Insect Biochemistry and Molecular Biology 30 (2000) 915–925

www.elsevier.com/locate/ibmb

Mini Review
Sunlight-activated insecticides: historical background and
mechanisms of phototoxic activity
*
Thameur Ben Amor, Giulio Jori
Department of Biology, University of Padova, Via U. Bassi 58B, Padova 35121, Italy

Received 21 January 2000; received in revised form 12 March 2000; accepted 14 March 2000

Abstract

Several photosensitizing agents, which are activated by illumination with sunlight or artificial light sources, have been shown to
be accumulated in significant amounts by a variety of insects when they are administered in association with suitable baits. The
subsequent exposure of such insects to UV/visible light leads to a significant drop in survival. Of the photosensitizers tested so
far, xanthenes (e.g. phloxin B) and porphyrins (e.g. haematoporphyrin) appear to be endowed with the highest photoinsecticidal
activity. In particular, porphyrins absorb essentially all the UV/visible light wavelengths in the emission spectrum of the sun; hence
they are active at very low doses. Thus, 1 h irradiation of Ceratitis capitata, Bactrocera oleae (also known as Dacus oleae) or
Stomoxys calcitrans which ingested a few nanomoles of porphyrin per fly with light intensities of the order of 1000 µE s⫺1 m⫺2
causes about 100% death in laboratory tests. Present evidence suggests that such photosensitizers act on the membranes of the
midgut with consequent feeding inhibition, as well as on the neuromuscular sheath. No apparent onset of photoresistance has been
observed. The rapid photobleaching of xanthenes and porphyrins when illuminated by visible light, as well as the lack of significant
toxicity of such compounds in the dark, minimizes the risk of an important environmental impact of such photoinsecticidal agents.
 2000 Elsevier Science Ltd. All rights reserved.

1. Introduction 1996). In addition, furocoumarines, upon photoexcit-

The use of photochemical processes as a tool to con-
trol the population of several types of insects has been
repeatedly examined in both laboratory experiments
(Heitz, 1987; Rebeiz et al., 1991) and field studies
(Pimprikar et al., 1980a; Lenke et al., 1987). Most inves-
tigations have been performed by using photoactivatable
polycyclic aromatic dyes that absorb near-UV light
wavelengths, including thiophenes, furocoumarines and
quinones (Robinson, 1983; Cunat et al., 1999). The use
of xanthene derivatives such as eosin and its analogues
absorbing selected light intervals in the visible spectral
range has also been proposed (Yoho et al., 1971;
Fondren and Heitz, 1978a; Heitz, 1997a). All of these
dyes require the presence of molecular oxygen to express
their phototoxic action; hence the overall photoinsectici-
dal process appears to be of the photodynamic type (Jori,
* Corresponding author. Tel.:+39-049-8276333; fax: +39-049-
8276344.
E-mail address: jori@civ.bio.unipd.it (G. Jori).
ation, can generate various types of addition products
with DNA bases, which often results in genotoxic effects
(Jori, 1985).
More recently, Rebeiz and co-workers proposed the
use of porphyrins as photoinsecticides (Rebeiz et al.,
1987). In particular, these authors tested protoporphyrin
IX and its Zn(II) derivative, which appear to be
especially promising photoinsecticidal agents since these
compounds absorb essentially all the UV–visible wave-
lengths, that is to say these molecules can be efficiently
excited by natural sunlight. Along the same line, we
recently demonstrated that haematoporphyrin is an
efficient phototoxin to several insects (Ben Amor et al.,
1998a). Subsequently, we extended our investigations to
some meso-substituted porphyrins (Ben Amor et al.,
1998b) in order to identify possible relationships
between the chemical structure and the photoinsecticidal
activity of this class of compounds. Ceratitis capitata, a
Mediterranean fruit fly, was selected as an experi-
mental model.
The present contribution is aimed at describing the
historical background and the present state of the art for

