A study on the lethal toxicity of aminocarb to freshwater crayfish and its in vivo metabolism

Adult crayfish (Orconetes limosus) were exposed to 0.1, 1.0, 5.0, 10.0, 25.0, 30.0, 40.0, 50.0 and 60.0 ppm of aminocarb in water at 15 degrees C under laboratory conditions for 144 h. No apparent behavioral changes were observed in crayfish exposed to 0.1, 1.0 and 5.0 ppm of aminocarb during the experiment. Symptoms of acute toxicity were apparent at concentrations greater than or equal to 10 ppm, and mortality occurred at and above 25 ppm. The LC50, 96 h, to adult crayfish was about 33 ppm. The parent compound and its metabolites, MA (4-methylamino-m-tolyl N-methylcarbamate) and AM (4-amino-M-tolyl N-methylcarbamate), were detected in crayfish 96 h after exposure to various concentrations of aminocarb. The primary metabolite detected was MA which accounted for 75% to 95% of the body residue. The highest total residue (Aminocarb + MA + AM) was 40 ppm detected in crayfish exposed to 60 ppm of aminocarb for 96 h.     

Distribution and persistence of aminocarb in stream water, sediment and fish after application of three Matacil formulations

Three aminocarb formulations, water-based Matacil 180F, oil-based Matacil 180F and oil-based Matacil 180D, were applied separately to the surface of a stream at 2.40 g a.i./min for 5 minutes. The highest aminocarb concentration in the top 1 cm of water was found 5 min after application of oil-based 180F (5000 ppb). The next highest concentration was 1862 ppb found in the top 1 cm layer of water 3 min after application of oil-based 180D, followed by 1306 ppb found 5 min after application of water-based 180F. Residue accumulations were first observed in sediment at 3 min and in fish tissues 0.5 h after application. The levels of accumulation were in the order of water-based 180F greater than oil-based 180F greater than oil-based 180-D. After the application of water-based 180F, the highest concentrations were 20.2 ppb in sediment after 5 min and 127 ppb in fish tissues after 3 h. Residues disappeared rapidly from all three matrices and could not be detected after a period of 3-25 h.     

Biochemical effects of clomazone herbicide on piava (Leporinus obtusidens)

This study aims to verify the effects of the clomazone concentration used in rice fields on acetylcholinesterase (AChE), thiobarbituric acid reactive substances (TBARS), protein carbonyl and catalase activity in tissues of piava (Leporinus obtusidens). LC(50)-96h was 5.0 mg L(-1) and the fish were exposed to 1/10 of LC(50)-96 h: 0.5 mg L(-1) of clomazone for 96 and 192h. The same parameters were also assayed after a recovery period of 192 h in clean water. AChE activity was reduced only in the brain and heart of fish exposed for 96 h. AChE activity was decreased in the brain, muscle and heart tissues after 192 h of exposure. After 192 h of recovery period, AChE activity remained diminished in brain and muscle and showed a decrease in eye. However, after 192 h of recovery, AChE activity in heart was recovered. Fish showed increased TBARS levels in brain at all experimental periods. TBARS levels decreased in liver and muscle tissues after 192 h of exposure. The increase in muscle TBARS persisted in fish transferred to clean water. Protein carbonyl in the liver was increased in all periods studied including the recovery period. Catalase activity was reduced during all periods. The present study demonstrates the occurrence of disorders in AChE, TBARS, protein carbonyl and catalase activity in piava. The results also show changes in fish after exposure to an environmentally relevant concentration of clomazone. Most effects observed persisted after the recovery period. Thus, these parameters may be used to monitor clomazone toxicity in fish.     

Organophosphorus pollutants (OPP) in aquatic environment at Damietta Governorate, Egypt: implications for monitoring and biomarker responses

