Investigation Of Hydrogen Sulfide Gas Content In A Chicken Farm In A Certain City And Its Effect On The Chicken Neutrophil Extranet

H₂S is a flammable and toxic gas widely existing in nature and the smell of colorless smelly eggs.It can be produced in many ways.With the development of economy and society,people pay more and more attention to the relationship between food safety and their own health,and then pay attention to the living environment problems in the process of raising livestock and poultry.As a common pollutant in livestock houses,H₂S has seriously endangered the safety of livestock and poultry products and endan-gered people’s health.H₂S exposure can stimulate the respiratory system,cause coughing,inflammation,and can also cause neutrophil damage,resulting in immune dysfunction.In 2004,the new neutrophil immune response mechanism（NETs）was discovered.It is one of the ways in which the body exerts its immune function.NETs is a network structure,which is composed of DNA,histones,myeloperoxidase（MPO）,neutrophil elastase（NE）,etc.H₂S is a common exogenous stimulus,but whether H₂S exposure will inhibit the formation of NETs through TOLL-like receptors and the PI3K/AKT pathway has not been clearly reported.H₂S is one of the main pollutants in the air of livestock and poultry houses.H₂S can cause great harm to the health of livestock and poultry.In order to explore the pollution degree of H₂S in livestock and poultry houses,we selected a chicken farm in a city in Northeast China.The number 10 of the farm was changed to a chicken house.According to the"Ambient Air Quality Standard"and other relevant regulations on the harmful gas content of livestock and poultry houses,the H₂S content of different heights and positions of the chicken house was detected by the H₂S detector.At the same time,in order to further study the effect of H₂S on chicken peripheral blood neutrophil extranets（NETs）.We selected200 broiler chickens to be raised under different concentrations of H₂S exposure.Divide 200 broiler chickens into two groups evenly.One group is kept in a normal environment without H₂S access.The H₂S concentration is 0.38 mg/m3.This group is the national standard group（group N）,and the other group is in the chicken house.H₂S,keep the H₂S concentration in the chicken house at 10 mg/m3 or more.This group is the H₂S group（HS group）.The two groups of chickens were provided with the same water,and the two groups of chickens were free to eat and collect water,and the feeding time was 28days.On the 28th day of feeding,10 chickens were randomly selected from each group of chickens,blood was collected,and neutrophils in the blood were extracted through a neutrophil isolation kit,and PMA was used to stimulate the production of NETs,and scan electrons Microscopic observation and fluorescence microscopic observation,observe the structure and morphology of NETs,detect the expres-sion of TIR4,TIR2,PI3K,AKT,MPO,NOX2 in TOLL-like receptors by fluorescence quantitative PCR（q RT-PCR）,and detect the protein by Werstern blot The expression of MPO,NE,AKT,PI3K,fluores-cence staining to detect the level of reactive oxygen species during respiratory burst,in order to clarify the impact of H₂S exposure on the peripheral blood NETs of chickens.The results showed that:（1）According to our measurement results of H₂S content and concentration in 10 chicken houses at different locations in a city in Heilongjiang Province,the H₂S concentration in the chicken house has exceeded the standard.The overall rate of H₂S concentration exceeding the standard rate is 30%,and the maximum value of the exceeding standard concentration is 35.69 mg/m3.（2）Through scanning electron microscope observation,fluorescence microscope observation,WB and other experimental results,it was found that under the condition of H₂S exposure,PMA-induced NETs production in N+PMA group was higher than HS+PMA group,and compared with N+PMA group,TLR4,TLR2,PI3K,AKT,NOX2,MPO m RNA expression of HS+PMA group de-creased.In summary,there are cases of chicken houses with H₂S exceeding the standard.Animal experi-ments have shown that H₂S can reduce the production of MPO and inhibit the production of NETs through the TOLL-like receptor pathway.This experiment clarified the mechanism of action of H₂S on NETs,and provided a data basis for the regulation of H₂S in poultry breeding.