Age and growth of Benthosema pterotum (Alcock, 1890) (Myctophidae) in the Oman Sea

Juvenile and adult specimens of Benthosema pterotum (skinnycheek lanternfish) were collected during several surveys conducted on the Iranian continental shelf of the Oman Sea. Age was estimated by enumeration of growth increments in sagittae otolith sections on the assumption of their daily deposition. Three distinct growth zones in otolith microstructure (central, middle, and external) were defined. These three zones presumably represent increments deposited during successive life history stages, characterized by a different migratory behavior and depth occurrence. The number of increments in the central zone of the B. pterotum otolith (26.8 on average) was thus far one of the lowest in myctophid species studied. A negative correlation between the number of increments in the central and middle zones was observed. This might suggest a functional relation between these two periods of early life history, when fewer larvae in the epipelagic layers may be compensated by a longer non‐migratory behavior of metamorphosis larvae and early juveniles. The maximum number of growth increments in B. pterotum otoliths, i.e. 315, indicated a short lifespan of probably <1 year. The relationship between otolith length and standard length was described by linear regression model (r² = 0.902), and between the body length and weight as an isometric growth in 274 juvenile and adult specimens (r² = 0.922). The growth model estimated only for juveniles and adults was curvilinear (SL = 1.150 × Age^0.616; r² = 0.868). Back‐calculated hatch dates might suggest a spawning season of B. pterotum from May to September, but due to the limited number of investigated specimens a prolonged spawning cannot be excluded. Further trials are needed to measure the population parameter dynamics.     

Chara hispida beds as a sink of nitrogen: Evidence from growth,nitrogen uptake and decomposition

Chara hispida forms dense beds (0.78–0.95 kg DW m⁻²) in Colgada Lake. The ability of Chara meadows to act as a nitrogen source or sink was evaluated by the following methods: (1) investigating Chara growth, (2) nitrogen incorporation and decomposition laboratory experiments and (3) relating experimental results to field conditions. Sediment oospores were germinated in large aquaria and observed growth rates were 0.001 m day⁻¹ (shoot length) and 0.0002 g day⁻¹ (dry weight). Nitrogen uptake rates were determined by addition of K¹⁵NO₃ during two different periods of Chara growth and the rates were 1.21 and 3.86 μM g DW⁻¹ h⁻¹ when charophytes were 166 days old (not sexually mature) and 323 days old (sexually mature), respectively. After the uptake experiments, the same charophytes were allowed to decompose within two types of litter bags (3 mm-pore litter bags and entire, non-porous plastic litter bags). Decomposition rates of Ch. hispida were 0.016 and 0.009 day⁻¹ in perforated and non-perforated bags, respectively, and fit a negative exponential model. The nitrogen release rate, calculated as the disappearance of N content from Chara tissues, was 0.012 day⁻¹ and there were no statistically significant differences between the values from the two different bag types. The dissolved organic nitrogen concentrations in aquarium and non-perforated litter bags waters increased linearly with time due to the leaching of soluble compounds from Chara. The rate of N loss from Chara tissues, total nitrogen and dissolved organic nitrogen release rates and the decrease in initial dry weight rate were all lower than the daily rate of Chara N uptake. By extrapolating laboratory data to field situations, we determined that approximately 38% of the N taken up by charophytes in Colgada Lake during the growth period is retained. Given the high charophyte biomass in the lake, its ability to incorporate nitrogen, its low decomposition rate and its ability to over-winter, we conclude that Chara beds could be acting as nitrogen sinks in this ecosystem.     

Seasonal growth differences of larval Hyporhamphus picarti (Hemiramphidae) in the Sine Saloum estuary,Senegal

The African halfbeak Hyporhamphus picarti (Hemiramphidae) is one of the most abundant species within the ichthyoplankton community of the Sine Saloum estuary (Senegal). A year‐round occurrence of larvae suggests that the Sine Saloum is an important spawning habitat for this species. Annual fluctuations in water temperature, however, can have severe impacts on the survival probabilities of marine fish larvae. To determine whether temperature has an effect on the growth of H. picarti during its larval development, larval age at length and somatic growth rates were investigated for two contrasting spawning seasons in 2014: February (cold season, 20.8°C) and June (warm season, 26.4°C). In both months H. picarti larvae were sampled at the mouth of the Saloum River using neuston nets. Sagittal otoliths’ increments were counted to estimate the larva age at a given standard length (SL). The age of larvae ranged between 2 and 22 days, with SL of 3.86–21.68 mm, respectively. In order to describe larval age at length during the contrasting spawning seasons, two distinctive Gompertz functions were applied. Accordingly, specimens sampled in June (0.94 ± 0.17 mm per day) exhibited significantly higher somatic growth rates than those sampled in February (0.60 ± 0.06 mm per day). These findings suggest that water temperature is an important factor influencing larval growth in H. picarti. Information concerning the early life stages of H. picarti are scarce and the results of the present study may contribute to a better understanding of the species’ biology and ecology.     

