The effect of relative humidity on the behaviour and development of Triatoma brasiliensis

Abstract The preference for relative humidity (RH) and suitability of different levels of this environmental parameter were investigated in the haematophagous bug Triatoma brasiliensis Neiva, 1911 (Hemiptera, Reduviidae). The hygropreference of T. brasiliensis was studied using a RH gradient and the effect of different RHs on the egg hatching, nymph mortality and moulting success was also analysed. The results show that egg hatching in first-instar nymphs of T. brasiliensis was lower at extreme RHs and, particularly, it was lowest at 9.3% RH. The survival of starved nymphs was not affected by RH, but the percentage of engorged nymphs and the ecdysis success of these nymphs once fed was diminished strongly by high humidity. Fourth-instar nymphs preferred to stay at the lowest RH during the first 5 days after feeding and during ecdysis. This preference changed markedly during starvation. Fifteen days after ecdysis, the bugs moved towards intermediate humidities, and 30 days after ecdysis they even preferred the most humid sectors of the gradient. Females preferred to lay eggs in dry environments, suggesting that they may not have a particular hygropreference for oviposition, but that they simply lay their eggs at the RHs where they prefer to stay.     

Effects of fluctuating moisture and temperature regimes on the infection potential of Beauveria bassiana for Rhodnius prolixus

The effect of both moisture and temperature on the infective potential of Beauveria bassiana to the Chagas' disease vector, Rhodnius prolixus, was studied under fluctuating regimes. At constant 25 degrees C, contaminated first-instar nymphs exposed to increasing daily periods of initial exposure to 97% RH, followed by transfer to reduced humidity (43, 53, 75, and 86% RH), showed a significant reduction in mortality when the 97% RH exposure time declined from 12 to 8 h per day. The duration of disease incubation depended on the daily 97% RH exposure time. Under fluctuating regimes of both humidity (97% RH versus 75% RH) and temperature (15/28, 20/25, 25/28, and 25/35 degrees C), first-instar mortality was affected by weather conditions, daily 97% RH exposure time (8, 12, and 16 h per day), and number of temperature and humidity fluctuations before transferring tested insects to constant unfavorable conditions. In most cases, at 12/12 h alternating cycles, high and rapid mortality required five cycles. Under these fluctuating regimes, fungus-induced mortality and mortality time were similarly affected in third- and fifth-instar nymphs by the daily 97% RH exposure time. Despite a lower susceptibility of older larval stages, mortality rates in insects exposed for at least 12 h per day at 97% RH remained very high except at 15 degrees C. Moisture and temperature regimes at 12/12 h cycling significantly affected the dose-mortality response in first-instar nymphs. The most favorable conditions consisted of 97%-20 degrees C combined with either 75%-25 degrees C or 43%-25 degrees C. Under less favorable alternating conditions (lower and higher temperatures) the amounts of inoculum required for killing 50% of first-instar nymphs were 10 or 20 times higher. From a vector control standpoint, daily high humidity appears to be the most crucial climatic constraint. B. bassiana has the potential to control R. prolixus populations with applications made during the rainy seasons when humidity is high.     

Evaluation of RbCl and CrCl₃ as markers of Triatoma brasiliensis (Hemiptera: Reduviidae) nymphs: persistence and influence of Rb and Cr on triatomine biology

In order to mark Triatoma brasiliensis, the vector of Chagas disease in Brazil, two chemical compounds, rubidium chloride (RbCl) and chromium chloride (CrCl?), were tested. First, 199 N2-N5 nymphs were fed on blood with 0.025M RbCl. Rb marker positivity ranged from 2.5% (N3)-70% (N2), with a maximum persistence of 98 days. Second, 265 N2-N5 nymphs were fed on blood containing 0.0015M CrCl?. Cr marker positivity ranged up to 93% (N5), with a maximum persistence of 119 days. Finally, we blood fed 213 T. brasiliensis to investigate whether CrCl? altered the biology of this insect. The developmental time of T. brasiliensis was unaltered, but the survival of the Cr-marked group was lower than that of the control group. Differences in the mean fecundity of the control (mean of 156.1) and experimental (mean of 135.6) groups were not statistically significant and 100% of the egg batches of females Cr-marked as nymphs were positive. In conclusion, CrCl? is a useful tool for marking T. brasiliensis nymphs due to its high positivity and persistence.     

