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Pathogens Associated with Sugarcane Borers, Diatraea spp. (Lepidoptera: Crambidae): A Review

Pathogens Associated with Sugarcane Borers, Diatraea spp. (Lepidoptera: Crambidae): A Review Hindawi Publishing Corporation International Journal of Zoology Volume 2012, Article ID 303589, 12 pages doi:10.1155/2012/303589 Review Article Pathogens Associated with Sugarcane Borers, Diatraea spp. (Lepidoptera: Crambidae): A Review 1 1 1 Vıct ´ or M. Hernandez-V ´ elazquez, ´ Laura P. Lina-Garcıa, ´ Veronica ´ Obregon-Bar ´ boza, 2 2 Adriana G. Trejo-Loyo, and Guadalupe Pena-Chor ˜ a Centro de Investigacion en Biotecnologıa, Universidad Autonoma del Estado de Morelos, Avenida Universidad No. 1001, ´ ´ ´ Colonia Chamilpa, 62210 Cuernavaca, MOR, Mexico Centro de Investigaciones Biolog ´ icas, Universidad Autonoma ´ del Estado de Morelos, Avenida Universidad No. 1001, Colonia Chamilpa, 62210 Cuernavaca, MOR, Mexico Correspondence should be addressed to V´ıctor M. Hernandez-V ´ elazquez, ´ vmanuelh@uaem.mx Received 14 June 2012; Accepted 29 August 2012 Academic Editor: Thomas Iliffe Copyright © 2012 V´ıctor M. Hernandez-V ´ elazquez ´ et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The objective of this paper was to analyze information related to entomopathogenic-associated Diatraea spp. Gaining a better understanding of the effects of these microorganisms will help in the development of successful microbial control strategies against stem borers that attack sugarcane plants. 1. Introduction also discuss the status of recent attempts to use pathogens in the field. The Diatraea spp. (Lepidoptera: Crambidae) complex is only found in the American continent, and it is the most impor- 2. Fungi tant group of stem borers that principally attack maize and sugarcane, as well as other gramineous crops, including rice, One promising field for research is the use of entomopatho- sorghum, and forage grasses [1]. The sugarcane borer (SCB) genic fungi as biological control agents of insect pests D. saccharalis Fab. is the most economically important pest in sugarcane plants. Approximately 80% of the etiological in South America [2, 3], whereas the neotropical corn stalk agents involved in insect diseases are fungi, which encompass borer (NCB) D. lineolata Walk is primarily found in Central 90 genera and more than 700 species [7]. A number of America [4], D. magnifactella Dyar and D. considerata fungi (Hypocreales: Clavicipitaceae) including B. bassiana, Heinrich are found in Mexico [1], and the southwestern corn B. brongniartii, M. anisopliae, P. fumosoroseus, Hirsutella sp., borer D. grandiosella Dyar is found in the United States [5]. Cylindrocarpon sp., and Nomuraea rileyi have been isolated Unfortunately, commercially available insecticides are from Diatraea spp. in the Americas from Argentina to not efficient for the control of Diatraea spp. for a variety of the USA (Table 1). Under certain climatic conditions, B. reasons, mainly because of the continuous presence of the bassiana has been reported to cause natural epizootics on D. host plants in fields throughout the year, the concomitant grandiosella [8]. presence of mature and immature forms of the insect, and The life cycle of entomopathogenic fungi in the arthro- the cryptic feeding habits of the insect [6]. An alternative pod hosts is initiated with the germination of conidia that strategy is integrated pest management with biological con- contacts the host integument and produces a germ tube trol as the first defense, which includes the use of parasitoids that penetrates the host through a combination of physical and entomopathogens. pressure and enzymatic degradation of the cuticle. The In this paper, we present a perspective on the attempts to fungus initially colonizes the host through a yeast phase. control Diatraea spp. using pathogens in the Americas. We Host death usually results from a combination of nutrient 2 International Journal of Zoology Table 1: Entomopathogens fungi from Diatraea spp. Host species Entomopathogen Country Reference D. grandiosella B. bassiana (1–8%) Mississippi, United States Inglis et al., 2000 [5] D. crambidoides D. grandiosella B. bassiana (epizootics) Texas High Plains, United States Knutson and Gilstrap, 1990 [8] B. bassiana D. saccharalis M. anisopliae Venezuela Zambrano et al., 2002 [9] P. fumosoroseus D. saccharalis M. anisopliae PE, Brazil Alves, et al. 2002 [10] Pinar del R´ıo, La Habana, D. saccharalis B. bassiana Matanzas, Villa Clara, Estrada et al., 2004 [11] Cienfuegos and Camaguey, Cuba B. bassiana M. anisopliae D. saccharalis Tucuman, ´ Argentina Yasen de Romero et al., 2008 [12] Nomuraea rileyi Iseria sp. M. anisopliae D. saccharalis Mexico Angel-Sahagun ´ et al., 2005 [13] (Ma2 and Ma3) B. bassiana ARSEF: 1489 Pernambuco 1832 Brazil 1834 Brazil 2629 Brazil 3020 Pernambuco 3858 Brazil D. saccharalis 5500 Pernambuco, Humber et al., 2009 [14] 5502 Brazil Brazil Santa Fe, Argentina Santa Fe, Argentina B. brigniartii D. saccharalis Sao Paulo, Brazil Humber et al., 2009 [14] ARSEF: 1830 Cylindrocarpon sp. D. saccharalis ARSEF: 8043 Colima, Mexico Humber et al., 2009 [14] M. anisopliae Colima, Mexico ARSEF: 3290 Colima, Mexico D. saccharalis 3291 Colima, Mexico Humber et al., 2009 [14] 3292 Colima, Mexico 3298 Colima, Mexico M. anisolpliae Hernandez ´ and Velazquez ´ D. magnifactella B. bassiana Mexico 2004 [15] Hirsutella sp. Marshal County, (Mississippi, D. grandiosella Nosema sp. isolate 167 Inglis et al. 2000 [5] USA) Oktibbeha County, (Mississippi, D. grandiosella Nosema sp. isolates 295, 504 Inglis et al. 2000 [5] USA) International Journal of Zoology 3 Table 1: Continued. Host species Entomopathogen Country Reference Washington County, D. grandiosella Nosema sp. isolates 181, 513, 522 Inglis et al. 2000 [5] (Mississippi, USA) Lastra and Gomez ´ 2000 D. saccharalis Nosema sp. Colombia [16] D. saccharalis Granulovirus (DsGV) Southern United States Pavan et al. (1983) [17] D. saccharalis Densovirus (DsDNV) Guadeloupe Meynadier et al. (1977) [18] Fonseca-Gonzalez et al. (2011) D. magnifactella B. thuringiensis Mexico [19] depletion, invasion of organs, and the action of fungal toxins. In general, these microorganisms live as parasites in Hyphae usually emerge from the cadaver. The mummified cells of the midgut epithelium, where they complete their corpse of the insect remains in the environment for several development, and the cycle starts when the infective states weeks and, in the case of stem borer, it keeps remains of microsporidia (spores) arrive at the digestive tube and protected inside the stem. Therefore, it is more common to colonize this region all the way to the excretory system. detect and isolate fungi compared to other pathogens that The spores germinate because of the acid intestinal pH, destroy the host, such as Bacillus thuringiensis (Bt), viruses, the microorganisms penetrate the midgut cells, and the or nematodes. intestinal activity is paralyzed between 14 to 21 days later Entomopathogenic fungi are widely distributed in all because the insect cannot assimilate nutrients. Nosema regions of the world; these species have wide genetic locustae infects the adipocytes of fat body, which interferes variation among the different isolates. Pathogenicity and with the adequate function of the insect’s intermediary virulence to differentspecies,aswellasenzymatic andDNA metabolism and competes with the insect for energetic characteristics, vary among different isolates [14, 20, 21]. reserves. Microsporidia produce effects that depend on the Therefore, it is important to evaluate as many geographic species and concentration; however, they generally produce isolates as possible from different areas to select the most weakness and eventually lead to death [31, 32]. suitable isolate based on its virulence and growth at high Only some species in this group have the possibility to temperatures. Several research groups have verified the be potentially relevant for natural or classical control. There pathogenicity and virulence of Hypocreales fungi, such as are many studies worldwide on the pathogenic effects of M. anisopliae and B. bassiana (Table 2), which have become the microsporidia N. pyrausta (Payllot) and Vairimorpha important biocontrol agents used for the microbial control of necatrix (Kramer) on borer organisms such as Ostrinia Diatraea spp. It is possible that a limited selection of available nubilalis (Hυ¨bner) (European corn borer), Lymantria dispar isolates of B. bassiana could identify highly virulent strains (Gypsy moth), and the grasshopper [5, 32]. In this group, from each of the different Diatraea species. only the microsporidium N. locustae has been registered as Field evaluations have been performed using M. aniso- a microbial insecticide for the control of grasshoppers in pliae and B. bassiana against several insect pests of sugar- grasslands [33]. cane, including D. saccharalis in Brazil. Application of M. In many areas of the USA and Europe, Nosema is the anisopliae at a rate of 1 × 10 spores per hectare caused main agent used for the control of grasshoppers; however, 58% mortality of D. saccharalis,and B. bassiana at 3.7 × 10 in Latin America, few studies have been performed with spores per milliliter reduced D. saccharalis damage by 45% Nosema [33]. For example, Inglis et al. [5]reportedsix (Alves et al., 1984 and 1985, cited in Legaspi et al. [22]). isolates of Nosema on larvae in the winter diapause stage For effectiveness in the field, it is important to consider the from D. grandiosella Dyer collected during 1998 in corn contact between the spores and the host, formulation, and stems from three locations in Mississippi, USA. However, the the virulence of the pathogens. frequency of infection in the field was very low (1, 3, and 15% in the counties of Marshal, Oktibbeha, and Washington, resp.), and no isolates were found in D. crambidoides (Grote) 3. Microsporidia (Table 3). Microsporidia (Eukaryota: Fungi) is the most ubiquitous When the mortality produced by the Nosema isolates group among insect populations [23, 24]. Microsporidia are was assessed in the laboratory using larvae that had stayed tiny unicellular organisms (from 2 to 40 μm in diameter), in environmental-like natural winter diapause conditions, that are opportunistic and obligate intracellular parasites variations in mortality between 0 and 55% were observed and attack different groups of invertebrate and vertebrate in larvae, and variations between 7 and 29% were observed animals. Microsporidia generally produce chronic diseases in pupae; homogenization of the dead larvae revealed a and reduce the physiological and reproductive ability of large amount of Nosema spores. However, in the surviving their hosts. Many species of microsporidia infect arthropods, adults, a large number of larvae were positive for the Nosema especially insects such as Lepidoptera and Coleoptera [23– spores when they were analyzed under light microscopy 25]. using staining techniques and electron microscopy. 4 International Journal of Zoology Table 2: Entomopathogenic-fungi bioassays on Diatraea spp. Host species Pathogen Bioassay Result Reference TL (conidia isolates Bb1 and 5): 4.3 days. Immersing third instar Mortality dry mycelium D. saccharalis B. bassiana larvae into a suspension of preparations: Arcas et al., 1999 [26] 10 conidia/mL 21.3% (Bb1) 82.5% (Bb5) at 7 days after inoculation LD in spores/mm 1st instar: 72.1 2nd instar: 384.3 Sprayed first, second and B. bassiana 3rd instar: 777.0 D. saccharalis third instar larvae with Legaspi et al., 2000 [27] Mycotrol strain GHA Mean days of survival 1mL 1st instar: 4.6 2nd instar: 4.8 3rd instar: 6.4 Dipped in conidial B. bassiana D. grandiosella suspension (dosage not Mortality from 10 to 21% Inglis et al., 2000 [5] Eight isolates specified) Third instar larvae sprayed B. bassiana ATCC 20872 D. saccharalis with suspension of TL 3.02 to 4.10 days Marques et al., 2000 [28] from Solenopsis invicta 10 conida/mL LC 5.6 × 10 yeast/mL Third instar larvae sprayed B. bassiana ATCC 20872 and D. sacccharalis with 3 mL of yeast-like cells Alves et al., 2002 [29] from Solenopsis invicta 4.8 × 10 conidia/mL or conidia B. bassiana 1cm larvae sprayed with CL 1.58 × 10 conidia/mL D. saccharalis Wenzel et al., 2006 [30] IBCB from Brazil 1mL at 6 days after inoculation Table 3: Entomopathogenic-microsporidia bioassays on Diatraea spp. Host specie Pathogen Bioassay Results Reference Surface of diet (squares), concentration not D. grandiosella Nosema sp isolate 167 Mortality of 1–15% Inglis et al., 2000 [5] mentioned. Larvae and pupae Surface of diet (squares), concentration not D. grandiosella Nosema sp isolate 295, 504 Mortality of 1–15% Inglis et al., 2000 [5] mentioned. Larvae and pupae Surface of diet (squares), Nosema sp isolate 181, 513, concentration not D. grandiosella Mortality of 1–15% Inglis et al., 2000 [5] 522 mentioned. Larvae and pupae Microsporidiosis LC = 1st instar larvae, Surface of 48.4 spores/mm D. saccharalis Vairimorpha necatrix Fuxa, 1981 [32] diet Gut damage LC = 8941 spores/mm 5-day-old larvae and Median infective dose of 2 2 3 pupae, leaf pieces ∼5cm × 10 spores per larva, D. grandiosella Nosema sp isolate 506 Inglis et al., 2003 [34] with 10 μl inoculum (10 to pupae small, decreased egg 10 spores) production In these experiments, the larvae were reared in the winter it is possible that Nosema can produce infection in natural diapause stage, and the temperature for Nosema was not conditions as the temperature inside the cane is higher than optimal for the development of infection; in the field during the external temperature. the winter, the prediapause larvae migrate to the base of Phoofolo et al. [35] observed a similar behavior, and the corn stem just below the surface of the soil [5], and they reported that the O. pyrausta infection by Nosema is International Journal of Zoology 5 chronic. Although no immediate mortality is produced, the Lastra and Gomez ´ [16] implemented a system to produce longevity and fecundity of the adults are reduced. Likewise, natural enemies of D. saccharalis in CENICANA (Clombian Phoofolo mentioned the possibility of a response to other Sugarcane Research Center, Colombia) and began their factors of mortality that regulate the population dynamics colony with larvae collected in the field. Their most signif- of borers, such as low temperature, the host plant, and icant result was the detection of one “protozoa,” possibly crowding, which have chronic effects on Nosema