Field Evaluation of Brazilian Isolates of Neozygites floridana (Entomophthorales: Neozygitaceae) for the Microbial Control of Cassava Green Mite in Benin,West Africa

Two Brazilian isolates and one Benin (indigenous) isolate of Neozygites floridana were released against the cassava green mite, Mononychellus tanajoa , in January 1999 in the Adjohoun district, Ouémé administrative region, Republic of Benin. Post-release monitoring conducted 8, 14, 22 and 36 weeks later showed very low mean infection rates on M. tanajoa by isolate (0.03-0.4%). However, 48 weeks after releases, mean infection rates increased noticeably to between 2.3 and 18.7%, and higher infection rates were observed for the Brazilian isolates compared with the indigenous one. The highest infection rate for the indigenous isolate was 4.5% while it reached over 30% for the Brazilian isolates (36.5 and 34.0%). Observations made to study dispersal from inoculated plants showed the absence of infected mites at 4 m from the inoculated plants in all fields 8 weeks after the releases, while they were already present on those at 2 m away. From the next monitoring, 14 weeks after the releases, infection was found at all three sampling positions (inoculated plants and plants at 2 and 4 m away). Only four mites with resting spores were found in over 460 000 mites examined. The highest infection levels were observed in December during 'harmattan' a period characterized by hot days and cool nights with high relative humidity.     

Seasonal and Habitat Variability in the Fungal Pathogens, Neozygites cf. floridana and Hirsutella thompsonii, Associated with Cassava Mites in Benin, West Africa

A survey of the pathogenic fungi associated with mites on cassava in Benin, West Africa, revealed both geographical and seasonal variation in the presence of Neozygites cf. floridana (Weiser and Muma) and Hirsutella thompsonii Fisher on Mononychellus tanajoa (Bondar) and Oligonychus gossypii (Zacher). Few dead and infected mites were found during the dry season, regardless of vegetation zone. In three of 30 surveyed sites, N. floridana was found infecting 1% of the dead M. tanajoa and 2% of the dead O. gossypii, while H. thompsonii was observed infecting 20% of the dead M. tanajoa in a single site. The frequency of sites having infected mites during the wet season was 3.5 times greater than that seen during the dry season. N. floridana infected 10% of the dead M. tanajoa and 19% of the dead O. gossypii on young leaves. Mites infected with N. floridana were found either in the coastal Southern Forest Mosaic (SFM) or in the Northern Guinea Savanna vegetation zones. N. floridana was rare in the low mite densities associated with mature leaves. H. thompsonii was found on 19% and 29% of the dead M. tanajoa on young and mature leaves respectively. All M. tanajoa infected with H. thompsonii on young leaves and mature leaves (75%) were found in the SFM. A single M. tanajoa was the only infected mite found in the Southern Guinea Savanna. Relatively few O. gossypii were infected with H. thompsonii. N. floridana and H. thompsonii were found together in three sites, but never on the same host. Phytoseiids were never found infected with either pathogen. In a regression analysis, the number of dead mites was significantly estimated from the total number of mites for both species, regardless of leaf age. The numbers of dead M. tanajoa on mature leaves were also estimated from the proportion infected with H. thompsonii. The numbers of infected mites on young leaves were estimated from their association with the SFM for M. tanajoa infected with H. thompsonii, and from total mites for O. gossypii infected with N. floridana. On mature leaves, infected mite numbers were estimated from the numbers of dead M. tanajoa infected with H. thompsonii. The merit of introducing more virulent or better adapted isolates of N. floridana to control M. tanajoa in Africa is discussed.     

Molecular detection and differentiation of Brazilian and African isolates of the entomopathogen Neozygites tanajoae (Entomophthorales: Neozygitaceae) with PCR using specific primers