0965-1748/00/$ - see front matter  2000 Elsevier Science Ltd. All rights reserved.
PII: S 0 9 6 5 - 1 7 4 8 ( 0 0 ) 0 0 0 7 2 - 2
916 T. Ben Amor, G. Jori / Insect Biochemistry and Molecular Biology 30 (2000) 915–925
the use of photoactivatable insecticides, as well as dis-
cussing the limitations, scope and potential of this novel
technique. The interest in this topic is enhanced by safety
and environmental considerations. The use of sunlight
as the promoter of the phototoxic activity is in line with
the growing trend to utilize and validate natural
resources. Moreover, a careful selection of the chemical
structure of the photosensitizing dye can modulate the
nature of the subcellular photodamaged sites, which is
quite important for optimizing the efficiency of the cyto-
cidal effects and minimizing the risk of selecting photor-
esistant insect species.
The photoinsecticides represent a possible alternative
to traditional chemical insecticides. The latter com-
pounds are known to cause several important problems,
including widespread toxicity to plants and animals and
in particular to humans, as well as the prolonged persist-
ence in the environment which may cause severe pol-
lution. As we will discuss in the present review, at least
some of these issues can be adequately addressed by the
use of photodynamic insecticides.
2. Historical background
The first scientific documentation that sunlight can be
toxic to biological systems was provided by Marcacci in
the late nineteenth century (Marcacci, 1888). This author
reported that the fermentation of plant alkaloids and
amphibian eggs becomes more important under
UV/visible light than in the dark. Shortly thereafter,
Raab (1900) observed that the presence of some exogen-
ously added visible light-absorbing compounds, such as
acridine orange, was necessary for sunlight to promote
the death of paramecia. Acridine orange and other dyes
with similar properties were defined as photosensitizers.
Raab’s observations were repeated with a variety of
multi- and unicellular organisms (Spikes, 1985). Finally,
Jodlbauer and Von Tappainer (1904) demonstrated that
the presence of oxygen is an essential requisite for pho-
tosensitization to occur. The combined effect of the three
elements, namely light, photosensitizer and oxygen, has
been termed photodynamic action (Blum, 1941).
While most organisms have developed specific protec-
tion or repair mechanisms to counteract the damaging
action of light, several attempts have been made to con-
trol the photoprocesses involving specific biological sys-
tems with the aim to obtain beneficial effects. Thus, pho-
totherapeutic techniques have been developed by taking
advantage of the property of some photosensitizing
agents to be accumulated in significant amounts by dis-
eased tissues, such as tumours, atheromas or psoriatic
plaques (Brown, 1997). At the same time, the photosen-
sitized inactivation of yeasts and bacteria is used for the
decontamination of microbially polluted waters by sun-
light (Merchat et al., 1996).
The possibility of using photoactivatable compounds
for controlling the pest population was first explored by
Barbieri (1928) who used a combination of xanthene
derivatives, including fluorescein, erythrosine and rose
bengal, against Anopheles and Aedes larvae. Barbieri
showed that light or photosensitizer alone had no detect-
able toxic effects on insects.
This research line underwent no further developments
until the early 1950s when Barbieri’s experiments on the
photosensitized killing of insects were repeated by
Schildmacher (1950), who concluded that the chemical
structure of the xanthene dyes exerts a profound influ-
ence on the degree of phototoxicity. Thus, the highest
photoinsecticidal activity is displayed by rose bengal fol-
lowed by eosin, erythrosine and fluorescein. The poten-
tial of xanthenes as promoters of lethal effects on insects
exposed to sunlight was explored in detail by Heitz and
co-workers (Heitz, 1987; Heitz, 1997a), who investi-
gated the role of various experimental parameters on the
photoinsecticidal efficacy of this class of compounds. Of
particular interest were the field studies based on several
combinations of bait and xanthene dyes (Carpenter et al.,
1981; Sakurai and Heitz, 1982). Xanthenes undergo
rapid photodegradation in aqueous media by analogy
with the well-known tendency to photobleach which is
typical of many polycyclic aromatics when exposed to
non-ionizing radiation (Spikes, 1992). This topic was the
subject of intense discussions at a recent congress of the
American Society of Photobiology (Heitz, 1997b).
At the same time, independent investigations perfor-
med by various laboratories demonstrated that near-UV-
or visible light-absorbing dyes belonging to different cat-
egories of organic compounds can develop a phototoxic
effect towards a variety of insects (Robinson, 1983). It
seems reasonable to propose that any photodynamically
active drug, which can be combined with a suitable bait
to facilitate its uptake by the target insect and avoid its
uptake by useful non-target insects, has the potential for
acting as an insecticide. In fact, UV/visible light can
penetrate to a depth of about 1 cm in most biological
tissues; as the extent of penetration is wavelength depen-
dent (Svaasand et al., 1990), it is possible to modulate
the volume of the photoinduced tissue damage by sel-
ecting a photosensitizing agent with specific absorption
properties. In this connection, porphyrin-based photoin-
secticides which have been recently developed (Rebeiz,
1992; Ben Amor et al., 1998a) are particularly interest-
ing since they exhibit a multiplicity of absorption bands
throughout the UV/visible spectrum; hence they can be
efficiently activated by either white light or selected
wavelength ranges depending on the desired extent of
photodamage.
 T. Ben Amor, G. Jori / Insect Biochemistry and Molecular Biology 30 (2000) 915–925
3. Mechanisms of photodynamic sensitization
Upon absorption of UV or visible photons, a photo-
sensitizer can be promoted to a variety of electronically
excited states. However, the efficiency of the photosensi-
tizing action is generally dependent on the photophysical
properties of the lowest excited triplet state (3Sens),
which is reached via intersystem crossing from the
initially formed excited singlet state (1Sens). The 3Sens
species is most often characterized by a lifetime in the
microsecond to millisecond range, hence it can play a
major role in diffusion-controlled processes (Carmichael
and Huh, 1986). Several deactivation pathways are poss-
ible for 3Sens; those which are of utmost importance
from a photobiological point of view can be schematized
as follows.
1. Electron transfer to or from a substrate with suitable
redox properties, e.g.
3
Sens⫹Sub→Sens·+⫹Sub·−
This pathway, defined as a type I mechanism (Jori and
Reddi, 1991), leads to the generation of radical inter-
mediates, which in turn can undergo further reactions
with other substrates, solvent molecules, or oxygen. The
latter process results in the formation of oxidized pro-
ducts. A particular case occurs when oxygen acts as an
electron acceptor:
3
Sens⫹O2→Sens·+⫹O2·−
The superoxide anion has a relatively low level of reac-
tivity. However, under certain experimental conditions,
it can be converted to very reactive and cytotoxic spec-
ies, such as the hydroxyl radical and hydrogen peroxide
(Bensasson et al., 1983).
2. Energy transfer to any substrate whose triplet state
energy lies at a lower level compared with the photosen-
sitizer triplet state; this pathway is defined as a type II
mechanism (Jori and Reddi, 1991):
3 or passive diffusion processes. Thus, cationic photosen-
Sens⫹Sub→Sens⫹3Sub
Most components of cells and tissues are not suitable
acceptors of electronic energy from 3Sens, since their
triplet states are too energetic. One notable exception is
represented by oxygen; this ubiquitous component of
biological systems can be readily promoted to its excited
singlet state, whose energy level lies at only 22.5 kcal
above the triplet ground state and which is endowed with
a high cytotoxicity:
3
Sens⫹3O2→Sens⫹1O2
The high reactivity of 1O2 is partly due to its long life-
time (3–4 µs in aqueous media, several tens of microse-
conds in lipid environments) which allows this species
to diffuse over relatively long distances before being
deactivated (Wasserman and Murray, 1989).
Both type I and type II photosensitization mechanisms
917
generate electrophilic species, hence the most photosen-
sitive targets are represented by electron-rich biomolec-
ules (Vilensky and Feitelson, 1999).
Table 1 shows that of the naturally occurring amino
acids only those which possess aromatic or sulphur-con-
taining side chains are readily photooxidized. Other rap-
idly attacked moieties include carbon–carbon double
bonds of unsaturated lipids and steroids, as well as the
heterocyclic ring of guanosine nucleotides. At a cellular
level the photosensitizing action is characterized by an
additional selectivity, since the overall photoprocess is
generally confined to the microenvironment of the pho-
tosensitizer owing to the tendency of the photogenerated
intermediates to react with a large variety of targets
(Moan et al., 1995). Thus, it appears essential to control
the subcellular distribution of the photosensitizing agent.
In this connection, a critical role is performed by the
chemical structure of the photosensitizer, and in parti-
cular by its degree of hydrophobicity (Jori and Reddi,
1993). This parameter is usually measured by the par-
tition coefficient between n-octanol and water. Thus,
moderately or highly lipophilic dyes (partition coef-
ficient ⬎8–10) become preferentially associated with the
cell membranes; at short incubation times the plasma
membrane represents the main binding site, while at
longer times significant photosensitizer concentrations
are recovered from other subcellular membranes includ-
ing the mitochondrial and lysosomal membranes, the
Golgi apparatus and the rough endoplasmic reticulum.
For in vivo administration, amphiphilic photosensitizers
(which are sufficiently water-soluble, yet are charac-
terized by the presence of a hydrophobic matrix facilitat-
ing the crossing of the lipid domains of cell membranes)
proved to be particularly useful. Hydrophilic photosensi-
tizers show a more complex pattern; although such com-
pounds, especially if electrically charged, undergo ionic
interactions with charged groups at the cell surface, the
possibility exists of their internalization by both active
sitizers are often localized in mitochondria, while anionic
dyes (e.g. carboxylated derivatives) are accumulated at
the lysosomal level (Spikes, 1994; Jori, 1996). Lastly,
some photosensitizers, such as furocoumarins, can reach
the cell nuclei and bind to the DNA bases (Armitage,
1998). The subcellular localization of a photosensitizer
determines the efficiency, as well as the mechanism of
photoinduced cell inactivation; in particular, it controls
the relative weight of apoptotic and random necrotic
pathways which are eventually responsible for cell death
(He et al., 1994)
4. Main classes of photoactivatable insecticides
Several photodynamic sensitizers have been shown to
act as efficient photoinsecticidal agents upon activation
918 T. Ben Amor, G. Jori / Insect Biochemistry and Molecular Biology 30 (2000) 915–925

