Weitere Untersuchungen über Chromosomenverhältnisse und DNS-Gehalt bei Anuren (Amphibia)

The karyotypes of the toad Bufo marinus L. (2n=22) and the frogs Limnodynastes tasmaniensis Gthr. (2n=24), Rana temporaria L., R. esculenta L. (both 2n=26) and R. arvalis Nills. (2n=24) were analysed in colchicine treated leukocyte and spermatogonial metaphases and/or embryonic and larval mitoses. The DNA content of Feulgen stained erythrocyte nuclei was measured microspectrophotometrically. Heteromorphic sex chromosomes are absent in all species. L. tasmaniensis has the lowest DNA content among these species. The south American toad B. marinus shows a karyotype similar to the other known toad species and contains the same amount of DNA as the European species B. calamita with the lowest DNA amount among the European toads. In southern German populations of R. temporaria besides animals with the “standard”-karyotype (2n=26) individuals with 1 or 2, in rare cases with 3 or 4 supernumerary chromosomes have been found. The supernumeraries are heterochromatic and smaller than the smallest chromosome of the “standard”-karyotype. If only 1 or 2 supernumerary chromosomes are present, they seem to show normal mendelian inheritance as a rule. The observation of a few tadpoles with intraindividual different numbers of supernumeraries points to the occurrence of unequal distribution of these chromosomes in individuals containing a higher number of supernumerary chromosomes. The karyotype of R. esculenta is very similar to the “standard”-karyotype of R. temporaria, but the chromosomes of R. esculenta are somewhat longer than those of R. temporaria. R. esculenta contains about 54% more DNA than R. temporaria in the erythrocyte nuclei, so that it must be assumed that all chromosomes of R. esculenta contain more DNA than their homologues in R. temporaria. R. arvalis possesses about 28% more DNA than R. temporaria. It is supposed that these interspecific differences in DNA content of the Rana species — as observed earlier in Bufo species — are not a consequence of differential polyteny but are caused during evolutionary processes by local increase in DNA in the chromosomes of R. esculenta and R. arvalis.     

The influence of anthropogenic factors on reproduction of <Emphasis Type="Italic">Rana temporaria</Emphasis> and <Emphasis Type="Italic">Rana arvalis</Emphasis>

Studies of spawning water bodies in the city of Moscow have shown that the urban populations of common (Rana temporaria) and moor (R. arvalis) frogs are small compared with suburban populations, and their individuals lead a hidden mode of life. The recorded increase in the fecundity of females from several populations of the city of Moscow may be accompanied by an increase or the preservation of the diameter of eggs as compared with the same indicator for suburban populations. The populations in which the females produced many small eggs died out during the study period. The most prosperous populations are urban populations of brown frogs whose females spawned eggs of various sizes. We consider the formation of these clutches as a manifestation of the bet-hedging strategy compensating for mortality in adverse and unstable environmental conditions.     

Age,growth and reproduction of the Iranian goby, <Emphasis Type="Italic">Ponticola iranicus</Emphasis> (Perciformes: Gobiidae) from the southern Caspian Sea basin

To estimate reproductive biology of Ponticola iranicus Vasil’eva et al., 2015, in Sefidroud River in the southern Caspian Sea basin age, sex ratio, fecundity, oocytes diameter, gonadosomatic, modified gonadosomatic and dobriyal indices were measured. Regression analyses were used to find relations between fecundity and fish size (length and weight), gonad weight, and age. Mature females and males were longer than 46 and 49 mm in total length, +1 in age. The Von Bertalanffy growth models were L _∞ = 23.00 cm (TL), K = 0.21, t ₀ =–0.25 for all specimens. Sex ratio was 1: 1.32 in favor of males. The average egg diameter was 0.96 mm. The spawning period extended from late February to late April. The averages of absolute and relative fecundity were calculated as 536.9 and 81.3, respectively. The absolute fecundity was significantly related to the body length and weight, as well as gonad weight.     

Intra-specific variation of sperm length in the malaria vector <Emphasis Type="Italic">Anopheles gambiae</Emphasis>: males with shorter sperm have higher reproductive success

Background

Intra-specific variation in sperm length influences male reproductive success in several species of insects. In males of the malaria vector Anopheles gambiae, sperm length is highly variable but the significance of this variation is unknown. Understanding what determines the reproductive success of male mosquitoes is critical for controlling malaria, and in particular for replacing natural populations with transgenic, malaria-resistant mosquitoes.

