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Background and Aim: African swine fever (ASF) is currently the most prevalent disease in swine. The disease is spreading throughout primary swine-producing countries with heavy losses in population and revenue. To date, no successful vaccines or medications have been reported. This study aimed to design and develop a blend of natural essential oils and test its efficacy against the ASF virus (ASFV) in swine. Materials and Methods: We attempted to develop a natural oil blend formulation (NOBF) and determine its efficacy against the ASFV. This study follows on from a previously published in vitro study that reported that the NOBF has anti ASFV properties. A study was designed using 21 healthy piglets of triple-cross (Landrace + Yorkshire + Durok) crossbred pathogen-free pigs with an average weight of 15 kg. The study consisted of NOBF-incubated, NOBF, positive control, and negative control groups. The NOBF groups were administered NOBF (80 mL/ton mixed in drinking water) beginning 10 days before the challenge and continuing throughout the experiment. The positive and negative control pigs consumed regular drinking water. The pigs were challenged by a sublethal dose of pure isolate ASFV strain Vietnam National University of Agriculture-ASFV-L01/HN/04/19 inoculation with 103.5 HAD50/dose through the intramuscular route. There were sic pigs in each group, three pigs directly IM challenged, and three pigs were considered cohoused pigs. Results: Both challenged (three) and cohoused (three) pigs in the positive control showed clinical signs of ASFV infection, as detected by real-time polymerase chain reaction (RT-PCR) in blood samples, oral swabs, and feces. There was 100% cumulative mortality, that is, both challenged and contact pigs died in the positive control group on day 20 of infection. No signs of infection or mortality were observed in the NOBF-incubated group. The challenged pigs in the NOBF-direct challenge group showed clinical signs and mortality, whereas no clinical signs or symptoms occurred in the cohoused pigs. The immunoglobulin G (IgG) level of the contact pigs was the highest in the treatment group and the lowest in the positive control group. The IgM level of the contact pigs in the treatment groups was the lowest, whereas that of the positive control was the highest. The RT-PCR test showed that the ASFV was deactivated in the NOBF-incubated group. The challenged and contact pigs of the positive control group had high Ct values. The challenged pigs of the NOBF group had high Ct values, whereas the contact pigs from the same group and those of the negative control were negative for the ASFV, determined by PCR, in all samples. The comparison of the challenged groups showed that the appearance of the virus was delayed by at least 2 days in the NOBF group compared to the positive control group. Conclusion: The results showed that NOBF can prevent the spread of the ASFV in a population. Moreover, NOBF can enhance the pig humoral immune system by enhancing IgG levels and reducing IgM levels. This study successfully demonstrated that NOBF is an anti-ASFV agent, which prevents horizontal transmission and enhances pig humoral immunity.

The Vibrio group of bacteria is considered highly pathogenic to shrimp larvae. Shrimp larvae are susceptible to Vibrio. It is difficult to eradicate Vibrio from the hatchery environment completely. Efforts were made to design an effective disinfectant with almost no side effects or residues. A formulation was developed by combining essential oils with antimicrobial properties to fight against pathogenic Vibrios, such as Vibrio harveyi and V. parahaemolyticus, in a hatchery environment. The anti-Vibrio formulation is a blend of Eucalyptus globulus, jasmine, and gardenia oils, designated as essential oil blend formulation (EOBF). The effective dose of EOBF was optimized on bacteria using TSA and on Vibrio using TCBS agar media. The optimized dose of 40 ppm EOBF was sprayed on the tank and culture area dry surfaces. The trial sites selected were the water filter tank, maturation tank, fry reservoir tank, quarantine and maturation tank, hatchery tank, spawning tank, rinsing tank, plankton tank, larvae rearing tank, packing tank, artemia tank, polychaetes (bucket and floor), squid (equipment and table), and freshwater tank of an experimental hatchery. The swab samples were collected after 3 hours, and a significant decrease in Vibrio, mostly green colonies, such as Vibrio harveyi and V. parahaemolyticus, was found after EOBF application. The obtained results showed that EOBF is an effective disinfectant against pathogenic Vibrios in shrimp hatcheries

Infectious Myonecrosis Virus (IMNV) was first reported in Indonesia in 2006. By 2009, it spread all over the significant shrimp farming areas in Indonesia. There was a significant drop in shrimp production due to IMNV outbreak. Several efforts were made to understand the nature, mode of disease progression, and minimize the virus load. The IMNV progression was divided into four stages. Chemicals, especially of chlorine origin, showed protection against IMNV. Tilapia came up with an effective biological remedy against IMNV. The blend of essential oils, pondguard showed significant protection against IMNV in the laboratory and farms.

