© Benaki Phytopathological Institute
        
        
          Toxicity of essential oils against
        
        
          Tribolium confusum
        
        
          31
        
        
          adult mosaics) being counted as dead. Also
        
        
          the larvae that survived the tests were kept
        
        
          until pupation to examine possible delayed
        
        
          mortality or morphological deformations.
        
        
          Four to seven doses were tested for each
        
        
          essential oil and developmental stage com-
        
        
          bination. Each dose was repeated three or
        
        
          four times. Due to differential toxicity, dos-
        
        
          es of the compounds tested ranged from
        
        
          0.27 to 165 μl/l of air depending on the es-
        
        
          sential oil, insect developmental stage, age
        
        
          and sex (based on preliminary tests). All ex-
        
        
          periments were carried out in an incubator
        
        
          at 26
        
        
          ±
        
        
          1°C, 65
        
        
          ±
        
        
          5% r.h. and 12 h light/12 h
        
        
          dark photoperiod.
        
        
          
            Statistical analysis
          
        
        
          Data obtained from each dose-response
        
        
          bioassay were subjected to probit analy-
        
        
          sis and LC
        
        
          50
        
        
          values and 95% confidence lim-
        
        
          its were estimated. The comparisons among
        
        
          LC
        
        
          50
        
        
          values were based on the overlap of
        
        
          confidence limits (Finney, 1971). All analyses
        
        
          were conducted using the statistical pack-
        
        
          age SPSS 14.0 (SPSS, 2004)
        
        
          
            Results
          
        
        
          
            Fumigant toxicity of essential oils
          
        
        
          The fumigant toxicity (LC
        
        
          50
        
        
          values) of the
        
        
          essential oils of
        
        
          L. hybrida,
        
        
          L. nobilis
        
        
          ,
        
        
          T. ori-
        
        
          entalis,
        
        
          C. sinensis
        
        
          ,
        
        
          C. limon
        
        
          and
        
        
          O. vulgare
        
        
          against immature and adult
        
        
          T. confusum
        
        
          is
        
        
          given in Tables 1, 2, 3, 4, 5 and 6, respective-
        
        
          ly. All but
        
        
          O. vulgare
        
        
          essential oils exhibited
        
        
          strong fumigant toxicity to all developmen-
        
        
          tal stages of
        
        
          T. confusum
        
        
          . LC
        
        
          50
        
        
          values ranged
        
        
          between 1.8 and 109 μl/l air depending on
        
        
          the essential oil and insect developmental
        
        
          stage, age and sex.
        
        
          The most susceptible stage of
        
        
          T. confu-
        
        
          sum
        
        
          to the vapours of
        
        
          L. hybrida
        
        
          essential oil
        
        
          was the 10-day-old larvae (LC
        
        
          50
        
        
          , 1.8 μl/l air),
        
        
          while the most tolerant was the 31-day-old
        
        
          larvae (LC
        
        
          50
        
        
          109.9 μl/l air) (Table 1). The esti-
        
        
          mated LC
        
        
          50
        
        
          values for female and male pu-
        
        
          pae were very close (37.3 and 38.7 μl/l air,
        
        
          respectively). An increase in the adults’ sus-
        
        
          ceptibility to the essential oil vapours with
        
        
          increasing age was recorded, with females
        
        
          being more tolerant than males, especially
        
        
          for 60-day-old adults, where the observed
        
        
          differences were statistically significant.
        
        
          The most susceptible stage to the va-
        
        
          pours of
        
        
          L. nobilis
        
        
          oil was that of the 10-day-
        
        
          old larvae, and the most tolerant that of the
        
        
          25-day-old larvae (Table 2). In the case of pu-
        
        
          pae, males were more susceptible than fe-
        
        
          males to the essential oil vapours. Also, adult
        
        
          susceptibility to the vapours decreased with
        
        
          increasing age, but no significant differenc-
        
        
          es between the sexes were observed.
        
        
          The10-day-old larvae were the most vul-
        
        
          nerable to the vapours of
        
        
          T. orientalis
        
        
          oil, fol-
        
        
          lowed by pupae and adults (Table 3). Nev-
        
        
          ertheless, the calculated LC
        
        
          50
        
        
          values for all
        
        
          stages (except those for the young larvae)
        
        
          could not provide a clear picture of the de-
        
        
          gree of susceptibility because in most cases
        
        
          the statistical analysis did not reveal signifi-
        
        
          cant differences.
        
        
          A similar case to the
        
        
          T. orientalis
        
        
          oil was
        
        
          recorded for
        
        
          C. sinensis
        
        
          and
        
        
          C. limon,
        
        
          with
        
        
          young larvae being more susceptible than
        
        
          older ones (Tables 4 and 5). A slight (but not
        
        
          statistically significant) decrease in suscepti-
        
        
          bility of adults to the citrus essential oil va-
        
        
          pours with increasing age was observed.
        
        
          The essential oil of
        
        
          O. vulgare
        
        
          showed a
        
        
          completely different effect from all the oth-
        
        
          er essential oils tested. In fact, with the ex-
        
        
          ception of 10-day-old larvae and pupae, its
        
        
          vapours did not provoke mortality even at
        
        
          doses up to 165 μl/l air (Table 6).
        
        
          
            Effect on surviving larvae
          
        
        
          The larvae that survived exposure to es-
        
        
          sential oils did not exhibit any noticeable
        
        
          delayed mortality or any kind of morpho-
        
        
          logical abnormalities (data not shown).
        
        
          
            Effect on pupae
          
        
        
          All the essential oils tested caused a
        
        
          greater or lesser proportion of deformations
        
        
          in
        
        
          T. confusum
        
        
          pupae. These morphological
        
        
          deformations were noted in both males and
        
        
          females, with pupae trapped in the pupari-
        
        
          um, individuals with an appearance interme-
        
        
          diate between pupae and adults (adultoids),