Thesis (Ph.D)--Stellenbosch University, 2000. / ENGLISH ABSTRACT: Copper oxychloride is a fungicide that is extensively used in vineyards in the Western Cape to treat and prevent fungal diseases. It is however not clear what the effects are on soil organisms, which play an important role in soil fertility, in South African soils. There is paucity of data linking results obtained in the laboratory to effects observed in the field, which will only become useful if a clear relation can be demonstrated. The aims of this study were to: ~ Determine the effects of copper oxychloride on field populations of earthworms and simultaneously monitor lysosomal membrane stability, measured as neutral red retention time (NRRT). ~ Validate experimental field studies by doing inventories of earthworm populations in long-term sprayed vineyards. ~ Determine the LC50 of copper oxychloride and simultaneously measuring NRRT, and linking them to the experimental field studies. ~ Conduct bioassays, burrowing activity- and soil-avoidance experiments to investigate their relations to earthworm population responses in the experimental field studies. Earthworms were sampled by hand-sorting in the field tests on treated and untreated field plots in the Western- (October 1998 - July 1999) and Northern Cape (April 1998 - October 1999). Soil samples and worms were analysed for copper contents and coelomocytes of live earthworms were used to conduct the neutral red retention assays. Acute toxicity tests were conducted over a period of 28 days during which the earthworms (Eisenia fetiday were exposed to different concentrations of copper oxychloride. Change in biomass and mortality were measured as endpoints, as well as NRRT. Bioassays, burrowing activity and soil-avoidance were conducted by exposing Aporrectodea caliginosa to grassland- and vineyard soil as well as grassland soil spiked with 60 J.1g.g-1copper in the form of copper oxychloride. Growth and mortality were recorded in the bioassays as well as copper concentrations In earthworm body tissues and substrates used over a period of28 days. Burrowing activity and soil-avoidance were determined by measuring the length of tunnels burrowed by A. caliginosa in soil profiles over a period of 4 days under different exposure regimes. Results from the field tests showed that spraying of copper oxychloride had a negative effect on earthworm populations at the prescribed application rates. NRR T in earthworms from the exposure plots was significantly (p<0.05) lower after just one spraying application. It was concluded that spraying copper oxychloride at prescribed application rates caused a decrease in field populations of earthworms and that NRRT was an early and reliable biomarker since it was indicative of later effects observed at the population level. Results obtained from the field inventory of earthworms in vineyards at Nietvoorbij, Robertson end Worcester confirmed data from the two field studies. The calculated LC50 of 882.78 I1g.g-1 for copper oxychloride and 519.40 I1g.g-1 for copper was ecologically relevant if a safety factor of 10 was applied. NRRT which manifested earlier than effects on biomass change in the acute toxicity tests, were significant when viewed against the background of responses of field populations of earthworms. From the bioassay experiments it was found that A. caliginosa exposed to copper oxychloride spiked soil had significantly (p<0.05) higher weight loss and mortality than those in grassland- and vineyard soil. This indicated that changes in biomass and mortality were indicative of population responses in the field and can be considered as ecologically relevant. Burrowing activity of A. caliginosa was significantly (p<0.05) lower in vineyard and copper oxychloride spiked soil than in grassland soil. Similarly in the soil avoidance experiments it was found that A. caliginosa avoided vineyard- and copper oxychloride contaminated soil. It is therefore concluded that burrowing activity and soil avoidance were ecologically relevant endpoints since they corresponded with population responses in the field. The study thus revealed that the long-term usage of copper oxychloride could have negative effects on earthworm populations. The spraying of copper oxychloride can have important implications on the sustainable use of agricultural soils since earthworms and other soil organisms play such an important role in soil fertility. The use of biomarkers and other ecotoxicological