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蝦苗廠藻類三級培養技術,看看育苗得下多少工夫

編輯:華泰來水產    發布時間:2017-11-06    瀏覽:2399次

對(dui)(dui)蝦種苗(miao)(miao)企業的(de)(de)(de)(de)(de)(de)生存主要(yao)是依靠蝦苗(miao)(miao)養(yang)殖效果來決定的(de)(de)(de)(de)(de)(de),蝦苗(miao)(miao)成活率的(de)(de)(de)(de)(de)(de)高低直接影響(xiang)著產業鏈的(de)(de)(de)(de)(de)(de)上(shang)上(shang)下(xia)下(xia)。單胞(bao)藻(zao)(zao)對(dui)(dui)于(yu)蝦苗(miao)(miao)早(zao)期(qi)是最適合的(de)(de)(de)(de)(de)(de)餌料,如(ru)(ru)果培育(yu)(yu)得當,不(bu)僅可(ke)以(yi)節約(yue)成本,減少育(yu)(yu)苗(miao)(miao)水(shui)體(ti)環境負荷,尤其是對(dui)(dui)于(yu)蝦苗(miao)(miao)的(de)(de)(de)(de)(de)(de)發育(yu)(yu)打下(xia)堅實的(de)(de)(de)(de)(de)(de)基礎(chu),提高成活率。結合個人的(de)(de)(de)(de)(de)(de)育(yu)(yu)苗(miao)(miao)感受,投喂(wei)單胞(bao)藻(zao)(zao)與(yu)不(bu)投喂(wei)的(de)(de)(de)(de)(de)(de)區別還是有(you)的(de)(de)(de)(de)(de)(de),比如(ru)(ru)苗(miao)(miao)的(de)(de)(de)(de)(de)(de)變(bian)態時間縮短(duan),活力(li)個頭大(da)小,水(shui)質均優于(yu)未投喂(wei)單胞(bao)藻(zao)(zao)的(de)(de)(de)(de)(de)(de)苗(miao)(miao),當然(ran)如(ru)(ru)果培育(yu)(yu)不(bu)得當,反作用也有(you),比如(ru)(ru)投喂(wei)有(you)害菌超標的(de)(de)(de)(de)(de)(de)藻(zao)(zao)液,藻(zao)(zao)類(lei)老化,投喂(wei)過量等。所(suo)以(yi)合理運用,方可(ke)收(shou)到(dao)理想效果。限(xian)于(yu)個人水(shui)平,描述方法的(de)(de)(de)(de)(de)(de)錯誤(wu)不(bu)足(zu)之處,懇請大(da)家指正。

一 、單胞藻純培養室操作描述

儀器使用

(1) 超凈工作臺

使(shi)用前(qian)提前(qian)開機,打開紫外線燈和風機開關(guan)20分鐘。超(chao)凈(jing)臺面上不(bu)存放不(bu)必要的物品,保(bao)持工作(zuo)(zuo)區內(nei)的凈(jing)化氣流(liu)不(bu)受(shou)干擾,工作(zuo)(zuo)時(shi)(shi)盡量(liang)避免作(zuo)(zuo)明顯(xian)干擾氣流(liu)的動(dong)作(zuo)(zuo);超(chao)凈(jing)臺保(bao)持光(guang)滑(hua)(hua)水(shui)平(ping),光(guang)滑(hua)(hua)便(bian)于用酒(jiu)精消毒布擦,水(shui)平(ping)是倒培養(yang)基時(shi)(shi)使(shi)培養(yang)皿(min)內(nei)平(ping)板厚度一致。工作(zuo)(zuo)時(shi)(shi)超(chao)凈(jing)臺周圍空氣流(liu)動(dong)要緩慢,雜菌盡量(liang)減少,所以要定期清(qing)潔(jie)消毒保(bao)種室內(nei),關(guan)閉門(men)窗,盡可能減少雜菌數量(liang);空氣過(guo)濾網定期清(qing)潔(jie)。

(2)真空(kong)泵(beng)和溶(rong)劑(ji)過濾(lv)器及過濾(lv)膜

此環節(jie)過濾只是(shi)為單胞藻純(chun)培(pei)養室(保種,提純(chun),擴培(pei)試管級)準備清潔無菌海水。其他級別的(de)培(pei)養無需此環節(jie)。

過(guo)(guo)濾(lv)(lv)膜使用(yong)0.2微米(mi)孔(kong)徑(jing),不(bu)可(ke)用(yong)手拿,用(yong)鑷(nie)子選取(qu)。真(zhen)空(kong)泵(beng)(beng)與溶劑過(guo)(guo)濾(lv)(lv)器之間通過(guo)(guo)合(he)適(shi)口徑(jing)的(de)橡(xiang)膠(jiao)管(guan)連接(jie)緊密,每次過(guo)(guo)濾(lv)(lv)要注意(yi)濾(lv)(lv)膜的(de)過(guo)(guo)濾(lv)(lv)效果(guo)(guo),防(fang)止(zhi)濾(lv)(lv)孔(kong)阻(zu)塞影響過(guo)(guo)濾(lv)(lv)效果(guo)(guo)。濾(lv)(lv)瓶內的(de)濾(lv)(lv)液(ye)不(bu)能沒過(guo)(guo)濾(lv)(lv)板(ban)出水口,否則濾(lv)(lv)液(ye)將(jiang)被抽吸入(ru)真(zhen)空(kong)泵(beng)(beng),導致真(zhen)空(kong)泵(beng)(beng)損(sun)壞。

應該使濾板(ban)內的營(ying)養液過濾盡后(hou)(無(wu)水滴(di))才(cai)能停泵。要定期注意(yi)真空泵內油(you)(you)量,注意(yi)加油(you)(you)。

(3) 恒溫鼓(gu)風干燥箱(xiang)

設(she)定(ding)溫(wen)度時(shi)(shi)。先按(an)控溫(wen)儀功能鍵SET進(jin)入(ru)溫(wen)度設(she)定(ding)狀態,此時(shi)(shi)SP設(she)定(ding)顯示(shi)(shi)一(yi)閃一(yi)閃,要(yao)按(an)移動(dong)位(wei)鍵配合加鍵或減鍵,設(she)定(ding)時(shi)(shi)先從(cong)分位(wei)數(shu)字設(she)定(ding),一(yi)直到百位(wei)數(shu),設(she)定(ding)結束(shu)需按(an)一(yi)下(xia)功能鍵SET,此時(shi)(shi)干燥箱進(jin)入(ru)升溫(wen)狀態,加熱(re)指示(shi)(shi)燈(deng)亮,當箱內溫(wen)度接近(jin)設(she)定(ding)溫(wen)度時(shi)(shi),加熱(re)指示(shi)(shi)燈(deng)閃動(dong)多(duo)次(ci),控制進(jin)入(ru)恒溫(wen)狀態。

使用注意事項(xiang):物品不能堆得太滿,更不能堵住出風口,以免影響溫度均勻上升。若是有牛皮紙和保鮮膜時,溫度不能超過180度。取出物品時,箱內外溫差不宜過高,防止玻璃器皿炸裂。橡膠制品不能放入。

(4)加濕器

配合生化培養箱使用。

水(shui)源(yuan)是蒸(zheng)餾水(shui);不(bu)應容(rong)器(qi)內帶水(shui)移動加(jia)濕器(qi);加(jia)水(shui)時應在容(rong)器(qi)上表示(shi)的滿(man)水(shui)線為止。