Brasiliensin: A novel intestinal thrombin inhibitor from Triatoma brasiliensis (Hemiptera: Reduviidae) with an important role in blood intake

Every hematophagous invertebrate studied to date produces at least one inhibitor of coagulation. Among these, thrombin inhibitors have most frequently been isolated. In order to study the thrombin inhibitor from Triatoma brasiliensis and its biological significance for the bug, we sequenced the corresponding gene and evaluated its biological function. The T. brasiliensis intestinal thrombin inhibitor, termed brasiliensin, was sequenced and primers were designed to synthesize double strand RNA (dsRNA). Gene knockdown (RNAi) was induced by two injections of 15mug of dsRNA into fourth instar nymphs. Forty-eight hours after the second injection, bugs from each group were allowed to feed on hamsters. PCR results showed that injections of dsRNA reduced brasiliensin expression in the anterior midgut by approximately 71% in knockdown nymphs when compared with controls. The reduction in gene expression was confirmed by the thrombin inhibitory activity assay and the citrated plasma coagulation time assay which showed activity reductions of approximately 18- and approximately 3.5-fold, respectively. Knockdown nymphs ingested approximately 39% less blood than controls. In order to confirm the importance of brasiliensin in blood ingestion, fourth instar nymphs were allowed to ingest feeding solution alone or feeding solution containing 15U of thrombin prior to blood feeding. Fifty-five percent less blood was ingested by nymphs which were fed thrombin prior to blood feeding. The results suggest that anticoagulant activity in the midgut is an important determinant of the amount of blood taken from the host. The role of anticoagulants during blood ingestion is discussed in the light of this novel insight.     

Effect of relative humidity on emergence and on dispersal and regrouping of first instar Nezara viridula (L.) (Hemiptera: Pentatomidae)

Laboratory studies with 1st instar of southern green stink bug, Nezara viridula (L.) indicated that relative humidity (RH) greatly affected nymph emergence and survivorship up to the 2nd instar, reaching the maximum value (approximately 90%) with RH of > 80%. At 60% RH, 60% of the nymphs emerged and survived, while with 0% RH only approximately 15% of eggs hatched, and most nymphs died. Emerged nymphs from egg masses placed in plastic boxes with a gradient of humidity remained on egg shells for ca. one day. After this period, they dispersed and regrouped on top of shells 6.8 +/- 0.67 times, until they abandoned the shells toward the source of humidity, avoiding the water-saturated areas. Duration taken for each rearrangement (dispersal + regroup) increased with time, with a range of approximately 26 min to 44 min. The mean duration of the grouping behavior on egg shells after each rearrangement decreased from approximately 102 min (1st) to 24 min (6th and last grouping). The rearrangement behavior of 1st instars on top of egg shells apparently compensates for the water loss of nymphs.     

Habitat requirements of the seabird tick, <Emphasis Type="Italic">Ixodes uriae</Emphasis> (Acari: Ixodidae), from the Antarctic Peninsula in relation to water balance characteristics of eggs,nonfed and engorged stages

The seabird tick Ixodes uriae is exposed to extreme environmental conditions during the off-host phase of its life cycle on the Antarctic Peninsula. To investigate how this tick resists desiccation, water requirements of each developmental stage were determined. Features of I. uriae water balance include a high percentage body water content, low dehydration tolerance limit, and a high water loss rate, which are characteristics that classify this tick as hydrophilic. Like other ticks, I. uriae relies on water vapor uptake as an unfed larva and enhanced water retention in the adult, while nymphs are intermediate and exploit both strategies. Stages that do not absorb water vapor, eggs, fed larvae and fed nymphs, rely on water conservation. Other noteworthy features include heat sensitivity that promotes water loss in eggs and unfed larvae, an inability to drink free water from droplets, and behavioral regulation of water loss by formation of clusters. We conclude that I. uriae is adapted for life in a moisture-rich environment, and this requirement is met by clustering in moist, hydrating, microhabitats under rocks and debris that contain moisture levels that are higher than the tick's critical equilibrium activity.     

Aggregation density and longevity correlate with humidity in first-instar nymphs of the cockroach (Blattella germanica L., Dictyoptera)

Nymphs and adults of the German cockroach, Blattella germanica, tend to aggregate. The effect of six different relative humidities (RH) on the aggregation behaviour of groups of 30 first instars was tested. The results show that grouping is denser under lower RHs than under higher humidities. The mean interindividual distances (nearest neighbour distances) proved to be inversely proportional to saturation deficiency (SD). The effect of aggregation behaviour in regulating water loss by evaporation is discussed. Under deprivation of food and drinking water survival times of the nymphs depend on RH. For five humidity levels tested the longevity (defined as MT(50)) proved to be inversely proportional to SD. The correlation is best fitted by a hyperbola.     

