Background and Aim: African swine fever is one of the severe pathogens of swine. It has a significant impact on production and economics. So far, there are no known remedies, such as vaccines or drugs, reported working successfully. In the present study, the natural oil blend formulation's (NOBF) efficacy was evaluated against ASFV in vitro using porcine alveolar macrophages (PAMs) cells of swine. Materials and Methods: The capacity of NOBF against the ASFV was tested in vitro. The NOBF combines Eucalyptus globulus, Pinus sylvestris, and Lavandula latifolia. We used a 2-fold serial dilution to test the NOBF formulation dose, that is, 105 HAD50/mL, against purified lethal dose of African swine in primary PAMs cells of swine. The PAM cells survival, real-time polymerase chain reaction (PCR) test, and hemadsorption (HAD) observation were performed to check the NOBF efficacy against ASFV. Results: The in vitro trial results demonstrated that NOBF up to dilution 13 or 0.000625 mL deactivates the lethal dose 105 HAD50 of ASFV. There was no HAD (Rosetta formation) up to dilution 12 or 0.00125 mL of NOBF. The Ct value obtained by running real-time PCR of the NOBF group at 96 h post-infection was the same as the initial value or lower (25), whereas the Ct value of positive controls increased several folds (17.84). Conclusion: The in vitro trial demonstrated that NOBF could deactivate the ASFV. The NOBF has the potential to act as anti-ASFV agent in the field. The next step is to conduct in vivo level trial to determine its efficacy.

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.

A bioassay trial was conducted to determine the efficacy of developed product, Pondguard as an anti-Acute Hepatopancreatic Necrosis Disease (AHPND) candidate. The Pondguard (Registration no. D 16060285-HBC) is consisting of natural oils, like, lavender oil, Eucalyptus oil and Pine oil. It maintains the immunity level of shrimp, which helps to protect shrimp from infectious diseases.

Natural guard liquid (NGL) is an immunomodulator consisting of an essential oil blend (lavender oil, eucalyptus oil, and pine oil) that can improve the immunity (IgG) of animals. The objective of this research was to assess the effectiveness of a mixture of NGL comprising of essential oil (eucalyptus, lavender, and pine oil) on growth performance, hematological profile, and antibody titer response to the Newcastle disease virus (NDV) in Lohman strain broiler chickens. A total of 400-day-old unsexed broiler chickens with an average weight of 42.48 ± 2.08 g were randomly distributed into four groups, each comprising two replications with 50 chicks. The control group, T0, received no essential oil, while other treatment groups, T1, T2, and T3, were administered NGL at 80, 100, and 200 ppm, respectively. The mixture was administered in drinking water for 30 days. The animals had received vaccinations at the Hatchery, including active NDV, inactive NDV, and Gumboro. Regular recordings were made for feed consumption, water intake, mortality, and body weight. Blood samples for routine hematological examination (hemoglobin, erythrocytes, packed cell volume, erythrocyte index, leucocytes, and differential leucocytes) and immune parameter (NDV antibody titer) assessment were collected at the onset as well as after 15 and 30 days of treatment. The hemoglobin levels, erythrocytes, total leukocytes, lymphocytes, and Heterophil-lymphocyte index showed significant differences in groups T1 and T2 compared to T0. While the NDV antibody titer showed a significant difference at T2 compared to T0, it was not significantly different at T1 and T3. The best results and performance was indicated in T2 (100 ppm), characterized by body weight (1,839 g), and feed conversion ratio (1.573). The hematological profile in the T2 group included hemoglobin (11.78 g/dL), total erythrocytes (2.82 ×106 u/L), total leukocytes (52.67 ×103 u/L]), and the NDV antibody titer (48 ± 10.20) in the fifth week. In conclusion, the optimum dose of NGL is 100 ppm, which significantly influences growth performance, hematological profile, and antibody titer against NDV. Keywords: Antibody titer, Broiler chicken, Growth performance, Hematological profile, Natural Guard Liquid

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. HIGHLIGHTS: The 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.

Impact of Natural Oil Blend Formulation (NOBF) on Calves during Transportation