Susceptibility of Eldana saccharina (Lepidoptera: Pyralidae), Busseola fusca and Sesamia calamistis (Lepidoptera: Noctuidae) to Bacillus thuringiensis Cry toxins and potential side effects on the larval parasitoid Cotesia sesamiae (Hymenoptera: Braconidae)

Laboratory trials were conducted to determine the efficacy of four Bacillus thuringiensis (Bt) Cry toxins at five different concentrations (0.016, 0.08, 0.4, 2, 10 μg/mL) for controlling three lepidopteran stem-borer species (i.e., the pyralid Eldana saccharina and the noctuids Busseola fusca and Sesamia calamistis) as well as to evaluate their indirect effect on the braconid larval parasitoid Cotesia sesamiae. In addition, larvae from the treatments above, after having been parasitized, were either fed a contaminated (group 1) or a toxin-free (group 2) diet and compared with a control (i.e., parasitized larvae which have never fed on Bt-toxin). All Bt Cry toxins induced larval feeding inhibition. Compared with the control, significant mortality resulted at all concentrations and for all species of Lepidoptera. Cry1Ab was the most toxic with 10 days post treatment mortalities ranging from 81% in B. fusca and S. calamistis to 100% in E. saccharina. In contrast, Cry1Ac had comparatively low toxicity particularly for B. fusca and S. calamistis (e.g., respectively, 54 and 74% mortality at 10 days at the highest concentration). In the toxin-treated group 1, percentages of C. sesamiae cocoon-producing moth larvae were higher compared to group 2 and the control, whereas clutch size was higher than in group 2 but similar to the control. In both groups, 0.016, 0.08 μg/mL CrylAc yielded lower female-biased sex ratios than the control. It was suggested that paralyses of the moth larvae caused by the toxin may have facilitated parasitization leading to higher parasitism and clutch size.     

Single-step PCR differentiation of Cotesia sesamiae (Cameron) and Cotesia flavipes Cameron (Hymenoptera: Braconidae) using polydnavirus markers

Cotesia sesamiae (Cameron) and Cotesia flavipes Cameron (Hymenoptera: Braconidae) are the main larval parasitoids of cereal stemborers in sub-Saharan Africa. Cotesia sesamiae is endemic to eastern and southern Africa, while C. flavipes was introduced into the region for biological control against the exotic lepidopteran Chilo partellus (Swinhoe) (Lepidoptera: Crambidae). The two are sibling parasitoids, difficult to distinguish morphologically. The introduced insect could potentially lead its African biotype to extinction because of their similar ecological niche. In order to distinguish the two species, multiplex primer-specific and PCR-RFLP tests were developed. Rapid identification of the two species was possible using primer-specific tests on DNA extracts as well as on pieces of tissue in a single PCR step followed by gel electrophoresis. The CRV1 gene of the polydnavirus, a symbiont to the wasps, was used as the marker. The results show that the morphological identifications, validated by molecular tests, are accurate in 93% of cases.     

Calyx fluid proteins of two Cotesia sesamiae (Cameron) (Hymenoptera: Braconidae) biotypes in Kenya: implications to biological control of the stem borer Busseola fusca (Fuller) (Lepidoptera: Noctuidae)

Cotesia sesamiae (Cameron) (Hymenoptera: Braconidae) is an indigenous larval endoparasitoid of Busseola fusca (Fuller) (Lepidoptera: Noctuidae) in sub-Saharan Africa. In Kenya, reports suggest that C. sesamiae occurs as two biotypes. Biotype avirulent to B. fusca gets encapsulated by haemocytes in this host and is unable to complete development. Biotype virulent to B. fusca is able to overcome immune defences. Factors present in the calyx fluid such as the PolyDNAviruses (PDV), venom and calyx fluid proteins have been implicated in the variation of C. sesamiae virulence against B. fusca. In the present study, calyx fluid proteins of the two C. sesamiae biotypes were compared using 2-D gel electrophoresis. More protein spots were observed in the virulent parasitoid calyx fluid, but some proteins were specifically observed in the avirulent parasitoid calyx fluid while others were observed in both. To study changes in proteins due to parasitism of B. fusca larvae by the two strains, SDS-PAGE gel were performed on fat body tissues and the haemolymph at three time points. Differences between the two strains were observed in both the fat body and haemolymph tissues. Parasitism-specific protein bands were detectable in fat body tissues of B. fusca larvae parasitized by the two C. sesamiae strains. These proteins were absent in unparasitized larvae. Implications for using C. sesamiae as a biocontrol agent of B. fusca in Africa are discussed.     