indicators can provide an early warning that soil organisms are under environmental stress. / AFRIKAANSE OPSOMMING: Die fungisied koperoksichloried word wyd gebruik in die Wes-Kaap om swamsiektes in wingerde te beheer en te voorkom. Dit is egter nie bekend wat die effek daarvan op Suid Afrikaanse grondbiota, wat 'n belangrike rol speel in grondvrugbaarheid, is nie. Daar is ook 'n tekort aan inligting wat die resultate van laboratoriumondersoeke in verband bring met veldstudies. Die doelstellings van die studie was om: ~ Die effek van koperoksichloried op erdwurmpopulasies in die veld te ondersoek en terselfdertyd membraanstabiliteit, as moontlike biomerker, gemeet as neutraal rooi retensietye (NRRT), te monitor. ~ Die geldigheid van eksperimentele veldstudies te toets deur ook grondanalises te doen in wingerde wat oor langtermyn met koperoksichloried bespuit is. ~ Die LC50 van koperoksichloried vir erdwurms te bepaal en terselfdertyd NRR T te meet asook om dié gegewens in verband te bring met die resultate van seisoenale veldstudies oor die uitwerking op erdwurmpopulasies. ~ Bio-evaluerings ("bioassays"), tonnelaktiwiteit- en vermydingseksperimente te onderneem en die verband tussen die toksiteitstoetse en populasieresponse, soos waargeneem in die veld, te ondersoek. Erdwurms is versamel deur handsortering tydens die veldtoetse in die Wes- (Oktober 1998 - Julie 1999) en Noord-Kaap (April 1998 - Oktober 1999) op kontrole en bespuite persele. Grondmonsters en erdwurms is spektrofotometries geanaliseer om koperinhoude te bepaal. Die selomosiete van lewende wurms is gebruik om NRR T te bepaal. Akute toksisiteitstoetse is uitgevoer oor 'n tydperk van 28 dae waartydens Eisenia fetida blootgestel is aan verskillende koperoksichloried konsentrasies. Veranderinge in biomassa en mortaliteit is bepaal asook NRRT. Bioevaluerings ("bioassays"), tonnelaktiwiteit- en vermydingseksperimente IS uitgevoer deur Aporrectodea caliginosa bloot te stel aan grasveld- en wingerdgrond asook grasveldgrond wat met koperoksichloried gekontamineer is. Groei en mortalitiet is bepaal in die "bioassays" asook koperkonsentrasies in die grond en erdwurm liggaamsweefsels oor 'n tydperk van 28 dae. Tonnelaktiwiteit en grondvermyding is bepaal deur die lengte van tonnels wat deur A. caliginosa gegrawe is te meet oor 'n tydperk van vier dae vir die verskillende blootgestelde groepe. Die resultate het aangedui dat koperoksichloriedbespuiting 'n negatiewe invloed het op erdwurmpopulasies teen die voorgeskrewe toedieningsprogram. NRRT in erdwurms van die blootstellingperseel, was beduidend (p<0.05) laer na 'n enkele bespuiting. Daar is verder bevind dat NRR T 'n betroubare en vroeë biomerker is, aangesien dit 'n aanduiding gegee het van latere effekte wat op populasievlak na vore getree het. Veldopnames in Nietvoorbij, Robertson en Worcester het die geldigheid van data verkry uit die veldstudies ondersteun. Die berekende LC50 van 882.78 ug.g" vir koperoksichloried en 519.40 ug.g" VIr koper was ekologies relevant indien 'n veiligheidsfaktor van 10 toegepas is. NRRT se ekologiese relevansie is bevestig deur dit te vergelyk met response wat in die veldtoetse waargeneem is. Deur bioevalueringseksperimente is bevind dat gewigsverlies en mortaliteit van A. caliginosa beduidend hoër was in koperoksichloried gekontamineerde grond as in die grasveld- (kontrole) en wingerdgronde. Veranderinge in biomassa en mortalitiet was aanduidend van populasieresponse soos waargeneem in die veldstudies en kan dus as ekologies relevante eindpunte beskou word. Tonnelaktiwiteit van A. caliginosa was beduidend (p<0.05) laer in wingerd- en koperoksichloried gekontamineerde grond as in grasveldgrond. Dieselfde is gevind in die grondvermydingstoetse waar A. caliginosa wingerd- en koperoksichloried gekontamineerde grond vermy het. Dit kan dus afgelei dat tonnelaktiwiteit en grondvermyding ook ekologies bruikbare eindpunte is aangesien dit verband hou met populasieresponse soos waargeneem in die veldstudies. Hierdie studie het getoon dat die herhaalde gebruik van koperoksichloried 'n nadelige invloed kan hê op erdwurmbevolkings. In die lig van die belangrike rol wat erdwurms en ander grondorganismes speel in grondvrugbaarheid kan die oormatige gebruik van hierdie fungisied ernstige implikasies inhou vir volhoubare benutting van landbougronde. Die gebruik van biomerkers en ander ekotoksikologiese eindpunte kan egter as vroeë waarskuwingsmetode dien dat die grondorganismes onder omgewingstres verkeer.

