| Black water fly(Hermetia illucens L)is a widely distributed scavenging insect,whose larvae mainly feed on animal feces,kitchen waste and decaying organic matter,and can intensively treat livestock and poultry feces.The treated black water fly larvae contain a large amount of protein and crude fat,which can be processed and produced as high-quality animal protein feed.At present,it is difficult to breed black water fly on a large scale,because the breeding technology of black water fly is not mature enough.The research and optimization of black water fly farming technology and the realization of large-scale black water fly farming can effectively alleviate the problem of environmental pollution caused by pig manure.In this paper,the larva of black water tabanus was taken as the research object,and pig-manure and water-manure were used as insect feed to breed the larvae of different ages,so as to study the effect of the addition ratio of pig-manure and water-manure on the weight gain,fecal-insect ratio and conversion rate of black water tabanus larvae.Using pig feed,bran and fermented soybean meal as nutrition additives,different proportions of pig manure and water manure were added to breed black water fly.The fecal ratio was 1:1.5 adding fresh pig manure and 3% fermented soybean meal to cultivate the black water fly,and the fecal ratio was 6.03:1.The fecal ratio was 1:0.8 adding water feces and 7.5% fermented soybean meal to cultivate the black water fly,and the fecal ratio of the larvae was 6.14:1.The fecal ratio was 1:2,adding pig manure and 25% bran to cultivate the black water fly,and the fecal ratio was 9.8:1.The fecal ratio of black water fly was 6.19:1 after adding pig manure and 20% pig feed to 1:3.The fecal ratio of black water fly larvae cultured with pig manure and 25% fermented soybean meal was 1:1,and the fecal ratio was 5.08:1.So fecula ratio of 1:1.5 add pig manure and 3% fermented soybean meal,fecula ratio of 1:1,1:2 add pig manure and 25% fermented soybean meal,worm dung pig and pig feed ratio of 1:3 added 10%,20%,than worm dung for 1:0.8 to add water manure and 7.5%fermented soybean meal formula black water fly larva breeding,such as to improve the weight of the larvae,dung)than and conversion rate has a significant effect.In order to explore the changes of CSFV and pseudorabies virus in the larva of black fly,the larva of black fly were inoculated by single injection,single oral PRV,CSFV and multiple oral PRV.After PRV and CSFV inoculation,the larvae of black water fly were detected by PCR and RT-PCR respectively,and the time difference between PRV and CSFV was detected and analyzed.20 gram black water fly larvae were injected with PRV in a single time,each larvae was injected with 2 μL PRV on an empty stomach,and 5 larvae were collected at intervals of 1 hour within 30 hours for PCR detection.The results showed that the 25 hours black water fly larvae had a specific band at 330 bp,and the size was consistent with the target gene band.In the single oral PRV group,1 m L of PRV and 15 m L of PBS were added to feed 20 gram of black water fly.After 1 hour,the black water fly was cleaned and then put on an empty stomach.Within 27 hours,5 larvae were collected at intervals of 1 hour for PCR detection.The results showed that the 22 hours black water fly larvae had a specific band at 330 bp,and the size was consistent with the target gene band.In the single oral PRV feed group,1 m L of PRV,5 gram of feed and 15 m L of PBS were added to feed 20 gram of black water fly.After 1 hour,the black water fly was cleaned on an empty stomach,and 5 larvae were collected at intervals of 1 hour within 22 hours for PCR detection.The results showed that the 14 hours black water fly larvae had a specific band at 330 bp,and the size was consistent with the target gene band.Multiple oral PRV group with 25 gram feed,5 m L PRV and PBS 45 m L raised 100 gram blackwater fly,then each interval 8 hours respectively collect article 5 larva and 3 gram slag with PCR detection,repeat adding virus and feed on each interval of 72 hours,virus after adding 4 times,collection of insect slag had specific bands,in the 330 bp size match the purpose gene bands;No specific bands were found in the collected black water fly larvae at 330 bp.15 gram black water fly larvae were injected with CSFV in a single time,each larvae was injected with 2 μL CSFV on an empty stomach,and 5 larvae were collected at intervals of 1 hour within 17 hours for RT-PCR detection.The results showed that 9 hours blackfly larvae had a specific band at 343 bp,and the size was consistent with the target gene band.In the single oral CSFV group,1 m L of CSFV and 15 m L of PBS were added to feed 15 gram of black water fly.After 1 hour,the black water fly was cleaned and then put on an empty stomach,and 5 larvae were collected at intervals of 1 hour within 12 hours for RT-PCR detection.The results showed that no specific bands were found in 343 bp of larvae collected within 12 hour.In the single oral CSFV feed group,1 m L of CSFV,5 gram of feed and 15 m L of PBS were added to feed 15 gram of black water fly.After 1 hour,the black water fly was cleaned on an empty stomach,and 5 larvae were collected at intervals of 1 hour within 10 hours for RT-PCR detection.The results showed that no specific bands were found in 343 bp of larvae collected within 12 hours.PCR results of larvae collected from PRV muscle injection group,oral venom group and oral venom feed group showed that PRV was carried by black water fly larvae for 25 hours,22 hours and 14 hours,respectively.RT-PCR results of larvae collected from the CSFV intrapersoneural injection group,the oral venom group and the oral venom feed group showed that the time of CSFV carried by the above three methods was 9 hours,0hour and 0 hour respectively.Black water fly larvae were given PRV for 4 times and CSFV for a single time,and they did not carry the virus in their bodies. |
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