應注意的事(shi)項:平(ping)時(shi)留意觀察水箱(xiang)的存水量,及時(shi)予以補充,遠離熱源,腐蝕物。

(5)蒸(zheng)汽消毒(du)器

容器加水時(shi)(shi),水位一(yi)定要超過電熱管,最(zui)好是到3個支撐點的(de)(de)下緣,如果連續使用(yong),應在每次消(xiao)(xiao)毒(du)前補足蒸發水量,不然少(shao)水會燒壞電熱管。蓋上的(de)(de)放(fang)氣軟管插入消(xiao)(xiao)毒(du)桶圓槽(cao)內(nei),對(dui)正蓋和主體的(de)(de)螺栓槽(cao),依次將相對(dui)方的(de)(de)翼型螺母予以旋緊(jin),使蓋與主體密(mi)合,加熱開始時(shi)(shi),應將放(fang)氣閥與安全(quan)閥的(de)(de)小手柄扳(ban)至垂直放(fang)氣方位,必(bi)須待有較急蒸汽噴出,即將小手柄扳(ban)回水平“關閉”方位。

當到(dao)達(da)所需(xu)壓力(li)和溫度(du),計算消毒時(shi)(shi)間,消毒畢后(hou),關閉電源,待壓力(li)指(zhi)針指(zhi)向0時(shi)(shi),將放(fang)(fang)氣(qi)閥的小手柄(bing)扳至(zhi)垂(chui)直放(fang)(fang)氣(qi)方位(wei),氣(qi)放(fang)(fang)完(wan)后(hou),打開容器蓋。物品消毒后(hou)要迅速干(gan)燥。

應注意滅菌液體時,不超(chao)過盛液體容器(qi)體積的(de)3/4,瓶塞(sai)不可用軟(ruan)木塞(sai)或未打孔(kong)的(de)橡(xiang)膠塞(sai)。滅菌器(qi)內冷空(kong)(kong)氣(qi)(qi)必(bi)(bi)須(xu)完全排除。因為空(kong)(kong)氣(qi)(qi)的(de)膨(peng)脹(zhang)壓(ya)(ya)大于水蒸(zheng)氣(qi)(qi)的(de)膨(peng)脹(zhang)壓(ya)(ya),當水蒸(zheng)氣(qi)(qi)含冷空(kong)(kong)氣(qi)(qi)時,壓(ya)(ya)力(li)表所表示的(de)壓(ya)(ya)力(li)是(shi)水蒸(zheng)氣(qi)(qi)壓(ya)(ya)力(li)和部(bu)分冷空(kong)(kong)氣(qi)(qi)壓(ya)(ya)力(li)之和,不是(shi)水蒸(zheng)氣(qi)(qi)的(de)實(shi)際壓(ya)(ya)力(li),而同一(yi)壓(ya)(ya)力(li)下的(de)實(shi)際溫度,含空(kong)(kong)氣(qi)(qi)的(de)蒸(zheng)汽低于飽(bao)和蒸(zheng)汽,這(zhe)樣導致達不了消毒效果(guo)。絕不可堵塞(sai)安全閥出氣(qi)(qi)孔(kong),必(bi)(bi)須(xu)留空(kong)(kong)位保證其暢通(tong)放氣(qi)(qi),發(fa)現超(chao)溫超(chao)壓(ya)(ya),立(li)即斷電。

蒸餾器

先關閉放水(shui)(shui)(shui)(shui)閥,打開進(jin)水(shui)(shui)(shui)(shui)閥,待溢水(shui)(shui)(shui)(shui)皮管有(you)水(shui)(shui)(shui)(shui)流(liu)出(chu),方可(ke)(ke)接通電源,此時可(ke)(ke)暫時關閉進(jin)水(shui)(shui)(shui)(shui)閥,待出(chu)水(shui)(shui)(shui)(shui)皮管有(you)蒸(zheng)餾水(shui)(shui)(shui)(shui)流(liu)出(chu),打開進(jin)水(shui)(shui)(shui)(shui)閥,制備(bei)蒸(zheng)餾水(shui)(shui)(shui)(shui)。定期清(qing)洗蒸(zheng)餾器內壁,除去水(shui)(shui)(shui)(shui)垢,不(bu)然影響蒸(zheng)餾水(shui)(shui)(shui)(shui)質量(liang)和使用效果。切不(bu)可(ke)(ke)關閉水(shui)(shui)(shui)(shui)閥制備(bei)蒸(zheng)餾水(shui)(shui)(shui)(shui),否則燒(shao)壞電熱管。

(6)鼓風機

定期(qi)更換(huan)吸氣口的過(guo)濾棉(mian),異常情況,請電工(gong)檢(jian)查。

(7)空(kong)調

保種(zhong)室若有生長藻種(zhong),需維持恒定溫度,反之關閉。一級(ji),二級(ji)空調視外(wai)界氣(qi)溫調整開關。

二 、衛生消毒

工具衛生消毒

1、濾(lv)水筒(tong)式過(guo)濾(lv)器(qi)。筒(tong)式過(guo)濾(lv)器(qi)的濾(lv)芯應用一次(ci)更換一次(ci),用過(guo)的濾(lv)芯浸泡在1%鹽酸溶(rong)液中消毒12h以上(shang)。然后(hou)用洗(xi)(xi)潔精清洗(xi)(xi),再(zai)用足量清水沖洗(xi)(xi)干凈,晾(liang)干備用。(濾(lv)芯規格應小于等(deng)于0.5微米)。

2、試(shi)管(guan)(規格10毫(hao)升到(dao)20毫(hao)升),錐形瓶(ping)(50毫(hao)升到(dao)200毫(hao)升),500ml錐形瓶(ping),2.5L瓶(ping)廣(guang)口(kou)瓶(ping),移液(ye)管(guan),移液(ye)槍(qiang)管(guan)/盒,玻(bo)璃氣(qi)管(guan),膠氣(qi)管(guan),氣(qi)石,溶(rong)劑(ji)過(guo)濾器,培養皿。用(yong)過(guo)后應(ying)當用(yong)清水(shui)清洗(xi)后浸(jin)泡(pao)于(yu)1% 鹽酸12h以上(溶(rong)劑(ji)過(guo)濾器除外,浸(jin)泡(pao)時相(xiang)近大(da)小器皿分類(lei)放不同(tong)的(de)消毒桶,規格相(xiang)差(cha)大(da),易(yi)(yi)導致相(xiang)互碰撞(zhuang)碎爛)然后用(yong)洗(xi)潔精清洗(xi)(用(yong)毛刷清洗(xi)試(shi)管(guan)底(di)部時,不可(ke)太用(yong)力(li),否(fou)則(ze)底(di)部易(yi)(yi)破裂),再用(yong)大(da)量清水(shui)沖洗(xi)干(gan)凈(jing),風干(gan)或曬(shai)干(gan),高壓滅(mie)菌后,放入(ru)恒溫干(gan)燥箱(膠氣(qi)管(guan),氣(qi)石除外)干(gan)燥備用(yong)。

3、聚乙烯藻袋(dai)用(yong)于二級藻類藻種掛袋(dai)培養(yang)或(huo)3級擴培(一般使用(yong)盛裝藻液12升(sheng)左右)。先用(yong)清水(shui)沖(chong)洗(xi)干(gan)凈內(nei)外,浸(jin)泡于1% 鹽酸12h以上,再用(yong)大(da)量(liang)清水(shui)沖(chong)洗(xi)干(gan)凈,風(feng)干(gan)或(huo)曬干(gan),備用(yong)。由(you)于藻袋(dai)近似密封性,對于防止(zhi)微生物(wu)污染有很(hen)好的效果。