A bacterial challenge assay was developed utilizing the earthworm, Lumbricus terrestris, in order to assess potential immunotoxic effects from exposure to specific polychlorinated biphenyl congeners. Earthworms were inoculated with Aeromonous hydrophila, establishing a 10-day LD50. In vitro assays for effects of PCBs on phagocytosis agreed with mammalian studies, demonstrating potent suppression of phagocytosis by the non-coplanar PCB congener 138 and no suppression by the coplanar congener 126. However, when the effects of the two PCB congeners were evaluated for suppression of resistance to a whole animal infection challenge assay, coplanar PCB 126 decreased the ability of L. terrestris to withstand infection while non-coplanar PCB 138 did not.

Le mercure suscite beaucoup d'attention de par les conséquences sanitaires et environnementales qu'il engendre. Son impact est particulièrement important dans la région amazonienne et en particulier en Guyane française où les teneurs en mercure sont naturellement élevées dans les sols, et s'ajoutent celles liées aux activités d'orpaillage. Si l'impact du mercure dans les milieux aquatiques a été largement étudié, peu d'études ont évalué son impact dans les milieux terrestres, où il est préférentiellement stocké. Dans ce contexte, les interactions entre les vers de terre, organismes ingénieurs des sols, et le mercure ont été étudiées dans des sols tropicaux, avec comme objectifs : i) d'évaluer l'abondance et la diversité des vers de terre in situ en lien avec le mercure, ii) d'évaluer expérimentalement la résistance au mercure d'une espèce de ver tropical présente sur le site d'étude et iii) d'évaluer l'impact en microcosmes de l'activité des vers de terre sur la mobilité et la répartition du mercure dans les sols. Ces objectifs ont été appréhendés au travers de trois volets, avec comme matériel d'étude des sols naturels (oxisols et sols hydromorphes) et des vers de terre tropicaux provenant d'un site orpaillé, situé dans les environs de Cacao, en Guyane française. Dans le premier volet, un échantillonnage standardisé des vers de terre (TSBF), a permis de proposer des espèces putatives (ou MOTUs) à partir de méthodologies de délimitation basées sur le barcode ADN. Ces MOTUs ont ensuite été identifiés morphologiquement, parmi lesquels Pontoscolex corethrurus, une espèce pantropicale considérée comme pérégrine. De plus, les caractéristiques des sols et les teneurs en mercure ont été déterminés afin de tenter d'expliquer la distribution des vers sur le site étudié. P. corethrurus se retrouve exclusivement dans l'oxisol, qui contient les concentrations les plus élevées de mercure total. Le second volet de ce travail a été consacré à l'étude de l'impact du mercure sur l'espèce tropicale P. corethrurus, à travers une approche écotoxicologique où le vers de terre a été exposé à différentes concentrations de mercure dans un oxisol dopé au laboratoire (HgCl2). Les résultats obtenus ont confirmé la résistance de cette espèce à des concentrations de mercure allant jusqu'à 20 µg g-1 de sol et ont révélé sa forte capacité bioaccumulatrice. Les résultats précédemment décrits ont ouvert la voie à un troisième volet où deux études en microcosmes se sont attachées à étudier l'impact des vers de terre sur la mobilité et la disponibilité du mercure (a) dans des conditions proches de celles rencontrées en conditions naturelles (fond géochimique), et (b) en présence d'une forte concentration pouvant être rencontrée sur les sites orpaillés (20 μg Hg g-1 de sol). Les résultats obtenus ont montré que les vers de terre agissent sur la mobilité du mercure directement par modifications physico-chimiques des sols (augmentation du pH et COD) et indirectement par stimulation des communautés bactériennes totales du sol. En conclusion, notre étude a montré que (1) P. corethrurus est une espèce résistante au mercure de par sa présence sur notre site d'étude et les résultats du test écotoxicologique mené au laboratoire, ce qui confirme l'idée d'un modèle tropical pour ce type d'étude ; (2) la bioaccumulation du mercure mesurée chez les vers de terre est importante et pourrait ainsi être responsable de l'entrée du mercure dans la chaîne terrestre trophique ; et (3) les vers de terre jouent un rôle dans la mobilité du mercure, modifiant sa partition dans les sols et diminuant les conséquences environnementales liées à sa biodisponibilité / Mercury is of major concern because of health and environmental problems that they cause. This toxic pollutant is particularly prevalent in tropical Amazonian regions and in French Guiana where soils are naturally rich in mercury and past and present gold mining activities occurred. While many studies have shown the impact of mercury in aquatic environments, to our knowledge only a few have focused on terrestrial ecosystems, where mercury is predominantly found. This doctoral research examined the interactions between earthworms, which are soil engineers, and mercury in a tropical soil from French Guiana. The objectives of the research were to: i) evaluate in situ earthworm diversity and distribution in soils related to mercury concentrations, ii) evaluate experimentally the impact of mercury on a tropical earthworm species present in our study site, and iii) evaluate the impact of earthworms on mercury mobility and availability in different soils. In order to achieve these objectives, three studies were conducted using natural tropical soils (oxisols and hydromorphic soils) and tropical earthworms sampled in a small catchment basin located near Cacao, in French Guiana. In the first part, earthworms were collected in the field using the standard TSBF method, and species delimitation was done based on DNA barcoding. Putative species were then morphologically identified, among them Pontoscolex corethrurus, a pantropical and peregrine species. Furthermore, soil characteristics and mercury concentrations were determined, and a first attempt was made to explain earthworm distribution in the field. P. corethrurus was found only in the oxisol, which contained highest mercury concentrations in the field. In the second part, an ecotoxicological study was done using a natural tropical soil in order to assess the impact of mercury on the identified tropical species: P. corethrurus. The results showed that P. corethrurus can resist up to high mercury concentrations ; it was not affected until 20 µg Hg g-1 soil and showed high capacity of Hg bioaccumulation. These results were used for the third part of the thesis. In the last part, two studies were conducted in controlled conditions in order to assess the impact of earthworms on mercury mobility and availability in soils. For these studies, conditions near natural conditions and also a high mercury concentration were applied to the soil (20 μg Hg g-1 soil). Results showed the impact of earthworm on mercury mobility and availability through different mechanisms: directly, through soil modifications (increase of soil pH and DOC), and indirectly, through stimulation of the soil microbial population. Furthermore, results confirmed the high capacity of earthworms for mercury bioaccumulation. In conclusion, we showed that (1) P. corethrurus is resistant to mercury, due to its presence in our field contaminated site and the results of the toxicity test done in the laboratory, thus confirming the use of a tropical species for these studies ; (2) mercury bioaccumulation measured in earthworms is important and thus could be responsible for its entry and biomagnification in the terrestrial food chain ; and (3) earthworms affect mercury mobility modifying its fractioning in soils and decreasing its availability