Intraspecific and Interspecific Competition Between Cotesia flavipes and Cotesia sesamiae (Hymenoptera: Braconidae), Gregarious Larval Endoparasitoids of Lepidopteran Stemborers

Super and multiple parasitism of Chilo partellus (Lepidoptera: Pyralidae) and Sesamia calamistis (Lepidoptera: Noctuidae) by Cotesia flavipes Cameron and Cotesia sesamiae (Cameron) (Hymenoptera: Braconidae) were investigated in the laboratory. Progeny production of Co. flavipes increased as a result of increasing the number of ovipositions, from one to three per one Ch. partellus host larva, then decreased as a result of four and five ovipositions per larva. Cocoon weight, sex ratio and emergence of the parasitoid progeny were not affected by superparasitism. Low progeny production of Co. sesamiae and poor survival of Ch. partellus host larvae were found as a result of superparasitism. When S. calamistis was the host, the duration of immature stages of Co. flavipes , parasitoid emergence, progeny production and sex ratio were not affected by superparasitism, but cocoon weight, adult longevity and the potential fecundity of adult females decreased. Superparasitism of S. calamistis by Co. sesamiae did not affect emergence, longevity or sex ratio of adult progeny of the parasitoid, but prolonged immature development, lowered cocoon weight and decreased potential fecundity of adult female progeny. Co. flavipes out-competed Co. sesamiae when Ch. partellus was parasitized by both species. The potential for local displacement of Co. sesamiae by Co. flavipes in areas dominated by Ch. partellus in East Africa is discussed.     

Multiple Parasitism by Cotesia sesamiae and Cotesia flavipes (Hymenoptera: Braconidae) on Busseola fusca (Lepidoptera: Noctuidae)

Busseola fusca (Fuller) is one of the most important pest of cereals in sub-Saharan Africa. Cotesia sesamiae (Cameron) is the predominant parasitoid attacking B. fusca larvae in many parts of Africa. An exotic parasitoid, Cotesia flavipes Cameron, was introduced into Kenya in 1993 for the control of Chilo partellus (Swinhoe). Laboratory studies indicated that although C. flavipes would search for, and attack B. fusca , it was not able to complete its development in this host. The aim of the present study was to investigate the outcome of multiple parasitism of B. fusca by the two Cotesia species. The study showed that when both parasitoid species stung a B. fusca larva at the same time, both parasitoids emerged from more than half of the host larvae, C. flavipes alone emerged from 17%, and C. sesamiae alone emerged from 9%. When the larvae were parasitized by C. sesamiae first, and then 2 h later by C. flavipes , and vice versa, most of the progeny were C. flavipes . However, when B. fusca larvae were stung by C. sesamiae three days before oviposition by C. flavipes , significantly more C. sesamiae emerged from the larvae. When C. flavipes oviposited first, no larvae produced C. flavipes only. The interaction of parasitoids and the host immune system, and the implications of these results for the biological control of stem borers in East Africa are discussed.     