The study was carried out from spring 1999 to spring 2001 to monitor the residue levels of organophosphorus pollutants (OPP) in aquatic environment of the drainage canal surrounding a pesticide factory at Damietta Governorate. Water, sediment, and fish samples were collected at six different seasonal periods. OPPs were analyzed by GLC and confirmed using GC-MS. Chlorpyrifos, chlorpyrifos-methyl, malathion, diazinon, pirimiphos-methyl and profenofos were detected in most samples. Chlorpyrifos was dominant in all water and sediment samples. It was ranged from 24.5 to 303.8 and 0.9 to 303.8 ppb in water and sediment samples, respectively. Diazinon level was slightly similar to chlorpyrifos in fish samples. Data based on the grand total concentration of OPP showed that the most polluted samples were collected either at spring 1999 or autumn 2000. They were 675.5 and 303.8 ppb in water samples and 43.0 and 52.2 ppb in fish collected at spring 1999 and autumn 2000, respectively. The obtained results are in parallel to that found in case of cholinesterase activity where the activity of both acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) was declined at these seasonal period. The activity levels of AChE and BuChE were found to be 77.18% and 59.67% of control at spring 1999 and 78.62% and 85.80% of control, at autumn 2000, respectively. Thus, AChE and BuChE could be used as biomarkers for tracing and biomonitoring OPP pollution.     

Sublethal effects of monocrotophos on locomotor behavior and gill architecture of the mosquito fish, Gambusia affinis

Subacute studies of monocrotophos [Dimethyl (E)-1-methyl-2-(methyl-carbamoyl) vinyl phosphate] on mosquito fish, Gambusia affinis, were carried out in vivo for 24 days to assess the locomotor behavior, structural integrity of gill, and targeted enzyme acetylcholinesterase (AChE, EC: 3.1.1.7) interactions. Monocrotophos (MCP) can be rated as moderately toxic to G. affinis, with a median lethal concentration (LC(50)) of 20.49 +/- 2.45 mgL(-1). The fish exposed to sublethal concentration of LC(10) (7.74 mgL(-1)) were under stress and altered their locomotor behavior, such as distance traveled per unit time (m min(-1)) and swimming speed (cm sec(-1)) with respect to the length of exposure. Inhibition in the activity of brain AChE and deformities in the primary and secondary lamellae of gill may have resulted in failure of exchange of gases. The maximum inhibition of 95% of AChE activity was observed on days 20 and 24.Morphological aberrations in the gills were also studied during exposure to the sublethal concentration of monocrotophos for a period ranging from 8 to 24 days. The extent of damage in gill was dependent on the duration of exposure. The findings revealed that inhibition in brain AChE activity and structural alteration in gill were responsible for altering the locomotor behavior of exposed fish.     

Thiobencarb toxicity and plasma AChE inhibition in the European eel

The acute toxicity of the herbicide thiobencarb (S-4-chlorobenzyl diethylthiocarbamate) was determined for the European eel (Anguilla anguilla). The 24, 48, 72 and 96 hours median lethal concentrations (LC50) were 25.7, 21.7, 17.0 and 13.2 mg/L, respectively. Fish were also exposed to a sublethal thiobencarb concentration (1/60 LC50-96 hr = 0.22 mg/L) during 96 hours in a flow-through system and then an elimination period of 192 hours in clean water was allowed. Eels were removed and blood samples taken out at each exposure time and recovery period in order to evaluate AChE activity. Thiobencarb induced significant inhibitory effects on plasma AChE activity of A. anguilla from the first contact time with the toxicant. This inhibition (under 50% activity) was maintained during the entire exposure period (96 hours) and even those animals transferred to clean water showed plasma AChE activity different from the controls. Differences between total and specific AChE activity were detected during the exposure period. Total AChE activity in the plasma from animals transferred to a medium free of toxicant recovered its normal value while specific AChE activity remained depressed (< 50%) until five days later.     

Simultaneous determination of fenitrothion and aminocarb in blueberry foliage and fruits: Application to the analysis of residues in field samples

Abstract

Samples of blueberry foliage and fruits were collected from spray blocks in Ontario after aerial application of fenitrothion and aminocarb at dosage rates of 210 g active ingredient (AI)/ha and 70 g AI/ha respectively. Residues were extracted from the samples by homogenizing with ethyl acetate, cleaned up by microcolumn chromatography using alumina as adsorbent, and analyzed by GLC‐AFID with a glass column packed with 1.5% OV‐17 and 1.95% OV‐210 on 80–100 mesh Chromosorb W‐HP. Average recoveries for fenitrothion and aminocarb from foliage at three fortification levels (1.0, 0.10 and 0.01 ppm) were respectively 99 and 96%. The corresponding values for the fruits were 99 and 95%. Foliage samples collected 1 h post‐spray contained on average 1.13 ppm of fe‐nitrothion and 1.14 ppm of aminocarb. However, residue levels reached below the detection limit (<0.01 ppm) in foliage collected 15 d after treatment. In addition, the fruit samples collected after 15 d post‐spray contained extremely low levels (0.03 ppm for fenitrothion and 0.02 ppm for aminocarb) of residues, and were barely above the detection limit.     