Induction of egg hatching by high humidity in the cicada Cryptotympana facialis

The timing of egg hatching in Cryptotympana facialis was examined in relation to short-term weather conditions. The brief underwater submergence, once a week, of dead twigs bearing cicada egg nests resulted in high hatching rates both at 25 degrees C and under outdoor conditions protected from rainfall. Under outdoor conditions with natural rainfall, most eggs hatched on rainy days. There was a significant positive correlation between the number of hatching eggs and daily relative humidity (RH). When eggs picked from twigs were exposed to various humidity levels, many eggs hatched quickly at higher humidity without direct contact with liquid water. Newly hatched nymphs showed a low tolerance to desiccation; at 81% RH at 25 degrees C, most of them died within 6 h. Under outdoor conditions, most nymphs died within 2 h on sunny days, whereas nymphs survived longer on rainy days. When newly hatched nymphs were released on dry ground, only 24% of them succeeded in burrowing into the soil, and many were killed by ants or desiccation. However, 92% of nymphs released onto wet ground successfully burrowed into it. The direct induction of hatching by high humidity ensures the survival and establishment in the soil of newly hatched nymphs in this species.     

Developmental and reproductive patterns of Triatoma brasiliensis infected with Trypanosoma cruzi under laboratory conditions

The aim of this work was to study the interaction between Trypanosoma cruzi-1 and Triatoma brasiliensis. A group of 1st instar nymphs was initially fed on T. cruzi-infected mice and a control group was fed on uninfected mice. From the second feeding onwards, both groups were otherwise fed on non-infected mice. The resulting adults were grouped in pairs: infected male/uninfected female, uninfected male/infected female, infected male and female and uninfected male/uninfected female. The infection affected only the 1st instar nymphs, which took significantly more time to reach the 2nd instar than uninfected nymphs. The differences in the molting time between the infected and uninfected nymphs from the 2nd to the 5th instars were not statistically significant. Both groups presented similar rates of nymphal mortality and reproductive performance was not significantly affected by infection in any of the treatments.     

Energetic consequences of repeated and prolonged dehydration in the Antarctic midge, Belgica antarctica

Larvae of the Antarctic midge, Belgica antarctica, routinely face periods of limited water availability in their natural environments on the Antarctic Peninsula. As a result, B. antarctica is one of the most dehydration-tolerant insects studied, surviving up to 70% loss of its body water. While previous studies have characterized the physiological effects of a single bout of dehydration, in nature larvae are likely to experience multiple bouts of dehydration throughout their lifetime. Thus, we examined the physiological consequences of repeated dehydration and compared results to larvae exposed to a single, prolonged period of dehydration. For the repeated dehydration experiment, larvae were exposed to 1-5 cycles of 24 h dehydration at 75% RH followed by 24 h rehydration. Each bout of dehydration resulted in 30-40% loss of body water, with a concomitant 2- to 3-fold increase in body fluid osmolality. While nearly 100% of larvae survived a single bout of dehydration, <65% of larvae survived five such cycles. Larvae subjected to multiple bouts of dehydration also experienced severe depletion of carbohydrate energy reserves; glycogen and trehalose content decreased with each successive cycle, with larvae losing 89% and 48% of their glycogen and trehalose, respectively, after five cycles of dehydration/rehydration. Larvae exposed to prolonged dehydration (99% RH for 10d) had 26% less water, 43% less glycogen, and 27% less lipid content than controls, but did not experience any mortality. Thus, both repeated and prolonged dehydration results in substantial energetic costs that are likely to negatively impact fitness.     

Performance of nymphal and adult southern green stink bug on an overwintering host plant and impact of nymph to adult food-switch

Performance of nymphs and adults of the southern green stink bug,Nezara viridula (L.) (Heteroptera: Pentatomidae) feeding on immature fruits of radish,Raphanus raphanistrum L. (Brassicaceae) was poor compared with feeding on immature soybean [Glycine max (L.) Merrill] pods, but a change from radish to soybean from nymph to adult alleviated some of the detrimental effects of this food on adult performance. Over 50% of nymphs died on radish, while only 15% died on soybean. To complete nymph development (stadia 2 through 5), male and female nymphs required 9.5 and 13.3 days longer on radish compared to soybean. Fresh body weight of 1-d-old adult females and males was ≅65% greater for those reared as nymphs on soybean than on radish, with females being significantly heavier than males on both foods. Survivorship and longevity of females, however, was greater for those fed on radish as nymphs and as adults, as compared to those that fed on soybean or radish followed by soybean. Males lived longer when fed exclusively on soybean. In general, reproductive performance and percentage gain in adult body weight was greatest for insects fed on soybean only, and were better for those whose food was switched from nymph to adult, as compared to those insects fed on radish solely.     