4、毛(mao)巾、保種(zhong)室,一(yi)級和二(er)級毛(mao)巾不(bu)可混用,所用毛(mao)巾后都(dou)應消毒清(qing)洗晾干(gan)后備用。

5、錫箔紙:除非粘上(shang)油污或污物以及破(po)損(sun),都(dou)應重復使用(yong)。

6、牛皮紙:定期拿(na)到烘箱(xiang)烘干。

7、定期更換空氣(qi)過(guo)(guo)濾(lv)器(qi)(用于1,2級培(pei)養的進(jin)氣(qi)過(guo)(guo)濾(lv))的濾(lv)芯,濾(lv)芯已(yi)油(you)化(hua)或沖(chong)散的棄(qi)去。

8、定(ding)期更換鼓風機進風口的(de)過濾棉。

9、對新購入的器皿(min)應先(xian)除(chu)去包裝(zhuang)沾染的污垢(gou)后,用洗(xi)潔精刷洗(xi),流(liu)水沖凈,再浸泡于1%的鹽(yan)酸12h,使(shi)游(you)離的堿性物(wu)質除(chu)去,再用清(qing)水沖洗(xi)干(gan)凈,風干(gan)或曬干(gan)備用。

室內衛生消毒

1、保種室每天早(zao)上用清水(shui)擦洗實驗臺(tai)面,地板(ban),墻壁,木(mu)架(jia)后(hou),開紫(zi)外燈(deng)(deng)消毒20分(fen)鐘,此時(shi)關(guan)掉空調,關(guan)掉日光燈(deng)(deng),由于紫(zi)外燈(deng)(deng)照(zhao)射會產生一(yi)定(ding)量的臭氧,對(dui)人體有害,所以消毒關(guan)閉(bi)紫(zi)外燈(deng)(deng)后(hou)人應(ying)隔(ge)一(yi)段(duan)時(shi)間再進去。進保種室換(huan)鞋。

2、一級和二級室每天(tian)清潔,接(jie)種前接(jie)種后(hou)承載(zai)物框(kuang)架都應用(yong)消毒(du)液消毒(du)。

3、門口消毒(du)池定(ding)期更換消毒(du)藥水(shui)(高(gao)錳酸鉀)。

4、常(chang)用消毒劑濃度 1%鹽酸(suan),次氯(lv)酸(suan)鈉20ppm(有效氯(lv)),碘(dian)液(1000ppm),酒精(jing)70-75%,高(gao)錳酸(suan)鉀1mg/L。

三 、接種

藻種固體培養基制備

所需燒杯,玻棒須(xu)清洗(xi)干凈,錐形瓶和(he)培養皿須(xu)消毒清洗(xi)滅菌(jun)烘干。將稱(cheng)好的(de)(de)純化瓊脂粉加(jia)(jia)入(ru)燒杯再加(jia)(jia)入(ru)一定(ding)量的(de)(de)海(hai)水(shui)(shui),加(jia)(jia)熱(必(bi)須(xu)不停攪拌(ban),以免瓊脂糊底燒焦)至(zhi)完(wan)全融化,加(jia)(jia)熱后的(de)(de)瓊脂培養基應當透明無(wu)顯(xian)著沉淀,若明顯(xian),可采用清潔的(de)(de)白色薄絨布或中間夾有薄層吸(xi)水(shui)(shui)棉的(de)(de)雙層紗布鋪在漏(lou)斗上趁(chen)熱過濾,融化畢(bi)后精確加(jia)(jia)入(ru)營養鹽(維(wei)生素B溶(rong)液須(xu)等培養基高壓(ya)滅菌(jun)后冷卻至(zhi)50攝氏度(du)以下再加(jia)(jia))。

培(pei)養基采用121攝氏(shi)(shi)度高壓蒸汽滅(mie)菌20至(zhi)30分鐘(zhong)即(ji)可,培(pei)養基滅(mie)菌后(hou)應立即(ji)取出(chu),冷卻至(zhi)50攝氏(shi)(shi)度以下(xia),即(ji)加維生素(su)B溶液。將盛裝培(pei)養基的錐形瓶和(he)培(pei)養皿放(fang)在超凈工作臺上,點燃酒精燈,右手托起錐形瓶底,將瓶口在酒精燈稍加燒,然(ran)后(hou)迅速將培(pei)養基倒入培(pei)養皿中,以鋪滿皿底為止(厚薄適(shi)中)。鋪放(fang)好(hao)后(hou),放(fang)置20-30分鐘(zhong),待培(pei)養基凝固后(hou),即(ji)開始(shi)操(cao)作。

若是做斜(xie)面培(pei)(pei)養基(ji),則(ze)加入維生素B溶液(ye)后,將盛裝(zhuang)培(pei)(pei)養基(ji)的錐形(xing)瓶和試管(guan)(guan)以(yi)及一根厚約(yue)1厘米(mi)的小(xiao)木板放在超凈工作(zuo)(zuo)臺上(shang)。分(fen)裝(zhuang)約(yue)為(wei)試管(guan)(guan)高(gao)度的1/3-2/3(視情況(kuang)操作(zuo)(zuo)),分(fen)裝(zhuang)量應以(yi)能(neng)形(xing)成2/3底(di)層和1/3斜(xie)面量為(wei)好,擺置成適當斜(xie)面,待其自然(ran)凝固。培(pei)(pei)養基(ji)制備好后,如發現破(po)裂,水分(fen)浸(jin)入,色澤異(yi)常等均應棄去.