Methods

A laboratory population of A. gambiae males was tested for intra-specific variation in sperm length. A full-sib quantitative genetic design was used to test for a genetic component of sperm length in A. gambiae males and estimate its heritability. This study also tested for a relationship between sperm length and male reproductive success in A. gambiae. Male reproductive success was measured as the proportions of inseminated and ovipositing females.

Results

There was intra-specific variation of sperm length in A. gambiae. There was no significant genetic variation in sperm length and its heritability was low (h² = 0.18) compared to other insects. Sperm length was correlated with male body size (measured as wing length). Males with short sperm had significantly higher reproductive success than males with long sperm and this was independent of body size.

Conclusion

This is the first study to demonstrate intra-specific variation in sperm length in A. gambiae and that males with short sperm have higher reproductive success. That sperm length influences female oviposition is important for any strategy considering the release of transgenic males.

     

Nucleolar organizing regions (NORs) of common vole chromosomes as nuclear markers of genome differentiation in data from a hybrid zone of two karyoforms, <Emphasis Type="Italic">arvalis</Emphasis> and <Emphasis Type="Italic">obscurus</Emphasis>

Contrasting chromosome location of nucleolar organizing regions (NORs), a poorly studied differentiating feature of two karyoforms of the 46-chromosome common voles, arvalis and obscurus, is demonstrated in the heterozygous karyotype of a natural hybrid from the arvalis/obscurus contact zone in European Russia. It was deduced from the karyotype analysis that the obscurus form was parental in this case and the arvalis form was defined as maternal via the markers of the mitochondrial genome. The similarities in the levels of chromosome polytypy and the lineage differentiation of the arvalis/obscurus and obscurus mitochondrial genomes according to the new and earlier published data is noticeable.     

Reproductive Behavior of the Bug <Emphasis Type="Italic">Molipteryx fuliginosa</Emphasis> Uhler (Heteroptera,Coreidae) in the South of the Russian Far East

Reproductive behavior of Molipteryx fuliginosa (Uhler) was investigated in Primorskii Territory of Russia. From 4 to 18 repeated copulations of one female lasting from 2 to 48 hours were recorded in cages. The behavior of ovipositing females and the stages of oviposition are described for the first time. The number of eggs laid between copulations varied from 1 to 13, the number of oviposition acts, from 4 to 11, and the total female fecundity, from 21 to 38 eggs. Caged females laid eggs on plants and also on dead substrates unsuitable for nymphal feeding, such as cloth, dry branches, and a wooden pole. Copulation of M. fuliginosa was also observed under natural conditions. The preferred mating places of M. fuliginosa in anthropogenically modified habitats and in small-leaved riparian forests were plants of Rubus idaeus L., R. caesius L., and Rubus sp. After mating, females migrated in search of places for oviposition. Single eggs were found on the following plants not known previously as hosts of this bug: Solanum lycopersicum L., Carex sp., Elytrigia repens (L.) Nevski, and Taraxacum officinale Wigg. The females seemed to lack selectivity in the choice of place for oviposition, which was not always associated with host plants, despite their abundance and availability.     

Nutritional symbionts of a putative vector, <Emphasis Type="Italic">Xyleborus bispinatus</Emphasis>, of the laurel wilt pathogen of avocado, <Emphasis Type="Italic">Raffaelea lauricola</Emphasis>

Ambrosia beetles subsist on fungal symbionts that they carry to, and cultivate in, their natal galleries. These symbionts are usually saprobes, but some are phytopathogens. Very few ambrosial symbioses have been studied closely, and little is known about roles that phytopathogenic symbionts play in the life cycles of these beetles. One of the latter symbionts, Raffaelea lauricola, causes laurel wilt of avocado, Persea americana, but its original ambrosia beetle partner, Xyleborus glabratus, plays an uncertain role in this pathosystem. We examined the response of a putative, alternative vector of R. lauricola, Xyleborus bispinatus, to artificial diets of R. lauricola and other ambrosia fungi. Newly eclosed, unfertilized females of X. bispinatus were reared in no-choice assays on one of five different symbionts or no symbiont. Xyleborus bispinatus developed successfully on R. lauricola, R. arxii, R. subalba and R. subfusca, all of which had been previously recovered from field-collected females of X. bispinatus. However, no development was observed in the absence of a symbiont or on another symbiont, Ambrosiella roeperi, recovered from another ambrosia beetle, Xylosandrus crassiusculus. In the no-choice assays, mycangia of foundress females of X. bispinatus harbored significant colony-forming units of, and natal galleries that they produced were colonized with, the respective Raffaelea symbionts; with each of these fungi, reproduction, fecundity and survival of the beetle were positively impacted. However, no fungus was recovered from, and reproduction did not occur on, the A. roeperi and no symbiont diets. These results highlight the flexible nature of the ambrosial symbiosis, which for X. bispinatus includes a fungus with which it has no evolutionary history. Although the “primary” symbiont of the neotropical X. bispinatus is unclear, it is not the Asian R. lauricola.     