infection. Nosema, identified as the causal agent for the diminution Therefore, although the pest did not die directly because of in larvae production. They observed refractile spores in the microsporidia infection, the population size was eventually fluids of the malpighi tubes or hemolymph using phase reduced. contrast microscopy in healthy adults. Macroscopically, the This effect has been previously observed by Fuxa [32], diseased larvae were dwarfed and white. when he evaluated the susceptibility of larvae in the first In addition, different pathogenic microorganisms have and third instars of six species of Lepidoptera, including D. been isolated from Diatraea sp. in experimentally trans- saccharalis, to the microsporidium Vairimorpha necatrix.He mitted infections in the laboratory; however, there is little described two routes of infection that resulted in mortality; information about the natural presence of entomopathogens one resulted from the chronic effects produced by the belonging to the microsporidia group in this species of borer. exposure of larvae to low doses of spores, which led to lethal It is important to perform a systematic search for infected septicemia (microsporidiosis) just before pupating, and the larvae, pupae, or adults of Diatraea, and studies need to other route was due to the intake of a large number of include microscopy techniques to complement the mortality spores, which apparently damaged the gut because of the bioassays because infections could be asymptomatic. As introduction of a large number of spore polar filaments. In many microsporidia do not produce insecticide activity thisstudy,Fuxa[32] concluded that this pathogen could be quickly and because many species have complex life cycles promising for the remaining five species, although not for D. that involve more than one host, very few attempts have been saccharalis, as he had observed direct mortality by damage to made to implement microsporidia and use them as agents of the gut and indirect mortality by septicemia and because a biological control. high concentration of spores was necessary. Inglis et al. [34] reported that one strain of Nosema 4. Nematodes (506), which had been previously isolated [5], can infect other species of insects in the Crambidae family, including O. Pathogenic nematodes of the Heterorhabditidae and Stein- nubilalis and D. crambidoides (Grote), but not other species ernematidae families live in the soil where they are parasitic of Noctuidae. However, they also observed that the infection to certain soil-dwelling insects. The free-living third juvenile can be transmitted transovarially, although at a very low stage (infective juveniles, IJs) locates a suitable host and frequency (a difference with other Nosema species that can be penetrates the host in different ways. Once inside the insect, transmitted frequently in the vertical rout, such as the genus the IJs initiate its development, the nutritional tract becomes Ostrinia). functional, and the symbiotic bacterium Xenorhabdus or Solter et al. [36] researched the vertical and horizontal Photorhabdus is released through the anus and begins to transmission of seven species of microsporidia, including multiply in the hemocoel, which kills the insect by septicemia two strains of Nosema sp. (isolated from their natural hosts and creates suitable conditions for the reproduction of the in the field D. saecharalis and Eoreuma loftini); although nematode. Nematodes feed on bacteria and the dead tissue five of these strains were transmitted at a low percentage of the host, and they pass through several generations until horizontally and vertically, two Nosema strains did not new IJs are produced, which emerge from the cadaver [37]. behave similarly. They have a ubiquitous distribution [37]; therefore, it is not In assessing the infectivity percentage under laboratory unusual to find them in the soil of sugarcane plantations conditions when applying high concentrations of spores, (Pizano et al., 1985 cited in Khan et al. [38, 39]). they observed very low mortality in the larvae of D. sacharalis However, there are few reports of nematodes infecting and E. loftini (2 and 4%, resp.), although the same strains stem borers of the Diatraea genus in natural and experi- produced high mortality at low concentrations in the other mental situations. In Costa Rica, several entomopathogenic species studied, including O. nubilalis. They explained the species, including nematodes, have been isolated from D. low mortality, even at high doses, as a function of low tabernella [40]; however, the author does not mention the horizontal and vertical transmission because few infective or nematode species. abnormal spores were produced. Khan et al. [38] reviewed the world bibliography on rice In this case, it is important to clarify that they could stem borers and found five reports of entomopathogenic obtain live infected larvae without mortality; however, nematodes associated with D. saccharalis,two reportsof they only measured mortality and did not measure other Steinernema (=Neoaplectana) glaseri in Brazil and three parameters, such as fecundity or larvae hatching in the next reports of Steinernema (=Neoaplectana) carpocapsae in the generation. As mentioned by Fuxa [32] and Inglis et al. [5], USA and Guadeloupe. However, in one study, they report infection does not always lead to mortality, and occasionally that the aim of the research was the use of D. saccharalis to the effects are observed long term and can be inferred from produce S. glaseri in controlled conditions because of its sus- a reduction in fecundity and susceptibility to other stress ceptibility to the nematode. Another study by Folegatti et al. situations. [41] used entomopathogenic nematodes and D. saccharalis 6 International Journal of Zoology Table 4: Entomopathogenic-nematode bioassays on Diatraea spp. Host specie Pathogen Bioassay Result Reference 100% mortality with 5000 Steinernema feltiae nematodes/larvae [Neoaplectana carpocapsae] Several concentrations for 100% mortality with 5000 Diatraea saccharalis Heterorhabditis heliothidis Sosa et al., 1993 [42] all species nematodes/larvae 30% mortality with 5000 S. glaseri nematodes/larvae S. feltiae 500 IJs/5 larvae in Petri S. rarum Adults and IJs produced. Diatraea saccharalis dishes with two moistened De Doucet et al., 1999 [43] Heterorhabditis Larval mortality >90% filter papers disks bacteriophora H. bacteriophora JPM4 Larvae were exposed to 25 100% larval mortality Diatraea saccharalis isolatedfromsoilinLarvae ± 5 IJs/100 μLfor 48 hin within 5 days. LT 2.1 d Molina et al., 2007 [44] MG, Brazil sand-filled Petri dishes and LT 4.4 d Heterorhabditis Diatraea saccharalis 500 mL/L applied to larvae 93% of larvae mortality Aguila et al., 2008 [45] bacteriophora in laboratory conditions to produce S. carpocapsae in vivo, Bohorova et al. [49] evaluated Cry1Aa, Cry1Ab, Cry1Ac, using larvae of the sugarcane borer as the host. Cry1B, Cry1C, Cry1D, Cry1E, and Cry1F Bt pure proteins We believe that pathogenic nematodes have good poten- against four species of Lepidoptera that are pests of maize, tial for the biological control of Diatraea species because including D. saccharalis. The proteins were diluted in water of their presence in sugarcane-producing regions (as men- and added to the diet at doses of 10 and 100 mg/mL of diet, tioned above) and because the species S. feltiae, S. glaseri, and mortality was recorded after seven days. D. saccharalis S. rarum, Heterorhabditis heliothis,and H. bacteriophora was susceptible to Cry1B protein at 10 mg/g, and the LC have demonstrated high infectivity in D. saccharalis [42–45] was 113.6 mg/g of meridic diet (Table 5). (Table 4)aswellasin E. loftini (some of them) [46, 47]in Twelve Bt strains were evaluated by Rosas Garc´ıa et al. experimental studies. We consider that performing a system- [50] at a dosages of 50 and 500 μg of total protein and spores atic search for infected larvae or pupae of Diatraea could be per milliliter against 2-day-old D. saccharalis larvae. The helpful in finding new nematode strains and species adapted strains used were HD1, HD2, HD9, HD29, HD37, HD59, to local environmental conditions and pest species that could HD133, HD137, and HD559, as well as the GM7, GM10, and be used in the future. GM34 native strains. The strains that killed more than 50% Although we could not find any report on field trials, of larvae were selected to obtain the LC . Strains HD133, we are aware that there have been mass rearings of H. HD559, GM7, GM10, and GM34 were toxic; however, GM34 bacteriophora in Cuba since 1987 to control soil pests, such was the most toxic with an LC of 33.21 μg/mL (Table 5). as D. saccharalis [45]. PCR analysis was performed to determine the cry1 genes of the toxic strains: HD133 cryAa, cry1Ab, cry1C; HD559 and GM7 cry1Aa, cry1Ab,and cry1B;GM10 cry1Aa, cry1Ab, 5. Bacteria cry1Ac,and cry1C;GM34 cry1Aa, cry1Ab,and cry1Ac. Bacillus thuringiensis (Bt) is a Gram-positive bacterium Gitahy et al. [51] evaluated a spore-crystal complex in that has been isolated from several sources, including soil, vitro from five native Bt strains (S48, S76, S90, S105, and water, phylloplane, and insect cadavers. Bt produces various S135) and HD1, which served as a positive control, on second instar larvae of D. saccharalis and mortality was recorded insecticidal crystal proteins during the onset of sporulation that are toxic to insects, acari, and nematodes [48]. The steps after 5 days. The strain S76 caused 100% mortality at 72 h, involved in the mode of action of the proteins after their HD1 caused 69% mortality, and 3% of the other native −1 strains caused mortality at 500 μgL of the spore-crystal ingestion are as follows: (a) solubilization of the crystals by the highly alkaline pH of the midgut, (b) activation of the complex. The LC values of the S76 and HD1 strains were proteins by proteases, (c) binding of the toxins to specific determined, and S76 was 11-fold more toxic (13.06 μg/L) receptors located on the microvilli membrane of the midgut than HD1 (143.88 μ/L) (Table 5). The S76 strain carries columnar epithelium cells, and (d) the insertion of the toxin cryAa, cry1Ab, cryAc, cry2Aa,and cry2Ab genes, which is similar to HD1. into the membrane, which forms a pore and induces cell lysis. There are no reports from the American continent There are other species of bacteria with potential insec- on the isolation of Bt strains from Diatraea spp. larvae ticide activity; Carneiro et al. [52]evaluated Photorhabdus temperata, which is a bacterium associated with Heterorhab- cadavers; however, some Bt strains and pure proteins have been evaluated against these pests. ditis entomopathogenic nematodes. Cells were injected with International Journal of Zoology 7 Table 5: Entomopathogenic-bacteria bioassays on Diatraea spp. Host species Pathogen Bioassay Result Reference Diet incorporated Diatraea saccharalis Bacillus thuringiensis LC 113.6 mg/g diet Bohorova et al., 1997 [49] spores-crystals Rosas-Garcia et al., 2004 D. saccharalis B. thuringiensis LC 33.21 μg/mL [50] Diet incorporated D. saccharalis B. thuringiensis LC 13.06 μg/L Gitahy et al., 2007 [51] spores-crystals complex Cells injected LD 16.2 bacterial cells D. saccharalis Photorhabdustemperata Carneiro et al., 2008 [52] LT 33.8 h Diet incorporated Diatraea grandiosella Bacillus thuringiensis LC 5.2 mg/g diet Bohorova et al., 1997 [49] spores-crystals Mean mortality on Bacillus thuringiensis 5-day-old larvae: 32, 31, 25, Immersing stem seedling in Biotrol BTB 183-25 15 out of 40. spores suspensions: 1.25 × Nutrilite Products Control mean mortality Sikorowski and Davis, 1970 8 7 7 Diatraea grandiosella 10 ,6.25 × 10 ,2.5 × 10 Incorporated, Kansas City, 0.01 [53] and 1.25 × 10 Missouri, USA Mean mortality on 10-day-old larvae: 15, 15, 12 and 6 out of 20. avolumeof10 μL of phosphate-buffered saline directly into these entomopathogenic viruses, OBs have independently the hemocoel of fourth instar D. saccharalis larvae, and the evolved as a protective mechanism to environmental factors, LD was16.2bacterial cellswithanLT of 33.8 h. which gives the viruses a great advantage as biological control 50 50 Sikorowski and Davis [53] determined the susceptibility agents [61]. Baculoviruses (BVs) and entomopoxviruses of 5- and 10-day-old D. grandiosella larvae with a Bt (EPVs: subfamily Entomopoxvirinae)havealargedouble- commercial product (Biotrol BTB 183-25) using 1.25 × 10 , stranded DNA genome, and cypoviruses (CPVs: family 7 7 7 6.25 × 10 ,2.5 × 10 , and 1.25 × 10 spores per milliliter Reoviridae, genera Cypovirus) contain segmented double- (Table 5) where two-inch stem seedlings were dipped for 30 stranded RNA viruses. min in spore suspensions. One 10-day-old or two 5-day- EPVs have been reported to infecting insects of a variety old larvae were allowed to feed on each stem. In addition, of orders, such as Coleoptera, Lepidoptera, Orthoptera, a known number of spores were placed or injected into 5- and Diptera. Some EPVs have two distinct OBs spheroids day-old stem seedlings at 1/4 inch sections. Stems dipped in and spindles. The spheroids occlude virions, whereas the water were used as controls. Mortality was recorded after spindles do not [62]. The most abundant proteinaceous 48 h. With 10-day-old larvae, five replicates with 20 larvae component of spheroids and spindles is proteins called for each treatment were used; for the 5-day-old larvae, two spheroidin and fusolin, respectively [63–66]. EPV fusolin is replicates with 40 larvae per treatment were used. With 5- an enhancing factor (EF) that increases BVs infection and and 10-day-old larvae at 1.25 × 10 spores per milliliter, a has been characterized as a chitin-binding protein [67]. The mean of 32 dead larvae out of 40 and 15 dead larvae out of 20 fusolin mechanism of action is similar to that of Calcofluor, were recorded (Table 5), respectively. Thus, it was concluded which facilitates BV infection by disrupting or preventing that this species is highly susceptible to B. thuringiensis. the formation of the peritrophic membrane [68]. EPVs are pathogenic but are scarcely virulent; infected larvae exhibit extreme longevity and take up to 70 days to die. However, 6. Viruses EPVs have potential as biological control agents for pest insects where BVs have not been isolated [61]. Because of Viruses that infect insects have received great attention as biological control agents because of their specificity on insect the activity of spindles, the EF of EPVs can be used as populations; they have little or no impact on the environ- a synergistic agent to increase BV infectivity or generate genetically modified organisms, such as BVs and transgenic ment and are an ecological friendly alternative to chemical pesticides [58]. Entomopathogenic viruses are grouped in 33 plants. genera within fifteen families [59]. However, only a few of CPVs have been mainly isolated from Lepidoptera these families, such as Baculoviridae, Poxviridae, and Reoviri- insects. OBs are dissolved in the midgut, and virions only dae, have potential as biological control agents and have been infect epithelial cells; therefore, CPVs are very pathogenic but act slowly and frequently to produce chronic infections [69]. successfully used in microbial control programs. A common characteristic among these entomopathogenic viruses is that At this time, no commercial bioinsecticides based on CPVs the virions (infective unit) are occluded within a crystalline have been developed. However, Caballero and Williams [61] suggest that their greatest potential as biological control protein matrix to form an occlusion body (OB) [60], which is a unique characteristic of viruses that infect insects. In agents is through inoculative or augmentative releases. 