待凝固后即進行操作,現一般采(cai)用平板劃線(xian)接種(zhong)和涂(tu)布接種(zhong)。

平板劃線接種

接(jie)(jie)種(zhong)(zhong)(zhong)(zhong)環(huan)在(zai)酒精(jing)燈(deng)(deng)火(huo)焰(yan)上(shang)(shang)(shang)充分燒紅后(hou),接(jie)(jie)種(zhong)(zhong)(zhong)(zhong)柄一邊(bian)轉動(dong)一邊(bian)慢慢來回通(tong)過火(huo)焰(yan)3次(ci),待冷卻(que)至(zhi)室溫方(fang)可挑(tiao)取藻(zao)種(zhong)(zhong)(zhong)(zhong)(為保證(zheng)藻(zao)種(zhong)(zhong)(zhong)(zhong)不(bu)被(bei)燙死(si),挑(tiao)種(zhong)(zhong)(zhong)(zhong)前可將(jiang)接(jie)(jie)種(zhong)(zhong)(zhong)(zhong)環(huan)接(jie)(jie)觸培(pei)養(yang)皿(min)(min)內(nei)壁(bi)或培(pei)養(yang)基(ji)),迅速(su)(su)接(jie)(jie)種(zhong)(zhong)(zhong)(zhong)至(zhi)新培(pei)養(yang)基(ji)(劃線(xian)可采用斜線(xian)法,方(fang)格法,曲線(xian)法,放射法等)。左(zuo)(zuo)手(shou)(shou)(shou)握住(zhu)平(ping)板,用大拇(mu)指(zhi)和食(shi)指(zhi)稍稍抬(tai)起皿(min)(min)蓋,同(tong)時(shi)靠近火(huo)焰(yan)周圍(wei),右(you)手(shou)(shou)(shou)持接(jie)(jie)種(zhong)(zhong)(zhong)(zhong)環(huan)伸入皿(min)(min)內(nei),在(zai)平(ping)板上(shang)(shang)(shang)一個區域作(zuo)(zuo)劃線(xian)(依方(fang)法不(bu)同(tong),動(dong)作(zuo)(zuo)不(bu)同(tong))。劃線(xian)時(shi)使接(jie)(jie)種(zhong)(zhong)(zhong)(zhong)環(huan)與(yu)平(ping)板表(biao)(biao)面成30-40度(du)角輕輕接(jie)(jie)觸,以腕力在(zai)表(biao)(biao)面作(zuo)(zuo)較快的滑動(dong)(注意不(bu)要劃破平(ping)板表(biao)(biao)面或伸進培(pei)養(yang)基(ji)內(nei))。接(jie)(jie)種(zhong)(zhong)(zhong)(zhong)環(huan)挑(tiao)種(zhong)(zhong)(zhong)(zhong)后(hou)和劃線(xian)完(wan)抽出時(shi)均不(bu)可碰及皿(min)(min)壁(bi)/口(kou),更(geng)不(bu)能碰其它物(wu)體和臺面。接(jie)(jie)種(zhong)(zhong)(zhong)(zhong)完(wan)后(hou),將(jiang)接(jie)(jie)種(zhong)(zhong)(zhong)(zhong)環(huan)從柄部至(zhi)環(huan)端逐漸通(tong)過酒精(jing)燈(deng)(deng)火(huo)焰(yan)滅菌(jun)(jun),不(bu)能直(zhi)接(jie)(jie)燒環(huan),以免(mian)殘留在(zai)接(jie)(jie)種(zhong)(zhong)(zhong)(zhong)環(huan)上(shang)(shang)(shang)的藻(zao)體或菌(jun)(jun)體爆濺污(wu)染空間。上(shang)(shang)(shang)述操作(zuo)(zuo)完(wan)成后(hou),即用無菌(jun)(jun)封口(kou)膜(mo)(mo)封培(pei)養(yang)皿(min)(min)口(kou)(膜(mo)(mo)使用前須(xu)用酒精(jing)消毒(du),紫(zi)外(wai)燈(deng)(deng)消毒(du),風干(gan)),封口(kou)時(shi),用左(zuo)(zuo)手(shou)(shou)(shou)大拇(mu)指(zhi)摁住(zhu)膜(mo)(mo)一邊(bian)末端,右(you)手(shou)(shou)(shou)大拇(mu)指(zhi)和食(shi)指(zhi)捏住(zhu)另一邊(bian)末端,迅速(su)(su)繞培(pei)養(yang)皿(min)(min)口(kou)繞一圈,封口(kou)。

涂(tu)布接(jie)種

所(suo)用(yong)種(zhong)液(ye)(ye)必須(xu)經過(guo)弧菌檢測合格(ge)后方可(ke)使用(yong)。涂布(bu)棒在火焰上(shang)慢慢來回(hui)過(guo)火焰3次,冷卻(不可(ke)驟(zou)冷),將(jiang)種(zhong)液(ye)(ye)用(yong)移液(ye)(ye)槍從試(shi)管吸出注入平(ping)板上(shang)面(槍管不可(ke)碰(peng)及(ji)試(shi)管外壁,更(geng)不能碰(peng)及(ji)其它物品或臺(tai)面),迅速用(yong)涂布(bu)棒在表面作來回(hui)左右的涂布(bu),讓藻液(ye)(ye)分布(bu)均勻,畢后,封口(kou)同平(ping)板劃線。

試管液體培養基接種

所用種液必須經過(guo)弧菌(jun)(jun)(jun)檢測(ce)合格后方(fang)可使(shi)用。火(huo)焰(yan)滅菌(jun)(jun)(jun)前試(shi)(shi)(shi)管(guan)(guan)口外壁不(bu)能(neng)有(you)水滴(易導(dao)致受熱不(bu)均而破裂(lie)),燒試(shi)(shi)(shi)管(guan)(guan)口時(shi),試(shi)(shi)(shi)管(guan)(guan)傾(qing)斜45度角,不(bu)斷移動(dong)試(shi)(shi)(shi)管(guan)(guan),使(shi)試(shi)(shi)(shi)管(guan)(guan)口受熱均勻,滅菌(jun)(jun)(jun)時(shi)間3秒左右(you)。滅菌(jun)(jun)(jun)后不(bu)能(neng)驟(zou)冷,防止試(shi)(shi)(shi)管(guan)(guan)破裂(lie)。畢(bi)后,用盛裝(zhuang)營養(yang)液150或(huo)200ml的錐形瓶(ping)(ping)(也要(yao)滅菌(jun)(jun)(jun)瓶(ping)(ping)口)向試(shi)(shi)(shi)管(guan)(guan)中(zhong)(zhong)傾(qing)倒營養(yang)液,每次只能(neng)拿一個試(shi)(shi)(shi)管(guan)(guan)(注:若幾個試(shi)(shi)(shi)管(guan)(guan)對比時(shi),可以同時(shi)拿在(zai)(zai)手里,用大拇(mu)指(zhi)和(he)(he)手掌),倒好后,同樣試(shi)(shi)(shi)管(guan)(guan)和(he)(he)錐形瓶(ping)(ping)口滅菌(jun)(jun)(jun),試(shi)(shi)(shi)管(guan)(guan)塞也要(yao)滅菌(jun)(jun)(jun),塞試(shi)(shi)(shi)管(guan)(guan)塞時(shi)不(bu)要(yao)移動(dong)試(shi)(shi)(shi)管(guan)(guan)迎(ying)接塞,以免試(shi)(shi)(shi)管(guan)(guan)在(zai)(zai)移動(dong)中(zhong)(zhong)納入可能(neng)含菌(jun)(jun)(jun)空氣,火(huo)焰(yan)旁(pang)為無菌(jun)(jun)(jun)。

所有試管分裝好后,即(ji)接種(zhong)(zhong)。用(yong)移(yi)液槍(qiang)(qiang)選取移(yi)液量(試管口滅菌),注入(ru)新培(pei)養(yang)(yang)基(ji),槍(qiang)(qiang)管不(bu)(bu)(bu)(bu)可碰(peng)及(ji)管外壁,更不(bu)(bu)(bu)(bu)能碰(peng)及(ji)其它物(wu)品或臺(tai)面,一(yi)支槍(qiang)(qiang)管只(zhi)能和同一(yi)種(zhong)(zhong)液使用(yong),選取另一(yi)試管種(zhong)(zhong)液時必須更換槍(qiang)(qiang)頭。接種(zhong)(zhong)畢(bi)后,選取合適光照溫(wen)度培(pei)養(yang)(yang)。由于試管培(pei)養(yang)(yang)是靜(jing)置培(pei)養(yang)(yang),而在靜(jing)置條件下,藻細胞(bao)會聚集在試管底(di)部,且藻細胞(bao)周(zhou)圍會形成(cheng)透過養(yang)(yang)分的屏障,營養(yang)(yang)鹽的攝入(ru)受阻,光照吸收不(bu)(bu)(bu)(bu)均(jun),為(wei)使營養(yang)(yang)鹽攝入(ru)正常,藻細胞(bao)受光均(jun)勻(yun),需(xu)要(yao)定時搖置試管(手先噴(pen)消(xiao)毒酒精,手腕使勁搖,不(bu)(bu)(bu)(bu)要(yao)擺臂,不(bu)(bu)(bu)(bu)能倒置搖)