Fecundity of the tropical catadromous eels <Emphasis Type="Italic">Anguilla bicolor bicolor</Emphasis>, <Emphasis Type="Italic">A. bengalensis bengalensis</Emphasis> and <Emphasis Type="Italic">A. marmorata</Emphasis>

Maturation is one of the most important ontogenetic transitions in an individual’s life. However, the reproductive ecology of the tropical anguillid eel genus Anguilla at the onset of oceanic spawning migration is poorly understood. To understand the reproductive ecology, the fecundity of the tropical eels Anguilla bicolor bicolor, A. bengalensis bengalensis and A. marmorata was examined using advanced migrating silver eels (Stage IV and V). A close linear relationship was found between total length and fecundity in A. bengalensis bengalensis. The fecundities of A. bicolor bicolor (0.55 to 4.96 × 10⁶), A. bengalensis bengalensis (0.33–1.72 × 10⁶) and A. marmorata (0.99 × 10⁶) were within the range of those observed in temperate eels.     

Life history traits of two dwarf cichlids species in the white waters of the Amazonian floodplain

The reproductive parameters are among the most important life history aspects of fishes influenced by environmental variation. During recent years, the main life history strategies of Amazonian fish species were defined mostly by a set of reproductive parameters. In the present work, we sought to describe important life history parameters, in particular on reproductive characteristics of Apistogramma agassizii and Apistogramma bitaeniata, found in floodplain lakes of the Brazilian Amazonia. The species presented a positive sexual dimorphism, and males were significantly bigger than females. For both sexes, four developmental phase of gonad maturation were detected, and based on those it was possible to identify mature, reproductive specimens throughout the entire period of the study. From the ovaries of mature females, fecundity and spawning type were determined. Low fecundity, short spawning periods, possibly separated only by few months, and total spawning are all good indications that A. agassizii and A. bitaeniata evolved an opportunistic strategy in their life history.     

Population Dynamic of the Mudskipper <Emphasis Type="Italic">Periophthalmus waltoni</Emphasis> Koumans, 1941 from the Bay of Hormozgan Province,Persian Gulf

We studied the population structure and reproductive biology of the mudskipper Periophthalmus waltoni Koumans, 1941, living on the mudflats of bay of Hormozgan province, Persian Gulf. The specimens were haphazardly and monthly collected from August 2008 to July 2009. A total of 2060 fishes was sampled, sexed 690 of which 221 (32%) were males, and 469 (68%) females. The total length (TL) and weight relationship demonstrated growth in both sexes was isometric with the function W = 0.0078TL^3.06. The asymptotic length (L∞) and growth coefficient (K) were estimated as 16.5 cm and 0.75 yr^–1, respectively. Age of the fish at zero length (to) was calculated–0.35 and the longevity was attained 3.65 year. The estimated Von Bertalanffy function was TL_(t) = 16.5[1–exp(–0.75(t + 0.35))]. Natural mortality (M) was estimated 1.74 yr^–1. Four cohorts were distinguished annuallywith mean of TL = 7.35, 10.17, 12.74 and 13.92 cm. The maximum recruitment rate was in January at 17.5%. The overall sex ratio (F: M) (1: 0.49) was significantly different than 1: 1. The peak of the gonadosomatic index (GSI) occurred in February. The size at onset of sexual maturity (LM₅₀)wasestimated at TL = 12 cm. The mean of absolute and relative fecundity were estimated as 3558 ± 2203 and 257 ± 101 eggs, respectively.     