Objective was to assess the toxicity of chromium (Cr) contaminated soil (CS) using the earthworm Lumbricus terrestris. Specific aims were to determine: (1) survival (LC50); .(2) immunotoxicity as indicated by lysozyme activity, coelomocyte counts, secretory (SR) and erythrocyte rosette (ER) formation, and phagocytosis; and (3) compare effects of CS exposure with those of Cr spiked artificial soil (AS) . CS Cr concentration was 8.78 mg/g with 98.2% being Cr^3+ and 1.8% being Cr^6+. Using 14 d AS protocol the LC50 was 6.49% CS: AS mixture. CS concentrations of 0.5, 1.0, 2.5 and 5.0% were sublethal, whereas 6.25, 12.5, 25, 50 and 100% CS were lethal. Sublethal exposure caused no immuno- modulation. Exposure to 50% CS: AS mixture for 5 d caused reduced SR and ER formation. Exposure to AS spiked with 0.27% Cr for 5 d resulted in immunomodulation equivalent to 50% CS: AS mixtures. Results indicated the CS to be acutely toxic.

No description available.

No description available.

There have been no landscape-scale studies on earthworm populations in Canada comparing vegetation types; previous studies on edge habitats have been conducted in agricultural systems. I examined the spatial variations of earthworm populations by measuring abundance based on regional municipality, vegetation type, and edge habitat. Earthworms were sampled throughout the season across a gradient of vegetation types including meadow, forest edge, and interior at a local-scale; and at the landscape level with vegetation types including meadow, deciduous forest, pine plantation and mixed forest. Regional effects were more significant than vegetation type likely due to a gradient of soil characteristics in southern Ontario; edges had intermediate earthworm abundance and a higher proportion of epigeic species. My research provides insight into the patterns of earthworm populations in southern Ontario and the possible effects of edge creation through landscape fragmentation. Field sampling of earthworm parasitoid cluster-flies (Calliphoridae: Pollenia) using synomones was also discussed.

There have been no landscape-scale studies on earthworm populations in Canada comparing vegetation types; previous studies on edge habitats have been conducted in agricultural systems. I examined the spatial variations of earthworm populations by measuring abundance based on regional municipality, vegetation type, and edge habitat. Earthworms were sampled throughout the season across a gradient of vegetation types including meadow, forest edge, and interior at a local-scale; and at the landscape level with vegetation types including meadow, deciduous forest, pine plantation and mixed forest. Regional effects were more significant than vegetation type likely due to a gradient of soil characteristics in southern Ontario; edges had intermediate earthworm abundance and a higher proportion of epigeic species. My research provides insight into the patterns of earthworm populations in southern Ontario and the possible effects of edge creation through landscape fragmentation. Field sampling of earthworm parasitoid cluster-flies (Calliphoridae: Pollenia) using synomones was also discussed.

