorded in a week after the new moon in October 2019 and January 2020. Spawning occurred between midnight and early morning, and the eggs hatched about 16 to 18 hours later. Throughout the incubation and larvae rearing period, water temperature was maintained at 30 to 31 °C. Water salinity, pH and DO were 31 to 33 ppt, 7.8 to 8, and 5 to 7 mg/l, respectively. Eggs diameter ranged from 0.550 to 0.603 mm. At 1 day after hatched (d AH), eyes of the larvae were formed but unpigmented and mouth structure was not developed. At 2 d AH, the pigmentation of eyes darkened, digestive tract formed, and anus and mouth opened. At 3 d AH, the lower jaw began to show movement, indicating that the ability of larvae to indulge in feeding. The buds of dorsal and ventral fins started to develop and elongate from 9 d AH and were covered with melanophores. At 25 d AH, the body of the larvae turned brownish and abdominal cavity was deeply pigmented, indicating the onset of metamorphosis. By the age of 30 d AH, the larvae completed development into juvenile stage. At 270 d AH, the juvenile reached maximum size of 21 cm in total length. This study was the first successful attempt at naturally spawning and hatchery rearing of S. javus into juvenile stage in the country. The development of a complete larval rearing protocol for S. javus could provide information of practical importance in hatchery breeding of other rabbitfish species.

Effects of Food Concentration and Photoperiod on Egg Production, Female Life Expectancy and Population Dynamics of the Paracalanid Copepod, Bestiolina amoyensis

The paracalanid copepod, Bestiolina amoyensis, is a widely distributed species occurring in subtropical inshore waters across the Pacific Ocean. Its small size, herbivorous feeding habit, and high adaptability make the species one of the most promising candidates as potential live feed for hatchery larval rearing. This study investigated effects of different feeding density of microalgae Isochrysis spp. (1 × 105, 2 × 105, 3 × 105, 4 × 105, and 5 × 105 cells ml–1) and photoperiod (8L:16D, 12L:12D, and 16L:8D) on productivity-related parameters, including egg production, female life expectancy and population dynamics of B. amoyensis. Results showed that total egg output over female lifespan, final population size and intrinsic rate of population of 12L:12D photoperiod treatments were always the highest among three photoperiod conditions, especially at the food concentration of 4 × 105 cells ml–1, indicating B. amoyensis had high reproductive performance and the population was in a more stable status. The number of nauplii from 4 × 105 cells ml–1 algal concentration treatment accounted for 75% of the population, and the ratio of females to males approaching 1:1 when photoperiod was 12L:12D; female life expectancy was 10.5 ± 0.6 days. In conclusion, our results showed that Isochrysis spp. is a suitable feed for B. amoyensis with an optimal concentration at 4 × 105 cells mL–1; the optimal photoperiod for B. amoyensis rearing is 12L:12D. The relatively long reproductive lifespan and high intrinsic population increase rate make B. amoyensis a good candidate to develop culture techniques for hatchery larval rearing.

ẢNH HƯỞNG CỦA MẬT ĐỘ VÀ TẦN SUẤT CHO ĂN ĐẾN HIỆU QUẢ ƯƠNG GIỐNG VÀ STRESS Ở CÁ LEO – Wallago attu (Bloch & Schneider, 1801)