The influence of temperature,photoperiod, and diet on development and reproduction in the four-spot lady beetle <Emphasis Type="Italic">Harmonia quadripunctata</Emphasis> (Pontoppidan) (Coleoptera,Coccinellidae)

This is the first report on the effects of temperature (from 17 to 30°C), photoperiod (from 10 to 16 h of light), and diet (the wheat aphid Schizaphis graminum and the green peach aphid Myzus persicae) on preimaginal developmental rates, adult body mass, preoviposition period, and female fecundity of the predatory lady beetle Harmonia quadripunctata (Pontoppidan) under the laboratory conditions. A constant temperature of 30°C was lethal to the embryos. Judging from the data obtained at 17, 20, and 25°C, the lower temperature threshold for total egg-to-adult development equaled 12.0°C and the respective sum of effective temperatures was 274 degree-days. Preimaginal development was somewhat faster under short-day conditions, the threshold for this quantitative photoperiodic response being approximately 14 h at the rearing temperatures of 20 and 25°C. At a temperature of 25°C, the mean body mass of newly emerged adults was greater for those individuals that during the larval stage had been fed on the green peach aphid (16.4 mg in males and 18.4 mg in females) than in those fed with the wheat aphid (13.8 mg in males and 15.3 mg in females). The preoviposition period (measured from adult emergence to the first egg laid) also depended on temperature and had a lower threshold of 13.4°C and a sum of effective temperatures of 152 degree-days. Maximum fecundity was observed at 25°C and 16-h day length; under these conditions, the oviposition period lasted over 100 d, and the average lifetime fecundity was about 800 eggs per female. Under short-day conditions, females of H. quadripunctata entered reproductive diapause. The photoperiodic threshold for this qualitative response at the temperatures of 20 and 25°C was about 14 h when fed on the peach aphid and 15 h when fed on the wheat aphid. Relatively small body size, low fecundity, and a strong photoperiodic response that hinders rapid adaptation to novel climates probably explain the fact that H. quadripunctata, in contrast to H. axyridis, has not become an aggressive invader.     

Autecology of a new species of carnivorous marsupial,the endangered black-tailed dusky antechinus (<Emphasis Type="Italic">Antechinus arktos</Emphasis>), compared to a sympatric congener,the brown antechinus (<Emphasis Type="Italic">Antechinus stuartii</Emphasis>)

The carnivorous marsupial Antechinus are one of the few mammal genera known to exhibit the phenomenon of semelparous reproduction, where all males die at the end of a frenetic annual mating period. The genus Antechinus has recently been revised, seeing four new species named and one existing subspecies raised to species status. Here, we present the first ecological assessment of one new species, the endangered black-tailed dusky antechinus (Antechinus arktos), based on a 2-year mark-recapture study of two proximate sites within the cloud forest of Springbrook National Park, south-east Queensland, Australia. We also present comparative ecological data from a sympatric congener, Antechinus stuartii. In total, 103 A. arktos (49 male; 54 female) and 2125 A. stuartii (1229 male; 896 female) captures were made over 16,630 trap nights at the two sites. The occurrence and synchrony of reproductive events observed in A. arktos closely parallel the semelparous reproductive strategy exhibited by all congeners studied to date. The A. arktos populations mate during mid-September, with birth of up to six young occurring during mid-October. The low abundance and fecundity, together with a limited and apparently retracting distribution, suggest A. arktos is under threat of climate-induced extinction in the coming decades.     

Life history strategies of the Scotia Sea icefish, <Emphasis Type="Italic">Chaenocephalus aceratus</Emphasis>, along the Southern Scotia Ridge

Reproductive capacity can influence distribution and abundance over large spatial scales through larval dispersal, even when adult stages remain isolated following settlement. We examined size distribution, reproductive traits and age structure in Scotia Sea icefish, Chaenocephalus aceratus, an abundant benthic species with a long larval pelagic phase found on continental shelves along the Southern Scotia Ridge. In particular, we compared life history strategies between fish caught during surveys undertaken off the South Orkney Islands (SOI) and South Shetland Islands (SSI). Results corroborated regional separation after settlement and suggested distinct life history strategies, in which fish from SOI invested much less in reproduction, and somewhat more in somatic growth earlier in their life history. Compared to SSI, body weight increased faster with length and absolute fecundity was 46 % lower and increased more slowly with size for SOI population. In addition, the proportion of spawning cohorts and L _∞ was lower and k higher for SOI. The differences appeared to be a phenotypic response to environmental conditions related to regional hydrography. Lower reproductive capacity around the SOI, and strong eastward flow in the large-scale circulation, suggests that the SSI may be more important in influencing distributions and abundance of icefish along the Southern Scotia Ridge.     