The master work presents the research of macrofauna (Lumbricidae) quantity, depending on the spieces of the plants grown and the hydrotermic modes. Research object: the soil macrofauna in the light granulometre composition of soils (in Akemnė district, Smiltinė village). Research objective: by field research to evaluate quantity of macrofauna (Lumbricidae) in the nothern part of Lithuania in the light granulometre composition of soils, in the ecological production farm, depending on the spieces of the plants grown and the hydrotermic modes. Research methods: the quantity of earthworms was established by digging 0.5 x 0.5 m and 0.25 m depth monolyths of soil, gathering earthworms, calculating, weitghting and measuring them. The research established that the quantity, weight, length of the earthworms depended on the species of plants grown, forecrops and hydrotermic modes. The most advantageous crops were the kitchen-garden, the garden, the potatoes. The best forecrops were the kitchen-garden. During the years of research strong reverse correlation links between the quantity of earthworms and the hydrotermic modes were established.

The effect of land management including undisturbed native forest, native grassland, sugarcane (preharvest burnt or green cane harvested), exotic forest (gum, pine or wattle), orchard crops (banana, orange and avocado) and grazed kikuyu grass pastures on soil organic matter status, size of the microbial biomass and size and composition of the earthworm community was studied. The study locality was in the tropical, northern part of KwaZulu-Natal near the town of Eshowe and sites were on a number of sugar estates in the area. Concentrations of soil organic C were notably high under kikuyu pasture, native forest and banana and lowest under burnt cane. Among the land uses, values followed the order: kikuyu pasture ≥ native forest > banana > native grassland = orange ≥ trashed cane = gum ≥ pine ≥ avocado > wattle ≥ burnt cane. Soluble C was considerably higher under kikuyu pasture than other land uses. Soils under native forest and banana also had relatively high values while lowest values were recorded under burnt cane. Values for microbial biomass C showed broadly similar trends with land use to soluble C. Very high values for microbial biomass C (> 2000 mg kg⁻¹ ) and microbial quotient (> 4.5 %) were recorded under kikuyu pasture, native forest, banana and orange whilst lowest values for microbial biomass C ( 250 - 750 mg kg⁻¹ ) and microbial quotient (1- 2 %) were found for soils under avocado, trashed and burnt sugarcane. Earthworm numbers followed the order: kikuyu pasture > native forest > banana > orange > wattle = pine = gum = trashed sugarcane ≥ native grassland ⁻¹ avocado > burnt sugarcane. Values for earthworm numbers and biomass were closely correlated. Earthworm numbers, microbial biomass C and soluble C were closely correlated with each other but none were significantly correlated with soil organic C content. Earthworm numbers were also positively correlated with soil pH and exchangeable Ca content. A total of 11 species of earthworm were collected from the sample sites. Over 80 % of the individuals collected were accidentally-introduced exotic species which originated from India, South America and West Africa. Most land uses supported between 5 and 7 species. Wattle forest and sugarcane, however, had only 2 or 3 species. Juveniles dominated the community under all land uses except kikuyu pasture and avocado where the majority of earthworms were adults. Epigeic species dominated the community under native forest and native grassland and this was also the case under avocado and gum. For the other land uses endogeic species predominated. The most numerous earthworm species present was Pontoscolex coreththrurus which was present under all the land uses. It is a peregrine, endogeic species originating from South America and is thought to have been introduced via India. The most common epigeic species was the Indian species Amynthas rodericensis which made up a particularly notable portion of the community under native and gum forests, avocado and banana. The third most numerous species was A. minimus, also from India, which is a polyhumic, endogeic species. It was particularly numerous under kikuyu pasture. In 8 out of 11 land uses, P. corethrurus, A. rodericensis and A. minimus coexisted together. Another polyhumic, endogeic species, Dichogastersaliens, which originates from West Africa, was present particularly under oranges, wattle and sugarcane. The only land use that contained mainly native species was native grassland where Tritogenia douglasi and Acanthodrilidae sp predominated. It was concluded that organic matter content, microbial biomass C, soluble C and the size and composition of earthworm communities in soils of the study area are greatly affected by land management practice. As is the case in most other parts of the world, the earthworm community under agricultural land management is dominated by accidentally introduced exotic species and these have also emigrated into soils under native vegetation; The role of these species in influencing soil chemical, physical and microbial properties, and thus soil fertility, deserves further studying. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2002.