Artificial diet and method using an artificial diet, for...

Animal husbandry – Entomological culture device

Reexamination Certificate

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Details

C426S002000

Reexamination Certificate

active

06293223

ABSTRACT:

TECHNICAL FIELD
The invention in one aspect is directed to a diet for mass rearing of insects that feed on hard, woody plant tissue. The invention in another aspect is directed to a method of mass rearing of insects that feed on hard, woody plant tissue, for example, to a method of mass rearing of the root-boring weevil
Hylobius transversovittatus
Goeze for use as biological control agent of
Lythrum salicaria
L. or to a method of mass rearing of the root-boring weevil
Cyphocleonus achates
Fahraeus for use as a biological control agent of
Centaurea maculosa
L.
BACKGROUND OF THE INVENTION
Purple loosestrife (
Lythrum salicaria
L., Lythraceae), a Eurasian perennial wetland plant, was introduced into North America in the early 1800's. Negative impacts on biodiversity of infested wetlands and the inability of chemical, physical or mechanical means to provide long-term control resulted in the importation and release of host-specific phytophagous insects from the native range of the plant, a classical biological weed control program. Of nine host-specific insects species considered as control agents, three (the two leaf feeders
Galerucella calmariensis
L., and
G. pusilla
Duftschmidt [Coleoptera: Chrysomelidae] and a root feeder
Hylobius transversovittatus
Goeze [Coleoptera: Curculionidae]) were introduced into North America in 1992. An additional species that attacks the flowers (
Nanophyes marmoratus
Goeze [Coleoptera: Curculionidae]) was introduced in 1994.
Because as much as 60% of the annual biomass production of
L. salicaria
is stored below-ground and rootstocks of over 1 kg fresh biomass are common in mature plants, the root-feeding weevil
Hylobius transversovittatus
is considered a very important one of the host-specific insect species introduced and a prime candidate as a biological control agent. Attack by
H. transversovittatus
reduces flowering period, plant height, and plant biomass and changes biomass allocation patterns resulting in reduced performance of purple loosestrife.
It has been rather difficult to establish viable field populations of
H. transversovittatus.
The low abundance of this species in Europe prevented field collections for release in North America. Adults for initial introductions in 1992 were offspring of weevils collected across Europe. Larvae were reared on potted plants (of purple loosestrife) in a common garden in Germany. A total of 1,331 adults and 30,830 eggs were released in nine states and two Canadian provinces through 1993 and initial establishment was confirmed. Rearing of
H. transversovittatus
has continued at Cornell University and to a lesser extent at facilities of the Bureau of Reclamation in Denver, since 1993. This has allowed the distribution of eggs for field release despite the low number of adults. Although well over 100,000 eggs were distributed by 1999, field establishment rates appear rather low.
Increasing the availability of adults for field release has been considered crucial to increase field populations of the weevil to avoid negative effects due to inbreeding in small populations which could jeopardize the biological control program targeting
L. salicaria.
SUMMARY OF THE INVENTION
One goal of the invention was to achieve field establishment of
H. transversovittatus
through mass rearing of larvae on an artificial diet and subsequent field release of adults and to produce individuals at lower costs and faster than would be possible from allowing natural population build-up in the field or relying on natural hosts in a common garden. This has resulted in the development of a diet for mass rearing of insects that feed on hard, woody plant tissue which includes the host plant part fed on by larvae of the insect and water and agar ingredients in amounts to provide a gel to hold the host plant part and other ingredients together in homogeneous admixture, and in the development of a method of mass rearing of insects that feed on hard, woody plant tissue including the development of a method for the mass rearing of the root-boring weevil
Hylobius transversovittatus
for use, for example, as a biological control agent of
Lythrum salicaria
and the development of a method of mass rearing of the root-boring weevil
Cyphocleonus achates
Fahraeus for use, for example, as a biological control agent of
Centaurea maculosa
L.
In one embodiment, the invention is directed to a diet for mass rearing of insects that feed on hard, woody plant tissue, comprising a homogeneous mixture of a first portion which is subjected to sterilization in an autoclave comprising water and agar in amounts to form a gel containing the other diet components and a nutrition providing effective amount of salt mix, and of a second portion which is not subjected to sterilization in an autoclave, said second portion comprising a larval development effective formulation compatible amount of ground host plant part fed on by larvae of the insects, a nutrition providing effective amount of vitamin mix, and a pathogen growth suppressing amount of antimicrobial agent(s).
In another embodiment, the invention is directed to a method of mass rearing of insects that feed on hard, woody plant tissue comprising the steps of: (a) placing first instar insect larvae on an artificial diet comprising ground host plant part fed on by the larvae; (b) incubating the larvae on the diet at a temperature of 15 to 30° C. and a relative humidity of 50 to 90%; and (c) retrieving adults as they emerge. The method of this embodiment can be used, for example, for mass rearing of the root boring weevil
Hylobius transversovittatus,
for use as a biological control agent of
Lythrum salicaria,
and in such case first instar
Hylobius transversovittatus
larvae are used in step (a), and the artificial diet of step (a) comprises ground purple loosestrife root. The method of this embodiment can also be used, for example, for mass rearing of the root-boring weevil
Cyphocleonus achates
Fahraeus, for use as a biological control agent of
Centaurea maculosa
L., and in such case first instar
Cyphocleonus achates
Fahraeus larvae are used in step (a), and the artificial diet of step (a) comprises ground
Centaurea maculosa
L. root or ground
Centaurea diffusa
root.
The term “host plant” is used herein to mean the plant species associated with larval development in the field. The insect species to which the invention herein pertains have developed exclusive relationships with a single or a few select plant species which are referred to by those skilled in the art as host plants. Larval development can only be completed on these host plants, and females will restrict their oviposition to host plants allowing larval development. Attacked plant parts can be stems, roots, wood, etc., depending on the species. In any rearing, the host plant part fed upon in the field by the larvae should be used in the diet.
The term “formulation compatible” is used herein to mean allowing formulation of a homogeneous mix and/or to allow pourability during formulation and positioning of the diet.
DETAILED DESCRIPTION
We turn now to the embodiment herein directed to a diet for mass rearing of insects that feed on hard, woody tissue. The diet is preferably for rearing of beetles including weevils, from larvae. The diet comprises a homogeneous mixture of a first portion which is subjected to sterilization in an autoclave comprising water and agar in amounts to form a gel containing the other diet components and a nutrition providing effective amount of salt mix, and of a second portion which is not subjected to sterization in an autoclave comprising a larval development effective formulation compatible amount of ground host plant part fed on by larvae of the insects, a nutrition providing effective amount of vitamin mix and a pathogen growth suppressing amount of antimicrobial agents.
Preferably, the diet has a first portion comprising by weight of the diet of 0.01 to 5% salt mix and a second portion comprising by weight of the diet of 0.1 to 40% ground ho

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