Neozygites tanajoae is an entomopathogenic fungus which has been used for biocontrol of the cassava green mite (Mononychellus tanajoa, CGM) in Africa. Establishment and dispersal of Brazilian isolates which have been introduced into some African countries in recent years to improve CGM control was followed with specific PCR assays. Two primer pairs, NEOSSU_F/NEOSSU_R and 8DDC_F/8DDC_R, were used to differentiate isolates collected from several locations in Brazil and from three countries in Africa, Benin, Ghana and Tanzania. The first primer pair enabled the species-specific detection of Neozygites tanajoae, while the second differentiated the Brazilian isolates from those of other geographical origin. PCR assays were designed for detection of fungal DNA in the matrix of dead infested mites since N. tanajoae is difficult to isolate and culture on selective artificial media. Our results show that all isolates (Brazilian and African) that sporulated on mummified mites were amplified with the first primer pair confirming their Neozygites tanajoae identity. The second pair amplified DNA from all the Brazilian isolates, but did not amplify any DNA samples from the African isolates. None of the two primers showed amplification neither from any of the non-sporulating mite extracts nor from the dead uninfected mites used as negative controls. We confirmed that the two primer pairs tested are suitable for the detection and differential identification of N. tanajoae isolates from Brazil and Africa and that they are useful to monitor the establishment and spread of the Brazilian isolates of N. tanajoae introduced into Benin or into other African countries for improvement of CGM biocontrol.     

Does Methyl Salicylate, A Component of Herbivore-induced Plant Odour, Promote Sporulation of the Mite-pathogenic Fungus Neozygites tanajoae?

Blends of volatile chemicals emanating from cassava leaves infested by the cassava green mite were found to promote conidiation of Neozygites tanajoae, an entomopathogenic fungus specific to this mite. Methyl salicylate (MeSA) is one compound frequently present in blends of herbivore-induced plant volatiles (HIPV) as well as that of mite-infested cassava. Here, we investigated the effect of methyl salicylate in its pure form on the production of pre-infective spores (conidia), and the germination of these spores into infective spores (capilliconidia), by a Brazilian isolate and a Beninese isolate of N. tanajoae. Mummified mites previously infected by the fungal isolates were screened under optimal abiotic conditions for sporulation inside tightly closed boxes with or without methyl salicylate diffusing from a capillary tube. Production of conidia was consistently higher (37%) when the Beninese isolate was exposed to MeSA than when not exposed to it (305.5 ± 52.62 and 223.2 ± 38.13 conidia per mummy with and without MeSA, respectively). MeSA, however, did not promote conidia production by the Brazilian isolate (387.4 ± 44.74 and 415.8 ± 57.95 conidia per mummy with and without MeSA, respectively). Germination of the conidia into capilliconidia was not affected by MeSA for either isolate (0.2%, 252.6 ± 31.80 vs. 253.0 ± 36.65 for the Beninese isolate and 4.2%, 268.5 ± 37.90 vs. 280.2 ± 29.43 for the Brazilian isolate). The effects of MeSA on the production of conidia were similar to those obtained under exposure to the complete blends of HIPV for the case of the Beninese isolate, but dissimilar (no promoting effect of MeSA) for the case of the Brazilian isolate. This shows that MeSA, being one compound out of many HIPV, can be a factor promoting sporulation of N. tanajoae, but it may not be the only factor as its effect varies with the fungal isolate under study.     

The role of resting spores in the survival of the mite-pathogenic fungus Neozygites floridana from Mononychellus tanajoa during dry periods in Brazil

Survival of pathogens during long periods of unfavorable conditions can be critical to their ecology and to their use in biological control. In northeastern Brazil, the mite pathogen Neozygites floridana must survive hot and dry conditions between wet seasons when it infects the cassava green mite Mononychellus tanajoa. We report on large numbers of mite cadavers bearing resting spores towards the end of epizootics in mid-1995. High within-leaf variability indicated that local factors may be important in determining resting spore formation. These spores remain in the host cadaver on a leaf until the cadaver breaks up, whereupon the spores fall freely to the soil, there to remain dormant. Laboratory simulation of field conditions led to ca. 25% of mycosed individuals bearing resting spores. Mummies (without resting spores) kept in hot and dry conditions showed little or no viability within 2 months, implying no role for survival over extended dry periods. It is proposed that resting spores form the principal means by which this pathogen survives the dry season in the study area. This has implications for its introduction to new areas in classical biological control.     

Development of Aerial Releases of Diachasmimorpha tryoni (Cameron) (Hymenoptera: Braconidae), a Parasitoid that Attacks the Mediterranean Fruit Fly, Ceratitis capitata (Weidemann) (Diptera: Tephritidae), in the Guatemalan Highlands

A braconid parasitoid, Diachasmimorpha tryoni (Cameron), was released from the air into Guatemalan coffee plantations that contained Mediterranean fruit flies, Ceratitis capitata (Weidemann). Parasitoid adults were chilled, placed in paper bags, and dropped from an altitude of 100 m and at an airspeed of ~ 130 km/h. Releases were made at four different densities over a period of two years. At the higher release rates parasitism levels reached as high as 84%. The feasibility of using a more technically sophisticated aerial release technique, the auger sterile-insect release machine utilized in C. capitata sterile-fly aerial eradication projects in California and Florida, was also examined. Chilled D. tryoni either alone or in combination with chilled, sterile C. capitata , were dropped over target areas and the released parasitoids examined for mortality and damage. Samples of released parasitoids were taken and tested for 'flight ability'; i.e. flight response after an opportunity to recover from chilling. There was no evidence of significant mortality due to aerial release, and the flight-ability of insects released at various rates and altitudes did not significantly differ from chilled controls that were not released from an airplane.     