Table 1
Main biological targets of photosensitized processes

Cell constituent Target Modified chromophore Main photoproducts

Protein Tryptophan Indole Hydroxyindoles, kynurenine

Tyrosine Phenol Quinones, melanin-type pigments
Histidine Imidazole Endoperoxide
Methionine Thioether Methionine sulphoxide
Cysteine Thiol Cystine, cysteic acid
Unsaturated lipids Oleic, linoleic, linolenic, Carbon–carbon double bond Allylic hydroperoxides,
arachidonic acids endoperoxides
Steroids Cholesterol Carbon–carbon double bond 5α- or 7α-hydroperoxide
Nucleic acids Guanosine Purine Ketone-type degradation products

with either natural or artificial UV or visible light wave-

lengths. A list of the main classes of photoinsecticides
is given in Table 2. Undoubtedly, xanthene dyes have
been most frequently and extensively studied largely due
to the systematic investigations carried out by Heitz and
co-workers (Heitz, 1987; Heitz, 1997a). Eventually,
phloxin B, a polyhalogenated fluorescein (spiro-
benzofuran-1(3H),(9H)-xanthen-3-one-2⬘,4⬘,5,7⬘-tetra-
bromo-4,5,6,7-tetrachloro-3⬘,6⬘-dihydroxy disodium
salt) has been developed for commercial use as a pestic-
ide (Dowell et al., 1997; Heitz, 1997a). While fluor-
escein can be considered as the parent compound of this
class of photosensitizers, several derivatives can be syn-
thetically prepared by the insertion of up to eight halogen
atoms into selected positions of the aromatic macrocycle
(Fig. 1A). All the xanthenes exhibit intense absorption
bands in the green region of the visible spectrum; the
maximal absorbance falls in the 480–550 nm interval,
while the exact position of the peak shifts to the red as
the number of the halogen substituents and their atomic
weight increase. These parameters also affect the photo- Fig. 1. Schematic chemical structure of (A) xanthene dyes and (B)
porphyrin dyes.
dynamic efficiency of xanthenes, since the presence of
heavy bromine or iodine atoms enhances the yield of

Table 2
Selected examples of photodynamic sensitizers which have been used as photoinsecticidal agents

