Factors Affecting Residual Dosages of Two Formulations of Bacillus thuringiensis subsp. israelensis Tested in the Same Stream During a 3-Year Experiment

Although many field trials have been conducted using Bacillus thuringiensis subsp. israelensis (Bti)-based formulations, most have been in rivers with different biotic and abiotic conditions thus rendering the evaluation of their performance very difficult. Recently, results of a threeyear experiment using a new field procedure brought new insight into the behavior and the performance (carry) of two liquid formulations of Bti, Teknar HP-D and Vectobac 1200L, tested in the same lotic environment and under similar abiotic and biotic conditions. Factors such as discharge, water temperature and the hyporheic zone were identified as elements affecting the downstream loss of activity of both products. However, to better understand the reduction of black fly mortality along a stream (measured by using gutters), data of residual dosages of both products (measured by laboratory assay with mosquito larvae) were compared with reduction of black fly mortality. Bti toxic activity was monitored from water samples taken at different distances downstream from an application point, and from probes driven into the hyporheic zone, to study the effects of abiotic factors on the loss of the toxic crystals. Results showed that the loss of dosage was exponential for both products but more crystals were recovered from Vectobac 1200L along the stream than from Teknar HP-D. However, the latter was more efficient, i.e. less toxins were needed to kill 50% of black fly larvae both in temperate (16°C) and warmer (19.5-22°C) water. Also, a rise in water temperature had a greater effect in the kill induced by Vectobac 1200L compared to Teknar HP-D. For the same residual dosages present at the stations, longer carries of toxin activity (higher mortalities) were obtained in warmer water. Finally, the hyporheic zone was identified as a major source of loss of activity of Bti products. Large stream discharges decreased the effect of the hyporheic zone and that was reflected in longer carry of the products.     

A New Field Procedure and Method of Analysis to Evaluate the Performance of Bacillus thuringiensis subsp. israelensis Liquid Formulations in Streams and Rivers

Many field tests have shown Bacillus thuringiensis subsp. israelensis ( Bti ) to be an effective simuliid larvicide. However, literature indicates that an effective evaluation and comparison of Bti -based formulations when tested in streams or rivers is difficult. Most field trials have been conducted in different rivers (different discharge, river profile, water temperature, suspended matter, larval species, etc.), thus rendering the evaluation of the performance of liquid formulations of Bti very arbitrary or even impossible. A new field procedure is proposed to evaluate the performance of liquid formulations of Bti in the same lotic environment and under similar abiotic and biotic conditions. The procedure, based on a system of gutters located on the bank of a stream, showed very good reproducibility of the mortality levels of the target pest(s) recorded at various distances (stations) along the stream over a two-year field trial, proving the efficacy of the system. The system allows either a single formulation or different formulations to be tested repeatedly in the same portion of a stream, thereby providing a more accurate evaluation of the performance of a Bti formulation or a much better comparison between different formulations. The use of the probit model (allowing comparison of slopes and intercepts) gives a reliable statistical value for the analysis of the results. Moreover, this system is not expensive and can be transferred easily to other streams or rivers.     

Influence of Stream Profile and Abiotic Factors on the Performance and Residual Dosages of Two Commercial Formulations of Bacillus thuringiensis subsp. israelensis During a 2-Year Experiment

Recently, results of a 3-year study showed the importance of abiotic factors such as water temperature, stream discharge and the hyporheic zone on the behaviour, performance and loss of the residual dosages (amount of the injected dosage left at each station) of two commercial liquid formulations of Bacillus thuringiensis subsp. israelensis ( Bti ), Teknar HP-D and Vectobac 1200L. Experiments were performed in the same stream and under environmental conditions that allowed comparisons of results between formulations. To improve our understanding of the influence of abiotic factors on the behaviour and the performance of these formulations, a 2-year experiment using the same products was conducted in another river characterized by a much different profile, higher discharges and colder temperatures. A field procedure based on a system of gutters located on the bank of the river already used in the previous 3-year experiment was utilized. Black fly larval mortalities were recorded along the river at different distances (stations) to evaluate the behaviour and the performance of the products while water samples taken at the stations were tested in the laboratory against mosquito neonate larvae to evaluate the residual dosages of Bti at each station. Results showed that higher residual dosages of the Vectobac 1200L were recovered compared to Teknar HP-D along the river. Although higher dosages were recovered, higher mortality was observed only for the Vectobac 1200 L in high discharge conditions. As seen before, the hyporheic zone (interstitial water between the streambed and groundwater) produced a major loss of the dosages in the first meters of the river. But because of the river profile, the hyporheic zone had a lesser effect on the loss of the dosages further in the river resulting in very long carries for both Teknar HP-D and Vectobac 1200L.     