Biological Control of Thrips on Ornamental Crops: Interactions Between the Predatory Mite Neoseiulus cucumeris (Acari: Phytoseiidae) and Western Flower Thrips, Frankliniella occidentalis (Thysanoptera: Thripidae), on Cyclamen

Difficulties in controlling outbreaks of Western flower thrips, Frankliniella occidentalis, have obstructed the widespread adoption of biological control in many ornamental crops. The efficacy of the predatory mite, Neoseiulus cucumeris, in controlling F. occidentalis on two cultivars of cyclamen was tested in glasshouse experiments. The establishment and development of F. occidentalis populations was compared in three treatment introductions of N. cucumeris (50, 200 and 350 mites m -2 per week) and an untreated control. F. occidentalis were sampled in the flowers over eight weeks and counted into different life stages. No differences were observed between the two cultivars. All treatments with the predator resulted in a decline in numbers of F. occidentalis compared to the untreated control. Although the proportion of first instar F. occidentalis was similar in all treatments, the level of control varied with the number of N. cucumeris introduced. Lower populations of F. occidentalis, combined with a more rapid decline in their numbers, were observed at the 200 and 350 mites m -2 rates. Numbers of F. occidentalis remained low in the 350 N. cucumeris m -2 rate and the proportion of second instar F. occidentalis in the samples was consistently lower than in the other treatments. Trap counts of adult F. occidentalis were strongly correlated with the numbers of both adult and total F. occidentalis in flower samples. High inoculative releases of N. cucumeris early in the flowering cycle followed by frequent low introductions of predators should provide a strong basis for preventative control of F. occidentalis and other thrips species on cyclamen.     

Laboratory Trials of the Efficacy of the Crown Weevil Mortadelo horridus (Coleoptera: Curculionoidae) for the Biological Control of Slender Thistles Carduus pycnocephalus and C. tenuiflorus in Southwestern Australia

The hypothesis that larval crown weevil (Mortadelo horridus Alonso-Zarazaga and Sanchez-Ruiz) can suppress vegetative growth, seed set and viability in the winter annual slender thistles (Carduus pycnocephalus L. and C. tenuiflorus Curt.) was tested in laboratory gardens. Different densities (eight and 16) of M. horridus larvae and eggs were inoculated onto slender thistle rosettes between one and four times. Controls received no eggs or larvae. Larval feeding destroyed the apical meristems of treated plants, producing multi-crowned rosettes and significant declines in leaf area in both species, irrespective of the number of larvae. Treated thistles bolted at the same time and had a 4-fold increase in stem number compared to controls. The stems of control plants were up to one-third taller and thicker than attacked plants. Relative to controls, the leaves of treated plants had significantly fewer spines, decreased leaf area and decreased leaf perimeter, with less deeply lobed leaf margins. The total number of capitula produced by control and experimental groups was not significantly different, but consideration of fertile capitula only revealed a reduction in fertile capitula production following weevil attack. This was most marked in C. pycnocephalus. Control plants also produced significantly wider capitula. No control plants died, compared to 17 and 26% of the attacked plants of C. pycnocephalus and C. tenuiflorus, respectively. Implications for biocontrol of slender thistles are discussed.     

