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Calyciflorus (Rotifera: Brachionidae} in Waste Water Frmil Food-Processing Industry in Mexico

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Calyciflorus (Rotifera: Brachionidae} in Waste Water Frmil Food-Processing Industry in Mexico Rey. Biol. Trop.,43(6):595c600, 1998 Population dynamics ofBrachionus calyciflorus (Rotifera: Brachionidae} in waste water frmIl food-processing industry in Mexico Raymundo Alfredo Arévalo Stevenson, S.S.S. Sarma* andS. Nandinil Carrera de Biología, UNAl\lCCarnpus.Iztacala, AP 314 CP 54000, Los Reyes, lztacala, Tlanepantla, Edo. de México, México. Telefax: +52 (5) 623 1212, E-Mail: [email protected] *Corresponding author. Receiyed 26-IX-1997. Correcled 15-IV-1998. Accepted 29-IVc1998. ADst¡:a,ct:Waste water from Mexico's largesl food processingindustrial sector (basedonmaize, nejayote water) was �sed ior growingBrachionus calyciflorus isolated from Lake Chapultepec inthe Federal District of Mexico.(D.F.). Nejayote waterwa s c(jllected from Colonia Providencia, D.F. Experiments wereconducted al 25°Cin 25 mI capacity "ialswith20 mIof medlUminto which we intr�ducedB. calyciflorus ataninitial density of � ind mI-l. Theexperi­ W\1ental designconsisteg a total 003 testves.sels (2 food óombinations X 5densities X 3 replicates = 30 plus 3 repli­ cates as colitrols tha! contaiped only¡¡lgae). Experi rp.entsW"re tenninated <ÚterdaY16,Wastewatetin original concen- i tr¡uÍoll did no! supportrQtifets.Jloweyer, whenqi1uted t? 5 con¡:entrations (ranging Crom 2% to 32% andpH adjusted . t07.0), rotifer density increase� \Y¡thincreasingconc¡:nt ration oí wastewater, Green a1gae (at COf¡stant density of 2 X 6 1 .o ceIIsmr � .' of Chlordla) in �Ombination w¡ih was�.waterresl}ltedi �a higher abundance 0f rotifers only al higher 1 (;oncentrations (above 8%}ofwaste water:The mrudmutn peakdensityof rotifers (238 ± 5.0 ind ml- ) was obtained at 16% dilution of waste water (\nd with addition.of Chlorella. Therate ofpopulation ¡ncrease per day (r) (mean±SD) varied from .0.355 ± .0. .059 to .0.457. ±0..o48 deper¡dingonfoodcom binatioll and concentration. R. otifer.,B.rachionus,waste water, Key .words: .popul:¡1i.(Í�, '"growt!)¡ ,�,, M' ,'<ex.ico. Maize, particularlyin the form oí tortilla (a p¡:¡rtlyhydrúlizesthe hard seedcpat andturnsthe kind of thin-plate like soft dry bread) ís one of seedsinto softeasilyq:u.shableil1\lterial to fmm the most important components in the dictof dough . for tort.illa man\lfacture. Dunng tbis the Mexican population, Tortitla industry process of boiling, anumber of organic (both ranges from large scale (up to 50 tons per day) particulate and.dissolved) sllbstances including to cottage levelproductlon (about 700 kgs per proteins, carbohydrates andlipids are lost into day)(Durán de Bazúa 1988): �orethan 95% of thewater, resulting in an organically rich medi­ the tortilla industry in. Mexico utilizes maize um (Pedroza1985). (and the re�t 4tilizes wbeat). For preparing tor­ Rotifers a.re known. to utílize particulate tilla bythe.ttaditipnal method caHed nixtamal­ organic matter directly (pQurriot 1965) anddis­ iZ\lüon (lime-steeping. ofcorn), the.maizeseeds solved organicmatter via bacteria and protozoa are pre-boiled in wate,rwitblime. This process (Arndt 1993). They thus form an important link DE 596 REVISTA BIOLOGIA TROPICAL in the aquatic food chain, particuIarly in trans­ selected a site (Street: Estado de Zacatecas and fering energy from lower to higher trophic lev­ Estado de MoreIos, Colonia Providencia) in the els. A number of workers have obtained high Federal Distríct of Mexico which regularly production of rotifers (up to 500 ind mrl) in receives the nejayote water. Every alternate day outdoor aquacultural practices using piggery waste water was collected from the sarue site wastes and liquid organic wastes (Jhingran and stored in a refrígerator for the experiments. 1991). We did not use waste water which was stored for Rotifers of the genus Brachionus have been more than 2 days. Since the initial pH of the widely used as starter food for rearing larval nejayote water was as high as 9.0 due to high fish and crustaceans in aquaculture (Lubzens et levels of lime (which does not support rotifer al. 1989), indicators of pollution (SIadecek growth (Mitchell and Joubert 1986», it was 1983) and as bioassay organisms (Snen and brought down to 7.0 by the addition of diluted Janssen 1995). They have also been included as HCl. This process resulted in sorne precipitation standard bioassay organisms by the American at the bottom of the vessel, which was siphoned Socíety of Testing and Materials in the USA off. Since Brachionus does not feed effectively (Anon. 1991). Since the waste water of tortilla on particles larger than 20 �m in size (Poumot industry (known as nejayote water) is rich in 1965), we filtered this water using 20!