Class Typical examples Targets References

Xanthenes Rose bengal House fly, Aedes larvae Carpenter et al. (1984)
Erythrosin B House fly, face fly Fairbrother et al. (1981)
Phloxin B Culex mosquito larvae Fondren et al. (1979)
Rhodamin 6G Fire ant, House fly Fondren and Heitz (1978b)
Heitz (1987)
Phenothiazines Methylene blue Yellow mealworms Lavialle and Dumortier (1978)
Cabbage butterfly
Furanocoumarins Xanthotoxin Black swallowtail larvae Cunat et al. (1999)
Angelicin Swallowtail butterfly Robinson (1983)
Thiophenes α-Terthienyl Aedes mosquito, Blackfly larvae Guillet et al. (2000)
Acridines Acridine red Acridine looper Robinson (1983)
Various Hypericin Fruit fly Armitage (1998)
Cercosporin Aedes mosquito larvae Bosca and Miranda (1998)
Benzopyrene Black fly larvae Cunat et al. (1999)
Polyacetylenes
 T. Ben Amor, G. Jori / Insect Biochemistry and Molecular Biology 30 (2000) 915–925
intersystem crossing to the reactive triplet state of the
dye (Jori and Reddi, 1991). Thus, other conditions being
the same, the greatest photosensitizing activity is dis-
played by tetraiodo xanthene derivatives, such as rose
bengal and erythrosin B. As shown in Fig. 2, both these
dyes are significantly more efficient than their tetra-
bromo analogue (eosin yellow) in photosensitizing the
killing of Musca domestica.
In general, xanthenes exert their phototoxic effects
through the generation of reactive oxygen species
(largely, singlet oxygen). However, one cannot rule out
the parallel occurrence of radical-involving processes
owing to the well-known photolability of covalent bonds
between halogens and aromatic rings (Spikes and Mack-
night, 1970). Xanthenes are typically localized in cell
membranes, so that at a molecular level xanthenes
mostly photosensitize the cross-linking of membrane
proteins and the formation of hydroperoxides from
unsaturated lipids, thereby markedly increasing the
osmotic fragility of cells (Pooler and Valenzeno, 1979).
Very similar considerations are found for acridines,
which efficiently absorb light wavelengths around 550
nm and largely act via the generation of activated oxygen
species (Rossi et al., 1981). One important difference
is represented by the tendency of acridines to yield a
heterogeneous subcellular distribution pattern, involving
both the partitioning in specific organelles such as lyso-
somes and the interaction with the phosphate groups in
double-stranded DNA (Briviba et al., 1997), which
enhances the possibility of photosensitized damage to
the genetic material.
Both furocoumarines and thiophenes preferentially
absorb near-UV light, which has a low penetration
power into most biological tissues (Anderson and Parr-
ish, 1992). Thus, these photosensitizing agents promote
the damage mainly at the level of superficial tissue lay-
Fig. 2. Effect of different xanthene dyes on the percent survival of
Musca domestica, upon exposure to sunlight after 12 h free access to
a bait with 0.125% photosensitizer concentration. Eosin yellow (×);
rose bengal (䊏); erythrosin B (쐌). Adapted from Fondren et al.
(1978, 1979).
919
ers. However, the photodamaged area can become quite
extensive since these compounds, once electronically
excited, can promote radical processes which signifi-
cantly amplify the initial damage through the induction
of chain reactions. Furocoumarines may also intercalate
among DNA bases with the formation of covalent pho-
toadducts and the consequent inhibition of cell repli-
cation (Armitage, 1998).
Lastly, phenothiazines (Boyle and Dolphin, 1996) and
hypericin (Bosca and Miranda, 1998) are characterized
by relatively intense absorption bands in the orange–red
spectral region. Both these classes of photosensitizers
therefore allow the direct damage of tissue compart-
ments located at depths of several millimetres below the
surface where light initially impinges. Moreover, several
constituents of these classes produce the highly reactive
singlet oxygen with a quantum yield greater than 0.6;
hence, the overall photodamage is most often confined
within the microenvironment of the photosensitizer. This
makes it important to control the biodistribution of such
photosensitizers as closely as possible.
5. Porphyric photoinsecticides
In recent years, increasing attention has been focused
on the photosensitizing properties of porphyrins (Fig.
1B) and their analogues, such as chlorins and phthalocy-
anines (Jori and Spikes, 1983). Porphyrins are natural
compounds or close derivatives of such compounds,
hence they are usually devoid of any appreciable intrin-
sic cytotoxicity in the absence of irradiation. Further-
more, porphyrins are characterized by specific features
which make them especially useful photosensitizers of
biological systems:
1. The ability to absorb essentially all the wavelengths
of the solar spectrum in the UV and visible range.
In particular, porphyrins exhibit an intense absorption
band (the Soret band) in the blue spectral region,
which represents the most intense component of the
sun’s emission around midday (Svaasand et al.,
1990). On the other hand, the red absorption bands
of porphyrins are useful at dawn and sunset, when
wavelengths greater than 600 nm represent an
important component of sunlight.
2. Several porphyrins yield long-lived triplet states with
a high quantum yield (⬎0.7) and therefore are quite
efficient photosensitizers. As a rule, the triplet state of
porphyrins is efficiently quenched by oxygen. Hence
porphyrins typically cause cell inactivation through
the generation of singlet oxygen (type II pathway),
even though radical transfer processes may also be
involved (Ricchelli et al., 1993). This circumstance
enhances the scope and potential of porphyrins as
photosensitizers, since they also express a high pho-
920 T. Ben Amor, G. Jori / Insect Biochemistry and Molecular Biology 30 (2000) 915–925
toactivity in biological systems which are charac-
terized by a low oxygen pressure.
3. The chemical structure of porphyrins can be modified
at different levels, including (i) the substituents pro-
truding from the peripheral positions of the pyrrole
rings or the meso-carbon atoms, (ii) the metal ions
possibly coordinated at the center of the tetrapyrrolic
macrocycle, and (iii) the ligands axial to the metal
ion. In this way, it is possible to modulate the phys-
ico-chemical properties of the porphyrin molecules
and control their partitioning among subcellular com-
partments.
4. Hydrophobic porphyrins are localized at the level of
the cell membranes including the plasma, mitochon-
drial and lysosomal membranes (Ricchelli and Jori,
1986). As a consequence, the genetic material is not
involved in the photoprocesses leading to cell death.
All the available evidence indicates that porphyrin
photosensitization of cells does not promote the onset
of mutagenic effects, thereby minimizing the risk of
selecting photoresistant cell clones (Bonnett and
Berenbaum, 1989).
5. The extraction and isolation of porphyrins from natu-
ral products, and their synthetic preparation (often by
modification of natural porphyrins), are relatively
simple procedures (Ricchelli et al., 1995). Hence, the
overall commercial cost of porphyrins can be fairly
low, of the order of 1–2 US$/g, which is a critical
factor for large-scale field utilization of porphyric
insecticides. It is to be underlined that the uptake of
nanomoles of porphyrin is sufficient to cause a rapid
mortality of several types of flies even under moder-
ate intensities of sunlight (Ben Amor et al., 2000).
6. Porphyrins undergo fast photobleaching in sunlight as
well as when exposed to artificial visible light sources
(Rotomskis et al., 1997). The photodegradation pro-
ducts do not appear to induce any appreciable toxic
or phototoxic effects in a variety of biological sys-
tems. Thus, the rapid disappearance of porphyrins