Effect of relative humidity and origin of isolates of Neozygites tanajoae (Zygomycetes: Entomophthorales) on production of conidia from cassava green mite, Mononychellus tanajoa (Acari: Tetranychidae), cadavers

Neozygites tanajoae is a very specialized fungus pathogenic to the cassava green mite (CGM), Mononychellus tanajoa, an important cassava pest introduced to Africa from the Neotropics. Conidial discharge from cadavers of CGM that died from infections with 14 isolates of N. tanajoae collected from diverse climates of Brazil was quantified to help select potential candidate strains for introduction to Africa. Studies aimed to identify isolates with lower requirements for relative humidity for sporulation and isolates that discharged more conidia during short periods of moisture. At 96 ± 0.5% RH, production of conidia was variable and even isolates from the Brazilian semi-arid region, e.g., Petrolina and Itaberaba, produced few conidia. Significant differences in the numbers of conidia produced by diverse Brazilian isolates were observed after 6, 9 and 12 h at 100% RH. At 100% RH, production of primary conidia increased considerably from an average of 57 ± 4 conidia at 6 h to 509 ± 37 conidia at 12 h. The isolate sporulating least (BIN21) discharged only 45.7% of the number of conidia produced by isolate BIN1, one of the isolates producing the most spores. Results from this study demonstrate that differences in production of conidia among isolates should be considered when selecting Neozygites isolates for new biological control introductions.     

Reproductive responses of Iphiseius degenerans and Neoseiulus teke (Acari: Phytoseiidae) to changes in the density of the cassava green mite, Mononychellus tanajoa (Acari: Tetranychidae)

We assessed the reproductive responses of adult female Iphiseius degenerans and Neoseiulus teke to increasing density of three stages of their prey, Mononychellus tanajoa, on cassava leaf discs under laboratory conditions. The oviposition rates increased with number of prey consumed per predator per day with a maximum of approximately two eggs per day for I. degenerans and four eggs per day for N. teke. The oviposition rate of N. teke was higher when consuming eggs than other prey stages. Neoseiulus teke was more efficient than I. degenerans in converting consumed prey into egg production. The data were adequately described by simple mathematical models.     

Phytoseiid mites from tropical fruit trees in Bahia State,Brazil (Acari,Phytoseiidae)

The cultivation of tropical fruit trees has grown considerably in the state of Bahia, northeastern Brazil. Some of these have been severely attacked by phytophagous mites, which are usually controlled by the use of chemical pesticides. However, there is today a growing interest for the adoption of less aggressive measures of pest control, as for example the use of predatory mites. Most of the plant-inhabiting predatory mites belong to the family Phytoseiidae. The objective of this paper is to report the phytoseiid species found in an intensive survey conducted on cultivated tropical fruit trees in fifteen localities of the southern coast of Bahia. Measurements of relevant morphological characters are provided for each species, to complement the understanding of the morphological variation of these species. Twenty-nine species of sixteen genera were identified. A key was elaborated to assist in the separation of these species. Fifteen species are reported for the first time in the state, raising to sixty-six the number of species of this family now known from Bahia. Seventy-two percent of the species collected belong to Amblyseiinae, followed by Typhlodrominae (21%) and Phytoseiinae (7%). The most diverse genus was Amblyseius. Amblyseiusoperculatus De Leon was the most frequent and abundant species. Studies should be conducted to evaluate the possible role of the most common predators as control agents of the phytophagous mites co-occurring with them.     

Formulation of Bacteria for Biological Weed Control Using the Stabileze Method

Two plant pathogenic Pseudomonas spp. were formulated using the 'Stabileze' method which involves the incorporation of bacteria in a water-absorbent starch matrix with oil and sucrose, then granulating the matrix with hydrated silica. In one experiment, P. syringae pv. tabaci formulated with the standard Stabileze formula was evaluated for storage viability at -15, 2 and 22°C. Bacteria stored for 1 year at -15 and 2°C lost only 0.2 and 0.5 log 10 colony forming units (CFU) g -1 respectively compared to a loss of log 10 3.5 CFU at 22°C. In a second experiment, the same pathogen was evaluated using variations of the formula with and without oil, and with and without sucrose. P.s. pv. tabaci formulated with sucrose and oil in combination, and sucrose and oil alone survived better than the formulation without oil or sucrose. A third experiment tested the effect of four levels of oil and four levels of sucrose (4 ×4 factorial) on survival of P.s. pv. tagetis over a 28 month period. Sucrose alone enhanced survival more than oil alone, and the beneficial effect of the sucrose was reduced when it was combined with oil. These experiments suggest that the Stabileze protocol is effective for stabilizing bacteria, but there are differences in response to different formulation components between species of bacteria.     