Respiratory and cuticular water loss in insects with continuous gas exchange: comparison across five ant species

Respiratory water loss (RWL) in insects showing continuous emission of CO(2) is poorly studied because few methodologies can measure it. Comparisons of RWL between insects showing continuous and discontinuous gas exchange cycles (DGC) are therefore difficult. We used two recently developed methodologies (the hyperoxic switch and correlation between water-loss and CO(2) emission rates) to compare cuticular permeabilities and rates of RWL in five species of ants, the Argentine ant (Linepithema humile) and four common native ant competitors. Our results showed that RWL in groups of ants with moderate levels of activity and continuous gas exchange were similar across the two measurement methods, and were similar to published values on insects showing the DGC. Furthermore, ants exposed to anoxia increased their total water loss rates by 50-150%. These results suggest that spiracular control under continuous gas exchange can be as effective as the DGC in reducing RWL. Finally, the mesic-adapted Argentine ant showed significantly higher rates of water loss and cuticular permeability compared to four ant species native to dry environments. Physiological limitations may therefore be responsible for restricting the distribution of this invasive species in seasonally dry environments.     

黑肩绿盲蝽耐饥饿能力的研究

【目的】为早期释放黑肩绿盲蝽Cyrtorhinus lividipennis提供理论依据,研究了黑肩绿盲蝽不同虫态耐受饥饿及低温的能力。【方法】室内条件下,比较了稻苗+水、无苗有水和无苗无水三种食物条件下黑肩绿盲蝽耐饥力大小。【结果】适温(26℃)下,黑肩绿盲蝽3龄若虫、5龄若虫和成虫在稻苗+水的存活时间显著高于无苗有水,无苗有水时的存活时间又显著高于无苗无水,表明稻苗对于黑肩绿盲蝽有营养作用,取食稻苗有助于提高黑肩绿盲蝽的耐饥力。黑肩绿盲蝽成虫耐饥力最强,3龄若虫和5龄若虫接近,有苗有水时,3龄若虫、5龄若虫和成虫的平均存活时间分别为2.10、2.22和4.25 d。在低温(15℃)下,水稻苗对黑肩绿盲蝽成虫的存活不利,有苗有水时雌雄成虫的存活时间均低于无苗有水处理,其中雌成虫两处理间的差异显著(P=0.001)。【结论】黑肩绿盲蝽对饥饿和低温有较强的耐受性,以成虫最强,水稻生长前期田间释放时以成虫为宜。     

Dehydration and viability of saffron crocus (Crocus sativus)

Saffron pollen reaches the shedding stage with a high water content which averages 58% under 18-20°C and RH 55-70%. Because of the precocious incision of anther stomium and the apertures of pollen exine, when the flower opens the pollen is soon exposed to dehydration. As a result, it goes from the highly hydrated stage to 15% water within 2 h from flower opening, and to the minimum 7% within 10 h. The transition to dry state results in a progressive loss of the plasmamembrane integrity, but this is restored after gradual rehydration. On average, 50% of the total pollen grains shows capacity for semi-vivo germination regardless of the dehydration degree. Pollen loses the competence for germination within 3 weeks from shedding. The results are discussed in comparison with fertile Crocus species.     

Revisiting Trypanosoma rangeli Transmission Involving Susceptible and Non-Susceptible Hosts

Trypanosoma rangeli infects several triatomine and mammal species in South America. Its transmission is known to occur when a healthy insect feeds on an infected mammal or when an infected insect bites a healthy mammal. In the present study we evaluated the classic way of T. rangeli transmission started by the bite of a single infected triatomine, as well as alternative ways of circulation of this parasite among invertebrate hosts. The number of metacyclic trypomastigotes eliminated from salivary glands during a blood meal was quantified for unfed and recently fed nymphs. The quantification showed that ~50,000 parasites can be liberated during a single blood meal. The transmission of T. rangeli from mice to R. prolixus was evaluated using infections started through the bite of a single infected nymph. The mice that served as the blood source for single infected nymphs showed a high percentage of infection and efficiently transmitted the infection to new insects. Parasites were recovered by xenodiagnosis in insects fed on mice with infections that lasted approximately four months. Hemolymphagy and co-feeding were tested to evaluate insect-insect T. rangeli transmission. T. rangeli was not transmitted during hemolymphagy. However, insects that had co-fed on mice with infected conspecifics exhibited infection rates of approximately 80%. Surprisingly, 16% of the recipient nymphs became infected when pigeons were used as hosts. Our results show that T. rangeli is efficiently transmitted between the evaluated hosts. Not only are the insect-mouse-insect transmission rates high, but parasites can also be transmitted between insects while co-feeding on a living host. We show for the first time that birds can be part of the T. rangeli transmission cycle as we proved that insect-insect transmission is feasible during a co-feeding on these hosts.     

