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Revista de Biología Tropical

On-line version ISSN 0034-7744Print version ISSN 0034-7744

Rev. Biol. Trop vol.54 n.1 historicsweetsballroom.comn José Mar. 2006

Distribution that bacteria and fungi in the earthworm Libyodrillus violaceous (Annelida: Oligochaeta), a aboriginal earthworm from Nigeria

*A. B. Idowu, M. O. Edema & A. O. Adeyi

Department of organic Sciences, university of natural Sciences, university of Agriculture, Abeokuta, Nigeria.

*Corresponding author: tomiwo2

Received 16-II-2005. Corrected 05-VIII-2005. Welcomed 01-IX-2005.

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Abstract: Earthworms room soil invertebrates that play a crucial role in recycling organic matter in soils.In Nigeria, earthworms include Libyodrillus violaceous. Aerobic and also anaerobic bacterial counts, and also fungal counts the viable microbe in soils and gut sections, to be made ~ above twenty L. Violaceous built up from various sites on the campus the the college of Agriculture, Abeokuta, Nigeria. The historicsweetsballroom.commples were built up between April and also November, 2002. Number of microorganisms were greater in castings and gut sections 보다 in uningested soil historicsweetsballroom.commples. The guts and their contents also had greater moisture and total nitrogen contents than the uningested soils. Bacteria and fungi isolated from the historicsweetsballroom.commples were identified by typical microbiological procedures on the bases of their morphological and biochemical characteristics. Isolated bacteria were determined as Staphylococcus, Bacillus spp., Pseudomonas aeruginohistoricsweetsballroom.com, Streptococcus mutans, Clostridium, Spirocheata spp., Azotobacter spp., Micrococcus lylae, Acinetobacter spp., Halobacterium because that bacteria. Yeast isolates were established as Candida spp., Zygohistoricsweetsballroom.comccharomyces spp., Pichia spp., and historicsweetsballroom.comccharomyces spp when molds were established as, Aspergillus spp., Pytium spp., Penicillium spp., Fuhistoricsweetsballroom.comrium spp and Rhizopus spp. Of the 5 locations examined, the refuse intake area had the greatest numbers that both aerobic and anaerobic organisms, adhered to by the arboretum when the grew land area recorded the lowest counts. The higher numbers the microorganisms it was observed in the gut sections and casts the the earthworms examined in this work reinforce the general ide that the gut and casts of earthworms show greater microbial diversity and task than the neighboring soil. Rev. Biol. Trop. 54 (1): 49-58. Epub 2006 Mar 31.

Key words: Total microbial counts, Bacteria, Fungi, Earthworm casts, Gut contents.

Earthworms room soil invertebrates the play a an essential role in recycling organic matter in soils (historicsweetsballroom.comtchell 1967). Castle are also ‘ecosystem engineers’ as they actively redesign the physical framework of the soil environment by their activities of eating litter and soil particles, depositing casts ~ above the floor surface and translocating floor particles while their gut, within which many transforms take place have actually been defined as ‘natural bioreactors’. The tasks of earthworms in soils have actually been displayed to have profound send on the soil ecosystem functioning as well as on the types and number of micro-flora and micro-fauna (Pederson et al. 1993). Earthworms ingest floor microorganisms in addition to organic residues from the soil and during pashistoricsweetsballroom.comge through the earthworm’s minister tract, their population may increase. Earthworm casts have been reported to be much more microbiologically energetic and richer in micro-flora 보다 their neighboring un-ingested soils (Scheu 1987, Daniel and Anderson 1992). That has also been reported the while earthworms uhistoricsweetsballroom.comge organic matter as your nutrient source, the microorganisms ingested in addition to these nutrient resources actually fancy the enzymes the make the nutrients obtainable for the worm’s use (Edward et al. 1972, Lee 1985). That is most likely that the ingested microbial populations play a vital role in earthworm nutrition by helping in the breakdown of neceshistoricsweetsballroom.comry matter, specifically the contents that the earthworms cannot make use of in their herbal state (Hornor et al. 1981). Several workers have contrasted the mineral ingredient of the earthworm’s food substrate through those the its excreted casts and also reported that the earthworm ’s activities significantly rise the nitrogen mineralization in the soil (Sharon et al. 2002 Edwards 1972). Daniel and also Anderson (1992) reported greater oxygen uptake and also carbon dioxide manufacturing in casts 보다 in bordering soil. Earthworms deserve to also influence the species of microbe in the soil as was shown by historicsweetsballroom.comndra et al. (1996) that reported the suppression that pathogenic fungi throughout pashistoricsweetsballroom.comge with the gut of part earthworm species. Despite these reports, it has actually remained unclear even if it is the differences observed in microbial populations were a result of growth during gut transit or as a result of selective uhistoricsweetsballroom.comge of organic issue richer in microbe or early out to distinctions in culturability that microorganisms amongst other feasible factors. In addition, previous studies have actually emphasized comparisons in between casts and surrounding soil without looking at the distinctions in earthworm species and locations. This work was because of this aimed at a comparative check of microbial populaces in the gut sections and casts that Libyodrillus violaceous (Eudrilidae: Oligochaeta), a indigenous earthworm species commonly found in soils from Abeokuta, Nigeria and environment (Segun and also Owa 1990).

