Chanathip Thammakarn

Inactivation of Avian Influenza Virus, Newcastle Disease Virus and Goose Parvovirus Using Solution of Nano-Sized Scallop Shell Powder

ABSTRACT Scallop shell powder produced by calcination process − the average diameter of the powder particles being 20 µm (SSP) − was further ground into nano-sized particles, with average diameter of 500 nm, here designated CaO-Nano. Solution of CaO-Nano could inactivate avian influenza virus within 5 sec, whereas the solution of SSP could not even after 1 hr incubation. CaO-Nano solution could also inactivate Newcastle disease virus and goose parvovirus within 5 sec and 30 sec, respectively. The virus-inactivating capacity (neutralizing index: NI>3) of the solution was not reduced by the presence of 20% fetal bovine serum. CaO-Nano solution seems to be a good candidate of materials for enhancement of biosecurity in farms.

Evaluation of sprayed hypochlorous acid solutions for their virucidal activity against avian influenza virus through in vitro experiments

Hypochlorous acid (HOCl) solutions were evaluated for their virucidal ability against a low pathogenic avian influenza virus (AIV), H7N1. HOCl solutions containing 50, 100 and 200 ppm chlorine (pH 6) or their sprayed solutions (harvested in dishes placed at 1 or 30 cm distance between the spray nozzle and dish) were mixed with the virus with or without organic materials (5% fetal bovine serum: FBS). Under plain diluent conditions (without FBS), harvested solutions of HOCl after spraying could decrease the AIV titer by more than 1,000 times, to an undetectable level (< 2.5 log10TCID50/ml) within 5 sec, with the exception of the 50 ppm solution harvested after spraying at the distance of 30 cm. Under the dirty conditions (in the presence of 5% FBS), they lost their virucidal activity. When HOCl solutions were sprayed directly on the virus on rayon sheets for 10 sec, the solutions of 100 and 200 ppm could inactivate AIV immediately after spraying, while 50 ppm solution required at least 3 min of contact time. In the indirect spray form, after 10 sec of spraying, the lids of the dishes were opened to expose the virus on rayon sheets to HOCl. In this form, the 200 ppm solution inactivated AIV within 10 min of contact, while 50 and 100 ppm could not inactivate it. These data suggest that HOCl can be used in spray form to inactivate AIV at the farm level.

Calcinated egg shell as a candidate of biosecurity enhancement material.

Calcinated egg shell (Egg-CaO), of which the main component is calcium oxide, was evaluated in the forms of powder and aqueous solutions for their efficacies as disinfectants against avian influenza virus (AIV), Newcastle disease virus (NDV), infectious bursal disease virus (IBDV), Salmonella Infantis and Escherichia coli. Egg-CaO powder inactivated these viruses within 3 min in the presence of 33% of fetal bovine serum (FBS). In Egg-CaO solutions, except AIV, all pathogens were inactivated within 1 hr, even in the presence of 5% of FBS. Without FBS, all pathogens, except AIV, were inactivated within 3 min, and AIV within 1 hr. In addition, persistence of virucidal activity against AIV and NDV of Egg-CaO powder was confirmed after exposure to sunlight for 2 weeks or resuspension with water for 7 times, simulating field harsh environments. Chick growth test was conducted to ensure the safety of the use of Egg-CaO powder in chicken cages and showed that it is safe to add Egg-CaO in litter or feed. In conclusion, Egg-CaO can be useful for the enhancement of biosecurity at farms.

Efficacy of scallop shell powders and slaked lime for inactivating avian influenza virus under harsh conditions

The efficacy and stability of scallop shell powder (SSP) were investigated, in terms of its capacity to inactivate avian influenza virus (AIV), and compared with slaked lime (SL). An environmental simulation was conducted by emulating sunlight and wet-dry conditions. The powders were collected at consecutive 2-week intervals under sunlight and upon every resuspension. These materials were tested by mixing them with AIV and incubating the mixture for 3 min or 20 h, followed by AIV titration. At the same time, a pH buffering test was conducted by neutralization with Tris-HCl. The results revealed that SSP and SL have high alkalinity and excellent ability to inactivate AIV. In a simulated harsh environment, SSP and SL retained a satisfactory ability to inactivate AIV within 20 h throughout the experimental procedure. However, SSP was able to inactivate AIV during a short contact period (3 min), even under harsh conditions, and it was more resistant than SL to neutralization.