8 International Journal of Zoology Table 6: Entomopathogenic-virus bioassays on Diatraea spp. Host species Pathogen Bioassay conditions Result Reference DL :1.32 × 4 and 7 day-old larvae were 3 ∗ 10 PIB /larvae (for Autographa californica fed strips of corn leaf D. grandiosella 4-day-old larvae) Davis and Sikorowsy, 1978 [54] MNPV (AcMNPV) soaked in different DL :1.32 × 10 PIB/larvae dilutions of virus. (for 7-day-old larvae). 3rd instar larvae feeding LD : 42 PIB/larva at 26 C Diatraea saccharalis individually on small LT :From29to63for Lastra and Gomez ´ D. saccharalis Granulosis Virus artificial diet discs treated 7 2 10 and10 PIB/larvae, et al., 1983 [16] (DsGV) with 2.7 μL of virus respectively. dilutions. 3rd instar larvae fed with LD :From7.9 × 10 to 5.3 Anticarsia gemmatalis artificial diet discs treated × 10 for 1 and 20 serial D. saccharalis MNPV (AgMNPV) with 5 different doses of Pavan and Ribeiro, 1989 [55] passages, respectively. PIB through 20 serial passages. 3rd instar larvae AgMNPV wt LT :From10to13for individually fed small 39 and 30 C, respectively artificial diet discs D. saccharalis Ribeiro and Pavan, 1994 [56] containing 10 PIB of virus From 9to29for 39 and and kept individually at 10 AgMNPV-D10 22 C, respectively different temperatures. 3rd instar larvae fed LT :From9to 16 for Trichoplusia ni MNPV individually with small 39 and 24 C, respectively (TnMNPV) wt artificial diet discs D. saccharalis Ribeiro and Pavan, 1994 [56] containing 10 PIB of virus and kept individually at 10 From 6to46for 39 and TnMNPV-D11 different temperatures. 17 C, respectively 3rd instar larvae fed on LD :From5.3 × formalin free diet disc (3 10 PIB/larvae (7A mm) inoculated with genotypic variant) to 8 × D. sacccharalis AgMNPV Ribeiro et al., 1997 [57] 2.7 μL of the viral solution 10 PIB/larvae (33B from 10 clonal isolates of genotypic variant) the AgMNPV-Ds20 PIB: Polyhedral inclusion bodies. BVs predominantly infect insects within the Lepidoptera control of these insect pests difficult. Degaspari et al. [77] order, which includes important agricultural insect pests have argued that chemical control of D. saccharalis in Brazil [70]. The Baculoviridae family includes two genera: nucle- is not economically feasible. To solve this problem, surveys to opolyhedrovirus (NPV), which forms large, polyhedric OBs isolate endemic entomopathogens of stem borer populations where many enveloped virions are occluded [71], and in maize and sugarcane crops have been developed. Granulovirus (GV), which forms small, granular OBs that Inglis et al. [5] developed an exhaustive survey to occlude only one enveloped virion each [72]. BVs are safe for isolate entomopathogens from the southern corn borer, humans and wildlife. Their specificity is usually very narrow D. grandiosella, and the southern corn stalk borer, D. and often is species specific. Because of their specificity crambidoides, and larvae in the diapause stage were collected and other characteristics, such as elevated virulence and from crops located in Mississippi and North Carolina. These pathogenicity, BVs are by far the most studied and extensively authors did not observe OBs in any of the collected larvae used as commercial biopesticides for the control of a variety and concluded that there are no naturally occurring viruses of insect pests in many countries around the world [60, 61, in these Diatraea populations [5]. According to Inglis et 73]. al. [5], Pavan and Ribeiro [55] mentioned that natural Stem borers of the Lepidoptera order attack gramineous populations of the SCB in Brazil do not exhibit endemic viral crops throughout the world [74, 75]. Diatraea stem borers pathogens. (DSB) are widely distributed in the Americas and attack a Currently, there are only two records of endemic ento- wide variety of host plants, including maize and sugarcane mopathogenic viruses isolated from Diatraea spp. larvae. [1]. Prediapause larvae of southwestern corn borers migrate Pavan et al. [17] isolated a GV from the sugarcane borer to the base of the stalk of the corn plant below the soil surface (SCB), D. saccharalis, from sugarcane crops in the southern to survive the winters [76]; cryptic habits make the chemical United States (Table 6). These authors developed bioassays International Journal of Zoology 9 with D. saccharalis third instar larvae (Table 6). External thank Mrs. Ingrid Masher for reviewing the paper and for symptoms of GV-infected larvae were similar to those editorial assistance. reported for other lepidopterous larvae, and the symp- toms and ultrastructures were determined using electronic References microscopy as well as replication of DsGV, which are typical [1] L. A. Rodriguez-Del-Bosque and J. W. Smith Jr., “Biological of GVs. The LD for the third instar larvae was 42.3 control of maize and sugarcane stemborers in Mexico: a OBs/larva (Table 6) with 14.5 and 123.6 OBs/larva as the review,” Insect Science and Its Application,vol. 17, no.3-4,pp. lower and upper limits at 95% probability, respectively [16]. 305–314, 1997. These authors stated that this work was the first of a series of [2] G. Serra and E. Trumper, “Estimacion ´ de incidencia de danos ˜ publications; however, there have been no additional studies provocados porlarvasde Diatraea saccharalis (Lepidoptera: produced to date, such as the molecular and biological Crambidae) en tallos de ma´ız mediante evaluacion ´ de signos characterization, of this GV, DsGV was introduced into ´ externos de infestacion,” Agrosciencia, vol. 23, no. 1, pp. 1–7, Brazil [16], and Moscardi [78] mentioned that this virus is 2006. [3] M. A. P. De Oliveira, E. J. Marques, V. Wanderley-Teixeira, currently being applied at a small scale as an experimental and R. Barros, “Effect of Beauveria bassiana (Bals.) Vuill. and product on sugarcane. Metarhizium anisopliae (Metsch.) Sorok. on biological charac- Because of their high pathogenicity combined with a teristics of Diatraea saccharalis F. (Lepidoptera: Crambidae),” limited host range, Densoviruses (DNVs) have potential as Acta Scientiarum, vol. 30, no. 2, pp. 220–224, 2008. effective insecticides [79]. Meynadier et al. [80] isolated [4] R. Reyes, “Sorghum stem borers in Central and South aDNV of D. saccharalis (DsDNV) from the Guadeloupe America,” in Proceedings of the International Workshop on sugarcane borer (Table 6). Kouassi et al. [81] tested the Sorghum Stem Borers, pp. 49–60, ICRISAT Center, Hyderabad, pathogenicity of DsDNV on its host. These authors observed India, 1987. that the infected larvae exhibited infection symptoms from [5] G. D. Inglis, A. M. Lawrence, and F. M. Davis, “Pathogens the fourth day postinfection, such as anorexia and lethargy associated with southwestern corn borers and southern corn followed by flaccidity and inhibition of molting and meta- stalk borers (Lepidoptera: Crambidae),” Journal of Economic morphosis. Larvae became paralyzed and stopped feeding Entomology, vol. 93, no. 6, pp. 1619–1626, 2000. [6] M.D.R.T.DeFreitas,E.L.DaSilva,A.D.L.Mendonc¸a after 7 days. The cumulative mortality of infected larvae et al., “The biology of Diatraea flavipennella (Lepidoptera: increased significantly and reached 60% after 12 days and Crambidae) reared under laboratory conditions,” Florida 100% at 21 days postinfection [81]. Although DNVs have Entomologist, vol. 90, no. 2, pp. 309–313, 2007. no potential for large scale use as a biological control agent [7] R.H.R.Destefano ´ , S. A. L. Destefano ´ , and C. L. Messias, because they have no OBs and are related to vertebrate “Detection of Metarhizium anisopliae var. anisopliae within pathogenic viruses, the genes involved with anorexia and infected sugarcane borer Diatraea saccharalis (Lepidoptera, paralysis could be used to produce transgenic BVs or plants. Pyralidae) using specific primers,” Genetics and Molecular Most BVs have a limited host range and may be species Biology, vol. 27, no. 2, pp. 245–252, 2004. specific in some cases, although there are several NPVs, such [8] A.E.Knutson andF.E.Gilstrap, “Seasonal occurrence of as Autographa californica MNPV, Anagrapha falcifera MNPV, Beauveria bassiana in the southwestern corn borer (Lepi- and Anticarsia gemmatalis MNPV, which have broader host doptera: Pyralidae) in the Texas High Plains,” Journal of the Kansas Entomological Society, vol. 63, no. 2, pp. 243–225, 1990. ranges within the Lepidoptera order [60]. Because of the [9] K. Zambrano, M. Dav ´ ila, and M. A. Castillo, “Deteccion ´ de scarcity of reports on natural isolates of entomopathogenic fragmentos de AND de hongos y su posible relacion ´ con la viruses from Diatraea spp. populations, cross-infectivity of s´ıntesis de prote´ınas de actividad entomopatogena, ´ ” Revista de AgMNPV, TnMNPV, and AgMNPV has been evaluated in D. la Facultad De Agronom´ıa (LUZ), vol. 19, pp. 185–193, 2002. saccharalis and D. grandiosella larvae (Table 6). [10] S. B. Alves, L. S. Rossi,R.B.Lopes,M.A.Tamai,and R. M. Although only two reports of entomopathogenic viruses Pereira, “Beauveria bassiana yeast phase on agar medium and isolated from Diatraea stem borers exist in the Americas, its pathogenicity against Diatraea saccharalis (Lepidoptera: entomopathogenic viruses, such as NPVs and GVs, have Crambidae) and Tetranychus urticae (Acari: Tetranychidae),” been reported to occur in Africa and Asia in cereal stem Journal of Invertebrate Pathology, vol. 81, no. 2, pp. 70–77, borers within the Crambidae family, including maize and sugarcane borers such as Chilo sp. [82], Chilo sacchariphagus [11] M. E. Estrada, M. Romero,M.J.Rivero, andF.Barroso, “Natural presence of Beauveria bassiana (Balsamo) Vuillemin [83, 84], Ch. infuscatellus [84, 85], Ch. partellus [86], and in the sugar cane (Saccharum sp. hybrid) in Cuba,” Revista Eldana saccharina [87]. As the Mexican territory is the origin Iberoamericana de Micologia, vol. 21, no. 1, pp. 42–43, 2004. of some Diatraea stem borer species, we hypothesize that a [12] M. G. Yasen de Romero, A. R. Salvatore, G. Lop ´ ez, and great diversity of pathogenic viruses in Diatraea populations E. Willink, “Presencia natural de hongos hyphomycetes en that attack sugarcane crops exists in Mexico. larvas invernantes de Diatraea ssaccharalis F. en cana ˜ de azucar ´ en Tucuman, ´ Argentina,” Revista Industrial y Agr´ıcola Acknowledgments de Tucuman ´ , vol. 85, no. 2, pp. 39–42, 2008. [13] C. A. Angel-Sahagun, ´ R. Lezama-Gutier ´ rez, J. Molina-Ochoa This study was financed in part by the project “Seleccion ´ de et al., “Susceptibility of biological stages of the horn fly, enemigos naturales de barrenadores de la cana ˜ de azucar ´ de Haematobia irritans, to entomopathogenic fungi (Hyphomy- Colima y Morelos con potencial como agentes de control cetes),” Journal of Insect Science (Online), vol. 5, no. 50, pp. 1–8, biolog ´ ico (SEP-PROMEP),” of Mexico. The authors also 2005. 10 International Journal of Zoology [14] R. A. Humber, K. S. Hansen, and M. M. Wheeler, Catalog of [28] E. J. Marques, S. B. Alves, and I. M. R. Marques, “Virulencia Species. ARS Collection of Entomopathogenic Fungal Cultures, de Beauveria bassiana (Bals.) Vuill. a Diatraea saccharalis (F.) USDA-ARS Biological Integrated Pest Management Research, (Lepidoptera: Crambidae) apos ´ armazenamiento de con´ıdios Ithaca, NY, USA, 2009. em baixa temperatura,” Anais da Sociedade Entomolog ´ ica do Brasil, vol. 29, no. 2, pp. 303–307, 2000. [15] V. M. Hernandez ´ Velazquez ´ and R. Lezama Gutier ´ rez, “Uso de [29] S. B. Alves, L. S. Rossi,R.B.Lopes,M.A.Tamai,and R. M. entomopatogenos ´ para el control biolog ´ ico de barrenadores Pereira, “Beauveria bassiana yeast phase on agar medium and del tallo,” in Taller Internacional Sobre Barrenadores Del Tallo ´ its pathogenicity against Diatraea saccharalis (Lepidoptera: De Cana ˜ De Azucar ´ ,L.A.Rodrıguez del Bosque, G. Vejar, Crambidae) and Tetranychus urticae (Acari: Tetranychidae),” and E. Cortez, ´ Eds., pp. 37–45, Sociedad Mexicana de Control Biolog ´ ico, Los Mochis, Sinaloa, Mexico, Noviembre de 2004. Journal of Invertebrate Pathology, vol. 81, no. 2, pp. 70–77, [16] B. Lastra andL.A.Gomez, ´ “Cr´ıa y produccion ´ masiva de [30] I. M. Wenzel, F. H. C. Gionetti, and J. E. M. Almeida, “Pato- insectosenunprogramadecontrol biolog ´ ico en cana ˜ de genicidade do isolade IBCB66 de Beauveria bassiana sbroca da azucar ´ ,” in 1er Curso Taller Internacional de Control Biolog ´ ico, ˜ ´ cana-de-acucar Diatraea saccharalis em condicoes de labo- pp. 335–340, Memorias, Bogota, ´ Colombia. rator ´ io,” Arquivos Do Instituto Biologico, vol. 72, no. 2, pp. 259– [17] O. H. O. Pavan, D. G. Boucias, L. C. Almeida, J. O. Gaspar, P. S. 261, 2006. M. Botelho, and N. Degaspari, “Granulosis Virus of Diatraea [31] C. E. Lange, “Niveles de esporulacion ´ experimentales y saccharalis (DsGV): pathogenicity, replication and ultrastruc- naturales de Nosema locustae (Microsporidia) en especies de ture,” in Proceedings of the International Congress of the tucuras y langostas (Orthoptera: Acridoidea) de la Argentina,” International Society of Sugar Cane Technologists (ISSCT ’83), RevistadelaSociedad Entomolog ´ ica Argentina, vol. 62, no. 1-2, vol. 2, pp. 644–659, La Havana, Cuba, 1983. pp. 15–22, 2003. [18] G. Meynadier, P. F. Galichet, J. C. Veyrunes, and A. Amargier, [32] J. R. Fuxa, “Susceptibility of Lepidopterous pests to two ´ ´ “Mise en evidence d’une densonucleose chez Diatraea saccha- types of mortality caused by the microsporidium Vairimorpha ralis [Lep.: Pyralidae],” Entomophaga, vol. 22, no. 1, pp. 115– necatrix,” Journal of Economic Entomology,vol. 74, no.1,pp. 120, 1977. 