from the environment strongly reduces the risk of
widespread or persistent contamination.
7. Several porphyrins are presently used as photothera-
peutic agents; toxicological studies have shown that
these dyes induce important damage to humans only
upon uptake of at least 100 mg/kg body weight, that
is far greater than the amount which is required for
generating an extensive toxicity to insects (Jori and
Reddi, 1991).
Two approaches have been devised for defining the
scope and potential of porphyric insecticides. One
approach involves the administration of a large excess
(some hundred milligrams) of 5- amino-levulinic acid
(ALA), which is a metabolic precursor of heme, the
prosthetic groups of hemoproteins (Rebeiz et al., 1988;
Rebeiz et al., 1990b). The excess ALA through a feed-
back mechanism inhibits the final step of the biosyn-
thetic pathway, namely the ferrochelatase-catalysed
insertion of the Fe2+ ion into the tetrapyrrolic macrocy-
cle. This leads to the accumulation of significant
amounts of protoporphyrin IX, a well-known photodyn-
amic agent. As a consequence, the protoporphyrin-
loaded cells become photosensitive, especially at the
level of mitochondria which represent the normal site of
heme biosynthesis. In general, the photosensitivity
develops after a time interval of 3–4 h from the exposure
of the insect to ALA. The extent of the protoporphyrin
accumulation can be enhanced by the presence of iron
chelating agents such as desferroxamine or phenanthro-
line (Rebeiz et al., 1990a).
A second strategy is based on the direct administration
of a photodynamically active porphyrin in combination
with a bait. Typically, 1 h exposure of the insects to
haematoporphyrin concentrations of the order of a
microgram per ml of bait are sufficient to obtain a 90–
100% decrease in the survival of widely diffused and
extremely noxious flies, such as Ceratitis capitata (fruit
fly), Bactrocera (Dacus) oleae (olive fly) and Stomoxys
calcitrans (stable fly) (Ben Amor et al. 1998a, 2000).
The kinetics of post-irradiation fly death after 1 h
irradiation with light intensities corresponding to an
early Autumn day at Mediterranean latitudes is shown
in Fig. 3. In general, the photosensitivity of porphyrin-
loaded insects persists for about 48 h after the adminis-
tration of the photosensitizer has been interrupted.
The chemical structure (Fig. 4) also has a profound
influence on the photoinsecticidal activity of porphyrins.
As shown in Fig. 5, highly water-soluble porphyrins,
such as tetracationic meso-substituted-N-methylpyridyl
porphine or tetraanionic meso-sulphonatophenyl por-
phine, are very inefficient photoinsecticidal agents (Ben
Fig. 3. Percent survival of three diptera, Ceratitis capitata (쐌), Bac-
trocera (Dacus) oleae (×) and Stomoxys calcitrans (䊏), after 1 h
irradiation with white light at a fluence rate of 1220 µE s⫺1 m⫺2. The
flies had been exposed to a bait containing 8 µM haematoporphyrin.
Some mortality of the flies was observed already during the exposure
to light (Ben Amor et al. 1998a, 2000).
 T. Ben Amor, G. Jori / Insect Biochemistry and Molecular Biology 30 (2000) 915–925
Fig. 4. Chemical structures of meso-substituted porphyrins: TMP,
meso-tetra(4N-methylpyridyl)porphine tetratosylate; DDP, meso-[di-
cis(4N-methylpyridyl)] cis-diphenylporphine ditosylate; TRP, meso-
tri(4N-methylpyridyl), monophenylporphine tritosylate; TCPP, meso-
tetra(4-carboxyphenyl)porphine tetrasodium salt. The counterions are
not indicated in the structures.
Fig. 5. Percent survival of Ceratitis capitata after 1 h exposure to
white light at a fluence rate of 1220 µE s⫺1 m⫺2 in the presence of
3.0 µM TCPP (왔), TRP (쐌), DDP (왖) and haematoporphyrin (HP)
(䊏) (Ben Amor et al., 1998b).
921
Amor et al., 1998b). Such inertness must be related to
the anatomical distribution of these porphyrins, since
both of them are accumulated in markedly large amounts
by the flies; moreover, the two porphyrins are efficient
generators of singlet oxygen and exhibit phototoxicity
towards a variety of biological systems both in vitro and
in vivo (Spikes, 1994). As has been observed for other
classes of photosensitizers, the photoactivity of porphy-
rins increases with hydrophobicity and is particularly
large in the case of amphiphilic derivatives, such as the
meso-cis-diphenyl, meso-cis-di-N-methylpyridyl-
porphine (Fig. 5); the latter is more phototoxic than the
tricationic analogue or the dianionic hematoporphyrin
even in the presence of a smaller uptake of the dicationic
porphyrin by the insects (Ben Amor et al., 1998b).
6. Factors affecting the photoinsecticidal activity
The efficiency of photodynamic sensitizers as insecti-
cidal agents is affected by a variety of experimental para-
meters. Two obvious factors controlling the photosensi-
tivity of insects are represented by the photosensitizer
dose and average light intensity. Typical examples of the
interplay between these two factors are shown in Fig. 6
for the action of a xanthene dye (eosin yellow) on the
house fly (Musca domestica, Fondren and Heitz, 1978b;
Carpenter and Heitz, 1980) and in Fig. 7 for the action
of a porphyrin dye (haematoporphyrin) on the fruit fly
(Ceratitis capitata, Ben Amor et al., 1998a). In both
cases the photoinsecticidal effect steadily increased with
increasing dose of photosensitizing agent in the bait and
no saturation effect was detected at the highest photosen-
sitizer concentration examined by the investigators.
Analogously, the rate and extent of the photosensitized
killing of insects appeared to increase with prolongation
of exposure to light, as well as with an increase in the
Fig. 6. Effect of the concentration of Eosin Yellow on the percent
survival of Musca domestica irradiated with natural and artificial light
for different time intervals. The fluence rate of the artificial light was
4300 foot candles. Data from Carpenter and Heitz (1980).
922 T. Ben Amor, G. Jori / Insect Biochemistry and Molecular Biology 30 (2000) 915–925
Fig. 7. Percent survival of Ceratitis capitata upon exposure to white
light at a fluence rate of 1220 µE s⫺1 m⫺2 in the presence of 8 µM
haematoporphyrin: irradiated control (䊊); 60 min exposure (䊏); 90
min exposure (쐌); 120 min exposure (왖). Data from Ben Amor et
al. (1998a).
light intensity. Such conclusions are likely to be of gen-
eral validity since essentially identical results were
obtained with other photosensitizers, including rhoda-
mines (Respicio and Heitz, 1981), rose bengal (Fondren
and Heitz, 1978a; Carpenter and Heitz, 1980) and meth-
ylene blue (Lavialle and Dumortier, 1978). These effects
have been interpreted as suggestive of an intrinsic
capacity of the insect to repair the photodamage; such
capacity would be more efficiently overcome as the light
intensity is enhanced (Fondren et al., 1978; Ben Amor
et al., 1998a,b).
It is interesting to note that several insects are insensi-
tive to even high light intensities in the absence of the
photosensitizer (see Ben Amor et al., 1998a and Fig. 7).
On the other hand, it is difficult to compare the relative
efficiency of different dyes since the overall photoinsec-
ticidal effect is dependent on the overlap between the
emission spectrum of the light source and the absorption
spectrum of the photosensitizer. Most frequently, the
photoinsecticides are used in open fields, hence they are
activated by sunlight. As shown in Figs. 6 and 7 and
reported by other authors (Clement et al., 1980; Heitz,
1987), natural sunlight is generally more efficient than
artificial sunlight, probably because of its significantly
greater intensity. From this point of view, as mentioned
earlier, porphyrins have the distinct advantage of absorb-
ing almost all the light wavelengths in the sun’s emission
spectrum. As a consequence, porphyrins are expected to
express an efficient photoinsecticidal action at lower