Storage Stability of Two Liquid Formulations of Bacillus thuringiensis subsp. israelensis and Effect of Freezing Over Time

Over the last two decades, many tests have been performed in the field to investigate the behaviour (persistence, carry, loss of activity, etc.) of different Bacillus thuringiensis subsp. israelensis ( Bti ) formulations. Depending on the experimental protocols, a single container of a formulation could be used more than once over time and field samples containing Bti may have to be frozen to preserve them for bioassays to be performed later. Thus, it is necessary to know how long a formulation could keep its level of efficacy and also the effects of time on frozen samples. Our results showed that the efficacy of two commercial liquid formulations of Bti (Teknar HP-D and Vectobac 1200L) when tested against Aedes triseriatus behaved differently over time when kept at room temperature. Teknar HP-D remained stable for the first two years and its LC 50 increased by 20% the third year. For Vectobac 1200L, although its larvicidal activity was better than that of Teknar HP-D every year, there was an increase in LC 50 by 22% the second year and by another 20% the third year for a total loss of activity of 46% over the three-year study. The efficacy of suspensions made with both formulations was greatly affected by freezing and the loss of efficacy increased over time. About half of the efficacy of Teknar HP-D was lost after one week of freezing and stayed at that level for three months, while with Vectobac 1200L, no significant effect of freezing was seen after one month, when compared to fresh material. However, both products showed similar efficacy after three or six months of freezing. Overall, the LC 50 s of both products had increased by a factor of about 2.5 after six months of freezing.     

Effects of Bacillus thuringiensis var. israelensis on Target and Nontarget Organisms: a Review of Laboratory and Field Experiments

Since the discovery of Bacillus thuringiensis var. israelensis (Bti) in 1976, extensive literature has proved its efficacy to control mosquitoes and black flies, of which many species are known as important vectors of diseases or simply as pests of humans and animals. Since 1978, Bti has been used in many countries on all continents and numerous studies have been made on target mosquitoes and black flies, as well as nontarget organisms (NTO). This review analyses the results of 75 studies on these organisms covering approximately 125 families, 300 genera and 400 species. Different factors such as species, instar, feeding behaviour and environmental parameters (larval density, water temperature, suspended matter etc.) may drastically affect the efficacy of the Bti products. This is addressed in detail by reviewing the main factors affecting mosquitoes as well as black flies. The results of a wide range of laboratory and field experiments using different target and nontarget species, various preparations and formulations of Bti and different biotic or abiotic factors are present in the literature, making the data difficult to compare on a common basis. Our analysis shows that, under different application conditions, the effects of Bti on target and nontarget organisms may be hard to predict. Although Bti has been proclaimed to be relatively highly specific, some studies show that some NTO are affected either by single or repeated Bti treatments. Present use against black flies seems ecologically acceptable. High frequencies of application and/or overdosages against mosquitoes may result in some persistence of the toxin crystals and ultimately this may have adverse effects on the food web. A long-term study (published in 1998) in mosquito habitats has shown that intensive Bti treatments over three years did in fact produce an impact on the food web in wetlands. This raises questions, for the first time, on Bti environmental specificity. The importance of this impact is discussed and the alternatives for practical pest control are considered. Some modifications of Bti use against mosquitoes, guided by research, is probably the best of these alternatives.