The Influence of Sire Presence on Survival and the Growth Rate of Juvenile Common Voles <Emphasis Type="Italic">Microtus arvalis</Emphasis>

Pup survival and growth rates were monitored to determine whether paternal care potentially contributes to reproductive success of male common voles Microtus arvalis Pallas, 1779. The subjects were 258 pups from 47 litters obtained from adult voles trapped in the wild. Reproducing pairs were formed and divided onto two groups in accordance with the age and parental experience of the reproducing individuals: group I included pairs with subadult males and nulliparous females; group II included pairs with adult males and multiparous females. Each of the groups was divided into two subgroups: in subgroups Iа (n = 13) and IIа (n= 11), litters were raised by females without sires; in subgroups Ib (n = 12) and IIb (n = 11), litters were raised by both parents. The litter size was recorded at parturition and again when the young were 12, 21, and 30 days old. Pups in each litter were weighed at parturition and also again when the young were 12, 21, and 30 days old. This study has shown that the presence of the sire may negatively affect both survival and growth rates of the pups, but this effect is dependent on the age of the reproducing individuals.     

Morphofunctional characterization of the reproductive system of the gastropods <Emphasis Type="Italic">Littorina brevicula,L. mandshurica</Emphasis>, and <Emphasis Type="Italic">Nucella heyseana</Emphasis> from uncontaminated and contaminated areas of Peter the Great Bay,Sea of Japan

The structure and state of the reproductive system were studied in the periwinkles Littorina brevicula and L. mandshurica from the coastal area of Vladivostok and in the dogwhelk Nucella heyseana from the protected area of Vostok Bay and from contaminated sites in Amursky Bay. These species were investigated for the presence of imposex and intersex (pathological changes caused by endocrine disruption under the influence of toxic substances). The pathological changes revealed in the gonads of Littorina were oocyte resorption and lipofuscin accumulation; an intersex condition was not found. Imposex specimens (with penis formation) were found among Nucella females from Amursky Bay. The relative penis length in females was 25% of the penis length in males. This kind of pathology was not found in Nucella from clean waters of Vostok Bay.     

Reproductive biology of <Emphasis Type="Italic">Astyanax lacustris</Emphasis> (Characiformes: Characidae) in the southern Pantanal floodplain,upper Paraguay River basin,Brazil

We studied the reproductive biology of Astyanax lacustris in the southern Pantanal, Brazil. A total of 382 fish were analyzed from February 2009 to January 2011. The sex ratio differed significantly from 1:1, with a larger proportion of females (χ² = 27.24; p < 0.001). Females achieve higher standard length than males (mean 72.30 mm and 56.51 mm, respectively), and possess negative allometric growth while males have positive allometric growth. Size at first maturation did not differ between sexes. The greatest reproductive activity occurred during the rainy season, when temperatures were highest (November to February). Total fecundity was significantly correlated with standard length (r_s = 0.74; p = 0.01), and spawn type was total. Then, the life history traits of A. lacustris are adjusted to seasonal variation in the Southern Pantanal.     

Dominance of a <Emphasis Type="Italic">Rhizopogon</Emphasis> sister species corresponds to forest age structure

Rhizopogon vesiculosus and Rhizopogon vinicolor are sister species of ectomycorrhizal fungi that associate exclusively with Douglas-fir (DF). They form tuberculate mycorrhizas and they can be easily distinguished using molecular tools. We are not aware of studies relating their relative abundance in forests with different age classes. Our objective was to determine whether a change in the number or relative abundance of R. vesiculosus and R. vinicolor tubercules and genotypes was related to a change in the percent of DF in a regenerating phase (<50 years old). R. vesiculosus and R. vinicolor were located by excavating tuberculate mycorrhizas from the forest floor. A DNA Alu1 digest was used to distinguish between the two species. Microsatellite markers were used to identify genotypes. The number of R. vesiculosus tubercules correlated positively with an increasing proportion of DF in a regenerating phase, while the number of R. vinicolor tubercules was similar across all forest age structures. The number of R. vesiculosus genotypes did not correlate with forest age structure, whereas the number of R. vinicolor genotypes showed a negative relationship with an increasing proportion of DF in a regenerating phase. When the numbers of R. vesiculosus tubercules and genotypes were expressed as a relative abundance of the two species, there was a positive correlation with an increasing proportion of DF in a regenerating phase for both genotypes and tubercules. Our results suggest that the degree of DF regeneration or ecosystem factors related to DF regeneration affect the population dynamics of R. vesiculosus and R. vinicolor differently.     