Water loss and metabolic water in starving Argas reflexus nymphs (Acari: Argasidae)

Basic components of the water balance were determined in A. reflexus unfed second instar nymphs kept at 30 degrees C and different relative humidities in darkness. At 2.5% and 32.5% RH, A. reflexus survived for 170+/-60 and 246+/-71 days, respectively. At 56%, 75.5% and 96.5% RH, the experiment was terminated after 168 days without any mortality. The initial water content of hydrated A. reflexus kept at 85% RH was 70.3+/-1.8% while the water content was close to 50% in ticks dehydrated at /=56% suggesting that A. reflexus increased its oxidative metabolism under dehydrating conditions and thus delayed lethal dehydration. Interestingly, the dry mass of dead ticks was almost identical irrespective of whether the ticks had previously been kept at 2.5% and 32.5% RH, i.e. irrespective of their different survival periods. This raises the question whether the ticks had died by dehydration or by exhaustion of energy reserves.     

Life-cycle and host specificity of Amblyomma tigrinum (Acari: Ixodidae) under laboratory conditions

Biological data of three generations of Amblyomma tigrinum in the laboratory are reported and the suitability of different host species for immature ticks are compared. Grouping the three generations, infestations by both the larval and nymphal stages were performed on chickens (Gallus gallus), wistar rats (Rattus norvegicus), rabbits (Oryctolagus cuniculus),wild mice (Calomys callosus), dogs (Canis familiaris) and opossums (Didelphis albiventris). Only dogs were used for infestations by adult ticks. Tick developmental periods were observed in an incubator at 27°C and RH 90%. The proportion of engorged larvae recovered from chickens (21.7% of the exposed larvae) was significantly larger (p<0.001) than those from the five mammal species used in the infestations (maximum of 3.1%). A significant larger (p<0.01) proportion of engorged larvae successfully molted after being fed on chickens than on mammal hosts. The proportion of engorged nymphs recovered from chickens (28.8% of the exposed nymphs) was significantly larger (p<0.001) than those from mammal hosts (range: 0–2.1%). Larvae showed similar feeding periods on exposure to different host species, except for those larvae fed on C. callosus, which showed significantly longer (p<0.001) feeding periods. Engorged larvae detachment peaked on the 5^th feeding day, followed by the 6^th day, on all hosts except for C. callosus. Larval premolt periods were similar for engorged ticks exposed to different host species, except for larvae fed on dogs, which showed significantly longer (p<0.001) premolt periods. Host detachment of engorged nymphs peaked on the 6^th feeding day on chickens. Although nymphal detachment on rats peaked on the 8^th day, only 15 nymphs were recovered from this host species. In a sample of 144 F₃ nymphs fed on chickens no significant difference (p>0.10) was found between the feeding or premolt periods of 82 males and 62 females, but female nymphs were significantly heavier (p<0.005) than male nymphs. Sixteen engorged females (61.5% of the exposed ticks) were recovered after being fed on dogs, and all these females laid viable eggs. Chickens, the only avian host, were the most suitable host when compared with the five mammal species. Dogs were demonstrated to be a suitable host for adults of A. tigrinum, which is consistent with, several reports of adult A. tigrinum ticks parasitizing dogs in different areas of South America. Our results reinforce that in these same areas avian species are the major hosts for immature stages of this tick species. This revised version was published online in July 2006 with corrections to the Cover Date.     