Age and growth of king and Spanish mackerel larvae and juveniles from the Gulf of Mexico and U.S. South Atlantic Bight

Synopsis Sagittal otoliths from 50 king mackerel 2.9–13.0 mm SL and 72 Spanish mackerel 2.8–22.0 mm SL collected off the southeast U.S. were examined whole at 400 × using a compound microscope-video system. Otoliths of both species had visible, presumably daily, growth increments as well as finer subdaily increments. Otolith growth was directly proportional to growth in standard length for king (r² = 0.91) and Spanish mackerel (r² = 0.86). Spanish mackerel were estimated to be 3–15 d old with a mean absolute growth rate (SL/number of growth increments) and 95% confidence interval of 1.15 ± 0.07 mm · d^–1. The least squares linear equation: SL = –1.30 + 1.31 (age in days), with r² = 0.67 and p < 0.001, described the relationship between length and age. There was a significant positive relationship between absolute growth rate and fish length. King mackerel were estimated to be 3–15 d old with a mean absolute growth rate of 0.89 ± 0.06 mm d^–1. The least squares linear equation: SL = 0.37 + 0.82 (age in days), with r² = 0.77 and p < 0.001, best described the relationship between length and age. The relationship between growth rate and fish length was not significant. The growth rate of king mackerel was slightly higher for fish from the Mississippi River plume than from all other locations combined, while Spanish mackerel growth rates were not significantly different.     

Compensatory population growth in <Emphasis Type="Italic">Daphniopsis tibetana</Emphasis> Sars (Crustacea: Cladocera) following starvation

We speculated that Daphniopsis tibetana may be adapted to short-term lack of food through a mechanism of population growth compensation. D. tibetana were collected from Lake Namuka Co, Tibet, and acclimated to diluted seawater in 2007. All larvae used in experiments were synchronized newborn (<24-h old) and of an average body length of 790 (770–810) μm. We evaluated the effect of continuous starvation for 1, 3, 5, 7, 9, and 11 days and intermittent periods of starvation of 0.5, 1, 1.5, and 2 days on the compensatory population growth effect in D. tibetana. The duration of starvation significantly influenced population growth, body length, and heart rate. After 1 and 3 days of continuous starvation and 0.5 days intermittent starvation, the age of first reproduction was 16.3 days and the reproduction frequency was 0.8 (10 days^?1), which were similar to the control group. In these two starved groups, lifetime fecundity values were 426.3 and 412.3, average life spans were 54.5 and 54.3 days, and intrinsic rates of increase (r _m) were 0.136 and 0.133 days^?1, respectively, which is significantly higher than the other starved groups and not different from the control group (P > 0.05). In the group with 7, 9, and 11 days of continuously starved groups, body lengths were significantly lower (P < 0.05) than those of controls. With increasing starvation time (1, 3, 5, and 7 days), heart rate progressively decreased (184, 178, 172, and 166 min^?1). Body length, measured at the end of the experiment, declined with increasing duration of intermittent starvation. After 2 days of intermittent starvation, final heart rate sharply decreased to 150 min^?1, which was highly significantly different from the other treatment groups. The results of this study conclude that D. tibetana is capable of complete compensatory population growth after short-term starvation.     

Otolith Microstructure of Larval Gymnocypris potanini Herzenstein from the Minjiang River in China

Synopsis Otolith microstructure of about 120 Gymnocypris potanini larvae from the Minjiang River in China was examined and analyzed. Larvae had multiple primordia in most lapilli and sagittae, while had only one primordium in a few specimens. There had only one nucleus in otoliths of the larvae, except for some few specimens with 2 nuclei. The transparence of many otoliths differed from center to edge, and part of them could be divided into inner low optically dense zone (LODZ) and outer optically dense zone (ODZ). Based on increment clarity, otoliths of this species could be classified into three types, which were otolith with subtile increments, otolith with almost identified increments, and otolith with fairly clear increments characterized by high contrast. The last two types of otolith accounted for 87.07% in lapilli and 94.46% in sagittae, respectively. Increment clarity of sagitta was higher than that of lapillus. Natural checks were identified in 32.50% lapilli and 48.33% sagittae. These checks primarily located in the first to sixth increment. According to the number of increments in otoliths, the age of this batch larvae was 14 – 22 days, birth date was on June 17 – 25, and average growth rate of body length was 0.8936 ± 0.08769 mm/d.     