三角瓶,500ml瓶,2.5L瓶液體培養基接種

三角(jiao)瓶接種需無(wu)菌操作。500ml瓶,2.5L瓶液體(ti)培養(yang)基接種均(jun)(jun)在室內(nei)衛生(sheng)條件下操作。3種瓶和(he)所需營(ying)養(yang)液及錫(xi)紙,移液管,洗耳球(qiu),氣(qi)管,氣(qi)石均(jun)(jun)需經消毒清洗,滅菌后使用。三角(jiao)瓶需定(ding)時搖置,500ml,2.5L瓶培養(yang)在通氣(qi)條件下培養(yang)。

二級聚乙烯藻袋液體培養基接種

氣(qi)管,氣(qi)石均需(xu)經消毒(du)(du)清洗(xi),滅菌后(hou)(hou)(hou)使用(yong)(yong),抽藻(zao)管前(qian)次用(yong)(yong)畢后(hou)(hou)(hou)應(ying)當(dang)用(yong)(yong)水(shui)(shui)泵將管內(nei)灌滿消毒(du)(du)鹽酸溶液(ye),今次用(yong)(yong)前(qian)需(xu)用(yong)(yong)海(hai)水(shui)(shui)或(huo)淡水(shui)(shui)沖(chong)洗(xi)干凈(jing)后(hou)(hou)(hou)使用(yong)(yong)。選取(qu)的(de)種液(ye)應(ying)當(dang)是無明顯沉淀,泡(pao)沫,原生(sheng)動(dong)物,附著物,色澤正(zheng)常(chang),藻(zao)細胞密度大,形態正(zheng)常(chang),弧(hu)菌檢測(ce)(ce)合格后(hou)(hou)(hou)選用(yong)(yong)。所用(yong)(yong)海(hai)水(shui)(shui)漂(piao)白水(shui)(shui)消毒(du)(du)有效(xiao)氯應(ying)足(zu)10ppm,曝氣(qi)12小時(shi),檢測(ce)(ce)余氯,加(jia)硫代硫酸鈉中和,方可使用(yong)(yong)。若使用(yong)(yong)稀釋(shi)的(de)漂(piao)白水(shui)(shui),應(ying)當(dang)天配制當(dang)天使用(yong)(yong),而未使用(yong)(yong)的(de)要(yao)在24小時(shi)后(hou)(hou)(hou)棄去。且稀釋(shi)的(de)漂(piao)白水(shui)(shui)必須加(jia)蓋放在避光的(de)容(rong)器(qi)內(nei)保存。由于漂(piao)白水(shui)(shui)會隨時(shi)間漸漸分解,一次性購(gou)入的(de)漂(piao)白水(shui)(shui)不應(ying)太多,不要(yao)過量儲存,以免影響(xiang)滅菌功效(xiao)。

1,2,3級接藻示意

試管→三角瓶→500ml瓶子(zi)→2500ml瓶子(zi)→20升(sheng)的(de)藻種聚(ju)乙烯(xi)袋(dai) →18升(sheng)的(de)擴培藻聚(ju)乙烯(xi)袋(dai)或者培藻池(chi)

相關說明

1、母(mu)液(ye)的配置(zhi)可以參考教科(ke)書上(shang)所寫(xie),然(ran)后(hou)(hou)結合當(dang)地的水質適當(dang)修改即可。氮,磷,鐵元素(su),維生素(su)B1,B12是(shi)必須使(shi)用,維生素(su)母(mu)液(ye)保存(cun)于低溫弱(ruo)光(guang)環境下,其(qi)中維生素(su)的添加需要先(xian)高壓滅(mie)菌(jun)后(hou)(hou)再(zai)加入母(mu)液(ye)中。其(qi)余營養鹽先(xian)配置(zhi)好后(hou)(hou),再(zai)滅(mie)菌(jun)。如果是(shi)硅(gui)藻類,還應加上(shang)硅(gui)酸鈉。

2、保種室所需(xu)營養液(母液千分之0.8的比例(li)添加)除(chu)需(xu)高(gao)壓(ya)滅菌(jun)外還(huan)必須經過真空泵(beng)抽濾(lv)后才能用,微(wei)孔濾(lv)膜直徑(jing)為50毫米,孔徑(jing)為0.2微(wei)米。

3、接種比例A:試管(guan)0.07—0.09(隨密度大小變化(hua)),10毫(hao)(hao)(hao)(hao)升(sheng)(sheng)試管(guan)加(jia)(jia)營養(yang)(yang)液7毫(hao)(hao)(hao)(hao)升(sheng)(sheng),藻(zao)種0.5毫(hao)(hao)(hao)(hao)升(sheng)(sheng),20毫(hao)(hao)(hao)(hao)升(sheng)(sheng)試管(guan)中營養(yang)(yang)液加(jia)(jia)12.5毫(hao)(hao)(hao)(hao)升(sheng)(sheng)營養(yang)(yang)液,1毫(hao)(hao)(hao)(hao)升(sheng)(sheng)藻(zao)種。B:三角(jiao)瓶(ping)0.09-0.11。在100毫(hao)(hao)(hao)(hao)升(sheng)(sheng),150毫(hao)(hao)(hao)(hao)升(sheng)(sheng),200毫(hao)(hao)(hao)(hao)升(sheng)(sheng),250毫(hao)(hao)(hao)(hao)升(sheng)(sheng)三角(jiao)瓶(ping)中分別加(jia)(jia)營養(yang)(yang)液,70,80,120,150毫(hao)(hao)(hao)(hao)升(sheng)(sheng)(可按一試管(guan)對(dui)應(ying)一個三角(jiao)瓶(ping)加(jia)(jia)入(ru)),C: 500毫(hao)(hao)(hao)(hao)升(sheng)(sheng)瓶(ping)(400毫(hao)(hao)(hao)(hao)升(sheng)(sheng)營養(yang)(yang)液)(按1個三角(jiao)瓶(ping)對(dui)應(ying)1個500毫(hao)(hao)(hao)(hao)升(sheng)(sheng)瓶(ping)加(jia)(jia)入(ru))D:2.5升(sheng)(sheng)瓶(ping)(2升(sheng)(sheng)營養(yang)(yang)液)(按1個500毫(hao)(hao)(hao)(hao)升(sheng)(sheng)瓶(ping)對(dui)應(ying)1個2.5升(sheng)(sheng)瓶(ping)加(jia)(jia)入(ru)),注(zhu)意:上述若發(fa)現藻(zao)生長情況不好,需要先鏡檢,判斷后(hou)再(zai)決定是(shi)否接種。

一(yi)般情況下3袋(dai)二級藻(zao)種應先擴培接(jie)種于容量400升(sheng)的桶(海水貯備及營養液(ye)提前配置好),然后立即進行二級藻(zao)袋(dai)擴種,每個聚乙烯袋(dai)加18升(sheng)左右藻(zao)水。

4、保(bao)種(zhong)培(pei)養基制備:按1.5%的(de)比(bi)例加瓊脂(zhi)粉,煮沸后加營(ying)養鹽(母(mu)液千分(fen)之(zhi)1添加),高(gao)壓滅菌(jun)(jun)。維生(sheng)素需高(gao)壓滅菌(jun)(jun)后培(pei)養基冷卻(que)至30攝氏度方可加入。(可用劃線(xian)法(fa),涂布法(fa)等分(fen)離培(pei)養藻(zao)種(zhong))。