99–102, 1981. [19] A. Fonseca-Gonzalez, ´ G. Pena-Chor ˜ a, A. Trejo-Loyo, L. Lina- [33] L. F. Solter and J. V. Maddox, “Microsporidia as classical Garc´ıa,L.A.Rodr´ıguez-Del Bosque, and V. M. Hernandez- ´ biological control agents: research and regulatory issues,” Velazquez, ´ “Virulencia de seis aislados de Bacillus thuringiensis Phytoprotection, vol. 79, no. 4, pp. 75–80, 1998. nativos delestado de Morelos y evaluados sobre Diatraea [34] G. D. Inglis, A. M. Lawrence, and F. M. Davis, “Impact of magnifactella,” in Memoria IV Congreso Nacional de Control a novel species of Nosema on the Southwestern corn borers Biolog ´ ico, pp. 31–34, Sociedad Mexicana de Control Biolog ´ ico, (Lepidoptera: Cambridae),” Journal of Economic Entomology, Monterrey, Nuevo Leon, ´ Mexic ´ o, Noviembre 2011. vol. 96, no. 1, pp. 12–20, 2003. [20] S. C. M. Leal,D.J.Bertioli,T.M.Butt, andJ.F.Peberdy, [35] M. W. Phoofolo, J. J. Obrycki, and L. C. Lewis, “Quantitative “Characterization of isolates of the entomopathogenic fungus assessment of biotic mortality factors of the european corn Metarhizium anisopliae by RAPD-PCR,” Mycological Research, borer (lepidoptera: crambidae) in field corn,” Journal of vol. 98, no. 9, pp. 1077–1081, 1994. Economic Entomology, vol. 94, no. 3, pp. 617–622, 2001. [21] S. Ali, Z. Huang, and S. Ren, “Media composition influences [36] L. F. Solter,J.V.Maddox, andC.R.Vossbrinck, “Physiological on growth, enzyme activity, and virulence of the entomopath- host specificity: a model using the European corn borer, ogen hyphomycete Isaria fumosoroseus,” Entomologia Experi- Ostrinia nubilalis (Hubner) ¨ (Lepidoptera: Crambidae) and mentalis et Applicata, vol. 131, no. 1, pp. 30–38, 2009. microsporidia of row crop and other stalk-boring hosts,” [22] J. C. Legaspi, T. J. Poprawski, and B. C. Legaspi Jr., “Laboratory Journal of Invertebrate Pathology, vol. 90, no. 2, pp. 127–130, andfieldevaluationof Beauveria bassiana against sugarcane stalkborers (lepidoptera: pyralidae) in the Lower Rio Grande [37] G. O. Poinar Jr., “Taxonomy and biology of Steinernematidae Valley of Texas,” Journal of Economic Entomology, vol. 93, no. and Heterorhabditidae,” in Entomopatogenic Nematodes in 1, pp. 54–59, 2000. Biological Control,R.Gaugler andH.K.Kaya, Eds.,pp. 23–61, [23] L. F. Solter and J. V. Maddox, “Microsporidia as classical CRC Press, 1990. biological control agents: research and regulatory issues,” [38] Z. R. Khan,J.A.Litsinger,A.T.Barrion,F.F.Villanueva, N. Phytoprotection, vol. 79, no. 4, pp. 75–80, 1998. J. Fernandez, and L. D. Taylo, World Bibliography of Rice Stem [24] N. Corradi and P. J. Keeling, “Microsporidia: a jour- Borers 1794–1990, International Rice Research Institute, 1991. ney through radical taxonomical revisions,” Fungal Biology [39] L. Dasrat, “Discovery of an indigenous entomopathogenic Reviews, vol. 23, no. 1-2, pp. 1–8, 2009. nematode and its pathogenicity to two sugar cane stem borers [25] J. Weiser, “Microsporidia and the society for invertebrate in Guyan,” http://wistonline.org/papers/proceedings/Paper16 pathology: a personal point of view,” Journal of Invertebrate .PDF. Pathology, vol. 89, no. 1, pp. 12–18, 2005. [40] F. Badilla, “Un programa exitoso de control biolog ´ ico de [26] J. A. Arcas, B. M. D´ıaz, and R. E. Lecuona, “Bioinsecticidal insectos plaga de la cana ˜ de azucar ´ en Costa Rica,” Manejo activity of conidia and dry mycelium preparations of two Integrado de Plagas y Agroecolog´ıa (Costa Rica), vol. 64, pp. 77– isolates of Beauveria bassiana against the sugarcane borer 87, 2002. Diatraea saccharalis,” Journal of Biotechnology,vol. 67, no.2- [41] M. E. Folegatti, S. Batista, P. R. Kawai, and P. S. Botelho, 3, pp. 151–158, 1999. “Nova metodologia para produc¸ao ˜ in vivo de Neoaplectana [27] J. C. LegaspiJr.,B.C.Legaspi, andR.R.Saldana, ˜ “Evalua- carpocapsae Weiser,” Nematologia Brasileira, vol. 12, pp. 76–83, tion of Steinernema riobravis (Nematoda: Steinernematidae) 1988. against the Mexican rice borer (Lepidoptera: Pyralidae),” [42] O. Sosa Jr., D. G. Hall, and W. J. Schroeder, “Mortality Journal of Entomological Science, vol. 35, no. 2, pp. 141–149, of sugarcane borer (Lepidoptera: Pyralidae) treated with 2000. entomopathogenic nematodes in field and laboratory trials,” International Journal of Zoology 11 Journal of the American Society of Sugar Cane Technology, vol. [56] H. C. T. Ribeiro and O. H. O. Pavan, “Effect of temperature 13, pp. 18–21, 1993. on the development of baculoviruses,” Journal of Applied [43] M. M. A. De Doucet, M. A. Bertolotti, A. L. Giayetto, and Entomology, vol. 118, pp. 316–320, 1994. M. B. Miranda, “Host Range, Specificity, and Virulence of [57] H. C. T. Ribeiro, O. H. O. Pavan, and A. R. Muotri, “Compar- Steinernema feltiae, Steinernema rarum,and Heterorhabditis ative susceptibility of two different hosts to genotypic variants bacteriophora (Steinernematidae and Heterorhabditidae) from of the Anticarsia gemmatalis nuclear polyhedrosis virus,” Argentina,” Journal of Invertebrate Pathology,vol. 73, no.3,pp. Entomologia Experimentalis et Applicata,vol. 83, no.2,pp. 237–242, 1999. 233–237, 1997. [44] J. P. M. Acevedo, R. I. Samuels, I. R. Machado, and C. Dolin- [58] S. R. Palli, T. R. Ladd, W. L. Tomkins et al., “Choristoneura ski, “Interactions between isolates of the entomopathogenic fumiferana entomopoxvirus prevents metamorphosis and fungus Metarhizium anisopliae and the entomopathogenic modulates juvenile hormone and ecdysteroid titers,” Insect nematode Heterorhabditis bacteriophora JPM4 during infec- Biochemistry and Molecular Biology, vol. 30, no. 8-9, pp. 869– tion of the sugar cane borer Diatraea saccharalis (Lepidoptera: 876, 2000. Pyralidae),” Journal of Invertebrate Pathology,vol. 96, no.2,pp. [59] C. M. Fauquet, M. A. Mayo, J. Maniloff,U.Desselberger, 187–192, 2007. andL.A.Ball, Eds., Virus Taxonomy, Eighth Report of the [45] P. J. Aguila, M. Vidal, M. Gonzalez, ´ L. A. Rodr´ıguez, and E. International Committee on Taxonomy of Viruses, Elsevier, San Mesa, “Criterios ecolog ´ icos en el manejo de plagas de la cana ˜ Diego, Calif, USA, 2005. de azucar ´ y cultivos varios una opcion ´ para lograr alimentos [60] J. S. Cory and H. F. Evans, “Viruses,” in Field Manual of Tech- sanos,” Agroecolog´ıa, vol. 3, pp. 51–53, 2008. niques in Invertebrate Pathology,L.A.Lacey andH.K.Kaya, [46] D. R. Ring and H. W. Browning, “Evaluation of entomopatho- Eds., pp. 149–174, Springer, Dordrecht, The Netherlands, genic nematodes against the Mexican rice borer (Lepidoptera: 2nd edition, 2007. Pyralidae),” Journal of Nematology, vol. 22, no. 3, pp. 420–422, [61] P. Caballero and T. Williams, “Virus Entomopatogenos, ´ ” in Control Biolog ´ ico de Plagas Agr´ıcolas,J.A.Jacas andA. [47] J. C. LegaspiJr.,B.C.Legaspi, andR.R.Saldana, ˜ “Evalua- Urbaneja, Eds., pp. 121–135, PHYTOMA-Espana, ˜ Navarra, tion of Steinernema riobravis (Nematoda: Steinernematidae) Espana, ˜ 2008. against the Mexican rice borer (Lepidoptera: Pyralidae),” [62] W. Mitsuhashi, H. Kawakita, R. Murakami et al., “Spindles of Journal of Entomological Science, vol. 35, no. 2, pp. 141–149, an entomopoxvirus facilitate its infection of the host insect by disrupting the peritrophic membrane,” Journal of Virology, vol. [48] E. Schnepf, N. Crickmore, J. Van Rie et al., “Bacillus thur- 81, no. 8, pp. 4235–4243, 2007. ingiensis and its pesticidal crystal proteins,” Microbiology and [63] M. Bergoin, J. C. Veyrunes, and R. Scalla, “Isolation and amino Molecular Biology Reviews, vol. 62, no. 3, pp. 775–806, 1998. acid composition of the inclusions of Melolontha melolontha [49] N. Bohorova, M. Cabrera, C. Abarca et al., “Susceptibility of poxvirus,” Virology, vol. 40, no. 3, pp. 760–763, 1970. four tropical lepidopteran Maize pests to Bacillus thuringiensis [64] R. L. Hall and R. W. Moyer, “Identification, cloning, and CryI-type insecticidal toxins,” Journal of Economic Entomology, sequencing of a fragment of Amsacta moorei entomopoxvirus vol. 90, no. 2, pp. 412–415, 1997. DNA containing the spheroidin gene and three vaccinia virus- [50] N. M. Rosas-Garc´ıa, B. Pereyra-Alfer ´ ez, K. A. Nin ˜o,L.J.Galn- related open reading frames,” Journal of Virology, vol. 65, no. Wong, and L. H. Morales-Ramos, “Novel toxicity of native and 12, pp. 6516–6527, 1991. HD Bacillus thuringiensis strains against to the sugarcane borer [65] D. Dall, A. Sriskantha, A. Vera, J. Lai-Fook, and T. Symonds, Diatraea saccharalis,” BioControl, vol. 49, no. 4, pp. 455–465, “A gene encoding a highly expressed spindle body protein of Heliothis armigera entomopoxvirus,” Journal of General [51] P. D. M. Gitahy,M.T.deSouza,R.G.Monnerat, E. D. B. Virology, vol. 74, no. 9, pp. 1811–1818, 1993. Arrigoni, and J. I. Baldani, “A Brazilian Bacillus thuringiensis [66] T. Hayakawa, J. Xu, and T. Hukuhara, “Cloning and sequenc- strain highly active to sugarcane borer Diatraea saccharalis ing of the gene for an enhancing factor from Pseudaletia (Lepidoptera: Crambidae),” Brazilian Journal of Microbiology, separata entomopoxvirus,” Gene, vol. 177, no. 1-2, pp. 269– vol. 38, no. 3, pp. 531–537, 2007. 270, 1996. [52] C. N. B. Carneiro,R.A.DaMatta,R.I.Samuels,and C. P. Silva, [67] Z. Li, C. Li, K. Yang et al., “Characterization of a chitin-binding “Effects of entomopathogenic bacterium Photorhabdus tem- protein GP37 of Spodoptera litura multicapsid nucleopolyhe- perata infection on the intestinal microbiota of the sugarcane drovirus,” Virus Research, vol. 96, no. 1-2, pp. 113–122, 2003. stalk borer Diatraea saccharalis (Lepidoptera: Crambidae),” [68] P. Wang and R. R. Granados, “Calcofluor disrupts the midgut Journal of Invertebrate Pathology, vol. 99, no. 1, pp. 87–91, defense system in insects,” Insect Biochemistry and Molecular Biology, vol. 30, no. 2, pp. 135–143, 2000. [53] P. Sikorowski andF.M.Davis,“Susceptibility of larvae of [69] T. Hukuhara and J. R. Bonami, “Reoviridae,” in Atlas of the southwestern corn borer, Diatraea grandiosella,to Bacillus Invertebrate Viruses,J.R.Adams andJ.R.Bonami, Eds.,pp. thuringiensis,” Journal of Invertebrate Pathology, vol. 15, no. 1, 393–434, CRC Press, Boca Raton, Fla, USA, 1991. pp. 131–132, 1970. [54] F. M. Davis and P. P. Sikorowsy, “Susceptibility of the South- [70] G. W. Blissard, B. Black, N. Crook, B. A. Keddie, R. Possee et al., “Family Baculoviridae,” in Virus Taxonomy: Seventh Report westerncornborer, Diatraea grandiosella Dyar (Lepidoptera: of the International Committee on Taxonomy of Virus,M.H.V. Pyralidae) to the baculovirus of Autographa californica,” Jour- van Regenmoertel, C. M. Fauquet, D. H. L. Bishop et al. et nal of the Kansas Entomological Society, vol. 51, no. 1, pp. 11– al., Eds., pp. 195–202, Academic Press, San Diego, Calif, USA, 13, 1978. [55] O. H. O. Pavan and H. C. T. Ribeiro, “Selection of a bac- ulovirus strain with a bivalent insecticidal activity,” Memor ´ ias [71] G. F. Rhormann, “Nuclear polyhedrosis viruses,” in Encyclope- do Insttituto Oswaldo Cruz, vol. 84, supplement 3, pp. 63–65, dia of Virology,R.G.Webster andA.Granoff, Eds., pp. 146– 1989. 152, Academic Press, London, UK, 2nd edition, 1999. 12 International Journal of Zoology [72] D. Winstanley and D. O. ’Reilly, “Granuloviruses,” in Encyclo- pedia of Virology,R.G.Webster andA.Granoff, Eds., pp. 140– 146, Academic Press, London, UK, 2nd edition, 1999. [73] B. Szewczyk, L. Hoyos-Carvajal, M. Paluszek, I. Skrzecz, and M. Lobo De Souza, “Baculoviruses—re-emerging biopesti- cides,” Biotechnology Advances, vol. 24, no. 2, pp. 143–160, [74] W. F. Jepson, A Critical Review of the World Literature of the Lepidopterous Stalk Borers of Graminaceous Crops, Common- wealth Institute of Entomology, London, UK, 1954. [75] J. W. SmithJr.,R.N.Wiedenmann, andW.A.Overholt, Parasites of Lepidopteran Stemborers of Tropical Gramineous Plants, ICIPE Science Press, Nairobi, Kenya, 1993. [76] G. M. Chippendale, “The southwestern corn borer, Diatraea grandiosella, case history of an invading insect,” Research Bulletin 1031, Missouri Agricultural Experiment Station, Columbia, Mo, USA, 1979. [77] N. Degaspari, P. S. M. Botelho, and N. Macedo, “Controle quimico da Diatraea saccharalis em cana-de-acucar na regiao ˜ Centro SuldoBrasil,” Boletim T´ecnico. IAA/PLANALSUCAR, vol. 3, pp. 1–16, 1981. [78] F. Moscardi, “Use of viruses for pest control in Brazil: the case of the nuclear polyhedrosis virus of the soybean caterpillar, Anticarsia gemmatalis,” Memor ´ ias do Insttituto Oswaldo Cruz, vol. 84, pp. 51–56, 1989. [79] G. Fedi ´ er ` e, “Epidemiology and pathology of Densovirinae,” in Parvovir Uses. From Molecular Biology to Pathology and Therapeutic Uses,S.Faisst andJ.Rommelaere, Eds.,pp. 1–11, Karger, Basel, Switzerland, 2000. [80] G. Meynadier, P. F. Galichet, J. C. Veyrunes, and A. Amargier, “Mise en ev ´ idence d’une densonucleose ´ chez Diatraea saccha- ralis [Lep.: Pyralidae],” Entomophaga, vol. 22, no. 1, pp. 115– 120, 1977. [81] N. Kouassi, J. X. Peng, Y. Li, C. Cavallaro, J. C. Veyrunes, and M. Bergoin, “Pathogenicity of Diatraea saccharalis densovirus to host insets and characterization of its viral genome,” Virologica Sinica, vol. 22, no. 1, pp. 53–60, 2007. [82] E. P. Steinhaus and G. A. Marsh, “Report of diagnosis of disease insects, 1951–1961,” Hilgardia, vol. 33, pp. 349–490, [83] U. K. Metha and H. David, “A granulosis virus disease of sugarcane internode borer,” Madras Agricultural Journal, vol. 67, pp. 616–619, 1970. [84] S. Easwaramoorthy and S. Jayaraj, “Survey of granulosis virus infection in sugarcane borers, Chilo infuscatellus Snellen and C. sacchariphagus indicus (Kapur) in India,” Tropical Pest Management, vol. 3, pp. 200–201, 1987. [85] S. Easwaramoorthy and H. David, “A granulosis virus of sugarcane shoot borer, Chilo infuscatellus Snell. (Lepidoptera: Crambidae),” Current Science, vol. 48, pp. 685–686, 1979. [86] S. Sethuraman and K. Narayanan, “Biological activity of Nucleopolyhedrovirus isolated from Chilo partellus (Swinhoe) (Lepidoptera: Pyralidae) in India,” Asian Journal of Experimen- tal Biological Sciences, vol. 1, pp. 325–330, 2010. [87] A. J. Cherry,C.J.Lomer,D.Djegui, andF.Schuethess, “Pathogen incidence and their potential as microbial control agents in IPM of maize stem borers in West Africa,” BioCon- trol, vol. 44, no. 3, pp. 301–327, 1999. 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Pathogens Associated with Sugarcane Borers, Diatraea spp. (Lepidoptera: Crambidae): A Review