Larval age, growth and mortality in the oceanic squid Sthenoteuthis pteropus (Cephalopoda, Ommastrephidae) from the eastern tropical Atlantic

An investigation was carried out on larvae of the oceanic tropicalsquid Sthenoteuthis pteropus in the equatorial Atlantic (230'N–7S;12W–830'E) The age of the larvae was calculated from thestatolith microstructure of 20 larvae; mortality was estimatedfrom the size structure of 1128 larvae. The larval stage lasts32–38 days. At ages ranging from 14 to 38 days. the dailyrelative growth rates of mantle length decrease from 7.5 to2.8% day^–1 and from 14–16 to 5.8% of body weightday^–1 At age 12–24 days, mortality rates were estimatedusing both raw catch data and corrected data accounting fornet avoidance. The mean value of raw mortality rates was 0.189,the corrected value was 0.158. During the proboscis division(transformation of the larva into juvenile) at age 25–35days, a sharp decrease in larval growth rates and a simultaneousincrease in mortality rates (raw 0.443, corrected 0.379) wereobserved.     

Daily growth increments in the larval otolith of the Japanese eel,Anguilla japonica

Early formation of otolith was studied on artificially hatched larvae of the Japanese eel,Anguilla japonica. Newly hatched larvae had a pair of sagittae which were flat and subelliptical with 8.3 μm in mean diameter. The diameter of the sagitta increased linearly with age. No growth increments were observed in the sagitta at hatching, while larvae which were 2, 4 and 6 days old had on average 2.1, 3.6 and 6.0 increments, respectively. The number of the increments (Y) and the age in days after hatching (X) showed a close linear relationship (Y=0.96X+ 0.06, r = 0.913, n = 40), suggesting daily deposition of sagittal increments. In 95 % of the field-caught elvers of this species, a distinct dark ring (check) with the diameter of 6–12 μm was found around the nucleus of the sagitta. This seems to be a “hatch check” deposited at hatching, since its diameter roughly agreed with that of the sagitta in the newly-hatched larvae. Possibly, the number of the increments outside the hatch check represents the age of the fish in days.     

Growth during the early life history of the Pacific tarpon,Megalops cyprinoides

The early growth of the Pacific tarpon,Megalops cyprinoides, was studied by larval otolith analysis and rearing of larvae and juveniles in the laboratory. Morphology of the sagitta, validation of sagittal daily increments, age at the start of metamorphosis, decrement of standard length in early metamorphosis, and growth under rearing conditions are described. The sagitta of fully-grown Pacific tarpon leptocephali were transparent and circular, with regular intervals between the neighboring rings becoming wider at the onset of metamorphosis. Alizarin complexone treatment of larvae confirmed the daily formation of the sagittal rings. Metamorphosis was estimated to start about one month after hatching. After drastic shrinkage during the first several days of metamorphosis, the body length more or less stabilized for one month and then resumed rapid growth. The early growth of Pacific tarpon was divided into four phases as follows: A) leptocephalus positive growth phase; B) leptocephalus negative growth phase; C) sluggish growth phase; and D) juvenile growth phase.     

Validation of daily otolith increments in larval and juvenile Chinese sucker, <Emphasis Type="Italic">Myxocyprinus asiaticus</Emphasis>

Otolith development was observed and the formation of daily growth increments in otoliths of Chinese sucker, Myxocyprinus asiaticus, was validated by monitoring known-age larvae and juveniles in the laboratory from 2003 to 2005. Otolith shape changed with larval and juvenile development, and there was an exponential relationship until a body length of 16 mm or so, and a linear relationship after a body length of 16 mm between otolith size and fish size. The first increment was identified in larvae 1 day after hatching. The regressed equations between daily age (D) and increment number in otoliths (N) were N = −0.64 + 0.96D in lapillus, and N = −0.31 + 0.98D in sagitta. The slopes were not significantly different than 1.0. This demonstrated that otolith increments in this species were formed daily and can be used for daily age determination.     