The white spot syndrome virus (WSSV) is lethal in penaeid shrimp. Successful efforts made to develop a natural oil blend formulation (NOBF) by blending Eucalyptus globulus, Pinus sylvestris, and Lavandula latifolia in different compositions with anti-WSSV properties. A bioassay challenge trial conducted using 1 g of specific pathogen-free Penaeus vannamei Boone, 1931 shrimp in 4 replicates of aquaria for each group. The NOBF dose of 0.4 ppm applied throughout the trial period by mixing in the aquarium water daily, starting seven days before the challenge. The efficacy of NOBF against WSSV was measured using the per os method of a challenge. The cumulative mortality in the positive control group reached 100 % after ten days of the challenge. The NOBF was applied in the commercial shrimp ponds to study its efficacy and palatability. A study in 8 ponds in a WSSV prone area in Indonesia, demonstrated no significant difference in crucial water quality parameters, especially on algae as natural food. The NOBF applied ponds had higher productivity (1.28 Kg of shrimp per m3) as compared to control (1.18 Kg of shrimp per m3). The FCR of NOBF group was lower (1.41) than control (1.53). The survival rate of the NOBF group was higher (86.96%) than control (80.36). The trial outcomes show that NOBF is safe and user-friendly, with properties to reduce pathogen load.

African swine fever is one of the severe pathogens of swine. It has a significant impact on production and on economics. So far, there are no known remedies, such as vaccines or drugs, reported. The natural oil blend formulation (NOBF) successfully tested against the African swine fever virus (ASFV) in in vitro conditions. The natural oil blend formulation (NOBF) combines Eucalyptus globulus, Pinus sylvestris, and Lavandula latifolia. We used a two-fold serial dilution to test the NOBF formulation dose. The in vitro trial results demonstrated that NOBF up to dilution 13 or 0.000625 ml deactivates the lethal dose 105HAD50 of ASFV. There was no hemadsorption (Rosetta formation) up to dilution 12 or 0.00125 ml of NOBF. The Ct value of the NOBF group at 96 hours post-infection was the same as the initial value or lower (25), whereas the Ct value of positive controls increased several folds (17.84). The in vitro trial demonstrated that NOBF can deactivate the African swine fever virus.HIGHLIGHTSThe natural oil blend formulation (NOBF) was formulated using three natural oils, i.e., Eucalyptus globulus, Pinus sylvestris, and Lavandula latifolia.The in vitro trial was conducted using porcine alveolar macrophages (PAMs) and further passaged in the PAMs; the stock used in the present study was that obtained after the 15th passage.The natural oil blend formulation (NOBF) showed protection against ASF virus up to a dilution of 13 or 0.000625 of a dilution of 16 or 0.000078 ml that was tried.The real-time PCR analysis showed that the virus did not replicate in the NOBF group, which implies that either ASFV growth was inhibited in the presence of NOBF or that it was inactivated.The in vitro trial outcome showed that NOBF has anti-ASFV properties.

Crayfish, commonly known as freshwater lobster, have been scientifically distributed in three genuses, which are as follows: - Cumberfish, Oronectes and Phalisaberus. Crayfish breeding and rearing techniques in China and some Adapted and practiced in South-East Asian countries. The proper ratio of broodstock female and male is 3: 1. Adult males are comparatively larger than females. The mating season of crayfish is usually from May to September. Larvae are stored in cultured ponds. Crayfish are cultivated in paddy fields with carp fish or eel fish in lotus and polyculture. The size of crayfish transported to the market is 20–60h in a period of 3 to 7 months, and is sold alive in most markets. Productivity varies from 750h to 3,000h per ha depending on the type of farming. It is possible that the farmer can earn up to 7000 to 14,000 rupees per day.