concentrations than many other dyes (see Fig. 7).
In some cases the degree of insect mortality was
shown to increase with increasing accessibility periods
of the insects to the photosensitizer-loaded bait before
exposure to light (Robinson, 1983). These findings were
confirmed by studies with porphyrins performed in our
laboratory (Ben Amor et al., 2000). However there
appeared to be little advantage in prolonging the “dark”
exposure of the photosensitizer beyond 24 h. Most
importantly, we observed that (at least in the case of
porphyrins) the insects display a significant photosensi-
tivity for about 48 h after having taken up the dye (Ben
Amor et al., 1998a,b).
A few experiments suggest that the pH of the formu-
lation in which the photosensitizer is offered to the
insects may play an important role. Thus the free acid
forms of xanthene dyes were found to be about tenfold
more effective as photosensitizers than the correspond-
ing salt derivatives against both Aedes (Pimprikar and
Heitz, 1984) and Culex (Carpenter et al., 1984; Respicio
et al., 1985) mosquito larvae. Since the carboxylic func-
tional groups of xanthenes have pK values around 5–5.5,
one can infer that a slightly acidic dietary preparation of
the photosensitizing agents should be preferred.
Lastly, some attempts have been made to protect
insects against photodynamic action, a typical oxidative
process, by the administration of antioxidizing agents,
such as carotenes, tocopherol and ascorbic acid
(Robinson and Beatson, 1985; Heitz, 1987). In all cases,
no appreciable protection was obtained, even though
these compounds are powerful inhibitors of photooxid-
ative reactions in vitro (Jori, 1996). The lack of photop-
rotection in the insects could reflect either a rapid meta-
bolization of the antioxidants or a different
biodistribution as compared with the photosensitizing
agents.
7. Mechanisms of the insecticidal action performed
by photodynamic sensitizers
The possibility of controlling pest population by
means of photodynamic action has been investigated so
far by using a variety of photosensitizers, target insects
and irradiation conditions. In spite of the large differ-
ences in the experimental protocols, a comparative
analysis of the whole set of results reported by the vari-
ous authors allows one to draw some general conclusions
regarding the mode of action of photodynamic-type
insecticidal agents.
These can be summarized as follows:
1. The membranes of the midgut wall appear to be
among the primary photodamaged sites
(Schildmacher, 1950; Robinson, 1983; Ben Amor et
al., 1998a), leading to feeding inhibition (Fondren et
al., 1978). Other lipoidal membranes, in particular the
neuromuscular sheath, become involved in the photo-
process as the photosensitizer diffuses to other sites
(Robinson, 1983). This is confirmed by the early pho-
toinactivation of enzymes such as acetylcholinesterase
(Callaham et al., 1977a; Ben Amor et al., 2000) which
represents the neurotransmitter enzyme. A gen-
 T. Ben Amor, G. Jori / Insect Biochemistry and Molecular Biology 30 (2000) 915–925
eralized oxidative modification of the membranes
takes place, as suggested by ultrastructural studies
(Callaham et al., 1977b).
2. Changes in membrane permeability are also demon-
strated by the presence of altered potassium levels in
the hemolymph (Weaver et al., 1976). The hemo-
lymph volumes decrease significantly upon photosen-
sitization and the hemocoel fluids undergo a rapid
transfer from the body cavity to the alimentary canal
with a consequent increase in crop volume.
3. Photosensitized insects also show large differences
from controls as regards weight, levels of water and
protein mass, suggesting the occurrence of a lethal
energy stress in the insect (Broome et al., 1976).
Other consequences reported for photosensitized flies
involve a lowered fecundity (Pimprikar et al., 1980b).
4. The photosensitized induction of physiological and
morphological abnormalities was detected at the lar-
val, pupal and adult stage (Pimprikar et al., 1979;
Fairbrother et al., 1981). Thus, adults deposit fewer
and less-viable eggs, treated eggs are less likely to
hatch, and larvae exhibit a strongly reduced prob-
ability of ultimate adult emergence. In particular,
there often appears to be an incomplete extrication of
the pupal stage from the larval cuticle (Pimprikar et
al., 1979), while several adults are stuck to the chitin
inner lining of the puparium (Fairbrother, 1978).
Moreover, pupae injected with the photosensitizer fre-
quently develop into adults that are especially suscep-
tible to photodynamic action (Sakurai and Heitz,
1982). In general, earlier instar larvae appear to be
more photosusceptible than later instar forms
(Heitz, 1987).
5. The possible onset of photoresistance in the xanthene-
sensitized house fly was studied (Respicio and Heitz,
1985). A wild strain of this insect developed a 48-
fold resistance after 32 generations that underwent
exposure to increasing levels of erythrosin plus light.
Upon removal of the selection pressures for 20 gener-
ations, the resistance remained at a fairly constant
level. No evident induction of photoresistance was
observed in porphyrin-phototreated flies (Ben Amor
et al., 2000). Thus, this specific aspect of the photoin-
secticidal action appears to be strictly connected with
the photosensitizer used and the irradiation con-
ditions. It is important that no cross-resistance was
detected for erythrosin-photosensitized flies that were
exposed to chemical pesticides, such as permethrin
and propoxur (Respicio and Heitz, 1985).
6. Some photodynamic sensitizers, including rose bengal
and other xanthenes, also exert a toxic effect towards
selected insects in the dark (Carpenter and Heitz,
1981). While this toxic mechanism occurs with adult
flies, it has been observed that boll weevils fed with
rose bengal during the larval development exhibit a
decrease in body weight, and in protein and lipid lev-
923
els (Broome et al., 1976). The dark process appears
to be markedly less efficient than the corresponding
light-induced reactions. Moreover, it seems to be pec-
uliar to xanthene dyes, since no similar effects were
observed with furocoumarines, α-terthienyl (Cunat et
al., 1999; Guillet et al., 2000) and porphyrins (Ben
Amor et al., 2000), at least at doses which are photo-
chemically active.
8. Conclusions
The photoactivatable insecticides, which act through
photodynamic pathways, clearly appear to possess sev-
eral favourable features and a broad scope of appli-
cations. As the mode of action of such photosensitizers
is more deeply elucidated, suitable strategies can be
developed to optimize their efficiency and control their
biodistribution at both the tissue and the cell level, ther-
eby orientating the photosensitizer towards those sites
(e.g. cell membranes) which are critical from a func-
tional and metabolic point of view, yet their modification
is associated with a minimal risk of inducing mutagenic
effects and promoting the onset of photoresistance. A
detailed control of the in vivo behaviour of ingested pho-
tosensitizers would also be of utmost importance for
obtaining a synergistic action between two or more sim-
ultaneously administered dyes; if the latter have different
absorption properties, a more efficient activation of the
photoinsecticides by sunlight can be achieved. Thus, the
photoinsecticidal action of porphyrins, which possess
relatively weak absorption bands in the red spectral
region, could be strengthened by their association with
tetrapyrrolic analogues, such as phthalocyanines and
chlorins, which display an intense absorbance in the
600–800 nm spectral range. In fact, a synergistic action
against the house fly has been observed upon the simul-
taneous administration of two xanthenes, namely fluor-