5、儀器的(de)清(qing)洗(xi)維護。溶(rong)劑過濾器使(shi)用(yong)后(hou)只用(yong)洗(xi)潔(jie)精和清(qing)水清(qing)洗(xi)。高(gao)壓(ya)滅菌(jun)后(hou)備(bei)(bei)用(yong)。其它玻(bo)璃儀器,氣管,氣石,移液槍筒使(shi)用(yong)后(hou)需要浸(jin)泡在百(bai)(bai)分之(zhi)(zhi)一到百(bai)(bai)分之(zhi)(zhi)十的(de)稀鹽酸溶(rong)液中至少半天,才(cai)能拿出清(qing)洗(xi),風(feng)干(gan),高(gao)壓(ya)滅菌(jun)后(hou)備(bei)(bei)用(yong)。

說明:10的2次(ci)方標示成10^2,以下數(shu)字的次(ci)方通用此表示法

三級藻生產

三(san)級藻系統包括用(yong)水處(chu)理以(yi)及配送,溫度與(yu)光照控(kong)制,藻種(zhong)的(de)供應等。為確保擴大培養(yang)成功,從接種(zhong)到(dao)收獲(huo)的(de)培養(yang)時間(jian)應該盡量縮短,以(yi)避免因(yin)為開放(fang)式培養(yang)隨培養(yang)規模擴大接觸外界感染源時間(jian)加長,三(san)級藻池滋生過多(duo)的(de)有害生物(wu),使三(san)級培養(yang)失(shi)敗。

1—1高(gao)水(shui)位低密(mi)(mi)(mi)度接種(接種密(mi)(mi)(mi)度為P×10^4cell/ml,P為接種密(mi)(mi)(mi)度萬位數(shu)值)(未特別(bie)說明藻(zao)水(shui)藻(zao)細胞(bao)密(mi)(mi)(mi)度就是(shi)cell/ml)(以(yi)下的推理是(shi)建立在三級(ji)(ji)藻(zao)第二次加水(shui)和收獲的密(mi)(mi)(mi)度為105級(ji)(ji))

若(ruo)一(yi)天(tian)(tian)(tian)三級(ji)(ji)(ji)濃(nong)縮(suo)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)需求量為(wei)(wei)(wei)A升(sheng),則(ze)總藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)細(xi)(xi)胞(bao)數(shu)(shu)目為(wei)(wei)(wei)K=A×10^3×(B×10^7)cell(K為(wei)(wei)(wei)A升(sheng)濃(nong)縮(suo)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)所含的(de)(de)(de)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)細(xi)(xi)胞(bao)數(shu)(shu)值(zhi),B為(wei)(wei)(wei)濃(nong)縮(suo)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)密度(du)(du)千(qian)萬位(wei)的(de)(de)(de)數(shu)(shu)值(zhi)),一(yi)般(ban)情況下二(er)(er)級(ji)(ji)(ji)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)種的(de)(de)(de)密度(du)(du)為(wei)(wei)(wei)C×10^6cell/ml(C為(wei)(wei)(wei)二(er)(er)級(ji)(ji)(ji)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)密度(du)(du)百萬位(wei)的(de)(de)(de)數(shu)(shu)值(zhi)),若(ruo)第(di)一(yi)天(tian)(tian)(tian)接種E噸藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)水,則(ze)一(yi)天(tian)(tian)(tian)需要二(er)(er)級(ji)(ji)(ji)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)種體積d=E×10^6×P×104/C×10^6=EP×10/C升(sheng),第(di)三天(tian)(tian)(tian)加(jia)(jia)水F噸,此(ci)時加(jia)(jia)水后(hou)(hou)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)水密度(du)(du)E×10^6×Y×10^5/(E+F)×10^6cell/ml(Y為(wei)(wei)(wei)第(di)三天(tian)(tian)(tian)加(jia)(jia)F噸水之前藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)水密度(du)(du)十萬位(wei)數(shu)(shu)值(zhi)),然后(hou)(hou)第(di)五天(tian)(tian)(tian)收獲時,藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)池(chi)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)水藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)細(xi)(xi)胞(bao)總數(shu)(shu)目應為(wei)(wei)(wei)H=(E+F)×10^5×Z×10^6(Z為(wei)(wei)(wei)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)池(chi)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)水密度(du)(du)十萬位(wei)數(shu)(shu),H為(wei)(wei)(wei)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)池(chi)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)水藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)細(xi)(xi)胞(bao)總數(shu)(shu)目),因為(wei)(wei)(wei)總需求量的(de)(de)(de)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)細(xi)(xi)胞(bao)數(shu)(shu)目應等(deng)于最后(hou)(hou)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)池(chi)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)水藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)細(xi)(xi)胞(bao)總數(shu)(shu)目,則(ze)K=H,則(ze)A×10^3×B×10^7=(E+F)×10^6×Z×10^5經過計算是A×B=(E+F)×10×Z。

舉例

若(ruo)一(yi)天需濃縮(suo)藻(zao)(zao)(zao)(zao)320升(sheng),則(ze)A=320,(一(yi)般情況下E和F的值(zhi)(zhi)接近(jin)),則(ze)因為K=H,所以320×B=20EZ,即320B=20EZ,E=16B/Z,即B/Z噸藻(zao)(zao)(zao)(zao)池藻(zao)(zao)(zao)(zao)水(shui)可以收(shou)集(ji)濃縮(suo)藻(zao)(zao)(zao)(zao)20升(sheng)(B為濃縮(suo)藻(zao)(zao)(zao)(zao)密(mi)度(du)千萬位的數(shu)值(zhi)(zhi),Z為藻(zao)(zao)(zao)(zao)池藻(zao)(zao)(zao)(zao)水(shui)密(mi)度(du)十萬位數(shu)),即若(ruo)一(yi)天需要濃縮(suo)藻(zao)(zao)(zao)(zao)A升(sheng),則(ze)每天應接種(zhong)E=AB÷20Z噸水(shui)。(Z和B的值(zhi)(zhi)應經過(guo)三級培養(yang)和收(shou)集(ji)藻(zao)(zao)(zao)(zao)水(shui)后才知道近(jin)似數(shu)值(zhi)(zhi))。

1—2高水位(wei)低密度接種(接種密度為(wei)(wei)P×10^4cell/ml,P為(wei)(wei)接種密度萬位(wei)數值(zhi))(未特別(bie)說明的藻(zao)水藻(zao)細胞密度就(jiu)是cell/ml)(以下的推(tui)理是建立(li)在三級藻(zao)第(di)二(er)次加水和收(shou)獲密度為(wei)(wei)10^6級)。