Sublethal Effects of Monocrotophos on Locomotor Behavior and Gill Architecture of the Mosquito Fish,Gambusia affinis

Subacute studies of monocrotophos [Dimethyl (E)-1-methyl-2-(methyl-carbamoyl) vinyl phosphate] on mosquito fish, Gambusia affinis, were carried out in vivo for 24 days to assess the locomotor behavior, structural integrity of gill, and targeted enzyme acetylcholinesterase (AChE, EC: 3.1.1.7) interactions. Monocrotophos (MCP) can be rated as moderately toxic to G. affinis, with a median lethal concentration (LC₅₀) of 20.49 ± 2.45 mgL^?1. The fish exposed to sublethal concentration of LC₁₀ (7.74 mgL^?1) were under stress and altered their locomotor behavior, such as distance traveled per unit time (m min^?1) and swimming speed (cm sec^?1) with respect to the length of exposure. Inhibition in the activity of brain AChE and deformities in the primary and secondary lamellae of gill may have resulted in failure of exchange of gases. The maximum inhibition of 95% of AChE activity was observed on days 20 and 24.

Morphological aberrations in the gills were also studied during exposure to the sublethal concentration of monocrotophos for a period ranging from 8 to 24 days. The extent of damage in gill was dependent on the duration of exposure. The findings revealed that inhibition in brain AChE activity and structural alteration in gill were responsible for altering the locomotor behavior of exposed fish.     

Evaluation of the susceptibility of the cultured shrimp Litopenaeus vannamei to vibriosis when orally exposed to the insecticide methyl parathion

The causes of disease in cultured shrimp are difficult to ascertain but there is evidence that disease is correlated with environmental factors. Crustaceans are particularly sensitive to insecticides due to their close phylogenetic relationship with insects. The objective of this study was to investigate whether there was an increased susceptibility of the shrimp Litopenaeus vannamei to Vibrio parahaemolyticus, when exposed to methyl parathion. The outline of the study was the following: An LC50 96 h was determined to methyl parathion orally offered to juvenile shrimp. Further experiments were carried out in order to determine a concentration that affected the shrimp (verified by measuring the acethylcholinesterase activity) while producing minimal mortalities. This sublethal concentration was used in a susceptibility experiment where methyl parathion was offered to shrimp which were later injected with V. parahaemolyticus in a dose expected to kill less than 15%. Probit analysis estimated a 96 h median lethal concentration (LC50) of 1.56 microg g(-1). Mortality and AChE activity showed a concentration-response relationship in the exposure treatments. The median inhibitory concentration (IC50) determined was 0.029 microg g(-1). These results suggested that a concentration of 0.1 microg g(-1) was appropriate for the bacteria-pesticide interaction test as it was able to elicit 11.1% mortality after 10 days of exposure, while producing an AChE inhibition of 57.12%. Cumulative mortalities were significantly increased (P<0.01) in the treatment that combined exposure to methyl parathion and V. parahaemolyticus (35.19%) in comparison with methyl parathion or V. parahaemolyticus alone (9.26% and 7.41%, respectively).     

Effect of clomazone herbicide on biochemical and histological aspects of silver catfish (Rhamdia quelen) and recovery pattern

The effects of the herbicide, clomazone, on acetylcholinesterase (AChE), catalase and TBARS formation in teleost fish (Rhamdia quelen) were studied. The fish were exposed to 0.5 or 1.0 mg L⁻¹ of clomazone for 12, 24, 48, 96 and 192 h. After 192 h of exposure period, fish were transferred to clean water and kept in the same for 192 h to study the recovery response. Same parameters as that of exposure period were assayed after 96 and 192 h of recovery period. Specific AChE activity was reduced in the brain and muscle after treatments, reaching a maximum inhibition of 47% in the brain and 45% in the muscle after 12 h of exposure. Fish exposed to clomazone increased TBARS production in the liver for all exposure periods. The brain presented elevated TBARS levels after 12, 24 and 48 h, but after 96 and 192 h, these levels decreased. The decrease of TBARS levels persisted in brain tissue after 96 h of recovery and returned to the control value after 192 h in clean water. Catalase activity was reduced for all periods of exposure. Histological analysis showed vacuolation in the liver after herbicide exposure. Some of the alterations observed were completely restored after recovery period.     