escein and rose bengal, whose absorption spectra are
shifted by about 60 nm (Carpenter et al., 1981).
The main advantage of sunlight-activatable photosen-
sitizers is certainly represented by their lack of toxicity
towards most biological systems in the dark, which mini-
mizes their impact on the environment. However, some
concerns still exist, especially as regards the induction
of phytotoxicity (Robinson, 1983; Heitz, 1987). Thus,
some tetrapyrroles have been found to be accumulated
in the leafy tissues of dicots, such as cotton and soybean
(Rebeiz et al. 1990b, 1991). Light exposure of such
plants is accompanied by severe damage to the leaves,
while the stems and the cotyledons remain unaffected.
The plants can recover from the initial damage by pro-
ducing new leaves. Thus, more detailed investigations
on this problem should be performed. Moreover, suitable
924 T. Ben Amor, G. Jori / Insect Biochemistry and Molecular Biology 30 (2000) 915–925
strategies for minimizing the uptake of such photosensit-
izing agents by non-target insects should be developed.
The concern about environmental effects of photoin-
secticidal agents could be alleviated by the sunlight-
induced photobleaching of the sensitizing agents. In fact,
while most phenothiazines and acridines are fairly pho-
tostable, both porphyrins and xanthenes undergo a fast
degradation of the aromatic macrocycle upon illumi-
nation with sunlight or equivalent artificial light sources,
with a consequent loss of absorption in the near-
UV/visible range (van Lier and Spikes, 1989; Spikes,
1992). This feature adds further value to the use of these
compounds as photoinsecticidal agents.
References
Anderson, R.R., Parrish, J.A., 1992. Optical properties of human skin.
In: Regan, J.D., Parrish, J.A. (Eds.), The Science of Photomedicine.
Plenum Press, New York, pp. 174–194.
Armitage, B., 1998. Photocleavage of nucleic acids. Chem. Rev. 98,
1171–1200.
Barbieri, A., 1928. Sensibilizadores fluorescentes como larvicidas.
Action fotodynamica de la luz. Riv. Malariol. 7, 456–463.
Ben Amor, T., Tronchin, M., Bortolotto, L., Verdiglione, R., Jori, G.,
1998a. Porphyrins and related compounds as photoactivatable
insecticides. 1. Phototoxic activity of haematoporphyrin toward
Ceratitis capitata and Bactrocera oleae. Photochem. Photobiol. 67,
206–211.
Ben Amor, T., Bortolotto, L., Jori, G., 1998b. Porphyrins and related
compounds as photoactivatable insecticides. 2. Phototoxic activity
of meso-substituted porphyrins. Photochem. Photobiol. 68, 314–
318.
Ben Amor, T., Bortolotto, L., Jori, G., 2000. Porphyrins and related
compounds as photoactivatable insecticides. 3. Laboratory and field
studies. Photochem. Photobiol. 71, 123–127.
Bensasson, R., Land, C., Truscott, R.G., 1983. Excited States and Free
Radicals in Biology and Medicine. Oxford University Press,
New York.
Blum, H.F., 1941. Photodynamic Action and Diseases Caused by
Light. Rheinold, New York. ACS Monograph Series.
Bonnett, R., Berenbaum, M., 1989. Porphyrins as photosensitizers. In:
Bock, G., Harnett, S. (Eds.), Photosensitizing Compounds: their
Chemistry, Biology and Clinical Use. Wiley, Chichester, pp. 40–
59.
Bosca, F., Miranda, M.A., 1998. Photosensitizing drugs containing the
benzophenone chromophore. J. Photochem. Photobiol., B: Biol. 43,
1–26.
Boyle, R.W., Dolphin, D., 1996. Structure and biodistribution of pho-
todynamic sensitizers. Photochem. Photobiol. 64, 469–485.
Briviba, K., Klotz, L.O., Sies, H., 1997. Toxic and signaling effects
of photochemically or chemically generated singlet oxygen in bio-
logical systems. Biol. Chem. 378, 1259–1265.
Broome, J.R., Callaham, M.F., Poe, W.E., Heitz, J.R., 1976. Biochemi-
cal changes in the boll weevil induced by rose bengal in the absence
of light. Chem. Biol. Interact. 14, 203–206.
Brown, S., 1997. Photodynamic therapy: the international scene. Int.
Photodyn. 1, 1–2.
Callaham, M.F., Palmertree, C.O., Broome, J.R., Heitz, J.R., 1977a.
Dye sensitized photoinactivation of the lactic dehydrogenase and
acetyl cholinesterase from the boll weevil, Anthonomus grandis.
Pestic. Biochem. Physiol. 7, 21–27.
Callaham, M.F., Lewis, L.R., Poe, W.E., Heitz, J.R., 1977b. Time
dependence of light-independent biochemical changes in the boll
weevil, Anthonomous grandis, caused by dietary rose bengal.
Environ. Entomol. 6, 668–673.
Carmichael, I., Huh, G.L., 1986. Triplet-triplet absorption spectra of
organic molecules in condensed phases. J. Phys. Chem. Ref. Data
15, 1–250.
Carpenter, T.L., Heitz, J.R., 1980. Light-dependent latent toxicity of
rose bengal to Culex pipiensquinque fasciatus. Environ. Entomol.
9, 533–537.
Carpenter, T.L., Heitz, J.R., 1981. Light-dependent and independent
toxicity of erythrosin B to Culex pipiensquinpuefasciatus. Environ.
Entomol. 10, 972–976.
Carpenter, T.L., Mundie, T.G., Ross, J.H., Heitz, J.R., 1981. Syner-
gistic effect of fluorescein on rose bengal-induced, light-dependent
toxicity. Environ. Entomol. 10, 953–957.
Carpenter, T.L., Respicio, N.C., Heitz, J.R., 1984. Comparative photo-
toxicity of soluble and insoluble forms of xanthene dyes against
Culex mosquito larvae. Environ. Entomol. 13, 1366–1370.
Clement, S.L., Schmidht, R.S., Szatmari-Goodman, G., Levine, E.,
1980. Activity of xanthene dyes against Black cutworm larvae. J.
Econ. Entomol. 73, 390–392.
Cunat, P., Primo, E., Sanz, I., Sarcera, N.D., March, N.A., Bowers,
W.S., Martinez Prado, A., 1999. Biocidal activity of some Spanish
Mediterranean plants. J. Agric. Food Chem. 38, 497–500.
Dowell, R.V., Wilson, W., Hennessy, M.K., 1997. Toxicity of Suredye
to beneficial insects. Photochem. Photobiol. 65S. Abstract TMP
D-6.
Fairbrother, T.E., 1978. Effects of xanthene dyes on face fly larvae
and effects of erythrosin B on ruminant digestion. PhD thesis, cited
by Robinson (1983).
Fairbrother, T.E., Essig, H.W., Combs, R.L., Heitz, J.R., 1981. Toxic
effects of rose bengal and erythrosin B on three life stages of the
face fly Musca autumnalis. Environ. Entomol. 10, 506–510.
Fondren, J.E., Heitz, J.R., 1978a. Xanthene dyes induced toxicity in the
adult face fly, Musca autumnalis. Environ. Entomol. 7, 843–846.
Fondren, J.E., Heitz, J.R., 1978b. Light intensity as a critical parameter
in the dye-sensitized photooxidation of the house fly, Musca dom-
estica. Environ. Entomol. 7, 891–894.
Fondren, J.E., Norment, B.R., Heitz, J.R., 1978. Dye-sensitized pho-
tooxidation in the house fly Musca domestica. Environ. Entomol.
7, 205–208.
Fondren, J.E., Norment, B.R., Heitz, J.R., 1979. Dye sensitized house
fly toxicity produced as a function of variable light sources.
Environ. Entomol. 8, 432–436.
Guillet, G., Harmatha, J., Waddell, T.G., Philogne Bernard, J.R., Arna-
son, J.T., 2000. Synergistic insecticidal mode of action between
sesquiterpene lactones and a phototoxin, α-terthienyl. Photochem.
Photobiol. 71, 115–122.
He, X.Y., Spikes, R.A., Thomson, S., Chung, L.W., Jacques, L., 1994.
Photosensitization with Photofrin induces programmed cell death
in carcinoma cell lines. Photochem. Photobiol. 59, 468–473.
Heitz, J.R., 1987. Development of photoactivated compounds as pes-
ticides. In: Heitz, J.R., Downum, K.R. (Eds.), Light Activated Pes-
ticides. ACS, Washington DC, pp. 1–21. ACS Symposium Series
339.
Heitz, J.R., 1997a. Historical perspectives of xanthene dyes as pestic-
ides. Photochem. Photobiol. 65S, Abstract TPM-D1.
Heitz, J.R., 1997b. Symposium on photosensitizing insecticides: phlox-
ine B comes of age. Photochem. Photobiol. 65S, abstracts TMP
D1, TMP D-2.
Jodlbauer, A., Von Tappainer, H., 1904. Über die Beteiligung der
Sauerstoffes bei der photodynamischem Wirkung fluorezierender
Stoffe. Münch. Med. Wochenschr. 26, 1139–1141.
Jori, G., 1985. Molecular and cellular mechanisms in photomedicine.
In: Bensasson, R.V., Jori, G., Land, E., Truscott, T.G. (Eds.), Pri-
mary Photoprocesses in Biology and Medicine. Plenum Press, New