若(ruo)一(yi)天(tian)(tian)三(san)級(ji)濃縮藻(zao)(zao)(zao)(zao)(zao)(zao)需(xu)(xu)求量(liang)為(wei)(wei)(wei)(wei)A升,則(ze)總(zong)(zong)(zong)藻(zao)(zao)(zao)(zao)(zao)(zao)細(xi)(xi)(xi)胞(bao)數(shu)目為(wei)(wei)(wei)(wei)K=A×10^3×(B×10^7)cell (K為(wei)(wei)(wei)(wei)A升濃縮藻(zao)(zao)(zao)(zao)(zao)(zao)所含的藻(zao)(zao)(zao)(zao)(zao)(zao)細(xi)(xi)(xi)胞(bao)數(shu)值(zhi)(zhi),B為(wei)(wei)(wei)(wei)濃縮藻(zao)(zao)(zao)(zao)(zao)(zao)密(mi)度(du)千萬(wan)位的數(shu)值(zhi)(zhi)),一(yi)般情(qing)(qing)況下(xia)二級(ji)藻(zao)(zao)(zao)(zao)(zao)(zao)種的密(mi)度(du)為(wei)(wei)(wei)(wei)C×10^6cell/ml(C為(wei)(wei)(wei)(wei)二級(ji)藻(zao)(zao)(zao)(zao)(zao)(zao)密(mi)度(du)百萬(wan)位的數(shu)值(zhi)(zhi)),若(ruo)一(yi)天(tian)(tian)接(jie)種E噸(dun)藻(zao)(zao)(zao)(zao)(zao)(zao)水(shui)(shui)(shui),則(ze)一(yi)天(tian)(tian)需(xu)(xu)要二級(ji)藻(zao)(zao)(zao)(zao)(zao)(zao)種體積(ji)d=E×10^6×P×10^4/C×10^6=EP×10/C升,第(di)三(san)天(tian)(tian)加水(shui)(shui)(shui)F噸(dun),此(ci)時(shi)加水(shui)(shui)(shui)后藻(zao)(zao)(zao)(zao)(zao)(zao)水(shui)(shui)(shui)密(mi)度(du)E×10^6×Y×10^6/(E+F) ×10^6 cell/ml(Y為(wei)(wei)(wei)(wei)第(di)三(san)天(tian)(tian)加F噸(dun)水(shui)(shui)(shui)之前藻(zao)(zao)(zao)(zao)(zao)(zao)水(shui)(shui)(shui)密(mi)度(du)百萬(wan)位數(shu)值(zhi)(zhi)),然后第(di)五天(tian)(tian)收(shou)獲(huo)時(shi),藻(zao)(zao)(zao)(zao)(zao)(zao)池(chi)藻(zao)(zao)(zao)(zao)(zao)(zao)水(shui)(shui)(shui)藻(zao)(zao)(zao)(zao)(zao)(zao)細(xi)(xi)(xi)胞(bao)總(zong)(zong)(zong)數(shu)目應為(wei)(wei)(wei)(wei)H=(E+F)×10^6×Z×10^6 (Z為(wei)(wei)(wei)(wei)藻(zao)(zao)(zao)(zao)(zao)(zao)池(chi)藻(zao)(zao)(zao)(zao)(zao)(zao)水(shui)(shui)(shui)密(mi)度(du)百萬(wan)位數(shu),H為(wei)(wei)(wei)(wei)藻(zao)(zao)(zao)(zao)(zao)(zao)池(chi)藻(zao)(zao)(zao)(zao)(zao)(zao)水(shui)(shui)(shui)藻(zao)(zao)(zao)(zao)(zao)(zao)細(xi)(xi)(xi)胞(bao)總(zong)(zong)(zong)數(shu)目),因為(wei)(wei)(wei)(wei)總(zong)(zong)(zong)需(xu)(xu)求量(liang)的藻(zao)(zao)(zao)(zao)(zao)(zao)細(xi)(xi)(xi)胞(bao)數(shu)目應等于(yu)最后藻(zao)(zao)(zao)(zao)(zao)(zao)池(chi)藻(zao)(zao)(zao)(zao)(zao)(zao)水(shui)(shui)(shui)藻(zao)(zao)(zao)(zao)(zao)(zao)細(xi)(xi)(xi)胞(bao)總(zong)(zong)(zong)數(shu)目,則(ze)K=H,則(ze) A×10^3×B×10^7 =(E+F)×10^6×Z×10^6經過計算是A×B=(E+F) ×100×Z, (一(yi)般情(qing)(qing)況下(xia)E和F的值(zhi)(zhi)接(jie)近(jin)),則(ze)近(jin)似AB=200EZ, 即(ji)若(ruo)一(yi)天(tian)(tian)需(xu)(xu)要濃縮藻(zao)(zao)(zao)(zao)(zao)(zao)A升,則(ze)每天(tian)(tian)應接(jie)種E=AB÷200Z噸(dun)水(shui)(shui)(shui)。(Z和B的值(zhi)(zhi)應經過三(san)級(ji)培養和收(shou)集藻(zao)(zao)(zao)(zao)(zao)(zao)水(shui)(shui)(shui)后才知道近(jin)似數(shu)值(zhi)(zhi))。

2—1低水位高密(mi)度接種(zhong)(接種(zhong)密(mi)度為P×10^5cell/ml,P為接種(zhong)密(mi)度十萬位數值)(未特別說明藻水藻細(xi)胞(bao)密(mi)度就是cell/ml)(以下(xia)的推(tui)理是建立在三級藻第二次,第三次加水和收獲(huo)的密(mi)度為105級)。

若一(yi)天(tian)(tian)(tian)三級(ji)濃縮(suo)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)需求(qiu)量為(wei)A升(sheng),則總藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)細(xi)胞數(shu)目(mu)為(wei)K=A×10^3×(B×10^7)cell (K為(wei)A升(sheng)濃縮(suo)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)所(suo)含的(de)(de)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)細(xi)胞數(shu)值,B為(wei)濃縮(suo)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)密(mi)(mi)度(du)(du)千萬(wan)(wan)位(wei)的(de)(de)數(shu)值),一(yi)般情況下二(er)(er)級(ji)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)種(zhong)的(de)(de)密(mi)(mi)度(du)(du)為(wei)C×10^6cell/ml(C為(wei)二(er)(er)級(ji)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)密(mi)(mi)度(du)(du)百(bai)萬(wan)(wan)位(wei)的(de)(de)數(shu)值),若第(di)一(yi)天(tian)(tian)(tian)接種(zhong)E噸(dun)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)水(shui)(shui)(shui)(shui),則一(yi)天(tian)(tian)(tian)需要二(er)(er)級(ji)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)種(zhong)體積d=E×10^6×P×10^5/C×10^6=EP×100/C升(sheng),第(di)二(er)(er)天(tian)(tian)(tian)加(jia)水(shui)(shui)(shui)(shui)F噸(dun),此時(shi)加(jia)水(shui)(shui)(shui)(shui)后藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)水(shui)(shui)(shui)(shui)密(mi)(mi)度(du)(du)E×10^6×Y×10^5/(E+F)×10^6cell/ml(Y為(wei)第(di)二(er)(er)天(tian)(tian)(tian)加(jia)F噸(dun)水(shui)(shui)(shui)(shui)之前(qian)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)水(shui)(shui)(shui)(shui)密(mi)(mi)度(du)(du)十(shi)萬(wan)(wan)位(wei)數(shu)值),然(ran)后第(di)三天(tian)(tian)(tian)加(jia)水(shui)(shui)(shui)(shui)G噸(dun)時(shi),藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)池(chi)(chi)(chi)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)水(shui)(shui)(shui)(shui)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)細(xi)胞密(mi)(mi)度(du)(du)應為(wei)J=(E+F)×10^6×Z×10^5/(E+F+G)×10^6(Z為(wei)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)池(chi)(chi)(chi)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)水(shui)(shui)(shui)(shui)密(mi)(mi)度(du)(du)十(shi)萬(wan)(wan)位(wei)數(shu)),第(di)四天(tian)(tian)(tian)或第(di)五天(tian)(tian)(tian)收獲時(shi),藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)池(chi)(chi)(chi)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)水(shui)(shui)(shui)(shui)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)細(xi)胞總數(shu)目(mu)應為(wei)H=(E+F+G)×10^6×W×10^5(W為(wei)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)池(chi)(chi)(chi)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)水(shui)(shui)(shui)(shui)密(mi)(mi)度(du)(du)十(shi)萬(wan)(wan)位(wei)數(shu),H為(wei)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)池(chi)(chi)(chi)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)水(shui)(shui)(shui)(shui)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)細(xi)胞總數(shu)目(mu)),因為(wei)總需求(qiu)量的(de)(de)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)細(xi)胞數(shu)目(mu)應等于(yu)最后藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)池(chi)(chi)(chi)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)水(shui)(shui)(shui)(shui)藻(zao)(zao)(zao)(zao)(zao)(zao)(zao)細(xi)胞總數(shu)目(mu)。