Effect of dietary chitosan on challenged <Emphasis Type="Italic">Dicentrarchus labrax</Emphasis> post larvae with <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

Effect of different dietary squilla chitosan (Csq) concentrations: 0 (control), 0.5, 1 and 2 g 100 g^–1 diets were studied for weaned sea bass (Dicentrarchus labrax) post larvae. Post larvae were challenged with Aeromonas hydrophila after 5 feeding days, in order to monitor the prophylactic effect on the Csq fed larvae. The experiment started with an average initial weight of 50 ± 2 mg and total length of 12 ± 2 mm for post larval stage (40 days post hatch; dph), then continued feeding diets for a period of 20 days. Larvae survival percentage (%), mean total length (TL), width (W), total weight (TW), total weight gain (TWG), average daily weight (ADW) and specific growth rate (SGR) were recorded as morphometric measurements representing growth compared to the control groups. The results revealed that 1g Csq 100 g^–1 diet at P < 0.05 was the most effective concentration that achieved higher survival percentages; 94.5 ± 0.5 and 74 ± 2.0%, increasing the specific growth rate by 7.22% and 5.77% for non challenged and challenged weaned larval groups, respectively. Otherwise, the control challenged group displayed the lowest performance in all assayed parameters with the coincidental decrease in the survival % and specific growth rates. Similarly, lower growth performance was also observed at 2 g 100 g^–1 diet. Thus, the incorporation of chitosan at a level of 1g in fish diet enhanced the performance and reduced the fish mortality under stress conditions.     

Multi-Level Effects of Low Dose Rate Ionizing Radiation on Southern Toad,Anaxyrus [Bufo] terrestris

Despite their potential vulnerability to contaminants from exposure at multiple life stages, amphibians are one of the least studied groups of vertebrates in ecotoxicology, and research on radiation effects in amphibians is scarce. We used multiple endpoints to assess the radiosensitivity of the southern toad (Anaxyrus [Bufo] terrestris) during its pre-terrestrial stages of development –embryonic, larval, and metamorphic. Toads were exposed, from several hours after oviposition through metamorphosis (up to 77 days later), to four low dose rates of ¹³⁷Cs at 0.13, 2.4, 21, and 222 mGy d^-1, resulting in total doses up to 15.8 Gy. Radiation treatments did not affect hatching success of embryos, larval survival, or the length of the larval period. The individual family variation in hatching success of embryos was larger than the radiation response. In contrast, newly metamorphosed individuals from the higher dose-rate treatments had higher mass and mass/length body indices, a measure which may relate to higher post-metamorphic survival. The increased mass and index at higher dose rates may indicate that the chronic, low dose rate radiation exposures triggered secondary responses. Additionally, the increases in growth were linked to a decrease in DNA damage (as measured by the Comet Assay) in red blood cells at a dose rate of 21 mGy d^-1 and a total dose of 1.1 Gy. In conclusion, the complex effects of low dose rates of ionizing radiation may trigger growth and cellular repair mechanisms in amphibian larvae.     

Do otolith increments allow correct inferences about age and growth of coral reef fishes?

Otolith increment structure is widely used to estimate age and growth of marine fishes. Here, I test the accuracy of the long-term otolith increment analysis of the lemon damselfish Pomacentrus moluccensis to describe age and growth characteristics. I compare the number of putative annual otolith increments (as a proxy for actual age) and widths of these increments (as proxies for somatic growth) with actual tagged fish-length data, based on a 6-year dataset, the longest time course for a coral reef fish. Estimated age from otoliths corresponded closely with actual age in all cases, confirming annual increment formation. However, otolith increment widths were poor proxies for actual growth in length [linear regression r ² = 0.44–0.90, n = 6 fish] and were clearly of limited value in estimating annual growth. Up to 60 % of the annual growth variation was missed using otolith increments, suggesting the long-term back calculations of otolith growth characteristics of reef fish populations should be interpreted with caution.     

Assessing the validity of stylets as ageing tools in Octopus pallidus

A method to age octopus was determined through the quantification of growth increments within the stylet structure (highly reduced internal shells). To validate the periodicity of the increments, stylets were analysed from known-age laboratory-raised Octopus pallidus. The animals, which ranged from 3 to 8 months old, were exposed either to a simulated natural or to constant temperature regime. Transverse stylet sections were embedded in thermo-plastic cement and ground until a thin section was achieved. A pre-hatch region and first post-hatch increment was identified in the stylet microstructure. The number of increments, across all ages and temperature treatments, was in remarkably close agreement to age (number of days), clearly demonstrating that stylet increments are deposited daily throughout the lifecycle of O. pallidus. Morphometric analyses of the stylet indicated that increments were laid down regularly during stylet growth and that stylets grew in concert with body size, further supporting daily periodicity. This study successfully validates daily increments in stylets and thus demonstrate a method to age octopus, therefore, potentially having critical implications for future octopus research and the effective management of stocks worldwide.     