Desiccation tolerance in fully developed embryos of two cicadas, Cryptotympana facialis and Graptopsaltria nigrofuscata

In two Japanese cicadas, Cryptotympana facialis and Graptopsaltria nigrofuscata , with different habitat distributions, fully developed embryos hatch in response to high humidity due to rainfall. Despite the advantage of hatching on rainy days, this trait burdens embryos with an extra period of desiccation until the unpredictable advent of rain. We compared the ability of the fully developed embryos of these cicadas to endure periods of low humidity. Eggs were exposed to a combination of different humidities (43% and 75% relative humidity, RH) and durations (0–15 days), and then transferred to an environment with 100% RH to stimulate hatching. In both species, total hatching rates decreased as duration increased, although there was no significant effect of humidity. In C. facialis , a considerable proportion of the eggs hatched during the desiccation period, and the hatching rate was higher at 75% RH than at 43% RH. After transfer to 100% RH, most hatching occurred within a day regardless of the desiccation level. In G. nigrofuscata , no nymphs hatched during the desiccation periods. However, more eggs required more than a day after transfer to 100% RH to hatch after desiccation at 43% RH than at 75% RH. Consequently, the overall proportion of timely hatching of eggs (eggs hatching within a day of moisture supply) was higher after desiccation at 43% RH in C. facialis , but it was higher after desiccation at 75% RH in G. nigrofuscata . These different physiological responses of the two species may reflect adaptation to habitat dryness.     

The importance of relative humidity and trophic resources in governing ecological niche of the invasive carabid beetle Merizodus soledadinus in the Kerguelen archipelago

Comprehensive studies to identify species-specific drivers of survival to environmental stress, reproduction, growth, and recruitment are vital to gaining a better understanding of the main ecological factors shaping species habitat distribution and dispersal routes. The present study performed a field-based assessment of habitat distribution in the invasive carabid beetle Merizodus soledadinus for the Kerguelen archipelago. The results emphasised humid habitats as a key element of the insect’s realised niche. In addition, insects faced food and water stress during dispersal events. We evaluated quantitatively how water availability and trophic resources governed the spatial distribution of this invasive predatory insect at Îles Kerguelen. Food and water stress survival durations [in 100%, 70%, and 30% relative humidity (RH) conditions] and changes in a set of primary metabolic compounds (metabolomics) were determined. Adult M. soledadinus supplied with water ad libitum were highly tolerant to prolonged starvation (LT₅₀ = 51.7 ± 6.2 d). However, food-deprived insect survival decreased rapidly in moderate (70% RH, LT₅₀ = 30.37 ± 1.39 h) and low (30% RH, LT₅₀ = 13.03 ± 0.48 h) RH conditions. Consistently, body water content decreased rapidly in insects exposed to 70% and 30% RH. Metabolic variation evidenced the effects of food deprivation in control insects (exposed to 100% RH), which exhibited a progressive decline of most glycolytic sugars and tricarboxylic acid cycle intermediates. Most metabolite levels were elevated levels during the first few hours of exposure to 30% and 70% RH. Augmented alanine and lactate levels suggested a shift to anaerobic metabolism. Simultaneously, peaks in threonine and glycolytic sugars pointed to metabolic disruption and a progressive physiological breakdown in dehydrating individuals. Overall, the results of our study indicate that the geographic distribution of M. soledadinus populations is highly dependent on habitat RH and water accessibility.     

Influence of diet on cold storage of the predator <Emphasis Type="Italic">Perillus bioculatus</Emphasis> (F.)

Cold storage of insects is a common practice used when transporting insects or to assist with efforts to increase the size of a colony. However, cold storage affects egg, nymphal and adult survival, fecundity, and egg viability. We evaluated the influence of nutrient source on the response of eggs, nymphs and adults of Perillus bioculatus when exposed to two temperatures most likely to be used for short and extended storage by comparing natural prey-fed insects to artificial diet-fed insects. The results of our study showed that the survival of eggs from either prey-fed or diet-fed insects declined at both 4 and 10°C as the length of storage exceeded one week. The survival of nymphs exposed to cold storage treatment was slightly higher than for eggs, with a similar response to time of storage. Unique to the nymphal stage was the superior performance of diet-fed insects over prey-fed insects. However, of the three developmental stages tested, the best survival was obtained with adults. Our results show that both prey-fed and diet-fed adults were able to withstand cold storage at 10°C for three weeks with very little loss of survival, fecundity or viability of eggs oviposited by those adults. Longer periods of storage resulted in a decline in survival, fecundity and egg viability and that decline was greatest at 4°C. Outcross mating of cold stored adults with adults held at 26°C indicated that adult females were more detrimentally affected by cold storage than males. There were differences in the response of P. bioculatus to cold storage compared to previously reported responses of P. maculiventris. However, the most notable difference is that P. maculiventris did better at almost all parameters than P. bioculatus. Collectively, these results demonstrate a nutrient influence on the response to cold storage at different developmental stages.