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Copyright © 2012 Víctor M. Hernández-Velázquez et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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Hindawi Publishing Corporation International Journal of Zoology Volume 2012, Article ID 303589, 12 pages doi:10.1155/2012/303589 Review Article Pathogens Associated with Sugarcane Borers, Diatraea spp. (Lepidoptera: Crambidae): A Review 1 1 1 Vıct ´ or M. Hernandez-V ´ elazquez, ´ Laura P. Lina-Garcıa, ´ Veronica ´ Obregon-Bar ´ boza, 2 2 Adriana G. Trejo-Loyo, and Guadalupe Pena-Chor ˜ a Centro de Investigacion en Biotecnologıa, Universidad Autonoma del Estado de Morelos, Avenida Universidad No. 1001, ´ ´ ´ Colonia Chamilpa, 62210 Cuernavaca, MOR, Mexico Centro de Investigaciones Biolog ´ icas, Universidad Autonoma ´ del Estado de Morelos, Avenida Universidad No. 1001, Colonia Chamilpa, 62210 Cuernavaca, MOR, Mexico Correspondence should be addressed to V´ıctor M. Hernandez-V ´ elazquez, ´ vmanuelh@uaem.mx Received 14 June 2012; Accepted 29 August 2012 Academic Editor: Thomas Iliffe Copyright © 2012 V´ıctor M. Hernandez-V ´ elazquez ´ et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The objective of this paper was to analyze information related to entomopathogenic-associated Diatraea spp. Gaining a better understanding of the effects of these microorganisms will help in the development of successful microbial control strategies against stem borers that attack sugarcane plants. 1. Introduction also discuss the status of recent attempts to use pathogens in the field. The Diatraea spp. (Lepidoptera: Crambidae) complex is only found in the American continent, and it is the most impor- 2. Fungi tant group of stem borers that principally attack maize and sugarcane, as well as other gramineous crops, including rice, One promising field for research is the use of entomopatho- sorghum, and forage grasses [1]. The sugarcane borer (SCB) genic fungi as biological control agents of insect pests D. saccharalis Fab. is the most economically important pest in sugarcane plants. Approximately 80% of the etiological in South America [2, 3], whereas the neotropical corn stalk agents involved in insect diseases are fungi, which encompass borer (NCB) D. lineolata Walk is primarily found in Central 90 genera and more than 700 species [7]. A number of America [4], D. magnifactella Dyar and D. considerata fungi (Hypocreales: Clavicipitaceae) including B. bassiana, Heinrich are found in Mexico [1], and the southwestern corn B. brongniartii, M. anisopliae, P. fumosoroseus, Hirsutella sp., borer D. grandiosella Dyar is found in the United States [5]. Cylindrocarpon sp., and Nomuraea rileyi have been isolated Unfortunately, commercially available insecticides are from Diatraea spp. in the Americas from Argentina to not efficient for the control of Diatraea spp. for a variety of the USA (Table 1). Under certain climatic conditions, B. reasons, mainly because of the continuous presence of the bassiana has been reported to cause natural epizootics on D. host plants in fields throughout the year, the concomitant grandiosella [8]. presence of mature and immature forms of the insect, and The life cycle of entomopathogenic fungi in the arthro- the cryptic feeding habits of the insect [6]. An alternative pod hosts is initiated with the germination of conidia that strategy is integrated pest management with biological con- contacts the host integument and produces a germ tube trol as the first defense, which includes the use of parasitoids that penetrates the host through a combination of physical and entomopathogens. pressure and enzymatic degradation of the cuticle. The In this paper, we present a perspective on the attempts to fungus initially colonizes the host through a yeast phase. control Diatraea spp. using pathogens in the Americas. We Host death usually results from a combination of nutrient 2 International Journal of Zoology Table 1: Entomopathogens fungi from Diatraea spp. Host species Entomopathogen Country Reference D. grandiosella B. bassiana (1–8%) Mississippi, United States Inglis et al., 2000 [5] D. crambidoides D. grandiosella B. bassiana (epizootics) Texas High Plains, United States Knutson and Gilstrap, 1990 [8] B. bassiana D. saccharalis M. anisopliae Venezuela Zambrano et al., 2002 [9] P. fumosoroseus D. saccharalis M. anisopliae PE, Brazil Alves, et al. 2002 [10] Pinar del R´ıo, La Habana, D. saccharalis B. bassiana Matanzas, Villa Clara, Estrada et al., 2004 [11] Cienfuegos and Camaguey, Cuba B. bassiana M. anisopliae D. saccharalis Tucuman, ´ Argentina Yasen de Romero et al., 2008 [12] Nomuraea rileyi Iseria sp. M. anisopliae D. saccharalis Mexico Angel-Sahagun ´ et al., 2005 [13] (Ma2 and Ma3) B. bassiana ARSEF: 1489 Pernambuco 1832 Brazil 1834 Brazil 2629 Brazil 3020 Pernambuco 3858 Brazil D. saccharalis 5500 Pernambuco, Humber et al., 2009 [14] 5502 Brazil Brazil Santa Fe, Argentina Santa Fe, Argentina B. brigniartii D. saccharalis Sao Paulo, Brazil Humber et al., 2009 [14] ARSEF: 1830 Cylindrocarpon sp. D. saccharalis ARSEF: 8043 Colima, Mexico Humber et al., 2009 [14] M. anisopliae Colima, Mexico ARSEF: 3290 Colima, Mexico D. saccharalis 3291 Colima, Mexico Humber et al., 2009 [14] 3292 Colima, Mexico 3298 Colima, Mexico M. anisolpliae Hernandez ´ and Velazquez ´ D. magnifactella B. bassiana Mexico 2004 [15] Hirsutella sp. Marshal County, (Mississippi, D. grandiosella Nosema sp. isolate 167 Inglis et al. 2000 [5] USA) Oktibbeha County, (Mississippi, D. grandiosella Nosema sp. isolates 295, 504 Inglis et al. 2000 [5] USA) International Journal of Zoology 3 Table 1: Continued. Host species Entomopathogen Country Reference Washington County, D. grandiosella Nosema sp. isolates 181, 513, 522 Inglis et al. 2000 [5] (Mississippi, USA) Lastra and Gomez ´ 2000 D. saccharalis Nosema sp. Colombia [16] D. saccharalis Granulovirus (DsGV) Southern United States Pavan et al. (1983) [17] D. saccharalis Densovirus (DsDNV) Guadeloupe Meynadier et al. (1977) [18] Fonseca-Gonzalez et al. (2011) D. magnifactella B. thuringiensis Mexico [19] depletion, invasion of organs, and the action of fungal toxins. In general, these microorganisms live as parasites in Hyphae usually emerge from the cadaver. The mummified cells of the midgut epithelium, where they complete their corpse of the insect remains in the environment for several development, and the cycle starts when the infective states weeks and, in the case of stem borer, it keeps remains of microsporidia (spores) arrive at the digestive tube and protected inside the stem. Therefore, it is more common to colonize this region all the way to the excretory system. detect and isolate fungi compared to other pathogens that The spores germinate because of the acid intestinal pH, destroy the host, such as Bacillus thuringiensis (Bt), viruses, the microorganisms penetrate the midgut cells, and the or nematodes. intestinal activity is paralyzed between 14 to 21 days later Entomopathogenic fungi are widely distributed in all because the insect cannot assimilate nutrients. Nosema regions of the world; these species have wide genetic locustae infects the adipocytes of fat body, which interferes variation among the different isolates. Pathogenicity and with the adequate function of the insect’s intermediary virulence to differentspecies,aswellasenzymatic andDNA metabolism and competes with the insect for energetic characteristics, vary among different isolates [14, 20, 21]. reserves. Microsporidia produce effects that depend on the Therefore, it is important to evaluate as many geographic species and concentration; however, they generally produce isolates as possible from different areas to select the most weakness and eventually lead to death [31, 32]. suitable isolate based on its virulence and growth at high Only some species in this group have the possibility to temperatures. Several research groups have verified the be potentially relevant for natural or classical control. There pathogenicity and virulence of Hypocreales fungi, such as are many studies worldwide on the pathogenic effects of M. anisopliae and B. bassiana (Table 2), which have become the microsporidia N. pyrausta (Payllot) and Vairimorpha important biocontrol agents used for the microbial control of necatrix (Kramer) on borer organisms such as Ostrinia Diatraea spp. It is possible that a limited selection of available nubilalis (Hυ¨bner) (European corn borer), Lymantria dispar isolates of B. bassiana could identify highly virulent strains (Gypsy moth), and the grasshopper [5, 32]. In this group, from each of the different Diatraea species. only the microsporidium N. locustae has been registered as Field evaluations have been performed using M. aniso- a microbial insecticide for the control of grasshoppers in pliae and B. bassiana against several insect pests of sugar- grasslands [33]. cane, including D. saccharalis in Brazil. Application of M. In many areas of the USA and Europe, Nosema is the anisopliae at a rate of 1 × 10 spores per hectare caused main agent used for the control of grasshoppers; however, 58% mortality of D. saccharalis,and B. bassiana at 3.7 × 10 in Latin America, few studies have been performed with spores per milliliter reduced D. saccharalis damage by 45% Nosema [33]. For example, Inglis et al. [5]reportedsix (Alves et al., 1984 and 1985, cited in Legaspi et al. [22]). isolates of Nosema on larvae in the winter diapause stage For effectiveness in the field, it is important to consider the from D. grandiosella Dyer collected during 1998 in corn contact between the spores and the host, formulation, and stems from three locations in Mississippi, USA. However, the the virulence of the pathogens. frequency of infection in the field was very low (1, 3, and 15% in the counties of Marshal, Oktibbeha, and Washington, resp.), and no isolates were found in D. crambidoides (Grote) 3. Microsporidia (Table 3). Microsporidia (Eukaryota: Fungi) is the most ubiquitous When the mortality produced by the Nosema isolates group among insect populations [23, 24]. Microsporidia are was assessed in the laboratory using larvae that had stayed tiny unicellular organisms (from 2 to 40 μm in diameter), in environmental-like natural winter diapause conditions, that are opportunistic and obligate intracellular parasites variations in mortality between 0 and 55% were observed and attack different groups of invertebrate and vertebrate in larvae, and variations between 7 and 29% were observed animals. Microsporidia generally produce chronic diseases in pupae; homogenization of the dead larvae revealed a and reduce the physiological and reproductive ability of large amount of Nosema spores. However, in the surviving their hosts. Many species of microsporidia infect arthropods, adults, a large number of larvae were positive for the Nosema especially insects such as Lepidoptera and Coleoptera [23– spores when they were analyzed under light microscopy 25]. using staining techniques and electron microscopy. 4 International Journal of Zoology Table 2: Entomopathogenic-fungi bioassays on Diatraea spp. Host species Pathogen Bioassay Result Reference TL (conidia isolates Bb1 and 5): 4.3 days. Immersing third instar Mortality dry mycelium D. saccharalis B. bassiana larvae into a suspension of preparations: Arcas et al., 1999 [26] 10 conidia/mL 21.3% (Bb1) 82.5% (Bb5) at 7 days after inoculation LD in spores/mm 1st instar: 72.1 2nd instar: 384.3 Sprayed first, second and B. bassiana 3rd instar: 777.0 D. saccharalis third instar larvae with Legaspi et al., 2000 [27] Mycotrol strain GHA Mean days of survival 1mL 1st instar: 4.6 2nd instar: 4.8 3rd instar: 6.4 Dipped in conidial B. bassiana D. grandiosella suspension (dosage not Mortality from 10 to 21% Inglis et al., 2000 [5] Eight isolates specified) Third instar larvae sprayed B. bassiana ATCC 20872 D. saccharalis with suspension of TL 3.02 to 4.10 days Marques et al., 2000 [28] from Solenopsis invicta 10 conida/mL LC 5.6 × 10 yeast/mL Third instar larvae sprayed B. bassiana ATCC 20872 and D. sacccharalis with 3 mL of yeast-like cells Alves et al., 2002 [29] from Solenopsis invicta 4.8 × 10 conidia/mL or conidia B. bassiana 1cm larvae sprayed with CL 1.58 × 10 conidia/mL D. saccharalis Wenzel et al., 2006 [30] IBCB from Brazil 1mL at 6 days after inoculation Table 3: Entomopathogenic-microsporidia bioassays on Diatraea spp. Host specie Pathogen Bioassay Results Reference Surface of diet (squares), concentration not D. grandiosella Nosema sp isolate 167 Mortality of 1–15% Inglis et al., 2000 [5] mentioned. Larvae and pupae Surface of diet (squares), concentration not D. grandiosella Nosema sp isolate 295, 504 Mortality of 1–15% Inglis et al., 2000 [5] mentioned. Larvae and pupae Surface of diet (squares), Nosema sp isolate 181, 513, concentration not D. grandiosella Mortality of 1–15% Inglis et al., 2000 [5] 522 mentioned. Larvae and pupae Microsporidiosis LC = 1st instar larvae, Surface of 48.4 spores/mm D. saccharalis Vairimorpha necatrix Fuxa, 1981 [32] diet Gut damage LC = 8941 spores/mm 5-day-old larvae and Median infective dose of 2 2 3 pupae, leaf pieces ∼5cm × 10 spores per larva, D. grandiosella Nosema sp isolate 506 Inglis et al., 2003 [34] with 10 μl inoculum (10 to pupae small, decreased egg 10 spores) production In these experiments, the larvae were reared in the winter it is possible that Nosema can produce infection in natural diapause stage, and the temperature for Nosema was not conditions as the temperature inside the cane is higher than optimal for the development of infection; in the field during the external temperature. the winter, the prediapause larvae migrate to the base of Phoofolo et al. [35] observed a similar behavior, and the corn stem just below the surface of the soil [5], and they reported that the O. pyrausta infection by Nosema is International Journal of Zoology 5 chronic. Although no immediate mortality is produced, the Lastra and Gomez ´ [16] implemented a system to produce longevity and fecundity of the adults are reduced. Likewise, natural enemies of D. saccharalis in CENICANA (Clombian Phoofolo mentioned the possibility of a response to other Sugarcane Research Center, Colombia) and began their factors of mortality that regulate the population dynamics colony with larvae collected in the field. Their most signif- of borers, such as low temperature, the host plant, and icant result was the detection of one “protozoa,” possibly crowding, which