Avoidance by olfaction in a fish, medaka (Oryzias latipes), to aquatic contaminants

Behavioral tests using a fish, medaka (Oryzias latipes), from which the olfactory organs were resected confirmed that fish avoid aquatic contaminants such as surfactants and pesticides mainly by olfaction. Control medaka (non-resected) exhibited significant avoidance to 10, 20 and 30 microg liter(-1) of LAS (sodium linear laurylbenzene sulfonate), 90 and 100 microg liter(-1) of fenitrothion (dimethyl 4-nitro-m-toryl phosphorothionate), and 500 microg liter(-1) of POE-ether (polyoxyethylene lauryl ether). By contrast, medaka with bilateral nose resections (BNRM) exhibited no avoidance to 10-50 microg liter(-1) of LAS, 90 and 100 microg liter(-1) of fenitrothion, and 500 microg liter(-1) of POE-ether. Although medaka with unilateral nose resection (UNRM) avoided some concentrations of these toxicants, the UNRM exhibited no avoidance to 10 and 40 microg liter(-1) of LAS and 90 microg liter(-1) of fenitrothion. That is, the avoidance response of the UNRM was less distinct than that of the controls. This difference may have been caused by the deterioration of detecting ability and the paralyzation or adaptation of olfaction.     

Residues of chlorpyrifos-methyl in balsam fir foliage, forest litter, soil, stream water, sediment and fish tissue after double aerial applications of Reldan

Chlorpyrifos-methyl was applied twice at 70 g A.I./ha by means of a fixed-wing aircraft to a mixed coniferous forest near Allardville, New Brunswick. Residue in balsam fir foliage was highest (1 ppm wet wt) 1 hr after spraying and rapidly declined to about 30% within 1 day, but persisted at a very low level (0.03 ppm wet wt) for 125 days. Current year's foliage contained a higher level of residue than old foliage. Chlorpyrifos-methyl persisted longer in forest litter than in soil. After 125 days, trace amounts (less than 6 ppb wet wt) were still found in litter but were not detected in soil. In stream water the residue dissipated very rapidly; more than 90% disappeared 3 hours after the second application and were not detected after 4 days. Low-level residue (less than 0.1 ppm wet wt) was present in the sediment and persisted for 10 days. Although brook trout and slimy sculpin captured in the stream within 3 days of the second application contained residues (less than 0.05 ppm fresh wt) none were detected in any fish captured, 9 and 47 days later.     

Impact of chemical exposure on the fish Pomatoschistus microps Krøyer (1838) in estuaries of the Portuguese Northwest coast

Juveniles of the estuarine fish Pomatoschistus microps were collected from autumn 2001 to summer 2002 in five stations along the Portuguese Northwest coast with different types and/or levels of environmental contamination: two reference sites with low levels of contamination (R1 and R2) and three differently impacted areas with higher levels of contamination. UI is located in an estuary under the influence of urban and industrial effluents, AA in a channel that receives intensive agriculture run-off and IE in a highly impacted industrial area. The activity of the enzymes acetylcholinesterase (AChE), lactate dehydrogenase (LDH), 7-ethoxyresorufin O-deethylase (EROD) and glutathione S-transferases (GST) were used as environmental biomarkers on P. microps. A significant seasonality effect on all the enzymatic activities was found, lower levels being registered in winter and spring on AChE, in autumn on LDH, and in winter on GST and EROD. The battery of biomarkers used was capable of discriminating sites with different types and/or levels of contamination, R1 and UI being the highest discriminated (91.7% and 66.7%, respectively). At R1 significantly lower levels of AChE and LDH were found, and EROD was significantly inducted at UI. Furthermore, IE presented higher levels of GST, and R2 and AA an inhibition of AChE in winter and spring. The results indicated that the battery of biomarkers used in this study seems to be a useful tool to distinguish between different types of environmental contamination in estuarine systems, and that P. microps is a suitable species to be used as bioindicator.     

