Abstract:
The Incomati River Estuary is prone to pollutants from anthropogenic activities such as agricultural and industrial activities in the upper catchments. The main aim of the study was to use free-living nematodes as pollution indicators in the Incomati River Estuary. The main objectives were to determine the relationship between nematodes assemblage and environmental variables, and to identify environmental factors that play a role in nematodes community structuring. Lastly, it was to identify free-living nematode genera that can be used as pollution indicators in the Incomati River Estuary. Four sites were selected following the salinity gradient of the Incomati River Estuary. Site E1 with a salinity range of 0-3NST (Oligohaline Zone), E2 with a salinity 3-5NST (Euhaline Zone), E3 with a salinity 6-18NST (Mesohaline Zone), and E4 with 20-27 NST (Polyhaline Zone) were selected and monitored bi-monthly from June 2017 to April 2018. Two sediments samples were collected per site during neap tide using a handheld perplex corer which was 1m long with a 3.6 diameter and 10cm penetration height. Plastic bottles with a height of 13cm and a diameter of 7cm were used to store the sediment samples. One of the two sediment samples was used for free-living nematodes, and the other bottle was used for the analysis of environmental factors. All environmental factors were analysed at Labserve Laboratory, Mbombela Town, Mpumalanga Province. Sediment particle size and organic matter analyses were done following the procedure set by Parker (1983) and Buchanan (1971) respectively. Metal analysis was done following the procedure used by Gyedu-Ababio et al.1999. Nutrients were done using different methods. For nitrates (NO3) analysis, a copper cadmium method by Bate and Heelas, 1975 was used, while a method designed by Strickland and Parson, 1972 was used to analyse orthophosphate. A mixed acid digestion procedure of Oles and Dean 1965 was followed for total phosphate. A method by Lorenzen and Jeffrey, 1980 was used for the analysis of chlorophyll-a. Heterotrophic bacteria analysis were done following a procedure by (Atlas, 1997). Nematodes were extracted using a method by Furstenberg et al.1978, with sucrose as a separating agent. Nematodes were counted following a procedure by Giere, 1993. Nematodes feeding types were investigated using Wieser, 1953 procedure. Different statistical packages including PRIMER version 6 were used to analyse the data. A Bray-Curtis Cluster analysis indicated a similarity between sites E1 (Oligohaline Zone) and E2 (Euhaline Zone), and between site E3 (Mesohaline Zone) and E4 (Polyhaline Zone) which was attributed to similar sediment particle sizes distribution within the sites. There was no significant difference (p>0.05) of sediments particle size between the sites. The highest concentration of metals was found at site E2 which was situated in the Euhaline Zone, whilst the second highest concentration was found at site E1 which was situated in the Oligohaline Zone. A PERMANOVA analysis indicated that there was a significant difference (p<0.05) of Metal concentration between sites sampled. The PCA analyses indicated that there was a positive correlation between Metals and Sediment Particle Size such as Granules. It was observed that sediment particle size and organic matter influenced the distribution of metals in the Estuary. The highest concentration of chlorophyll-a and nitrates (NO3) were found at site E3 which was situated in the Mesohaline Zone, and the second highest was found at site E4 which was situated in the Polyhaline Zone. There was a positive correlation between Heterotrophic bacteria and environmental factors such as zinc, fine sand, very fine sand and mud. This indicated that certain metals and sediment particles size played a role in structuring food source for meiofauna, especially nematodes. The number of free-living nematodes were found to decrease towards site E1. This indicated that salinity influenced the diversity and density of free-living nematodes in the estuary. Site E2 had the lowest diversity and richness followed by site E1. The lower diversity, richness and Maturity Index at site E2 and E1 indicated that these sites were under stress. A Bray-Curtis Cluster analysis indicated that there was a spatio-temporal variation of diversity and density of free-living nematodes in the estuary. All four nematodes feeding types were found in the Estuary and feeding type 1B was the dominant feeding type at the sites, followed by feeding type 2A. The highest number of feeding type 1B (non-selective deposit feeders) was identified at site E2. The life strategy characterisation (colonizer- persisters) indicated that site E2 was dominated by colonizer and intermediate genera (c-p 2 and 3), which indicated that the site was under stress. The study found that genera such as Terschellingia and Theristus were pollution indicators because they were found in higher abundance at a site that was mostly polluted by metals, organic matter, and total phosphate. Further studies in other River Estuaries in South Africa and SADC should be undertaken to add to the findings of the current study.