have chronic effects on Nosema infection. Nosema, identified as the causal agent for the diminution Therefore, although the pest did not die directly because of in larvae production. They observed refractile spores in the microsporidia infection, the population size was eventually fluids of the malpighi tubes or hemolymph using phase reduced. contrast microscopy in healthy adults. Macroscopically, the This effect has been previously observed by Fuxa [32], diseased larvae were dwarfed and white. when he evaluated the susceptibility of larvae in the first In addition, different pathogenic microorganisms have and third instars of six species of Lepidoptera, including D. been isolated from Diatraea sp. in experimentally trans- saccharalis, to the microsporidium Vairimorpha necatrix.He mitted infections in the laboratory; however, there is little described two routes of infection that resulted in mortality; information about the natural presence of entomopathogens one resulted from the chronic effects produced by the belonging to the microsporidia group in this species of borer. exposure of larvae to low doses of spores, which led to lethal It is important to perform a systematic search for infected septicemia (microsporidiosis) just before pupating, and the larvae, pupae, or adults of Diatraea, and studies need to other route was due to the intake of a large number of include microscopy techniques to complement the mortality spores, which apparently damaged the gut because of the bioassays because infections could be asymptomatic. As introduction of a large number of spore polar filaments. In many microsporidia do not produce insecticide activity thisstudy,Fuxa[32] concluded that this pathogen could be quickly and because many species have complex life cycles promising for the remaining five species, although not for D. that involve more than one host, very few attempts have been saccharalis, as he had observed direct mortality by damage to made to implement microsporidia and use them as agents of the gut and indirect mortality by septicemia and because a biological control. high concentration of spores was necessary. Inglis et al. [34] reported that one strain of Nosema 4. Nematodes (506), which had been previously isolated [5], can infect other species of insects in the Crambidae family, including O. Pathogenic nematodes of the Heterorhabditidae and Stein- nubilalis and D. crambidoides (Grote), but not other species ernematidae families live in the soil where they are parasitic of Noctuidae. However, they also observed that the infection to certain soil-dwelling insects. The free-living third juvenile can be transmitted transovarially, although at a very low stage (infective juveniles, IJs) locates a suitable host and frequency (a difference with other Nosema species that can be penetrates the host in different ways. Once inside the insect, transmitted frequently in the vertical rout, such as the genus the IJs initiate its development, the nutritional tract becomes Ostrinia). functional, and the symbiotic bacterium Xenorhabdus or Solter et al. [36] researched the vertical and horizontal Photorhabdus is released through the anus and begins to transmission of seven species of microsporidia, including multiply in the hemocoel, which kills the insect by septicemia two strains of Nosema sp. (isolated from their natural hosts and creates suitable conditions for the reproduction of the in the field D. saecharalis and Eoreuma loftini); although nematode. Nematodes feed on bacteria and the dead tissue five of these strains were transmitted at a low percentage of the host, and they pass through several generations until horizontally and vertically, two Nosema strains did not new IJs are produced, which emerge from the cadaver [37]. behave similarly. They have a ubiquitous distribution [37]; therefore, it is not In assessing the infectivity percentage under laboratory unusual to find them in the soil of sugarcane plantations conditions when applying high concentrations of spores, (Pizano et al., 1985 cited in Khan et al. [38, 39]). they observed very low mortality in the larvae of D. sacharalis However, there are few reports of nematodes infecting and E. loftini (2 and 4%, resp.), although the same strains stem borers of the Diatraea genus in natural and experi- produced high mortality at low concentrations in the other mental situations. In Costa Rica, several entomopathogenic species studied, including O. nubilalis. They explained the species, including nematodes, have been isolated from D. low mortality, even at high doses, as a function of low tabernella [40]; however, the author does not mention the horizontal and vertical transmission because few infective or nematode species. abnormal spores were produced. Khan et al. [38] reviewed the world bibliography on rice In this case, it is important to clarify that they could stem borers and found five reports of entomopathogenic obtain live infected larvae without mortality; however, nematodes associated with D. saccharalis,two reportsof they only measured mortality and did not measure other Steinernema (=Neoaplectana) glaseri in Brazil and three parameters, such as fecundity or larvae hatching in the next reports of Steinernema (=Neoaplectana) carpocapsae in the generation. As mentioned by Fuxa [32] and Inglis et al. [5], USA and Guadeloupe. However, in one study, they report infection does not always lead to mortality, and occasionally that the aim of the research was the use of D. saccharalis to the effects are observed long term and can be inferred from produce S. glaseri in controlled conditions because of its sus- a reduction in fecundity and susceptibility to other stress ceptibility to the nematode. Another study by Folegatti et al. situations. [41] used entomopathogenic nematodes and D. saccharalis 6 International Journal of Zoology Table 4: Entomopathogenic-nematode bioassays on Diatraea spp. Host specie Pathogen Bioassay Result Reference 100% mortality with 5000 Steinernema feltiae nematodes/larvae [Neoaplectana carpocapsae] Several concentrations for 100% mortality with 5000 Diatraea saccharalis Heterorhabditis heliothidis Sosa et al., 1993 [42] all species nematodes/larvae 30% mortality with 5000 S. glaseri nematodes/larvae S. feltiae 500 IJs/5 larvae in Petri S. rarum Adults and IJs produced. Diatraea saccharalis dishes with two moistened De Doucet et al., 1999 [43] Heterorhabditis Larval mortality >90% filter papers disks bacteriophora H. bacteriophora JPM4 Larvae were exposed to 25 100% larval mortality Diatraea saccharalis isolatedfromsoilinLarvae ± 5 IJs/100 μLfor 48 hin within 5 days. LT 2.1 d Molina et al., 2007 [44] MG, Brazil sand-filled Petri dishes and LT 4.4 d Heterorhabditis Diatraea saccharalis 500 mL/L applied to larvae 93% of larvae mortality Aguila et al., 2008 [45] bacteriophora in laboratory conditions to produce S. carpocapsae in vivo, Bohorova et al. [49] evaluated Cry1Aa, Cry1Ab, Cry1Ac, using larvae of the sugarcane borer as the host. Cry1B, Cry1C, Cry1D, Cry1E, and Cry1F Bt pure proteins We believe that pathogenic nematodes have good poten- against four species of Lepidoptera that are pests of maize, tial for the biological control of Diatraea species because including D. saccharalis. The proteins were diluted in water of their presence in sugarcane-producing regions (as men- and added to the diet at doses of 10 and 100 mg/mL of diet, tioned above) and because the species S. feltiae, S. glaseri, and mortality was recorded after seven days. D. saccharalis S. rarum, Heterorhabditis heliothis,and H. bacteriophora was susceptible to Cry1B protein at 10 mg/g, and the LC have demonstrated high infectivity in D. saccharalis [42–45] was 113.6 mg/g of meridic diet (Table 5). (Table 4)aswellasin E. loftini (some of them) [46, 47]in Twelve Bt strains were evaluated by Rosas Garc´ıa et al. experimental studies. We consider that performing a system- [50] at a dosages of 50 and 500 μg of total protein and spores atic search for infected larvae or pupae of Diatraea could be per milliliter against 2-day-old D. saccharalis larvae. The helpful in finding new nematode strains and species adapted strains used were HD1, HD2, HD9, HD29, HD37, HD59, to local environmental conditions and pest species that could HD133, HD137, and HD559, as well as the GM7, GM10, and be used in the future. GM34 native strains. The strains that killed more than 50% Although we could not find any report on field trials, of larvae were selected to obtain the LC . Strains HD133, we are aware that there have been mass rearings of H. HD559, GM7, GM10, and GM34 were toxic; however, GM34 bacteriophora in Cuba since 1987 to control soil pests, such was the most toxic with an LC of 33.21 μg/mL (Table 5). as D. saccharalis [45]. PCR analysis was performed to determine the cry1 genes of the toxic strains: HD133 cryAa, cry1Ab, cry1C; HD559 and GM7 cry1Aa, cry1Ab,and cry1B;GM10 cry1Aa, cry1Ab, 5. Bacteria cry1Ac,and cry1C;GM34 cry1Aa, cry1Ab,and cry1Ac. Bacillus thuringiensis (Bt) is a Gram-positive bacterium Gitahy et al. [51] evaluated a spore-crystal complex in that has been isolated from several sources, including soil, vitro from five native Bt strains (S48, S76, S90, S105, and water, phylloplane, and insect cadavers. Bt produces various S135) and HD1, which served as a positive control, on second instar larvae of D. saccharalis and mortality was recorded insecticidal crystal proteins during the onset of sporulation that are toxic to insects, acari, and nematodes [48]. The steps after 5 days. The strain S76 caused 100% mortality at 72 h, involved in the mode of action of the proteins after their HD1 caused 69% mortality, and 3% of the other native −1 strains caused mortality at 500 μgL of the spore-crystal ingestion are as follows: (a) solubilization of the crystals by the highly alkaline pH of the midgut, (b) activation of the complex. The LC values of the S76 and HD1 strains were proteins by proteases, (c) binding of the toxins to specific determined, and S76 was 11-fold more toxic (13.06 μg/L) receptors located on the microvilli membrane of the midgut than HD1 (143.88 μ/L) (Table 5). The S76 strain carries columnar epithelium cells, and (d) the insertion of the toxin cryAa, cry1Ab, cryAc, cry2Aa,and cry2Ab genes, which is similar to HD1. into the membrane, which forms a pore and induces cell lysis. There are no reports from the American continent There are other species of bacteria with potential insec- on the isolation of Bt strains from Diatraea spp. larvae ticide activity; Carneiro et al. [52]evaluated Photorhabdus temperata, which is a bacterium associated with Heterorhab- cadavers; however, some Bt strains and pure proteins have been evaluated against these pests. ditis entomopathogenic nematodes. Cells were injected with International Journal of Zoology 7 Table 5: Entomopathogenic-bacteria bioassays on Diatraea spp. Host species Pathogen Bioassay Result Reference Diet incorporated Diatraea saccharalis Bacillus thuringiensis LC 113.6 mg/g diet Bohorova et al., 1997 [49] spores-crystals Rosas-Garcia et al., 2004 D. saccharalis B. thuringiensis LC 33.21 μg/mL [50] Diet incorporated D. saccharalis B. thuringiensis LC 13.06 μg/L Gitahy et al., 2007 [51] spores-crystals complex Cells injected LD 16.2 bacterial cells D. saccharalis Photorhabdustemperata Carneiro et al., 2008 [52] LT 33.8 h Diet incorporated Diatraea grandiosella Bacillus thuringiensis LC 5.2 mg/g diet Bohorova et al., 1997 [49] spores-crystals Mean mortality on Bacillus thuringiensis 5-day-old larvae: 32, 31, 25, Immersing stem seedling in Biotrol BTB 183-25 15 out of 40. spores suspensions: 1.25 × Nutrilite Products Control mean mortality Sikorowski and Davis, 1970 8 7 7 Diatraea grandiosella 10 ,6.25 × 10 ,2.5 × 10 Incorporated, Kansas City, 0.01 [53] and 1.25 × 10 Missouri, USA Mean mortality on 10-day-old larvae: 15, 15, 12 and 6 out of 20. avolumeof10 μL of phosphate-buffered saline directly into these entomopathogenic viruses, OBs have independently the hemocoel of fourth instar D. saccharalis larvae, and the evolved as a protective mechanism to environmental factors, LD was16.2bacterial cellswithanLT of 33.8 h. which gives the viruses a great advantage as biological control 50 50 Sikorowski and Davis [53] determined the susceptibility agents [61]. Baculoviruses (BVs) and entomopoxviruses of 5- and 10-day-old D. grandiosella larvae with a Bt (EPVs: subfamily Entomopoxvirinae)havealargedouble- commercial product (Biotrol BTB 183-25) using 1.25 × 10 , stranded DNA genome, and cypoviruses (CPVs: family 7 7 7 6.25 × 10 ,2.5 × 10 , and 1.25 × 10 spores per milliliter Reoviridae, genera Cypovirus) contain segmented double- (Table 5) where two-inch stem seedlings were dipped for 30 stranded RNA viruses. min in spore suspensions. One 10-day-old or two 5-day- EPVs have been reported to infecting insects of a variety old larvae were allowed to feed on each stem. In addition, of orders, such as Coleoptera, Lepidoptera, Orthoptera, a known number of spores were placed or injected into 5- and Diptera. Some EPVs have two distinct OBs spheroids day-old stem seedlings at 1/4 inch sections. Stems dipped in and spindles. The spheroids occlude virions, whereas the water were used as controls. Mortality was recorded after spindles do not [62]. The most abundant proteinaceous 48 h. With 10-day-old larvae, five replicates with 20 larvae component of spheroids and spindles is proteins called for each treatment were used; for the 5-day-old larvae, two spheroidin and fusolin, respectively [63–66]. EPV fusolin is replicates with 40 larvae per treatment were used. With 5- an enhancing factor (EF) that increases BVs infection and and 10-day-old larvae at 1.25 × 10 spores per milliliter, a has been characterized as a chitin-binding protein [67]. The mean of 32 dead larvae out of 40 and 15 dead larvae out of 20 fusolin mechanism of action is similar to that of Calcofluor, were recorded (Table 5), respectively. Thus, it was concluded which facilitates BV infection by disrupting or preventing that this species is highly susceptible to B. thuringiensis. the formation of the peritrophic membrane [68]. EPVs are pathogenic but are scarcely virulent; infected larvae exhibit extreme longevity and take up to 70 days to die. However, 6. Viruses EPVs have potential as biological control agents for pest insects where BVs have not been isolated [61]. Because of Viruses that infect insects have received great attention as biological control agents because of their specificity on insect the activity of spindles, the EF of EPVs can be used as populations; they have little or no impact on the environ- a synergistic agent to increase BV infectivity or generate genetically modified organisms, such as BVs and transgenic ment and are an ecological friendly alternative to chemical pesticides [58]. Entomopathogenic viruses are grouped in 33 plants. genera within fifteen families [59]. However, only a few of CPVs have been mainly isolated from Lepidoptera these families, such as Baculoviridae, Poxviridae, and Reoviri- insects. OBs are dissolved in the midgut, and virions only dae, have potential as biological control agents and have been infect epithelial cells; therefore, CPVs are very pathogenic but act slowly and frequently to produce chronic infections [69]. successfully used in microbial control programs. A common characteristic among these entomopathogenic viruses is that At this time, no commercial bioinsecticides based on CPVs the virions (infective unit) are occluded within a crystalline have been developed. However, Caballero and Williams [61] suggest that their greatest potential as biological control protein matrix to form an occlusion body (OB) [60], which is a unique characteristic of viruses that infect insects. In agents is through inoculative or augmentative releases. 