Egg size and the evolution of phenotypic plasticity in larvae of the echinoid genus Strongylocentrotus

Planktotrophic larvae grow by utilizing energy obtained from food gathered in the plankton. Morphological plasticity of feeding structures has been demonstrated in multiple phyla, in which food-limited larvae increase feeding structure size to increase feeding rates. However, before larvae can feed exogenously they depend largely on material contained within the egg to build larval structures and to fuel larval metabolism. Thus, the capacity for plasticity of feeding structures early in development may depend on egg size. Using the congeneric sea urchins Strongylocentrotus franciscanus and S. purpuratus, which differ in egg volume by 5-fold, I tested whether the degree of expression of feeding structure (larval arm length) plasticity is correlated with differences in the size of the egg. I experimentally manipulated egg size of S. franciscanus (the larger-egged species) by separating blastomeres at the 2-cell stage to produce half-sized larvae. I reared half-size and normal-size larvae under high and low food treatments for 20 days. I measured arm and body lengths at multiple ages during development and calculated the degree of plasticity expressed by larvae from all treatments. Control and unmanipulated S. franciscanus larvae (from ∼ 1.0 nl eggs) had significantly longer arms relative to body size and a significantly greater degree of plasticity than half-sized S. franciscanus larvae (from < 0.18 nl eggs), which in turn expressed a significantly greater degree of plasticity than S. purpuratus larvae (from ∼ 0.3 nl eggs). These results indicate that egg size affects larval arm length plasticity in the genus Strongylocentrotus; larger eggs produce more-plastic larvae both in an experimental and a comparative context. However, changes in egg size alone are not sufficient to account for evolved differences in the pattern of plasticity expressed by each species over time and may not be sufficient for the evolutionary transition from feeding to non-feeding.     

Effects of aeration rate and feed on growth,productivity and nutrient composition of black soldier fly (Hermetia illucens L.) larvae

Black soldier fly larvae (Hermetia illucens L.) are widely cultivated as bioconversion agents. These larvae are known to be highly sensitive to changes in the external environment, such as temperature and relative humidity. Therefore, a rearing hive equipped with an instrumentation system known as the Modular Fly Hive was designed to cultivate black soldier larvae. This study aimed to determine the effects of aeration rate (0 – 0.84 m³/s.) and feed type (coconut endosperm waste and soybean curd) on growth and productivity of black soldier fly cultivated in Modular Fly Hives as well as well as feed digestibility and nutrient composition of the larval biomass. Increasing aeration rate decreased the average temperature and relative humidity within the hives. The results showed that the larvae demonstrated a sigmoidal growth curve for all types of feed and the type of feed significantly affects the average wet weight and length of the larvae. The greatest dry biomass productivity (52.85 ± 3.85 g/m²/day) was observed when the larvae were cultivated using soybean curd residue with an aeration rate of 0.84 m³/s, while the lowest dry biomass productivity (35.27 ± 9.72 g/m²/day) was observed when the larvae were cultivated using coconut endosperm waste without aeration. Proximate analysis revealed that the larval biomass had total protein, lipid, ash and carbohydrate content in the range of 37.20 – 48.60%, 9.61 – 20.02%, 4.80 – 6.40%, 33.86 – 38.70%, respectively. Amino acid in the biomass were dominated by glutamic acid (11.11–12.30%), aspartic acid (8.25 – 10.35%), leucine (8.09 – 8.57%), and lysine (6.74 – 8.14%). Lipid isolated from the larval biomass were mainly composed of lauric acid (28.35–61.68%), linoleic acid (6.27 – 30.29%), palmitic acid (7.62 – 15.23%), and myristic acid (5.05 – 14.34%).     