Effects of methylisocyanate on soil microflora and the biochemical activity of soils

Methylisocyanate at 500, 1000 and 2500 microg ml(-1) h(-1) markedly affected the fungal propagules in treated soils. Immediately after exposure to gas, both bacteria and actinomycetes were appreciably reduced, although by the seventh day, their populations had gradually increased. Increased soil respiration was evident at 500 microg ml(-1) of methylisocyanate, whereas, inhibition of respiration occurred at 1000 and 2500 microg ml(-1) h(-1). Methylisocyanate adversely affected soil nitrification; inhibition increased with increasing concentration. Up to 2500 microg ml(-1) h(-1), it stimulated ammonification, but the NH(4)-N level gradually declined with increased incubation.     

Trout farm effluents: characterization and impact on the receiving streams

Effluents from three rainbow trout (Oncorhynchus mykiss) farms located in Northern Portugal were characterized and their impact on the receiving streams was evaluated. Mean fish productions in the studied fish farms were 15, 55 and 500 t of trout per year, respectively. The feeding water was abstracted from Fornelo, Inha and Coura Rivers, at flow rates ranging from 1.2 (15 t year(-1) fish farm) to 4.8 litre s(-1) per ton annual fish production (500 t year(-1) fish farm). As the water flows through the farms, net variations in the chemical characteristics were observed: a mean reduction in the dissolved oxygen (DO) concentration between 0.7 and 2.4 mg litre(-1); mean increases between 1.9 and 3.2 mg CaCO3 litre(-1) for total alkalinity, between 0.9 and 14 mg litre(-1) for BOD5, between 0.27 and 1.46 mg litre(-1) for ammonia nitrogen (NH4-N), between 0.060 and 0.579 mg litre(-1) for soluble phosphorus (PO4-P) and less than 16 mg litre(-1) for suspended solids; variations in the pH value and nitrate nitrogen concentration were not statistically significant (p<0.05). At the 500 t year(-1) fish farm it was also possible to detect net increases of total hardness (3.2 mg CaCO3 litre(-1)), electric conductivity (19 mS cm(-1)) and permanganate value (3.6 mgO2 litre(-1)). At the other farms net variations in these parameters were not significant. Net mass flow variations reported to the annual fish production are presented. The DO mass flow decreased, on average, between 255 and 549 g t(-1) of fish per day. The mean daily BOD5 increase ranged from 353 to 1510 g t(-1) of fish. The corresponding ranges for the other parameters were 105-157 g t(-1) for NH4-N, 24-62 g t(-1) for PO4-P, 348-1035 g CaCO3 t(-1) for total alkalinity and 224 x 10(6)-506 x 10(6) t(-1) for mesophilic bacteria. Daily net variations of suspended solids, total hardness, electric conductivity and permanganate value were below 1753 g t(-1), 342 g CaCO3 t(-1), 2081 mS cm t(-1) and 392 gO2 t(-1), respectively. Longitudinal concentration profiles for the most relevant parameters show the impact of the effluent discharges on the physico-chemical and bacteriological river water quality downstream from the trout farms. Analyzing the situations from a purely chemical point of view, the polluted stretches were 3, 5 and 12 km long downstream from the effluent discharges, respectively. The microbiological contamination extended over longer distances.     

Mortality response and LC50 values for juvenile and adult crayfish, Procambarus clarkii exposed to Thiodan (insecticide), Treflan, MSMA, Oust (herbicides) and Cutrine-Plus (algicide)

Toxicities of three herbicides (Treflan, MSMA and Oust), an algicide (Cutrine-plus) and an insecticide (Thiodan) to juvenile (3.0-3.4 cm) and adult (9.0-10.0 cm) crayfish, Procambarus clarkii, were determined by 96 h static bioassays. Freshly prepared 0.01% and 1.0% aqueous stock solutions of these pesticides were diluted to desired concentrations. Mortalities were recorded each day up to 96 h. Aged tap-water was used for preparing all test solutions. Per cent mortalities were analyzed for linear regression and LC(50) values were computed by probit analysis. LC(50) values for juvenile P.clarkii in the descending order of toxicity were: 24 ppb Thiodan, 13 ppm Treflan, 101 ppm MSMA, 461 ppm Cutrine-plus, 12,174 ppm Oust; and for adults these values were: 423 ppb Thiodan, 26 ppm Treflan, 1019 ppm MSMA and 2945 ppm Cutrine-plus. No LC(50) values for adult crayfish could be computed for Oust herbicide which caused no mortalities up to 60,000 ppm concentration. Comparing the overall toxicities of these pesticides, Thiodan was the most toxic to all crayfish; followed by Treflan, MSMA, Cutrine-plus and Oust. Oust was more than 1/2 million times less toxic than Thiodan to juvenile crayfish. Published LC(50) values indicate that freshwater crayfish are more tolerant than marine decapods, Crangon septemspinosa and Mysidopsis bahia, to Thiodan insecticide and Treflan herbicide.     