8 International Journal of Zoology Table 6: Entomopathogenic-virus bioassays on Diatraea spp. Host species Pathogen Bioassay conditions Result Reference DL :1.32 × 4 and 7 day-old larvae were 3 ∗ 10 PIB /larvae (for Autographa californica fed strips of corn leaf D. grandiosella 4-day-old larvae) Davis and Sikorowsy, 1978 [54] MNPV (AcMNPV) soaked in different DL :1.32 × 10 PIB/larvae dilutions of virus. (for 7-day-old larvae). 3rd instar larvae feeding LD : 42 PIB/larva at 26 C Diatraea saccharalis individually on small LT :From29to63for Lastra and Gomez ´ D. saccharalis Granulosis Virus artificial diet discs treated 7 2 10 and10 PIB/larvae, et al., 1983 [16] (DsGV) with 2.7 μL of virus respectively. dilutions. 3rd instar larvae fed with LD :From7.9 × 10 to 5.3 Anticarsia gemmatalis artificial diet discs treated × 10 for 1 and 20 serial D. saccharalis MNPV (AgMNPV) with 5 different doses of Pavan and Ribeiro, 1989 [55] passages, respectively. PIB through 20 serial passages. 3rd instar larvae AgMNPV wt LT :From10to13for individually fed small 39 and 30 C, respectively artificial diet discs D. saccharalis Ribeiro and Pavan, 1994 [56] containing 10 PIB of virus From 9to29for 39 and and kept individually at 10 AgMNPV-D10 22 C, respectively different temperatures. 3rd instar larvae fed LT :From9to 16 for Trichoplusia ni MNPV individually with small 39 and 24 C, respectively (TnMNPV) wt artificial diet discs D. saccharalis Ribeiro and Pavan, 1994 [56] containing 10 PIB of virus and kept individually at 10 From 6to46for 39 and TnMNPV-D11 different temperatures. 17 C, respectively 3rd instar larvae fed on LD :From5.3 × formalin free diet disc (3 10 PIB/larvae (7A mm) inoculated with genotypic variant) to 8 × D. sacccharalis AgMNPV Ribeiro et al., 1997 [57] 2.7 μL of the viral solution 10 PIB/larvae (33B from 10 clonal isolates of genotypic variant) the AgMNPV-Ds20 PIB: Polyhedral inclusion bodies. BVs predominantly infect insects within the Lepidoptera control of these insect pests difficult. Degaspari et al. [77] order, which includes important agricultural insect pests have argued that chemical control of D. saccharalis in Brazil [70]. The Baculoviridae family includes two genera: nucle- is not economically feasible. To solve this problem, surveys to opolyhedrovirus (NPV), which forms large, polyhedric OBs isolate endemic entomopathogens of stem borer populations where many enveloped virions are occluded [71], and in maize and sugarcane crops have been developed. Granulovirus (GV), which forms small, granular OBs that Inglis et al. [5] developed an exhaustive survey to occlude only one enveloped virion each [72]. BVs are safe for isolate entomopathogens from the southern corn borer, humans and wildlife. Their specificity is usually very narrow D. grandiosella, and the southern corn stalk borer, D. and often is species specific. Because of their specificity crambidoides, and larvae in the diapause stage were collected and other characteristics, such as elevated virulence and from crops located in Mississippi and North Carolina. These pathogenicity, BVs are by far the most studied and extensively authors did not observe OBs in any of the collected larvae used as commercial biopesticides for the control of a variety and concluded that there are no naturally occurring viruses of insect pests in many countries around the world [60, 61, in these Diatraea populations [5]. According to Inglis et 73]. al. [5], Pavan and Ribeiro [55] mentioned that natural Stem borers of the Lepidoptera order attack gramineous populations of the SCB in Brazil do not exhibit endemic viral crops throughout the world [74, 75]. Diatraea stem borers pathogens. (DSB) are widely distributed in the Americas and attack a Currently, there are only two records of endemic ento- wide variety of host plants, including maize and sugarcane mopathogenic viruses isolated from Diatraea spp. larvae. [1]. Prediapause larvae of southwestern corn borers migrate Pavan et al. [17] isolated a GV from the sugarcane borer to the base of the stalk of the corn plant below the soil surface (SCB), D. saccharalis, from sugarcane crops in the southern to survive the winters [76]; cryptic habits make the chemical United States (Table 6). These authors developed bioassays International Journal of Zoology 9 with D. saccharalis third instar larvae (Table 6). External thank Mrs. Ingrid Masher for reviewing the paper and for symptoms of GV-infected larvae were similar to those editorial assistance. reported for other lepidopterous larvae, and the symp- toms and ultrastructures were determined using electronic References microscopy as well as replication of DsGV, which are typical [1] L. A. Rodriguez-Del-Bosque and J. W. Smith Jr., “Biological of GVs. The LD for the third instar larvae was 42.3 control of maize and sugarcane stemborers in Mexico: a OBs/larva (Table 6) with 14.5 and 123.6 OBs/larva as the review,” Insect Science and Its Application,vol. 17, no.3-4,pp. lower and upper limits at 95% probability, respectively [16]. 305–314, 1997. These authors stated that this work was the first of a series of [2] G. Serra and E. Trumper, “Estimacion ´ de incidencia de danos ˜ publications; however, there have been no additional studies provocados porlarvasde Diatraea saccharalis (Lepidoptera: produced to date, such as the molecular and biological Crambidae) en tallos de ma´ız mediante evaluacion ´ de signos characterization, of this GV, DsGV was introduced into ´ externos de infestacion,” Agrosciencia, vol. 23, no. 1, pp. 1–7, Brazil [16], and Moscardi [78] mentioned that this virus is 2006. [3] M. A. P. De Oliveira, E. J. Marques, V. Wanderley-Teixeira, currently being applied at a small scale as an experimental and R. Barros, “Effect of Beauveria bassiana (Bals.) Vuill. and product on sugarcane. Metarhizium anisopliae (Metsch.) Sorok. on biological charac- Because of their high pathogenicity combined with a teristics of Diatraea saccharalis F. (Lepidoptera: Crambidae),” limited host range, Densoviruses (DNVs) have potential as Acta Scientiarum, vol. 30, no. 2, pp. 220–224, 2008. effective insecticides [79]. Meynadier et al. [80] isolated [4] R. Reyes, “Sorghum stem borers in Central and South aDNV of D. saccharalis (DsDNV) from the Guadeloupe America,” in Proceedings of the International Workshop on sugarcane borer (Table 6). Kouassi et al. [81] tested the Sorghum Stem Borers, pp. 49–60, ICRISAT Center, Hyderabad, pathogenicity of DsDNV on its host. These authors observed India, 1987. that the infected larvae exhibited infection symptoms from [5] G. D. Inglis, A. M. Lawrence, and F. M. Davis, “Pathogens the fourth day postinfection, such as anorexia and lethargy associated with southwestern corn borers and southern corn followed by flaccidity and inhibition of molting and meta- stalk borers (Lepidoptera: Crambidae),” Journal of Economic morphosis. Larvae became paralyzed and stopped feeding Entomology, vol. 93, no. 6, pp. 1619–1626, 2000. [6] M.D.R.T.DeFreitas,E.L.DaSilva,A.D.L.Mendonc¸a after 7 days. The cumulative mortality of infected larvae et al., “The biology of Diatraea flavipennella (Lepidoptera: increased significantly and reached 60% after 12 days and Crambidae) reared under laboratory conditions,” Florida 100% at 21 days postinfection [81]. Although DNVs have Entomologist, vol. 90, no. 2, pp. 309–313, 2007. no potential for large scale use as a biological control agent [7] R.H.R.Destefano ´ , S. A. L. Destefano ´ , and C. L. Messias, because they have no OBs and are related to vertebrate “Detection of Metarhizium anisopliae var. anisopliae within pathogenic viruses, the genes involved with anorexia and infected sugarcane borer Diatraea saccharalis (Lepidoptera, paralysis could be used to produce transgenic BVs or plants. Pyralidae) using specific primers,” Genetics and Molecular Most BVs have a limited host range and may be species Biology, vol. 27, no. 2, pp. 245–252, 2004. specific in some cases, although there are several NPVs, such [8] A.E.Knutson andF.E.Gilstrap, “Seasonal occurrence of as Autographa californica MNPV, Anagrapha falcifera MNPV, Beauveria bassiana in the southwestern corn borer (Lepi- and Anticarsia gemmatalis MNPV, which have broader host doptera: Pyralidae) in the Texas High Plains,” Journal of the Kansas Entomological Society, vol. 63, no. 2, pp. 243–225, 1990. ranges within the Lepidoptera order [60]. Because of the [9] K. Zambrano, M. Dav ´ ila, and M. A. Castillo, “Deteccion ´ de scarcity of reports on natural isolates of entomopathogenic fragmentos de AND de hongos y su posible relacion ´ con la viruses from Diatraea spp. populations, cross-infectivity of s´ıntesis de prote´ınas de actividad entomopatogena, ´ ” Revista de AgMNPV, TnMNPV, and AgMNPV has been evaluated in D. la Facultad De Agronom´ıa (LUZ), vol. 19, pp. 185–193, 2002. saccharalis and D. grandiosella larvae (Table 6). [10] S. B. Alves, L. S. Rossi,R.B.Lopes,M.A.Tamai,and R. M. Although only two reports of entomopathogenic viruses Pereira, “Beauveria bassiana yeast phase on agar medium and isolated from Diatraea stem borers exist in the Americas, its pathogenicity against Diatraea saccharalis (Lepidoptera: entomopathogenic viruses, such as NPVs and GVs, have Crambidae) and Tetranychus urticae (Acari: Tetranychidae),” been reported to occur in Africa and Asia in cereal stem Journal of Invertebrate Pathology, vol. 81, no. 2, pp. 70–77, borers within the Crambidae family, including maize and sugarcane borers such as Chilo sp. [82], Chilo sacchariphagus [11] M. E. Estrada, M. Romero,M.J.Rivero, andF.Barroso, “Natural presence of Beauveria bassiana (Balsamo) Vuillemin [83, 84], Ch. infuscatellus [84, 85], Ch. partellus [86], and in the sugar cane (Saccharum sp. hybrid) in Cuba,” Revista Eldana saccharina [87]. As the Mexican territory is the origin Iberoamericana de Micologia, vol. 21, no. 1, pp. 42–43, 2004. of some Diatraea stem borer species, we hypothesize that a [12] M. G. Yasen de Romero, A. R. Salvatore, G. Lop ´ ez, and great diversity of pathogenic viruses in Diatraea populations E. Willink, “Presencia natural de hongos hyphomycetes en that attack sugarcane crops exists in Mexico. larvas invernantes de Diatraea ssaccharalis F. en cana ˜ de azucar ´ en Tucuman, ´ Argentina,” Revista Industrial y Agr´ıcola Acknowledgments de Tucuman ´ , vol. 85, no. 2, pp. 39–42, 2008. [13] C. A. Angel-Sahagun, ´ R. Lezama-Gutier ´ rez, J. Molina-Ochoa This study was financed in part by the project “Seleccion ´ de et al., “Susceptibility of biological stages of the horn fly, enemigos naturales de barrenadores de la cana ˜ de azucar ´ de Haematobia irritans, to entomopathogenic fungi (Hyphomy- Colima y Morelos con potencial como agentes de control cetes),” Journal of Insect Science (Online), vol. 5, no. 50, pp. 1–8, biolog ´ ico (SEP-PROMEP),” of Mexico. The authors also 2005. 10 International Journal of Zoology [14] R. A. Humber, K. S. Hansen, and M. M. Wheeler, Catalog of [28] E. J. Marques, S. B. Alves, and I. M. R. Marques, “Virulencia Species. ARS Collection of Entomopathogenic Fungal Cultures, de Beauveria bassiana (Bals.) Vuill. a Diatraea saccharalis (F.) USDA-ARS Biological Integrated Pest Management Research, (Lepidoptera: Crambidae) apos ´ armazenamiento de con´ıdios Ithaca, NY, USA, 2009. em baixa temperatura,” Anais da Sociedade Entomolog ´ ica do Brasil, vol. 29, no. 2, pp. 303–307, 2000. [15] V. M. Hernandez ´ Velazquez ´ and R. Lezama Gutier ´ rez, “Uso de [29] S. B. Alves, L. S. Rossi,R.B.Lopes,M.A.Tamai,and R. M. entomopatogenos ´ para el control biolog ´ ico de barrenadores Pereira, “Beauveria bassiana yeast phase on agar medium and del tallo,” in Taller Internacional Sobre Barrenadores Del Tallo ´ its pathogenicity against Diatraea saccharalis (Lepidoptera: De Cana ˜ De Azucar ´ ,L.A.Rodrıguez del Bosque, G. Vejar, Crambidae) and Tetranychus urticae (Acari: Tetranychidae),” and E. Cortez, ´ Eds., pp. 37–45, Sociedad Mexicana de Control Biolog ´ ico, Los Mochis, Sinaloa, Mexico, Noviembre de 2004. Journal of Invertebrate Pathology, vol. 81, no. 2, pp. 70–77, [16] B. Lastra andL.A.Gomez, ´ “Cr´ıa y produccion ´ masiva de [30] I. M. Wenzel, F. H. C. Gionetti, and J. E. M. Almeida, “Pato- insectosenunprogramadecontrol biolog ´ ico en cana ˜ de genicidade do isolade IBCB66 de Beauveria bassiana sbroca da azucar ´ ,” in 1er Curso Taller Internacional de Control Biolog ´ ico, ˜ ´ cana-de-acucar Diatraea saccharalis em condicoes de labo- pp. 335–340, Memorias, Bogota, ´ Colombia. rator ´ io,” Arquivos Do Instituto Biologico, vol. 72, no. 2, pp. 259– [17] O. H. O. Pavan, D. G. Boucias, L. C. Almeida, J. O. Gaspar, P. S. 261, 2006. M. Botelho, and N. Degaspari, “Granulosis Virus of Diatraea [31] C. E. Lange, “Niveles de esporulacion ´ experimentales y saccharalis (DsGV): pathogenicity, replication and ultrastruc- naturales de Nosema locustae (Microsporidia) en especies de ture,” in Proceedings of the International Congress of the tucuras y langostas (Orthoptera: Acridoidea) de la Argentina,” International Society of Sugar Cane Technologists (ISSCT ’83), RevistadelaSociedad Entomolog ´ ica Argentina, vol. 62, no. 1-2, vol. 2, pp. 644–659, La Havana, Cuba, 1983. pp. 15–22, 2003. [18] G. Meynadier, P. F. Galichet, J. C. Veyrunes, and A. Amargier, [32] J. R. Fuxa, “Susceptibility of Lepidopterous pests to two ´ ´ “Mise en evidence d’une densonucleose chez Diatraea saccha- types of mortality caused by the microsporidium Vairimorpha ralis [Lep.: Pyralidae],” Entomophaga, vol. 22, no. 1, pp. 115– necatrix,” Journal of Economic Entomology,vol. 74, no.1,pp. 120, 1977. 