The effect of neem (Azadirachta indica A. Juss) oil on oviposition, development and reproductive potentials of Sesamia calamistis Hampson (Lepidoptera: Noctuidae) and Eldana saccharina Walker (Lepidoptera: Pyralidae)

Abstract 1 The effect of different treatments of neem oil (0, 0.075, 0.1 and 0.15 mL/plant) and their persistence (0, 5 and 10 days after application; DAA) on the oviposition behaviour and the bionomics of the noctuid Sesamia calamistis and the pyralid Eldana saccharina were tested in laboratory and greenhouse experiments. 2 For most variables, no difference was found between DAA, showing that the treatments had a long‐term effect, and pooled analyses across DAA were performed. Compared with the control (0 mL/plant), mean reductions due to neem in numbers of egg batch and eggs laid were 70 and 88% for S. calamistis and 50 and 49% for E. saccharina, respectively, but no differences were found among neem concentrations. 3 For both species, larval and pupal development time was shortest in the control and longest with the highest oil concentration. Immature survival, larval weight and fecundity were highest in the control and similar in the neem treatments. 4 No differences were found in sex ratios. Egg viability was highest in the control (approximately 87%) and lowest (72%) with the highest oil concentration. As a result of lower fecundity and longer developmental time, on average, the intrinsic rates of increase in the neem treatments were 30% lower than in the control. 5 In view of the low oviposition rates, immature survival, fecundity and egg viability in the neem treatments, and the relatively high persistence of neem oil, it can be expected that the reduction in densities of the two borers species in the field will be considerable.     

Biology of Cotesia ( = Apanteles) telengai (Hymenoptera: Braconidae) on its natural host Agrotis segetum (Lepidoptera: Noctuidae)

The host suitability of Agrotis segetum Denis & Schiff., A. ipsilon Hufn., Spodoptera littoralis Boisd, S. exigua Hub., Mythimna loreyi Duponchel and Mamestra oleracea L. for the gregarious braconid Cotesia ( = Apanteles) telengai Tobias was determined under laboratory conditions. The parasitoid only completed its development in larvae of A. segetum. The percentage of successfully parasitized larvae and the mean duration of C. telengai egg‐larval period were inversely related to the age of the host or host instar at parasitization. The mean number of parasitoids which emerged per parasitized larva was positively correlated with the larval age. The sex ratio was consistently high (ca. eight males to one female), independent of the host instar parasitized, as compared to 1:1 as observed frequently in field populations of this wasp. The females of C. telengai were active and produced offspring at temperatures of 15, 20, 25 and 30° C. However, the mean percentage of parasitized larvae increased from 13.1 to 72% and the mean progeny per parasitoid female increased from 14.7 to 129.4 parasitoids, both significant, when the experimental temperature was raised from 15 to 30°C, while their mean development time decreased from 75.5 to 19.2 days. At 25°C, the virgin and mated females continued oviposition until days 16 and 17, with a lifetime total of progeny of 397.6 (SD ±224.7) and 611.1 (SD± 128.8) parasitoids respectively, reaching a maximum of 64.3 and 99.2 on day 2 respectively.     

Release and establishment of Cotesia flavipes Cameron (Hymenoptera: Braconidae) an exotic parasitoid of Chilo partellus (Swinhoe) (Lepidoptera: Crambidae) in East and Southern Africa

Cotesia flavipes Cameron (Hymenoptera: Braconidae) was first imported into Kenya in 1991 from Pakistan for control of Chilo partellus (Swinhoe) (Lepidoptera: Crambidae). First releases were made at the Kenya coast in 1993 and a few recoveries of the parasitoid were made the following year. Additional foreign exploration for C. flavipes was conducted in central India in 1996 and 1998, which resulted in additional importation of the parasitoid for subsequent releases in eastern and southern Africa. Region-wide releases commenced with releases in Mozambique in 1996, Somalia in 1997 and Uganda in 1998. By 2005, many releases had been made in 10 countries in East and Southern Africa with establishment being reported in all of them except Eritrea but including Ethiopia where releases had never been made. This paper describes the progress made in the release and establishment of C. flavipes in East and Southern Africa and quantifies the rate of spread from the initial release sites in Kenya.     