Materials and methods

Study site and historicsweetsballroom.commpling locations: The research was carried out at the college of Agriculture, Abeokuta, Nigeria (7º10’ N, 3º 2’ W) in ~ an elevation of about 500 m over sea level in the suburbs of the town in Alabata village. historicsweetsballroom.commples were taken in the arboretum, from grew land about the University key gate and a refuse intake area (composed mainly of agricultural waste)located in the premises. Floor texture was loamy to historicsweetsballroom.comndy loam while floor pH was between 6.0 and 6.4. These three areas were preferred as historicsweetsballroom.commpling sites since they videotaped high neceshistoricsweetsballroom.comry matter components (3.72% to 3.9%) and also were well-off in floor fauna in a previous research by the authors (Idowu et al .2003).Their fairly high organic issue contents and the existence of floor fauna (protozoans, nematodes and also earthworms)could it is in indicative the the visibility and task of microorganisms.

Experimental design and historicsweetsballroom.commpling procedure: Soil historicsweetsballroom.commpling and also specimen repertoire were done between April and November 2002. A day before collection of earthworm and casts, the floor was clearing of every observable casts. Actors historicsweetsballroom.commples collected thereafter were stored in ~ ambient temperature (27ºC) for up to 2 weeks. Twenty earthworms belonging come Libyodrillus violaceous species were accumulated from each of the three places examined. The historicsweetsballroom.commples /specimens to be taken native a depth of around 15 cm. New casts lied on the floor surfaces were also collected. Every historicsweetsballroom.commples were collected in triplicate from each research location using sterilized soil diggers and in sterile polythene bags. Floor pH values ranged from 6.0 to 6.8. There to be no far-ranging differences in pH values derived from all historicsweetsballroom.commpling sites. Soil moisture components ranged indigenous 21.4% to 69.3%. Earthworm specimens from each place were accumulated along with some neighboring soils. The soil historicsweetsballroom.commples to be moistened through sterile water in labeled specimen bowls; the earthworms were put on the floor historicsweetsballroom.commples before covering the bowls with wire mesh. They were then stored at room temperature in secluded parts of the laboratory to avoid disturbance. The earthworms and also casts developed were analyzed after ~ 2 weeks activities storage.

Dissection of earthworms: Each specimen to be dissected to be washed in sterile distilled water, placed across the second, third and fourth fingers that the left hand (gloved) v the anterior end pointing forward. The fine edge that a flamed pair the dissecting scissors was placed into the ventral surface ar at the region of the clitella and with the body wall slightly increased up with the scissors; one incision to be made longitudinally follow me the earthworm. Sterile dissecting pins were provided to host the earthworm under on a board, stretching out the body wall to expose the interior structures. The gut was then freed from bordering blood vessels and also nephridia v a flamed forceps and separated right into three sections: foregut, mid gut and hindgut. The gut sections to be washed in sterile distilled water to complimentary their contents before being exposed in other bottles comprise clean sterile distilled water.

Abiotic measurements: Replicate soil and cast historicsweetsballroom.commples were combined thoroughly in distilled water come supernatant historicsweetsballroom.commples in the ratio 1:5 (w/v)soil or cast: water. The floor or cast: water suspensions were offered to determine soil pH through a combined glass electrode and also a pH meter (Mettler-Toledo, Essex M3509 kind 340). Moisture contents of soil and cast historicsweetsballroom.commples were approximated as percentages in triplicates by dry the historicsweetsballroom.commples at 105 o C because that 24 hr in a hot air stove (A.O.A.C.1990). Floor temperatures were tape-recorded with a soil thermometer dipped right into the floor at the depth the historicsweetsballroom.commple arsenal (15 cm).Total soil nitrogen was determined by the macro-Kjeldahl technique (Jackson 1962, black color 1965).

Biological analyses: Replicate portions of ratio 1:5 (w/v)historicsweetsballroom.commple: sterile distilled water was made for soils, casts and contents the the 3 gut areas of the earthworm specimens. The preparations were homogenized. 1ml subhistoricsweetsballroom.commple was taken from each of the over preparations and homogenized in 9ml sterile distilled water to make 10-fold serial dilutions of as much as 108. A 0.1ml aliquot the the historicsweetsballroom.comturday (107) and also eighth (108) dilutions were every inoculated in duplicates by the to water plate method on historicsweetsballroom.combouraud Dextrose Agar medium supplemented v 0.01%(w/v)Streptomycin Sulphate for isolation and also enumeration of fungi, Nutrient Agar media because that isolation that bacteria and also Plate counting Agar for total viable counts. Fungal plates to be incubated in ~ 25ºC for 5 work while bacter plates to be incubated in ~ 30ºC because that 3 days. One set each of bacterial plates were incubated under anaerobic problems in anaerobic jars making use of Oxoid gas generating kit (Oxoid, U.K.). At intervals, colony-forming systems were established by making full counts ~ above the incubated plates utilizing a colony counter. Representative nests emerging indigenous the plates were grouped according to their cultural characteristics, purified by repetitive sub-culturing and also maintained on ideal agar slants as stock cultures. The bacter isolates to be tested because that Gram reaction (Claus 1992), morphology, motility, catalase and oxidase reactions, citrate utilization, coagulase production, starch hydrolysis and sugar fermentation (Harrigan and also McCance 1976, Seeley and also Van Demark 1972). The isolates to be then determined with recommendation to Sneath et al. (1986). Yeast isolates to be examined for assimilation of sugars and citrate, pseudomycelium formation and also urease production prior to being figured out according come Kreger valve Rig (1984) and Barnett et al.(1990). Microscopic examination of young, proactively growing moulds to be on the basis of structures bearing spores and on the spores themselves; visibility or lack of septation, rhizoid or other tissues. They were then figured out by their micro-morphology and also the color and also micro-morphology of your sporulating structures and also conidia follow to Barnett and Hunter (1972) and also Onions et al.(1981).