Inactivation of bacteria on surfaces by sprayed slightly acidic hypochlorous acid water: in vitro experiments

The capacity of slightly acidic hypochlorous acid water (SAHW), in both liquid and spray form, to inactivate bacteria was evaluated as a potential candidate for biosecurity enhancement in poultry production. SAHW (containing 50 or 100 ppm chlorine, pH 6) was able to inactivate Escherichia coli and Salmonella Infantis in liquid to below detectable levels (≤2.6 log10 CFU/ml) within 5 sec of exposure. In addition, SAHW antibacterial capacity was evaluated by spraying it using a nebulizer into a box containing these bacteria, which were present on the surfaces of glass plates and rayon sheets. SAHW was able to inactivate both bacterial species on the glass plates (dry condition) and rayon sheets within 5 min spraying and 5 min contact times, with the exception of 50 ppm SAHW on the rayon sheets. Furthermore, a corrosivity test determined that SAHW does not corrode metallic objects, even at the longest exposure times (83 days). Our findings demonstrate that SAHW is a good candidate for biosecurity enhancement in the poultry industry. Spraying it on the surfaces of objects, eggshells, egg incubators and transport cages could reduce the chances of contamination and disease transmission. These results augment previous findings demonstrating the competence of SAHW as an anti-viral disinfectant.

Inhibition of infectious bursal disease virus transmission using bioceramic derived from chicken feces

Bioceramic powder (BCX), at pH 13.0, derived from chicken feces, was evaluated for its efficacy to inactivate virus and inhibit virus horizontal transmission by fecal-oral route, using infectious bursal disease virus (IBDV) vaccine strain D78 as a challenge virus. Three 1-week-old SPF chicks were vaccinated per os and used as seeder birds. Six hours later, 3 sentinel 1-week-old SPF chicks were introduced into the same cage. Results revealed that BCX had excellent efficacy to inactivate IBDV within 3 min. Treating IBDV contaminated litter in the cage with BCX could prevent transmission of IBDV to new sensitive chicks completely. Further, transmission of IBDV to the sentinel chicks was significantly inhibited by adding BCX to litter and chicken feed. These data suggest that BCX at pH 13, derived from chicken feces, has excellent efficacy to inactivate IBDV, which can be applied in bedding materials for preventing viral transmission during production round. It is a good material that can effectively be used for enhancing biosecurity system in poultry farms.

Aerosol Disinfection Capacity of Slightly Acidic Hypochlorous Acid Water Towards Newcastle Disease Virus in the Air: An In Vivo Experiment

SUMMARY Existence of bioaerosol contaminants in farms and outbreaks of some infectious organisms with the ability of transmission by air increase the need for enhancement of biosecurity, especially for the application of aerosol disinfectants. Here we selected slightly acidic hypochlorous acid water (SAHW) as a candidate and evaluated its virucidal efficacy toward a virus in the air. Three-day-old conventional chicks were challenged with 25 doses of Newcastle disease live vaccine (B1 strain) by spray with nebulizer (particle size <3 &mgr;m in diameter), while at the same time reverse osmosis water as the control and SAHW containing 50 or 100 parts per million (ppm) free available chlorine in pH 6 were sprayed on the treated chicks with other nebulizers. Exposed chicks were kept in separated cages in an isolator and observed for clinical signs. Oropharyngeal swab samples were collected from 2 to 5 days postexposure from each chick, and then the samples were titrated with primary chicken kidney cells to detect the virus. Cytopathic effects were observed, and a hemagglutination test was performed to confirm the result at 5 days postinoculation. Clinical signs (sneezing) were recorded, and the virus was isolated from the control and 50 ppm treatment groups, while no clinical signs were observed in and no virus was isolated from the 100 ppm treatment group. The virulent Newcastle disease virus (NDV) strain Sato, too, was immediately inactivated by SAHW containing 50 ppm chlorine in the aqueous phase. These data suggest that SAHW containing 100 ppm chlorine can be used for aerosol disinfection of NDV in farms.

Virucidal Properties of Bioceramic Derived from Chicken Feces pH 13 and its Stability in Harsh Environments

SUMMARY Bioceramic derived from chicken feces (BCX) is a material produced by a sintering process for the purpose of use in animal farms to control livestock infectious diseases. In the present study, BCX at pH 13 was evaluated for the durability of its virucidal activity in simulated field conditions. First it was shown that BCX had activity toward Newcastle disease virus, infectious bursal disease virus, and goose parvovirus within 3 min and toward avian influenza virus (AIV) within 1 hr. BCX was further tested by keeping it under simulated harsh environmental conditions with sunlight for several weeks as well as by repeatedly soaking it with water and drying under sunlight many times. After sampling every 2 consecutive weeks and every 2 (of 9) consecutive resuspensions, BCX was evaluated for its efficacy against AIV. Evaluation under the harsh conditions illustrated that BCX could retain its satisfactory efficacy toward AIV throughout 7 wk and through 9 resuspensions. It is hence concluded that BCX is an excellent material for applying in livestock farming as a trapping disinfectant, due to its efficacy to inactivate various viruses, and that this efficacy is prolonged even under harsh environmental conditions.