Seasonal Fluctuations of Noctuid Stemborer Egg Parasitism in Southern Benin with Special Reference to Sesamia calamistis Hampson (Lepidoptera: Noctuidae) and Telenomus spp. (Hymenoptera: Scelionidae) on Maize

Parasitism of noctuid stemborer eggs was assessed in monthly surveys in maize fields in southern Benin, from May 1995 to October 1996, and on alternative hosts, i.e. wild grasses, during the dry season from November 1995 to February 1996. Sesamia calamistis was the most prevalent stemborer species, accounting for 98% of the larvae identified from subsamples reared on ear pieces. Egg batch densities ranged between 0.02-0.15 per plant, with lowest densities found during high rainfall periods. 77% of all egg batches were found to be parasitized The egg parasitoids were the scelionids Telenomus busseolae, T. isis , and the trichogrammatid Lathromeris ovicida , accounting for 55.6, 41.6 and 2.8% of total counts, respectively. Parasitism increased in the course of the cropping season and reached peaks of 92 and 86% during the second growing season of 1995 and 1996, respectively. This coincides with the time when farmers have planted a second crop and during a stage when the plants were most attractive and susceptible to S. calamistis attacks. During the dry season, relatively high parasitism was found on maize and wild grasses in inland valleys, and it was concluded that those habitats play an important role in stabilizing the system for both the pest and its natural enemies. As a result, in the Dahomey Gap, which includes parts of Benin, Togo and Ghana, S. calamistis is usually not of economic importance. Although T. busseolae is ubiquitous in Africa, T. isis was only reported from West Africa, including Cameroon. It is recommended that the geographic range of T. isis should be expanded and include East and Southern Africa, where another host, Busseola fusca , is the most important noctuid stemborer pest on cereals.     

Olfactory responses of Cotesia flavipes (Hymenoptera: Braconidae) to target and non-target Lepidoptera and their host plants

The attraction of Cotesia flavipes Cameron to volatiles from a range of non-target lepidopteran larvae and their host plants (grasses and trees) or food substrate (honeycomb) was evaluated using a Y-tube olfactometer. The non-target host larvae used in the study included Galleria mellonella (L.), Charaxes cithaeron Felder, Bombyx mori L., and Eldana saccharina Walker. The target insects, Chilo partellus (Swinhoe) and Chilo orichalcociliellus (Strand), were used as controls. Host plants included Afzelia quanzensis Welw., Morus alba L., Cyperus papyrus L., Pennisetum purpureum Schumach, and Zea mays L. The response of C. flavipes to volatiles from the non-target larvae and their food was variable. Attraction to uninfested maize was not significantly different from uninfested plants of non-target hosts or honeycomb. Only maize and honeycomb were preferred over clean air. C. partellus infested maize plants were significantly more attractive than M. alba, A. quanzensis, and honeycomb infested with their herbivores. Infested maize and C. papyrus were more attractive than uninfested ones. When odors from naked larvae were tested, C. flavipes preferred odors from C. partellus larvae over those of E. saccharina and C. cithaeron and larvae of C. partellus and G. mellonella were preferred to clean air. The implications of these findings for biological control and its effect on non-target organisms are discussed.     

Self-, Intra-, and Interspecific Host Discrimination in Telenomus busseolae Gahan and T. isis Polaszek (Hymenoptera: Scelionidae), Sympatric Egg Parasitoids of the African Cereal Stem Borer Sesamia calamistis Hampson (Lepidoptera: Noctuidae)

Oviposition behavior, intra- and interspecific host discrimination, and super- and multiparasitism by the scelionids Telenomus busseolae and T. isis were studied using batches of eggs of the noctuid stalk borer Sesamia calamistis as the host. Both Telenomus species were able to discern eggs already parasitized. As a result, self-superparasitism was only 4.0% for T. busseolae and 5.8% for T. isis. Likewise, intraspecific superparasitism was avoided by both species and was significantly higher for T. busseolae than T. isis, and higher when parasitized eggs were offered immediately (0 h) and after 48 h than after 24 h; apparently, the recognition of parasitized eggs at 24 and 48 h was based on the presence of parasitoid larvae rather than a specific marking substance. Multiparasitism was avoided if the female had a choice between unparasitized and parasitized eggs. In a choice experiment, it was 10.2 and 2.5% for T. busseolae and T. isis, respectively. In a nonchoice experiment, multiparasitism did not vary between species and time treatment; it was low, varying between 9.6 and 24.1%. In the 0-h treatment, T. busseolae always outcompeted T. isis, accounting for 63.4 or 91.7% of the offspring, when T. isis or T. busseolae, respectively, was the first species. In the 24-h treatment, the first species emerged more often than the second did but the host egg mortality, i.e., eggs from which neither borer larvae nor parasitoid emerged, was >40%. Whereas T. busseolae is ubiquitous in Africa, T. isis has never been reported from eastern Africa, and it has been proposed for introduction against the prevailing noctuid pest Bussoela fusca. Based on earlier life table and host suitability studies and on the present findings, it is concluded that T. isis would establish in the midaltitudes but not the highlands of eastern Africa.     