Ambient ozone and crop loss: establishing a cause-effect relationship

This paper provides the results of a retrospective mathematical analysis of the US NCLAN (National Crop Loss Assessment Network) open-top chamber data. Some 77% of the 73 crop harvests examined, showed no statistically significant yield differences between NF (non-filtered open-top chamber) and AA (chamberless, ambient air) treatments (no easily discernable chamber effects on yield). However, among these cases only seven acceptable examples showed statistically significant yield reductions in NF compared to the CF (charcoal filtered open-top chamber) treatment. An examination of the combined or cumulative hourly ambient O3 frequency distribution for cases with yield loss in NF compared to a similar match of cases without yield loss showed that the mean, median and the various percentiles were all higher (>/= 3 X) in the former in contrast to the latter scenario. The combined frequency distribution of hourly O3 concentrations for the cases with yield loss in NF were clearly separated from the corresponding distribution with no yield loss, at O3 concentrations > 49 ppb. Univariate linear regressions between various O3 exposure parameters and per cent yield losses in NF showed that the cumulative frequency of occurrence of O3 concentrations between 50 and 87 ppb was the best predictor (adjusted R2 = 0.712 and p = 0.011). This analysis also showed that the frequency distribution of hourly concentrations up to 87 ppb O3 represented a critical point, since the addition of the frequency distributions of > 87 ppb O3 did not improve the R2 values. In fact as the frequency of hourly O3 concentrations included in the regression approached 50-100 ppb, the R2 value decreased substantially and the p value increased inversely. Further, univariate linear regressions between the frequencies of occurrence of various O3 concentrations between 50 and 90 ppb and: (a) cases with no yield difference in NF and (b) cases with yield increase in NF compared to the CF treatment (positive effect) provided no meaningful statistical relationship (adjusted R2 = 0.000) in either category. These results support the basis that additional evaluation of the frequency of occurrence of hourly O3] concentrations between 50 and 87 ppb for cases with the yield reductions could provide a meaningful ambient O3 standard, objective or guideline for vegetation.     

Cholinesterase activity and effects of its inhibition by neurotoxic drugs in Dictyostelium discoideum

Recently, we found acetylcholinesterase (AChE) activity in the ciliate protozoan Paramecium primaurelia. As in the slime mould Dictyostelium discoideum the presence of a serine esterase was found with strong sequence identity to Torpedo AChE, we extended to D. discoideum the investigation on the characterization and possible functions of cholinesterases (ChEs). In amoeboid cells, histochemical, biochemical, and electrophoresis analyses evidenced both a ChE activity able to hydrolyze the substrate PrTChI, and AChE (E.C. 3.1.1.7.) activity similar to Electrophorus electricus AChE. Conversely, butyrylcholinesterase activity was nearly absent, according to our previous results on P. primaurelia. Moreover, the possibility to utilize D. discoideum in a bioassay for the pre-chemical screening both of moist environments and fresh waters, in relation to the occurrence of the neurotoxic organophosphate drugs, such as "basudin", inhibiting ChE activity, was investigated. Exposure to basudin inhibited propionylcholinesterase (PrChE) activity in a dose-dependent manner in the range 10(-1)-10(-7) M (60% at 10(-4) M), without any significant effect on AChE activity. PrChE activity was inhibited slightly by 10(-5) M eserine, and reduced significantly both by 10(-5) M iso-OMPA and BW284C51, classically used to discriminate the different ChE molecular forms. The effects on cell morphology, cell density, and differentiation were evaluated in cultures exposed to PrTChI 10(-5) M or basudin 10(-4) M for a three-day period. The PrTChI-exposed sample exhibited cell morphology, cell density, ability to aggregate, and to form fruiting bodies similar to the control; whereas, the basudin-exposed sample showed anomalies in cell morphology and lower cell density than the control, together with inability to aggregate.