Feeding and growth kinetics of the planktotrophic larvae of the spionid polychaete Polydora ciliata (Johnston)

We studied the effect of food concentration on the feeding and growth rates of different larval developmental stages of the spionid polychaete Polydora ciliata. We estimated larval feeding rates as a function of food abundance by incubation experiments with two different preys, presented separately, the cryptophyte Rhodomonas salina (ESD = 9.7 µm) and the diatom T.weissflogii (ESD = 12.9 µm). Additionally, we determined larval growth rates and gross growth efficiencies (GGE) as a function of R. salina concentration.P.ciliata larvae exhibited a type II functional response. Clearance rates decreased continuously with increasing food concentration, and ingestion rates increased up to a food saturation concentration above which ingestion remained fairly constant. The food concentration at which feeding became saturated varied depending on the food type, from ca. 2 µg C mL^− 1 when feeding on T. weissflogii to ca. 5 µg C mL^− 1 when feeding on R. salina. The maximum carbon specific ingestion rates were very similar for both prey types and decreased with increasing larval size/age, from 0.67 d^− 1 for early larvae to 0.45 d^− 1 for late stage larvae. Growth rates as a function of food concentration (R. salina) followed a saturation curve; the maximum specific growth rate decreased slightly during larval development from 0.22 to 0.17 d^− 1. Maximum growth rates were reached at food concentrations ranging from 2.5 to 1.4 µg C mL^− 1 depending on larval size. The GGE, estimated as the slope of the regression equations relating specific growth rates versus specific ingestion rates, were 0.29 and 0.16 for early and intermediate larvae, respectively. The GGE, calculated specifically for each food level, decreased as the food concentration increased, from 0.53 to 0.33 for early larvae and from 0.27 to 0.20 for intermediate larval stages.From an ecological perspective, we suggest that there is a trade-off between larval feeding/growth kinetics and larval dispersal. Natural selection may favor that some meroplanktonic larvae, such as P.ciliata, present low filtration efficiency and low growth rates despite inhabiting environments with high food availability. This larval performance allows a planktonic development sufficiently long to ensure efficient larval dispersion.     

Growth and otolith microstructure of cod (Gadus morhua L.) larvae

Cod larvae from Irish Sea stocks were reared under differentgrowth conditions, and the otolith growth and increment formationexamined in sagittae and lapilli. Otolith increments were firstdeposited around the time of hatching and increment counts,on average, reflected larval age. The growth rate of fish larvaereared in different sized tanks was significantly different(P < 0.001), but there was no detectable effect on incrementformation. Otolith size was independent of larval size for individuals<5 mm in length. In larvae >6 mm, larger, faster growingindividuals had larger and faster growing otoliths.     

Verification of daily growth increment formation in saury otoliths by rearing larvae from hatching

Naturally spawned eggs of the Pacific saury,Cololabis saira, were collected in the field and reared in a tank to examine daily periodicity of growth increment formation in the otolith. Larvae were 6.9 mm in knob length at hatching. Their otoliths (sagittae) were 31 μm in radius and had 3–6 faint concentric rings. They started feeding within two days and grew at a rate of 1.1 mm/day on average through larval and juvenile stages feeding on rotifers,Artemia nauplii, and artificial diets. Otolith growth increments showed a concentric pattern with a distance of 3.5–5.0 μm between two adjacent increments. The number of growth increments was almost equal to a known age in days plus 4 or 5. A regression line of number of increments (N) on known age in days (D) between 0–30 days after hatching was N = 4.81 + 1.01D, which shows that one increment was deposited per day.     

A novel cysteine proteinase inhibitor from seeds of Enterolobium contortisiliquum and its effect on Callosobruchus maculatus larvae

This study focused on the characterization of a novel cysteine proteinase inhibitor from Enterolobium contortisiliquum seeds targeting the inhibition of the growth of Callosobruchus maculatus larvae, an important cosmopolitan pest of the cowpea Vigna unguiculata during storage. The inhibitor was isolated by ion-exchange besides of size exclusion chromatography. EcCI molecular mass is 19,757 Da, composed of two polypeptide chains. It strongly inhibits papain (Ki_app 0.036 nM) and proteinases from the midguts of C. maculatus (80 μg mL^?1, 60% inhibition). The inhibitory activity is reduced by 40% after a heat treatment at 100 °C for 2 h. The protein displayed noxious activity at 0.5% and 1% (w/w) when incorporated in artificial seeds, reducing larval mass in 87% and 92%, respectively. Treatment of C. maculatus larvae with conjugated EcCI-FIT and subsequent biodistribution resulted in high fluorescence intensity in midguts and markedly low intensity in malpighian tubules and fat body. Small amounts of labeled proteins were detected in larvae feces. The detection of high fluorescence in larvae midguts and low fluorescence in their feces indicate the retention of the FITC conjugated EcCI inhibitor in larvae midguts. These results demonstrate the potential of the natural protein from E. contortisiliquum to inhibit the development of C. maculatus.