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Several studies on concordance between macrophyte and macroinvertebrate communities were carried out for decades while any investigation on co-occurrence of single pair of taxa of these two groups was never addressed. Our main aim was to verify the existence of co-occurrence of single macrophyte–macroinvertebrate pair in a Mediterranean river of central Italy. We compared the co-occurrence frequencies of different plant and animal taxa with those expected by chance through the null model approach. Specifically, we proposed two main co-occurrence patterns of macrophyte and macroinvertebrate taxa in terms of aggregation (when groups co-occur more frequently than expected) and segregation (when groups co-occur less frequently than expected). This approach was supported by correlations and analysis of covariance in order to identify the most significant co-occurring pairs. In this study we collected a total of 123 taxa (50 macrophytes + 73 macroinvertebrates) in October 2008 and 102 (42+60) in June 2009. We obtained a total of 379 significantly correlating macrophyte-macroinvertebrate pairs. Amongst all these correlating pairs, 83.7% of them correlated in October session while 90.3% in June. All the correlating pairs showed a high relationships with human alterations in both the sampling sessions. Our results confirmed that direct correlations there are between macrophytes and macroinvertebrates, highlighting how these two biological groups co-occur mainly following the aggregative model in both sampling sessions. Therefore, it was possible to observe significant relationships between macrophyte and macroinvertebrate pairs along an entire river ecosystem. It allows us to demonstrate that there are some prospective for trying to increase the knowledge on macrophytes-macroinvertebrates co-occurrence and for suggesting to use only single pair of co-occurring taxa in the monitoring running water quality. Aware that more investigations ought to be performed for confirming our proposal, the findings of the present study may be suggested as a new monitoring tool since it represents an investigation approach easy in the application, feasible and rapid in the execution, and within the European Water Framework Directive freshwater management policies. In addition, a such tool does not need a detailed taxonomical expertise since its applicability is restricted only to the few taxa indicated as representatives of a certain water quality condition.