-lm mesh organic matter, we aimed at testing its suitabili­ and the supernatant was diluted as required. This ty for growing rotifers. process also helped in removing most ciliates in the waste water. Once adjusted, the pH of the medium did not vary over a 24 h periodo Based MA'IERIALS AND METHODS on prelíminary tests it was observed that B. caly­ ciflorus did not grow well directly on the waste The rotifer Brachionuscalyciflorus (average water at its original concentration. We therefore adult length excluding spines = 185 ± 12 !-1m) diluted this water (using EPA medium) to vari­ was originally isolated from Lake Chapultepec ous concentrations up to 32% (= 68% dilution) (Mexico City) and successfully cultured in the which permitted the rotifers to grow for a rea­ laboratory using the single-celled green algae sonable periodo To estimate the growth of B. (Chlorella vulgaris, average cell diarueter: 5.48 calyciflorus, we chose 5 concentrations of the ± 1.21 !-1m) as the exclusive food (Sarma et al. waste water viz. 2, 4, 8, 16 to 32%. Growth of 1997). Chlorella was mass cultured using Bold the rotifer on the waste water was tested, both Basal medium (Borowitzka and Borowitzka with and without an additional food source 1988). (Chlorella at 2 X 106 cells m¡-l). Our stock as well as mass cultures of rotifers For experiments, we used 25 mI capacíty were maintained in EPA medium (Anon. 1985) transparent vials containing 20 mI of the waste androutinely fed daily the green algae at a densi­ water at the desired concentration and combina­ ty of approximately 2 X 106 cells mrl . Although tion of waste water. AH experiments were con­ in our routine cultures we were able to obtain ducted in thermostatically controlled water­ rotifers at a density of about 100 ind rnl-I; we baths set at 25 oc. The initial pH of the medium generally maintained the population below 50 ind was adjusted to 7.0. For each dilution of waste mr 1 in order to reduce thepossibility of male pro­ water, we used 3 replicates. In combinations duction. For regular feeding as well as for exper­ where alga was used as an addítional food 6 ¡ments, we used log phase algae, centrifuged at source, íts concentratíon was kept at 2 X 10 4000 rpm, rinsed in distiUed water and resus­ cells m¡-l. Thus, the experimental design con­ pended in EPA medium. The density of algae was sisted of 33 test vesseIs (2 food combinations X estimated using a haemocytometer. 5 densities X 3 replicates = 30 plus 3 replicates For obtaining a constant source of waste as control which contained only algae at the water from tortilla processing industry, we same density). lnto each of the test vessels, we AREVALO eral.: Population dynamicsófBráchionus calyciflorus 597 introduced B. calyciflorus at a density of 1 ind mI-l. The initial popuJation ofrotifers, counted 80 Control (0%) individuaUy, consisted of actively reproducing females of mixed age-group obtained from a 60 mass-culture tank during the exponential phase of their growth. The test vessels were main­ tained in diffuse and continuous fluorescent 40 iIlumination. Aeration was provided to the test vessds in arder to keep the food-párfiCles úHhe test medium in suspension. For counting rotifers, we used one of the two methods: a) whole count when the density of o rotifers was less than 5 ind mr1 or, b) aliquot O 2 4 6 8 10 12 14 16 subsamples of 1-5 mI volume whenthedensity 50 was greater than 5 ind mI-l. Fore�ch replicate, � 2% .- we counted at least 3 subsample5j.Following the . inoculatíon, we estimatedthe popuJation densi­ E 40 -t:i ty every day until most replicates completed §. one population Thus, the experiment was 30 cyc1e. �U) c: tenni�iltedafter day16. Everyday, after esti­ Q) "O matihg".the'populatioll density, rotifersftom' all c: 20 o repli2&,eswere transferred to fresh rnediUm :; "5 with appropriately diluted ""aster water and a. o 10 foodcombination. o.. For,estimating the pppulation density, vie counted onIy Iive female rotifers.Population dens1ty of rotifers was expressed,as num:ber per mLFor estimating the populationgrowth I-a,te (r)of totifers, we used the following fórmula: 1'= Nc nNo)/t (In l . where No :¡nitial population den:sity Nt = ¡:>opulatlon densit)' afrerthe,time t t= tirtte in �ays RESULTS 2 4 6 810 12 14 16 In general the density of Brachionus caly­ Time (days) ciflorus increased with incteasing concentra­ HonoLwaste water in themedil.ltn. The popu­ latíon density of the rotifers grownin waster water with additíon of algae showed a similar Fig.l. Population growth. of ¡he .totifer B. calyciflorus in trend(Figs 1 & 2). Anincrease in the proP9r­ relationto differentGoncentratlons ofwaste waterand in the presence andahsence ofthealgae ehlarel/a(at 2 X I06cells tion . of w�ste water in the mediu1l1 beyond mI-l). Shown ate the mean±SE values based on tbree replí· 16% did not suppqrt high population density cate recordings. Concentration of nejayote water is rotifers. of expressed in percentage. 598 REVISTA DE BIOLOGIA TROPICAL 140 300 8% - Presence � A 120 ";" = Absence _____ Presence E 250 ___ 100 Absence "'Óc: ;;;.. 80 � 200 c: (\) 60 "t:I .§ 150 40 (¡j "S c.
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