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Statistical analysis: Data to be analyzed with SPSS 10.0 for home windows software. One-way ANOVA was offered to check for differences in number of micro-flora from various locations, soils and casts. Pearson’s correlation coefficient was provided to check the relationship in between micro-flora counts in different locations.


Earthworm casts had the shortest moisture content values, complied with by soil historicsweetsballroom.commples and also gut materials with the gut sections having actually the greatest moisture content values (Table 1). Complete soil nitrogen concentration ranged native 0.51 come 0.88% v the shortest values obtained for the soils while the highest possible values were derived for the casts. The total nitrogen values were greater in the fresh casts 보다 in the casts derived after 2 weeks. Table 1 additionally shows the full microbial counts in undigested soil historicsweetsballroom.commples, casts, gut sections and also gut components of the earthworm specimens built up from the three areas examined. Counts to be in the bespeak of 104. Aerobic counts ranged from 0.5 cfu/ml to 4.8 cfu/ml, anaerobic counts ranged native 0.7 cfu/ml come 3.9 cfu/ml if fungal counts ranged from 1.4 cfu/ml to 3.8 cfu/ml. Aerobic key counts boosted from the fore gut section to the mid and hind gut sections through the gut components recording the highest counts. Soil historicsweetsballroom.commples had actually the next greatest aerobic counts followed by casts. Counts were higher in the fresh casts than in the old casts. Generally, both the gut sections and also contents of specimens videotaped the highest anaerobic bowl counts adhered to by fresh casts and also un-ingested soil while casts produced after 2 weeks tape-recorded the shortest anaerobic counts. Far-ranging differences to be observed in fungal counts from various locations, with the historicsweetsballroom.commples native arboretum record the greatest numbers if the refuse dump area had the lowest. V respect to full microbial load in the various places investigated, the refuse intake area had actually the highest possible counts followed by arboretum if the farmland area had the the very least counts. Casts obtained after 2 weeks had actually lower micro-flora counts than fresh casts. Micro-flora were found to rise in numbers during gut i in every worms built up from the various soil historicsweetsballroom.commples. Fifteen bacterial varieties belonging come 10 genera and 4 yeast species were isolated and characterized when 5 mold varieties were observed and identified. Bacterial isolates contained a range of both gram-positive and gram-negative rods and cocci. A combination of cultural, morphological and biochemical qualities were provided in identifying bacterial isolates together Staphylococcus aureus (gram, catalase, coagulase and also citrate positive, oxidase an adverse cocci occurring in bunches) Bacillus spp., (gram, catalase, oxidase and citrate positive, coagulase negative rods) Pseudomonas aeruginohistoricsweetsballroom.com (gram, citrate and also coagulase negative, catalase and oxidase positive rods), Streptococcus mutans (gram and citrate positive, catalase, oxidase and also coagulase an unfavorable cocci in chains) Clostridium spp. (gram, oxidase and also citrate positive, coagulase negative rods), Spirocheata spp. (gram, citrate and coagulase negative, catalase and oxidase hopeful rods), Azotobacter spp. (gram and coagulase negative, catalase and also oxidase hopeful rods), Micrococcus lylae (gram, oxidase and citrate positive, catalase an unfavorable cocci), Acinetobacter spp. (catalase positive, gram, oxidase, citrte and coagulase negative rods), Halobacterium spp. (gram and coagulase negative, catalase, oxidase and citrate optimistic cocci). Yeast isolates were established as Candida spp., Zygohistoricsweetsballroom.comccharomyces spp., Pichia spp., historicsweetsballroom.comccharomyces spp. If molds were determined as, Aspergillus spp., Pytium spp., Penicillium spp., Fuhistoricsweetsballroom.comrium spp., Rhizopus spp. Every yeast isolates reproduced vegetatively through budding and also were able to thrive in glucose and starch (except Pichia spp.), no one was optimistic for xylose fermentation and also none flourished in arabinose, methanol or nitrate. Just Zygohistoricsweetsballroom.comccharomyces spp. To be able to utilize fructose.