A Preliminary Study On The Ammonia Content Of Chicken Farms In A City And The Effect Of Ammonia On Neutrophils

The living environment of livestock and poultry is closely related to the quality of livestock and poultry products and the health of the breeders.When livestock and poultry are raised,some harmful gases such as ammonia,hydrogen sulfide,carbon monoxide,etc.will be produced.Investigations have shown that the excretion of livestock and poultry and the decomposition of uric acid in the litter will produce a large amount of ammonia gas,and the ammonia gas in the air will The health of livestock and brooders has a serious impact.As an immune cell in the body,neutrophils are the first line of defense against external stimuli and inflammation,and can respond positively to various inflammatory responses caused by external stimuli.This test evaluated the ammonia pollution of chicken farms in a certain city of Heilongjiang Province.The method was to use the ammonia gas detector to detect the ammonia concentration of chicken farms in different locations in the city.Then we further explored the effect of ammonia on peripheral blood neutrophils and neutrophil nets in chickens.We raised 200 Ross-308 broilers at different ammonia concentrations and divided 200 broilers into two groups.One group was housed in an ammonia atmosphere at 0.38 mg/m3 and the other group was housed in an ammonia atmosphere at 22.77 mg/m3.Both groups of chickens were free to eat and drink,and the test period was 28 days.After 28 days,10 chickens were randomly selected from each group.Peripheral blood was collected,and neutrophils were isolated from chicken peripheral blood neutrophil isolation kit.Then,neutrophils were stimulated with PMA,and neutrophils were observed before and after PMA stimulation.Morphology of cells and neutrophils,ROS production,and subsequent detection of genes involved in neutrophil recruitment and inflammation(MYD88,TRAF6,TLR4,TLR2,PI3 K,AKT,NF-?B,NOX-2,MPO),heat shock proteins(HSP27,HSP40,HSP60,HSP70,HSP90),cytokines(IL-1?,IL-2,IL-4,IL-6,IL-10),glucose metabolism levels(HK2,HK1,LDHB2,SDHB,LDHA,PFK,and PK)and the expression levels of inflammatory corpuscle levels(NLRP3,caspase-1),in order to elucidate the ammonia-to-chicken peripheral blood neutrality from the perspective of metabolism and inflammation.The effects of granulocytes.The results showed that:(1)The ammonia content of chicken houses in a city in northeast China was measured.The ammonia concentration exceeded the standard rate of 30.77% and the maximum value was 40.38 mg/m3.(2)m RNA levels of HSP27,HSP40,HSP60,HSP70,HSP90,IL-1?,IL-2,IL-4,IL-6,IL-10,NLRP3,and caspase-1 in the ammonia group compared with the national standard group The expression showed an upward trend,and the m RNA expression of the three groups in the ammonia group showed a downward trend after PMA stimulation.For the m RNA expression levels of HK2,HK1,LDHB2,SDHB,LDHA,PFK and PK,the ammonia group was lower than the national standard group.It indicates that ammonia can cause dysfunction of neutrophil glucose metabolism and immune function.(3)Under the scanning electron microscope,the amount of neutrophil extracellular nets in the ammonia group after PMA stimulation was lower than that in the national standard group.The results of fluorescent staining also confirmed this conclusion.The results of ROS showed that the ROS level in the ammonia group was higher than that in the national standard group under normal conditions.After PMA stimulation,the ROS levels in the two groups increased,but the national standard group was higher than the ammonia group.Compared with the national standard group,the m RNA expression levels of MYD88,TRAF6,TLR4,TLR2,PI3 K,AKT,NF-?B,NOX-2 and MPO in the ammonia group were increased,and the m RNA of the ammonia group was compared with the national standard group after PMA stimulation.The amount of expression is reduced.This indicates that ammonia can cause respiratory bursts in neutrophils,and the formation of MPO is reduced by the TOLL-like receptor pathway,which in turn affects the production of neutrophil extracellular traps.In summary,there is a situation in the city where the ammonia in the chicken house exceeds the standard.Animal experiments have shown that ammonia can reduce immune function by affecting the m RNA expression levels of neutrophil heat shock proteins,cytokines,glucose metabolism and inflammatory bodies in chicken peripheral blood.It can also affect the neutrophil respiratory burst and MPO production,and inhibit the production of neutrophil extracellular nets through the TOLL-like receptor pathway.This experiment clarified the mechanism of action of ammonia on chicken peripheral blood neutrophils and neutrophil extracellular nets,and provided a theoretical basis for the regulation of ammonia in poultry breeding.