99–102, 1981. [19] A. Fonseca-Gonzalez, ´ G. Pena-Chor ˜ a, A. Trejo-Loyo, L. Lina- [33] L. F. Solter and J. V. Maddox, “Microsporidia as classical Garc´ıa,L.A.Rodr´ıguez-Del Bosque, and V. M. Hernandez- ´ biological control agents: research and regulatory issues,” Velazquez, ´ “Virulencia de seis aislados de Bacillus thuringiensis Phytoprotection, vol. 79, no. 4, pp. 75–80, 1998. nativos delestado de Morelos y evaluados sobre Diatraea [34] G. D. Inglis, A. M. Lawrence, and F. M. Davis, “Impact of magnifactella,” in Memoria IV Congreso Nacional de Control a novel species of Nosema on the Southwestern corn borers Biolog ´ ico, pp. 31–34, Sociedad Mexicana de Control Biolog ´ ico, (Lepidoptera: Cambridae),” Journal of Economic Entomology, Monterrey, Nuevo Leon, ´ Mexic ´ o, Noviembre 2011. vol. 96, no. 1, pp. 12–20, 2003. [20] S. C. M. Leal,D.J.Bertioli,T.M.Butt, andJ.F.Peberdy, [35] M. W. Phoofolo, J. J. Obrycki, and L. C. Lewis, “Quantitative “Characterization of isolates of the entomopathogenic fungus assessment of biotic mortality factors of the european corn Metarhizium anisopliae by RAPD-PCR,” Mycological Research, borer (lepidoptera: crambidae) in field corn,” Journal of vol. 98, no. 9, pp. 1077–1081, 1994. Economic Entomology, vol. 94, no. 3, pp. 617–622, 2001. [21] S. Ali, Z. Huang, and S. Ren, “Media composition influences [36] L. F. Solter,J.V.Maddox, andC.R.Vossbrinck, “Physiological on growth, enzyme activity, and virulence of the entomopath- host specificity: a model using the European corn borer, ogen hyphomycete Isaria fumosoroseus,” Entomologia Experi- Ostrinia nubilalis (Hubner) ¨ (Lepidoptera: Crambidae) and mentalis et Applicata, vol. 131, no. 1, pp. 30–38, 2009. microsporidia of row crop and other stalk-boring hosts,” [22] J. C. Legaspi, T. J. Poprawski, and B. C. Legaspi Jr., “Laboratory Journal of Invertebrate Pathology, vol. 90, no. 2, pp. 127–130, andfieldevaluationof Beauveria bassiana against sugarcane stalkborers (lepidoptera: pyralidae) in the Lower Rio Grande [37] G. O. Poinar Jr., “Taxonomy and biology of Steinernematidae Valley of Texas,” Journal of Economic Entomology, vol. 93, no. and Heterorhabditidae,” in Entomopatogenic Nematodes in 1, pp. 54–59, 2000. Biological Control,R.Gaugler andH.K.Kaya, Eds.,pp. 23–61, [23] L. F. Solter and J. V. Maddox, “Microsporidia as classical CRC Press, 1990. biological control agents: research and regulatory issues,” [38] Z. R. Khan,J.A.Litsinger,A.T.Barrion,F.F.Villanueva, N. Phytoprotection, vol. 79, no. 4, pp. 75–80, 1998. J. Fernandez, and L. D. Taylo, World Bibliography of Rice Stem [24] N. Corradi and P. J. Keeling, “Microsporidia: a jour- Borers 1794–1990, International Rice Research Institute, 1991. ney through radical taxonomical revisions,” Fungal Biology [39] L. Dasrat, “Discovery of an indigenous entomopathogenic Reviews, vol. 23, no. 1-2, pp. 1–8, 2009. nematode and its pathogenicity to two sugar cane stem borers [25] J. Weiser, “Microsporidia and the society for invertebrate in Guyan,” http://wistonline.org/papers/proceedings/Paper16 pathology: a personal point of view,” Journal of Invertebrate .PDF. Pathology, vol. 89, no. 1, pp. 12–18, 2005. [40] F. Badilla, “Un programa exitoso de control biolog ´ ico de [26] J. A. Arcas, B. M. D´ıaz, and R. E. Lecuona, “Bioinsecticidal insectos plaga de la cana ˜ de azucar ´ en Costa Rica,” Manejo activity of conidia and dry mycelium preparations of two Integrado de Plagas y Agroecolog´ıa (Costa Rica), vol. 64, pp. 77– isolates of Beauveria bassiana against the sugarcane borer 87, 2002. Diatraea saccharalis,” Journal of Biotechnology,vol. 67, no.2- [41] M. E. Folegatti, S. Batista, P. R. Kawai, and P. S. Botelho, 3, pp. 151–158, 1999. “Nova metodologia para produc¸ao ˜ in vivo de Neoaplectana [27] J. C. LegaspiJr.,B.C.Legaspi, andR.R.Saldana, ˜ “Evalua- carpocapsae Weiser,” Nematologia Brasileira, vol. 12, pp. 76–83, tion of Steinernema riobravis (Nematoda: Steinernematidae) 1988. against the Mexican rice borer (Lepidoptera: Pyralidae),” [42] O. Sosa Jr., D. G. Hall, and W. J. Schroeder, “Mortality Journal of Entomological Science, vol. 35, no. 2, pp. 141–149, of sugarcane borer (Lepidoptera: Pyralidae) treated with 2000. entomopathogenic nematodes in field and laboratory trials,” International Journal of Zoology 11 Journal of the American Society of Sugar Cane Technology, vol. [56] H. C. T. Ribeiro and O. H. O. Pavan, “Effect of temperature 13, pp. 18–21, 1993. on the development of baculoviruses,” Journal of Applied [43] M. M. A. De Doucet, M. A. Bertolotti, A. L. Giayetto, and Entomology, vol. 118, pp. 316–320, 1994. M. B. Miranda, “Host Range, Specificity, and Virulence of [57] H. C. T. Ribeiro, O. H. O. Pavan, and A. R. Muotri, “Compar- Steinernema feltiae, Steinernema rarum,and Heterorhabditis ative susceptibility of two different hosts to genotypic variants bacteriophora (Steinernematidae and Heterorhabditidae) from of the Anticarsia gemmatalis nuclear polyhedrosis virus,” Argentina,” Journal of Invertebrate Pathology,vol. 73, no.3,pp. Entomologia Experimentalis et Applicata,vol. 83, no.2,pp. 237–242, 1999. 233–237, 1997. [44] J. P. M. Acevedo, R. I. Samuels, I. R. Machado, and C. Dolin- [58] S. R. Palli, T. R. Ladd, W. L. Tomkins et al., “Choristoneura ski, “Interactions between isolates of the entomopathogenic fumiferana entomopoxvirus prevents metamorphosis and fungus Metarhizium anisopliae and the entomopathogenic modulates juvenile hormone and ecdysteroid titers,” Insect nematode Heterorhabditis bacteriophora JPM4 during infec- Biochemistry and Molecular Biology, vol. 30, no. 8-9, pp. 869– tion of the sugar cane borer Diatraea saccharalis (Lepidoptera: 876, 2000. Pyralidae),” Journal of Invertebrate Pathology,vol. 96, no.2,pp. [59] C. M. Fauquet, M. A. Mayo, J. Maniloff,U.Desselberger, 187–192, 2007. andL.A.Ball, Eds., Virus Taxonomy, Eighth Report of the [45] P. J. Aguila, M. Vidal, M. Gonzalez, ´ L. A. Rodr´ıguez, and E. International Committee on Taxonomy of Viruses, Elsevier, San Mesa, “Criterios ecolog ´ icos en el manejo de plagas de la cana ˜ Diego, Calif, USA, 2005. de azucar ´ y cultivos varios una opcion ´ para lograr alimentos [60] J. S. Cory and H. F. Evans, “Viruses,” in Field Manual of Tech- sanos,” Agroecolog´ıa, vol. 3, pp. 51–53, 2008. niques in Invertebrate Pathology,L.A.Lacey andH.K.Kaya, [46] D. R. Ring and H. W. Browning, “Evaluation of entomopatho- Eds., pp. 149–174, Springer, Dordrecht, The Netherlands, genic nematodes against the Mexican rice borer (Lepidoptera: 2nd edition, 2007. Pyralidae),” Journal of Nematology, vol. 22, no. 3, pp. 420–422, [61] P. Caballero and T. Williams, “Virus Entomopatogenos, ´ ” in Control Biolog ´ ico de Plagas Agr´ıcolas,J.A.Jacas andA. [47] J. C. LegaspiJr.,B.C.Legaspi, andR.R.Saldana, ˜ “Evalua- Urbaneja, Eds., pp. 121–135, PHYTOMA-Espana, ˜ Navarra, tion of Steinernema riobravis (Nematoda: Steinernematidae) Espana, ˜ 2008. against the Mexican rice borer (Lepidoptera: Pyralidae),” [62] W. Mitsuhashi, H. Kawakita, R. Murakami et al., “Spindles of Journal of Entomological Science, vol. 35, no. 2, pp. 141–149, an entomopoxvirus facilitate its infection of the host insect by disrupting the peritrophic membrane,” Journal of Virology, vol. [48] E. Schnepf, N. Crickmore, J. Van Rie et al., “Bacillus thur- 81, no. 8, pp. 4235–4243, 2007. ingiensis and its pesticidal crystal proteins,” Microbiology and [63] M. Bergoin, J. C. Veyrunes, and R. Scalla, “Isolation and amino Molecular Biology Reviews, vol. 62, no. 3, pp. 775–806, 1998. acid composition of the inclusions of Melolontha melolontha [49] N. Bohorova, M. Cabrera, C. Abarca et al., “Susceptibility of poxvirus,” Virology, vol. 40, no. 3, pp. 760–763, 1970. four tropical lepidopteran Maize pests to Bacillus thuringiensis [64] R. L. Hall and R. W. Moyer, “Identification, cloning, and CryI-type insecticidal toxins,” Journal of Economic Entomology, sequencing of a fragment of Amsacta moorei entomopoxvirus vol. 90, no. 2, pp. 412–415, 1997. DNA containing the spheroidin gene and three vaccinia virus- [50] N. M. Rosas-Garc´ıa, B. Pereyra-Alfer ´ ez, K. A. Nin ˜o,L.J.Galn- related open reading frames,” Journal of Virology, vol. 65, no. Wong, and L. H. Morales-Ramos, “Novel toxicity of native and 12, pp. 6516–6527, 1991. HD Bacillus thuringiensis strains against to the sugarcane borer [65] D. Dall, A. Sriskantha, A. Vera, J. Lai-Fook, and T. Symonds, Diatraea saccharalis,” BioControl, vol. 49, no. 4, pp. 455–465, “A gene encoding a highly expressed spindle body protein of Heliothis armigera entomopoxvirus,” Journal of General [51] P. D. M. Gitahy,M.T.deSouza,R.G.Monnerat, E. D. B. Virology, vol. 74, no. 9, pp. 1811–1818, 1993. Arrigoni, and J. I. Baldani, “A Brazilian Bacillus thuringiensis [66] T. Hayakawa, J. Xu, and T. Hukuhara, “Cloning and sequenc- strain highly active to sugarcane borer Diatraea saccharalis ing of the gene for an enhancing factor from Pseudaletia (Lepidoptera: Crambidae),” Brazilian Journal of Microbiology, separata entomopoxvirus,” Gene, vol. 177, no. 1-2, pp. 269– vol. 38, no. 3, pp. 531–537, 2007. 270, 1996. [52] C. N. B. Carneiro,R.A.DaMatta,R.I.Samuels,and C. P. Silva, [67] Z. Li, C. Li, K. Yang et al., “Characterization of a chitin-binding “Effects of entomopathogenic bacterium Photorhabdus tem- protein GP37 of Spodoptera litura multicapsid nucleopolyhe- perata infection on the intestinal microbiota of the sugarcane drovirus,” Virus Research, vol. 96, no. 1-2, pp. 113–122, 2003. stalk borer Diatraea saccharalis (Lepidoptera: Crambidae),” [68] P. Wang and R. R. Granados, “Calcofluor disrupts the midgut Journal of Invertebrate Pathology, vol. 99, no. 1, pp. 87–91, defense system in insects,” Insect Biochemistry and Molecular Biology, vol. 30, no. 2, pp. 135–143, 2000. [53] P. Sikorowski andF.M.Davis,“Susceptibility of larvae of [69] T. Hukuhara and J. R. Bonami, “Reoviridae,” in Atlas of the southwestern corn borer, Diatraea grandiosella,to Bacillus Invertebrate Viruses,J.R.Adams andJ.R.Bonami, Eds.,pp. thuringiensis,” Journal of Invertebrate Pathology, vol. 15, no. 1, 393–434, CRC Press, Boca Raton, Fla, USA, 1991. pp. 131–132, 1970. [54] F. M. Davis and P. P. Sikorowsy, “Susceptibility of the South- [70] G. W. Blissard, B. Black, N. Crook, B. A. Keddie, R. Possee et al., “Family Baculoviridae,” in Virus Taxonomy: Seventh Report westerncornborer, Diatraea grandiosella Dyar (Lepidoptera: of the International Committee on Taxonomy of Virus,M.H.V. Pyralidae) to the baculovirus of Autographa californica,” Jour- van Regenmoertel, C. M. Fauquet, D. H. L. Bishop et al. et nal of the Kansas Entomological Society, vol. 51, no. 1, pp. 11– al., Eds., pp. 195–202, Academic Press, San Diego, Calif, USA, 13, 1978. [55] O. H. O. Pavan and H. C. T. Ribeiro, “Selection of a bac- ulovirus strain with a bivalent insecticidal activity,” Memor ´ ias [71] G. F. Rhormann, “Nuclear polyhedrosis viruses,” in Encyclope- do Insttituto Oswaldo Cruz, vol. 84, supplement 3, pp. 63–65, dia of Virology,R.G.Webster andA.Granoff, Eds., pp. 146– 1989. 152, Academic Press, London, UK, 2nd edition, 1999. 12 International Journal of Zoology [72] D. Winstanley and D. O. ’Reilly, “Granuloviruses,” in Encyclo- pedia of Virology,R.G.Webster andA.Granoff, Eds., pp. 140– 146, Academic Press, London, UK, 2nd edition, 1999. [73] B. Szewczyk, L. Hoyos-Carvajal, M. Paluszek, I. Skrzecz, and M. Lobo De Souza, “Baculoviruses—re-emerging biopesti- cides,” Biotechnology Advances, vol. 24, no. 2, pp. 143–160, [74] W. F. Jepson, A Critical Review of the World Literature of the Lepidopterous Stalk Borers of Graminaceous Crops, Common- wealth Institute of Entomology, London, UK, 1954. [75] J. W. SmithJr.,R.N.Wiedenmann, andW.A.Overholt, Parasites of Lepidopteran Stemborers of Tropical Gramineous Plants, ICIPE Science Press, Nairobi, Kenya, 1993. [76] G. M. Chippendale, “The southwestern corn borer, Diatraea grandiosella, case history of an invading insect,” Research Bulletin 1031, Missouri Agricultural Experiment Station, Columbia, Mo, USA, 1979. [77] N. Degaspari, P. S. M. Botelho, and N. Macedo, “Controle quimico da Diatraea saccharalis em cana-de-acucar na regiao ˜ Centro SuldoBrasil,” Boletim T´ecnico. IAA/PLANALSUCAR, vol. 3, pp. 1–16, 1981. [78] F. Moscardi, “Use of viruses for pest control in Brazil: the case of the nuclear polyhedrosis virus of the soybean caterpillar, Anticarsia gemmatalis,” Memor ´ ias do Insttituto Oswaldo Cruz, vol. 84, pp. 51–56, 1989. [79] G. Fedi ´ er ` e, “Epidemiology and pathology of Densovirinae,” in Parvovir Uses. From Molecular Biology to Pathology and Therapeutic Uses,S.Faisst andJ.Rommelaere, Eds.,pp. 1–11, Karger, Basel, Switzerland, 2000. [80] G. Meynadier, P. F. Galichet, J. C. Veyrunes, and A. Amargier, “Mise en ev ´ idence d’une densonucleose ´ chez Diatraea saccha- ralis [Lep.: Pyralidae],” Entomophaga, vol. 22, no. 1, pp. 115– 120, 1977. [81] N. Kouassi, J. X. Peng, Y. Li, C. Cavallaro, J. C. Veyrunes, and M. Bergoin, “Pathogenicity of Diatraea saccharalis densovirus to host insets and characterization of its viral genome,” Virologica Sinica, vol. 22, no. 1, pp. 53–60, 2007. [82] E. P. Steinhaus and G. A. Marsh, “Report of diagnosis of disease insects, 1951–1961,” Hilgardia, vol. 33, pp. 349–490, [83] U. K. Metha and H. David, “A granulosis virus disease of sugarcane internode borer,” Madras Agricultural Journal, vol. 67, pp. 616–619, 1970. [84] S. Easwaramoorthy and S. Jayaraj, “Survey of granulosis virus infection in sugarcane borers, Chilo infuscatellus Snellen and C. sacchariphagus indicus (Kapur) in India,” Tropical Pest Management, vol. 3, pp. 200–201, 1987. [85] S. Easwaramoorthy and H. David, “A granulosis virus of sugarcane shoot borer, Chilo infuscatellus Snell. (Lepidoptera: Crambidae),” Current Science, vol. 48, pp. 685–686, 1979. [86] S. Sethuraman and K. Narayanan, “Biological activity of Nucleopolyhedrovirus isolated from Chilo partellus (Swinhoe) (Lepidoptera: Pyralidae) in India,” Asian Journal of Experimen- tal Biological Sciences, vol. 1, pp. 325–330, 2010. [87] A. J. Cherry,C.J.Lomer,D.Djegui, andF.Schuethess, “Pathogen incidence and their potential as microbial control agents in IPM of maize stem borers in West Africa,” BioCon- trol, vol. 44, no. 3, pp. 301–327, 1999. 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