Comparative assessment of the thermal tolerance of spotted stemborer,Chilo partellus (Lepidoptera: Crambidae) and its larval parasitoid,Cotesia sesamiae (Hymenoptera: Braconidae)

Under stressful thermal environments, insects adjust their behavior and physiology to maintain key life‐history activities and improve survival. For interacting species, mutual or antagonistic, thermal stress may affect the participants in differing ways, which may then affect the outcome of the ecological relationship. In agroecosystems, this may be the fate of relationships between insect pests and their antagonistic parasitoids under acute and chronic thermal variability. Against this background, we investigated the thermal tolerance of different developmental stages of Chilo partellus Swinhoe (Lepidoptera: Crambidae) and its larval parasitoid, Cotesia sesamiae Cameron (Hymenoptera: Braconidae) using both dynamic and static protocols. When exposed for 2 h to a static temperature, lower lethal temperatures ranged from ?9 to 6 °C, ?14 to ?2 °C, and ?1 to 4 °C while upper lethal temperatures ranged from 37 to 48 °C, 41 to 49 °C, and 36 to 39 °C for C. partellus eggs, larvae, and C. sesamiae adults, respectively. Faster heating rates improved critical thermal maxima (CT_max) in C. partellus larvae and adult C. partellus and C. sesamiae. Lower cooling rates improved critical thermal minima (CT_min) in C. partellus and C. sesamiae adults while compromising CT_min in C. partellus larvae. The mean supercooling points (SCPs) for C. partellus larvae, pupae, and adults were ?11.82 ± 1.78, ?10.43 ± 1.73 and ?15.75 ± 2.47, respectively. Heat knock‐down time (HKDT) and chill‐coma recovery time (CCRT) varied significantly between C. partellus larvae and adults. Larvae had higher HKDT than adults, while the latter recovered significantly faster following chill‐coma. Current results suggest developmental stage differences in C. partellus thermal tolerance (with respect to lethal temperatures and critical thermal limits) and a compromised temperature tolerance of parasitoid C. sesamiae relative to its host, suggesting potential asynchrony between host–parasitoid population phenology and consequently biocontrol efficacy under global change. These results have broad implications to biological pest management insect–natural enemy interactions under rapidly changing thermal environments.     

Experimental taxonomy of Bulinus (Gastropoda: Planorbidae): the West and North African species reconsidered, based upon an electrophoretic study of several enzymes per individual

Horizontal starch gel electrophoresis was used to score mobilities of seven different enzymes in species of Bulinus from North and West Africa. An account of the intra- and inter-population variation observed was given. Based on the data obtained it was suggested that enzymic data should be used as taxonomic characters in parallel with morphological and anatomical characters. It is suggested that enzymic characters could be used to place taxa into synonymy if the data are collected in such a way that one can write up an enzyme profile for the single individual. The nine taxa of Bulinus known to occur in North and West Africa were revised, the taxon B. jousseaumei was synonymized with B. globosus , and the two taxa B. guernei and B. rohlfsi were synonymized with B. truncatus. As a result of the revision only six species could be recognized as valid from the area, these being B. forskalii, B. globosus, B. senegalensis, B. truncatus, B. ugandae and B. umbilicatus. Finally, some enzymic characters are suggested that may be used for a reliable identification of these morphologically very difficult taxa.