Reproductive interference between two serious pests,oriental fruit flies <Emphasis Type="Italic">Bactrocera carambolae</Emphasis> and <Emphasis Type="Italic">B. dorsalis</Emphasis> (Diptera: Tephritidae), with very wide but partially overlapping host ranges

Bactrocera carambolae and B. dorsalis (Diptera: Tephritidae) are extremely destructive pests of fruits and vegetables in the Asia-Pacific region. Earlier reports have described that B. carambolae and B. dorsalis, respectively, use mainly star fruit and mango, suggesting a certain level of host partitioning which can be ascribed neither to differences in larval food qualities nor host-specific parasitoid mortality. This study specifically examined reproductive interference (antagonistic sexual interaction) between B. carambolae and B. dorsalis as a potential factor strongly affecting their host partitioning. We observed mating behaviors, especially interspecific courtships and mating, by cohabiting the conspecific and heterospecific pairs together. Consequently, we quantified their effects on the reproductive success of females. Males of both species frequently courted their own females, but they also courted females of other species. Courtship refusal by females was not selective in males of either species. This incomplete discrimination of both sexes led to frequent occurrences of interspecific sexual interactions in both species, but only B. carambolae females showed reduced reproductive success. These results suggest that B. dorsalis, superior in reproductive interference, can occupy high-quality mango, whereas B. carambolae, inferior in reproductive interference, must use low-quality star fruit.     

Mating system in a population of the wasp <Emphasis Type="Italic">Polistes dominulus</Emphasis> (Christ) (Hymenoptera,Vespidae)

The specific features of mating females and males of different morphs collected from a population of the wasp P. dominulus in Southern Ukraine were studied. Data analysis of mating individuals in experiments indicated a random mate choice by size and assortative mating by coloration. The most actively copulating males in the experiments had light-colored mesopleuron, sternite, and fore and middle coxae. They were similar in the color pattern and size to resident males, which defend their territories in the habitat. Most P. dominulus females mated only once. In their coloration, those females represented the majority of the individuals in the population. The color variability of the foundresses was shown to change in a cyclic pattern. The possibility that such variability may result from selective or assortative mating is discussed. The color and pattern traits of P. dominulus females and males could serve as markers of their reproductive strategies and their reproductive success on the whole.     

Complex effects of Ayurvedic formulation: <Emphasis Type="Italic">Guduchi</Emphasis> and <Emphasis Type="Italic">Madhuyashti</Emphasis> on different components of life history may elude the elixir effect

Formulations from the traditional Indian medicine, Ayurveda, have long been considered to have potent life-style-enhancing effects, possibly by their effect(s) on key life-history attributes. Although several studies have reported beneficial effects of these formulations on different components of life history, few have investigated their concurrent influence on various life-history traits. Here, we report the results of an investigation showing the effect of two well-known Ayurvedic formulations, Guduchi and Madhuyashti, on fecundity and longevity of Drosophila melanogaster. Flies were either grown (i.e., larval exposure) and/or maintained (i.e., adult exposure) on standard food supplemented with 0.5% Guduchi or 0.5% Madhuyashti. It was observed that the longevity of adult flies of both sexes was not affected on feeding Guduchi food, but fecundity of the females was greatly enhanced. Fecundity was also found to be affected by the adult food and whether their mates were grown on Guduchi or normal food. Madhuyashti, on the other hand, significantly reduced mean longevity and had a stimulatory effect on female fecundity. This fecundity enhancing effect however seemed to be mediated through its effect on the males. Interestingly, much of these effects interacted with age in a complex way, making it difficult to generalize the overall effect of these formulations on the reproductive output of the flies. Our study underlines the importance of evaluating the interacting effects of these (and similar) formulations on a range of life-history traits in a holistic way to understand their utility better.