則(ze)K=H,則(ze)A×10^3×B×10^7=(E+F+G)×10^6×W×10^5經過計算是A×B=(E+F+G) ×10×W,

(一般情況(kuang)下(xia)E,F,G的(de)值接近(jin)),則(ze)近(jin)似(si)AB=30EW, 即若一天需要濃縮藻A升,則(ze)每天應接種E=AB÷30W噸水(shui)。(W和(he)B的(de)值應經過三級培(pei)養和(he)收(shou)集藻水(shui)后才知(zhi)道近(jin)似(si)數值)。

2—2低水(shui)位(wei)高密度(du)接(jie)(jie)種(接(jie)(jie)種密度(du)為(wei)P×10^5cell/ml,P為(wei)接(jie)(jie)種密度(du)十萬位(wei)數值)(未特別說明(ming)藻水(shui)藻細胞密度(du)就是cell/ml)(以下的推理是建立在三(san)級藻第二次,第三(san)次加水(shui)和收獲的密度(du)為(wei)10^6級)。

若一(yi)(yi)天(tian)(tian)(tian)三(san)級濃(nong)縮(suo)(suo)藻(zao)(zao)需求量為(wei)(wei)(wei)(wei)A升,則(ze)總(zong)藻(zao)(zao)細(xi)(xi)胞(bao)數(shu)(shu)(shu)目為(wei)(wei)(wei)(wei)K=A×10^3×(B×10^7)cell (K為(wei)(wei)(wei)(wei)A升濃(nong)縮(suo)(suo)藻(zao)(zao)所(suo)含的(de)(de)藻(zao)(zao)細(xi)(xi)胞(bao)數(shu)(shu)(shu)值(zhi),B為(wei)(wei)(wei)(wei)濃(nong)縮(suo)(suo)藻(zao)(zao)密(mi)度(du)(du)(du)千萬(wan)(wan)位(wei)(wei)(wei)的(de)(de)數(shu)(shu)(shu)值(zhi)),一(yi)(yi)般(ban)情況(kuang)下(xia)二級藻(zao)(zao)種(zhong)(zhong)的(de)(de)密(mi)度(du)(du)(du)為(wei)(wei)(wei)(wei)C×10^6cell/ml(C為(wei)(wei)(wei)(wei)二級藻(zao)(zao)密(mi)度(du)(du)(du)百萬(wan)(wan)位(wei)(wei)(wei)的(de)(de)數(shu)(shu)(shu)值(zhi)),若一(yi)(yi)天(tian)(tian)(tian)接(jie)種(zhong)(zhong)E噸藻(zao)(zao)水(shui)(shui),則(ze)一(yi)(yi)天(tian)(tian)(tian)需要二級藻(zao)(zao)種(zhong)(zhong)體(ti)積(ji)d=E×10^6×P×10^5/C×10^6=EP×100/C升,第(di)二天(tian)(tian)(tian)加(jia)水(shui)(shui)F噸,此時加(jia)水(shui)(shui)后(hou)(hou)藻(zao)(zao)水(shui)(shui)密(mi)度(du)(du)(du)E×10^6×Y×10^6/(E+F)×10^6cell/ml(Y為(wei)(wei)(wei)(wei)第(di)二天(tian)(tian)(tian)加(jia)F噸水(shui)(shui)之前藻(zao)(zao)水(shui)(shui)密(mi)度(du)(du)(du)百萬(wan)(wan)位(wei)(wei)(wei)數(shu)(shu)(shu)值(zhi)),然后(hou)(hou)第(di)三(san)天(tian)(tian)(tian)加(jia)水(shui)(shui)G噸時,藻(zao)(zao)池藻(zao)(zao)水(shui)(shui)藻(zao)(zao)細(xi)(xi)胞(bao)密(mi)度(du)(du)(du)應(ying)為(wei)(wei)(wei)(wei)J=(E+F)×10^6×Z×10^6/(E+F+G) ×10^6 (Z為(wei)(wei)(wei)(wei)藻(zao)(zao)池藻(zao)(zao)水(shui)(shui)密(mi)度(du)(du)(du)百萬(wan)(wan)位(wei)(wei)(wei)數(shu)(shu)(shu)),第(di)四(si)天(tian)(tian)(tian)或(huo)第(di)五天(tian)(tian)(tian)收獲時,藻(zao)(zao)池藻(zao)(zao)水(shui)(shui)藻(zao)(zao)細(xi)(xi)胞(bao)總(zong)數(shu)(shu)(shu)目應(ying)為(wei)(wei)(wei)(wei)H=(E+F+G)×10^6×W×10^6 (W為(wei)(wei)(wei)(wei)藻(zao)(zao)池藻(zao)(zao)水(shui)(shui)密(mi)度(du)(du)(du)百萬(wan)(wan)位(wei)(wei)(wei)數(shu)(shu)(shu),H為(wei)(wei)(wei)(wei)藻(zao)(zao)池藻(zao)(zao)水(shui)(shui)藻(zao)(zao)細(xi)(xi)胞(bao)總(zong)數(shu)(shu)(shu)目),因為(wei)(wei)(wei)(wei)總(zong)需求量的(de)(de)藻(zao)(zao)細(xi)(xi)胞(bao)數(shu)(shu)(shu)目應(ying)等于(yu)最后(hou)(hou)藻(zao)(zao)池藻(zao)(zao)水(shui)(shui)藻(zao)(zao)細(xi)(xi)胞(bao)總(zong)數(shu)(shu)(shu)目,則(ze)K=H,則(ze)A×10^3×B×10^7=(E+F+G)×10^6×W×10^6經過(guo)(guo)計算是(shi)A×B=(E+F+G) ×10×W。(一(yi)(yi)般(ban)情況(kuang)下(xia)E,F,G的(de)(de)值(zhi)接(jie)近),則(ze)近似(si)AB=300EW, 即若一(yi)(yi)天(tian)(tian)(tian)需要濃(nong)縮(suo)(suo)藻(zao)(zao)A升,則(ze)每(mei)天(tian)(tian)(tian)應(ying)接(jie)種(zhong)(zhong)E=AB÷300W噸水(shui)(shui)。(W和B的(de)(de)值(zhi)應(ying)經過(guo)(guo)實(shi)地三(san)級培養(yang)和收集藻(zao)(zao)水(shui)(shui)后(hou)(hou)才知(zhi)道(dao)近似(si)數(shu)(shu)(shu)值(zhi))。

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