Nhằm xác định ảnh hưởng của mật độ và tần suất cho ăn đến sinh trưởng và tỉ lệ sống cá Leo Wallago attu giai đoạn ương giống, thí nghiệm được tiến hành với 5 mức mật độ (1, 2, 4, 8 và 16 con/L) và các tần suất cho ăn (1, 2, 3, 4 và 5 lần/ngày). Kết quả nghiên cứu cho thấy mật độ ương và tần suất cho ăn đã ảnh hưởng có ý nghĩa thống kê đến tỉ lệ sống của cá thí nghiệm. Tỉ lệ sống của cá giảm ở mật độ ương nuôi cao (p<0,05). Tỉ lệ ăn thịt đồng loại giảm ở các nghiệm thức mật độ nuôi thấp. Tần suất cho cá ăn có tương quan thuận với tỉ lệ sống, tỉ lệ sống của cá tăng trên 50% khi tăng số lần cho ăn lên 3 – 5 ngày/lần. Tỉ lệ ăn thịt đồng loại của cá có mối tương quan nghịch với tần suất cho ăn. Nồng độ cortisol trong máu cá được xem như chỉ thị đối với mức độ stress của cá. Kết quả của nghiên cứu này cho thấy rằng nồng độ cortisol của cá Leo ở mật độ nuôi cao và tần suất cho ăn 1 và 2 lần/ngày cao hơn so với những nghiệm thức còn lại. Do vậy, mức độ stress của cá Leo có thể được xem như một trong những yếu tố chính dẫn đến hiện tượng ăn thịt lẫn nhau và làm giảm tỉ lệ sống đối với loài cá này ở giai đoạn giống. Những kết quả của nghiên cứu này có thể cung cấp thông tin hữu ích nhằm nâng cao hiệu quả của việc ương nuôi cá Leo giống thông qua quản lý mật độ nuôi và tần suất cho ăn. ABSTRACT In order to elucidate the effects of stocking density and feeding frequency on the growth and survival of Wallago attu, post-hatch larvae were stocked at different densities (1, 2, 4, 8 and 16 individuals per liter) and fed with (1, 2, 3, 4 and 5-times per day). The results indicate that stocking density and feeding frequency had significant influences on the survival rate of the larvae (p<0.05). In particular, the survival rate of larvae decreased at trials with high stocking density (p<0.05) due to the cannibalism. The percentage of cannibalism decreased in treatments with low stocking densities. Regarding effects of feeding frequency, the proportionality between the survival rate of larvae and high feeding frequency was observed in this study (p<0.05). The survival rate of larvae reached 50% when fish were fed 3 -5 times per day. The cannibalism had negative relationship with frequency of daily feeding. The cortisol concentration in fish blood is considered as an indicator of stress levels of fish. The results of this study indicated that cortisol concentration of W. attu reared at high stocking density and fed 1-2 times/day was higher than that in other treatments. Therefore, stress levels of W. attu could be one of the main factors causing cannibalism and reduction of survival rate at larval stages. This study provided useful information to enhance the effectiveness of larval rearing of W. attu via controlling stocking density and feeding frequency.

Aedes albopictus life table: environment, food, and age dependence survivorship and reproduction in a tropical area

Background Environmental conditions affect the biology of mosquito vectors. Aedes albopictus is a major vector of many important diseases including dengue, Zika, and chikungunya in China. Understanding the development, fecundity, and survivorship of Ae. albopictus mosquitoes in different environmental conditions is beneficial for the implementation of effective vector control measures. Methods Aedes albopictus larval and adult life-table experiments were conducted under natural conditions in indoor, half-shaded, and fully shaded settings, simulating the three major habitat types in Hainan Province, a tropical island in the South China Sea. Temperature, humidity, and light intensity were recorded daily. Larval rearing used habitat water and tap water, with and without additional artificial food. Development time, survivorship, pupation rate, and adult emergence rates were monitored. Adult mosquito survivorship and fecundity were monitored daily and reproductive rates were determined, and age-dependent survivorship and reproduction were analyzed. Results The pupation time and male and female emergence times were significantly shorter in indoor conditions than in shaded and half-shaded conditions for both tap water with added food and habitat water with added food groups. For habitat water with added food, the shaded environment had the lowest pupation rate among the settings. For tap water with added food group, the shaded environment had the lowest pupation rate. The mean survival time of females was 27.3 ± 0.8 days in the indoor condition, which was significantly longer than that in the half-shaded (18.4 ± 0.6 days) and shaded (13.8 ± 1.2 days) conditions. Adult mortality was age-dependent, and the rate of change in mortality with age was not significantly different among different environmental conditions. The mean net replacement rate (R0) of female mosquitoes showed no significant difference among the three conditions, whereas the per capita intrinsic growth rate (r) in the shaded condition was 42.0% and 20.4% higher than that in the indoor and half-shaded conditions, respectively. Female daily egg mass was also age-dependent in all the settings, decaying exponentially with age. Conclusions Our results imply that half-shaded conditions are likely the best natural condition for adult emergence and female reproduction, and food supply is crucial for larval development and pupation. The results provide new avenues for integrated mosquito management in indoor and outdoor areas, especially in half-shaded areas. Graphical Abstract