食品中桔霉素控制方法的研究进展

桔霉素(Citrinin,CIT)是由青霉、曲霉和红曲霉属产生的一种具有肾毒性的真菌毒素。许多食品和饲料中均含有桔霉素,污染范围十分庞大。桔霉素可与其他真菌毒素发生协同作用,如展青霉素(Patulin,PAT)、赭曲霉毒素(Ochratoxin,OTA)等,从而增强其毒性作用,对人及动物健康造成更大的危害。现阶段常用的控制手段主要有物理、化学和生物方法,均取得了一定的成就。本文简单介绍了桔霉素的毒性及污染状况,对桔霉素控制方法的研究进展进行了综述。     

东方胎球蚧的生物学特性及防治

东方胎球蚧在长春年生1代,以2龄若虫固着在二年生枝条的芽腋、分枝和不光滑处群集越冬。3月末至4月初开始活动为害。越冬若虫为害盛期为4月下旬至6月初。此时虫体明显膨大。5月下旬至6月上旬为成虫期和卵盛期。7月初起当年若虫自介壳下爬出分散转移到叶片、叶炳和嫩枝上为害,老虫共2龄。调查未见雄虫,均行孤雌生殖。主要天敌有黑缘红飘虫和一种寄生性小峰。当前生产上可在若虫越冬后为害盛期前和当年若虫出壳分散期。因地制宜地用有机磷或合成菊酯类杀虫剂进行常规喷雾。防治效果均较好。     

两种植绥螨的同类相残和集团内捕食作用

巴氏新小绥螨(Neoseiulus barkeri)和黄瓜新小绥螨(N.cucumeris)是两种多食性植绥螨,主要捕食叶螨和蓟马等,目前在我国广泛应用于农业生物防治中.本文研究了这两种植绥螨种内的同类相残(cannibalism)和种间的集团内捕食作用(intraguild predation)以及相互之间的攻击强度,以明确两者之间的相互关系,为合理构建天敌组合及评估生物防治的作用提供依据.结果显示:两种植绥螨对同种或异种幼螨的捕食量最大,其次是若螨,而对卵的捕食量极低.两种植绥螨对异种幼螨或若螨的捕食量均极显著高于对同种幼螨或若螨的捕食量.可见,无其他猎物存在情况下,两种植绥螨同时发生时更倾向于发生种间的集团内捕食.而巴氏新小绥螨对异种幼螨或若螨的捕食量均高于黄瓜新小绥螨对异种幼螨或若螨的捕食量,并且巴氏新小绥螨和黄瓜新小绥螨相比,巴氏新小绥螨对异种幼螨的攻击性更强,因此当这两种植绥螨发生集团内捕食时,巴氏新小绥螨是潜在的集团内捕食者,而黄瓜新小绥螨是潜在的集团内猎物.     

Production of Bacillus thuringiensis Berliner var. kurstaki Grown in Alternative Media

Agro - industrial residues and by - products available in southeastern Brazil were used as ingredients for low - cost culture media for liquid fermentation of Bacillus thuringiensis var. kurstaki. Highest spore yield was obtained with a medium containing cheese whey , soya bean milk and molasses (WSM) . Crystals and spores were produced in all media and potency of the final product was highest for nutrient broth + yeast extract medium (NBY) . There was no correlation between the number of spores in the fermented media and the potency of the preparations . Considering all three factors , the potencies , costs and yields of the final products , lowest relative cost was obtained with BMM medium ( Bombyx mori pupae + molasses) . NBY and WSM had intermediate relative cost approximately nine times higher than BMM . The cost analysis suggests that BMM medium should be preferred for local production of B. thuringiensis var . kurstaki in comparison to other media tested . The results also demonstrate the importance of considering yields , cost and potency of the B. thuringiensis preparations in selecting the production medium .     

Functional responses of Iphiseius degenerans and Neoseiulus teke (Acari: Phytoseiidae) to changes in the density of the cassava green mite, Mononychellus tanajoa (Acari: Tetranychidae)

The functional responses of protonymph and adult female Iphiseius degenerans and Neoseiulus teke to increasing density of three stages of their prey, the cassava green mite (CGM), Mononychellus tanajoa, were studied on excised cassava leaf discs under laboratory conditions. The responses obtained were predominantly sigmoid type III curves with the highest plateau when both stages of I. degenerans and N. teke were preying on CGM eggs. In all cases, the predation rate of the former species exceeded that of the latter. The empirical data were fitted by four different models. From the models, the attack coefficient (a) and handling time (T _h) were estimated. For a given predator stage (protonymph or adult female), the predator's attack coefficient declines and handling time increases as the prey gets larger. For a given prey stage, the predator's attack coefficient increases and handling time decreases as the predator stage becomes larger.