York, pp. 349–355.
Jori, G., 1996. Photosensitizers: approaches to enhance the selectivity
 T. Ben Amor, G. Jori / Insect Biochemistry and Molecular Biology 30 (2000) 915–925
and efficiency of photodynamic action. J. Photochem. Photobiol.,
B: Biol. 36, 87–93.
Jori, G., Reddi, E., 1991. Second generation photosensitizers for the
photodynamic therapy of tumours. In: Douglas, R.H., Moan, J.,
Ronto, G. (Eds.), Light in Biology and Medicine. Plenum Press,
London, pp. 253–266.
Jori, G., Reddi, E., 1993. The role of lipoproteins in the delivery of
tumour-targeting photosensitizers. Int. J. Biochem. 25, 1369–1375.
Jori, G., Spikes, J.D., 1983. Photobiochemistry of porphyrins. In:
Smith, K.C. (Ed.), Topics in Photomedicine. Plenum Press, New
York, pp. 183–319.
Lavialle, M., Dumortier, B., 1978. Effect photodynamique du bleu de
methylène sur les larves de Pieris brassicae. C. R. Acad. Sci., Ser.
D 287, 875–878.
Lenke, L.A., Koehler, P.G., Patterson, R.S., Feger, M.B., Eickhoff, A.,
1987. Field development of photooxidative dyes as insecticides. In:
Heitz, J.R., Downum, K.R. (Eds.), Light Activated Pesticides.
ACS, Washington DC, pp. 156–167. ACS Symposium Serves 339.
Marcacci, A., 1888. Sur l’action des alcaloides dans le règne végétale
et animal. Arch. Ital. Biol. 9, 2–4.
Merchat, M., Spikes, J.D., Bertoloni, G., Jori, G., 1996. Studies on
the mechanism of bacteria photosensitization by meso-substituted
cationic porphyrins. J. Photochem Photobiol., B: Biol. 35, 149–157.
Moan, J., Peng, Q., Iani, V., Berg, K., Konshaug, M., Nesland, J.M.,
1995. Biodistribution, pharmacokinetic and in vivo fluorescence
spectroscopic studies of photosensitizers. In: Ehrenberg, B., Jori,
G., Moan, J. (Eds.), Photochemotherapy: Photodynamic Therapy
and Other Modalities. SPIE, Bellingham, USA, pp. 234–250.
Pimprikar, G.D., Heitz, J.R., 1984. Unusually high insecticidal activity
of insoluble free acid forms of xanthene dyes to Aedes mosquito
larvae. J. Miss. Acad. Sci. 29, 77–80.
Pimprikar, G.D., Norment, B.R., Heitz, J.R., 1979. Toxicity of rose
bengal to various instars of Culex pupiens quinquefasciatus and
Aedes triseriatus. Environ. Entomol. 9, 856–859.
Pimprikar, G.D., Fondren, J.E., Heitz, J.R., 1980a. Small and large
scale field tests of erythrosine B for house fly control in caged layer
chicken houses. Environ. Entomol. 9, 53–58.
Pimprikar, G.D., Noe, B.L., Norment, B.R., Heitz, J.R., 1980b. Ovi-
cidal, larvicidal and biotic effects of xanthene derivatives in the
house fly Musca domestica. Environ. Entomol. 9, 785–788.
Pooler, J.P., Valenzeno, D.P., 1979. The role of singlet oxygen in the
photooxidation of excitable cell membranes. Photochem. Photobiol.
30, 581–589.
Raab, O., 1900. Über die Wirkung fluoresceinder Stoffe and Infusor-
ien. Zeit für Biol. 39, 524–546.
Rebeiz, C.A., 1992. Porphyric insecticides. J. Photochem. Photobiol.,
B: Biol. 18, 97–114.
Rebeiz, C.A., Montazaer-Zouhour, A., Mayasich, J.M., Tipathy, B.C.,
Wu, S.M., Rebeiz, C.C., 1987. Porphyric insecticides. In: Heitz,
J.R., Downum, K.R. (Eds.), Light Activated Pesticides. ACS,
Washington, DC, pp. 295–328. ACS Symposium Series 339.
Rebeiz, C.A., Juvick, J.A., Rebeiz, C.C., 1988. Porphyric insecticides. I.
Concepts and phenomenology. Pestic. Biochem. Physiol. 30, 11–27.
Rebeiz, C.A., Juvick, J.A., Rebeiz, C.C., Bowton, C.E., Sut, L.J.,
1990a. Porphyric insecticides. I. Phenanthroline, a potent porphyric
insecticide modulator. Pestic. Biochem. Physiol. 36, 201–220.
Rebeiz, C.A., Reddy, K.N., Nandihalli, U.B., Velu, J., 1990b. Tetrapyr-
role dependent photodynamic herbicides. Photochem. Photobiol.
52, 1099–1117.
Rebeiz, C.A., Nandihalli, U.B., Reddy, K.N., 1991. Tetrapyrrole
dependent photodynamic herbicides and chlorophyll biosynthesis
modulators. In: Baker, N.R., Percival, M.B. (Eds.), Herbicides.
Elsevier, Amsterdam, pp. 173–208.
Respicio, N.C., Heitz, J.R., 1981. Comparative toxicity of rhodamine B
and rhodamine G to the house fly Musca domestica. Bull. Environ.
Contam. Toxicol. 27, 274–277.
Respicio, N.C., Heitz, J.R., 1985. Cross resistance of erythrosine B-
925
resistant house flies to different pesticides. J. Econ. Entomol. 79,
315–317.
Respicio, N.C., Carpenter, T.L., Heitz, J.R., 1985. Toxicity to Culex
mosquito larvae of non-dispersible and dispersible formulations of
fluorescein and erythrosine B. J. Econ. Entomol. 78, 30–34.
Ricchelli, F., Jori, G., 1986. Distribution of porphyrins in the various
compartments of unilamellar liposomes of dipalmitoyl-phosphatid-
ylcholine as probed by fluorescence spectroscopy. Photochem. Pho-
tobiol. 44, 151–158.
Ricchelli, F., Gobbo, S., Jori, G., Moreno, G., Salet, C., 1993. Photo-
sensitization of mitochondria by liposome-bound porphyrins. Pho-
tochem. Photobiol. 58, 53–58.
Ricchelli, F., Gobbo, S., Jori, G., Moreno, G., Salet, C., 1995. Tem-
perature-induced changes in fluorescence properties as a probe of
porphyrin microenvironment in lipid menbranes. I. The partition of
protoporphyrin and haematoporphyrin in liposomes. Eur. J.
Biochem. 233, 159–164.
Robinson, J.R., 1983. Photodynamic insecticides: a review of studies
on photosensitizing dyes as insect control agents, their practical
application, hazards and residues. In: Heitz, J.R. (Ed.). Residue
Reviews, vol. 88. Springer Verlag, Berlin, pp. 69–100.
Robinson, J.R., Beatson, E.P., 1985. Effect of selected antioxidants on
the phototoxicity of erythrosin B toward house fly larvae. Pestic.
Biochem. Physiol. 24, 375–383.
Rossi, E., Van De Vorst, A., Jori, G., 1981. Competition between the
singlet oxygen and electron transfer mechanisms in the sensitized
photooxidation of l-tryptophan and tryptamine in aqueous cellular
dispersions. Photochem. Photobiol. 34, 447–452.
Rotomskis, R., Streckyte, G., Bagdonas, S., 1997. Phototransformations
of photosensitizers. 2. Photoproducts formed in aqueous solution of
porphyrins. J. Photochem. Photobiol., B: Biol. 39, 172–175.
Sakurai, H., Heitz, J.R., 1982. Growth inhibition and photooxidative
toxicity in the house fly Musca domestica, caused by xanthene dyes
in larval growth medium and after injection. Environ. Entomol. 11,
467–470.
Schildmacher, H., 1950. Über Photosensibilisierung von Stechmücken-
larven durch fluoreszierende Farbstoffer. Biol. Zentralbl. 69,
468–477.
Spikes, J.D., 1985. In: Bensasson, R.V., Jori, G., Land, E., Truscott,
T.G. (Eds.), Primary Processes in Biology and Medicine. Plenum
Press, New York, pp. 209–227.
Spikes, J.D., 1992. Quantum yield and kinetics of the photobleaching
of haematoporphyrin, Photofrin II, tetra(4-sulfonatophenyl) por-
phine and uroporphyrin. Photochem. Photobiol. 55, 797–808.
Spikes, J.D., 1994. Photobiology of porphyrins. In: Doiron, D.R.,
Gomer, C.J. (Eds.), Porphyrin Localization and Treatment of
Tumours. Alan R. Liss, New York, pp. 19–39.
Spikes, J.D., Macknight, M.L., 1970. Dye-sensitized photooxidation of
proteins. Ann. N. Y. Acad. Sci. 171, 149–162.
Svaasand, L.O., Martinelli, E., Gomer, G.J., Profio, A.E., 1990. Optical
characteristics of tumours in the visible and near-infrared. Proc.
SPIE 1203, 2–21.
van Lier, J.E., Spikes, J.D., 1989. In: Bock, G., Harnett, S. (Eds.),
Photosensitizing Compounds: their Chemistry, Biology and Clini-
cal Use. Wiley, Chichester, pp. 17–32.
Vilensky, A., Feitelson, J., 1999. Reactivity of singlet oxygen with
tryptophan residues and with melittin in liposome systems. Photo-
chem. Photobiol. 700, 841–846.
Wasserman, H.H., Murray, R.W., 1989. Singlet Oxygen. Academic
Press, New York.
Weaver, J.E., Butler, L., Yoho, T.P., 1976. Photodynamic action in
insects: volumetric change in the hemolymph and crop contents of
dye-treated, light-exposed cockroaches. Environ. Entomol. 5,
840–843.
Yoho, T.P., Butler, L., Weaver, J.E., 1971. Photodynamic effects of
light on dye-fed house flies: preliminary observations of mortality.
J. Econ. Entomol. 64, 972–973.