SUSCEPTIBILITY OF DESERT LOCUST, SCHISTOCERCA GREGARIA (ORTHOPTERA: ACRIDIDAE) TO BACILLUS CEREUS ISOLATED FROM EGYPT

Examination was done at preliminary bracketing bioassay on one old 4th nymphal instar of desert locust. Results showed that two isolates, namely NDL1 and NDL2 were having highly potentiality as entomopathogenic bioagents. Thirty isolates were isolated from dead/ infected nymphs of desert locust occurred in raring cages at Department of Locust and Grasshoppers Research, Plant Protection Research Institute, Agricultural Research Center, Dokki, Giza, Egypt. Molecular identification of isolated bacteria was done using universal primers of 16s rRNA, followed by DNA sequencing. Nucleotides were blasted at (https://www.ncbi. nlm.nih.gov /genbank/) to recognize that NDL1 and NDL2 isolates were two different isolates of Bacillus cereus with a high similarity (100%).  Susceptibility of 4th nymphal instar of Schistocerca gregaria (Forskal) to the isolated B. cereus was determined using two bioassay procedures, Leaf-dip and per os. The insecticidal activity of both isolates against locust nymph in leaf dipping showed that NDL2 was more efficient than NDL1. However, the opposite trend was observed in using per os.  Both Isolates have the potential to be a successful biocidal agent to control desert locust

Development of a microslide aglutination assay with the aid if an inexpensive projection microscope

A microslide agglutination assay was developed involving the mixing of 2.5 microl each of antiserum and a cell suspension of Listeria monocytogenes. Cell agglutination in the final volume of 5.0 microl was visually observed at a direct magnification of 22 x on the projection screen of an inexpensive 20 US dollar projection microscope. The procedure has the advantage of increasing by a factor of 20 the number of agglutination assays that can be performed with a given volume of antiserum with the use of an inexpensive optical projection system

Development of a new lysis solution for releasing genomic DNA from bacterial cells for DNA amplification by polymerase chain reaction

A new lysis solution designated TZ, consisting of 2.0% Triton X-100 plus 2.5 mg sodium azide/ml in 0.1 M Tris-HCl buffer at pH 8.0, yielded higher levels of genomic DNA from Escherichia coli O157:H7 cells compared with a number of other commonly used cell lysis methods. Ethidium bromide stained DNA bands resulting from PCR amplification of target DNA from 100 CFU of E. coli O157:H7 were readily detected following electrophoresis of agarose gels. In contrast, conventional cell lysis methods failed to detect target DNA from 100 CFU after PCR amplification. The new solution was highly effective for lysing cell suspensions of Salmonella enteritidis, Pseudomonas putida, Lysteria monocytogenes and Psychrobacter immobilis

Detection, enumeration, and RAPD analysis of Listeria monocytogenes isolates in fish derived from retail sources in Western Massachusetts

Fish were sampled over a 24-month period from two major supermarket retail outlets in Hadley, Massachusetts, USA, designated A and B, for the incidence of Listeria monocytogenes and numbers of the organism present per 100 g of tissue. Fifteen species of fish were represented. Seventy-four samples out of a total of 320 were confirmed by PCR as yielding L. monocytogenes. From retail source A, a total of 171 samples yielded 59 (34.5%) that were positive for the presence of L. monocytogenes. In contrast, from retail source B, a total of 149 samples yielded 15 (10.0%) that were positive. Only six samples (3.5%) from retail source A had MPN counts of L. monocytogenes in the range of 100 to 1,000 per 1,000 g. Only two samples (1.3%) from retail source B had counts of L. monocytogenes from 100 to 1,000 per 100 g. A total of 221 strains of L. monocytogenes were derived from the MPN cultures, 164 from retail source A, and 57 from retail source B. All 221 strains were subjected to RAPD analysis using three random primers. Primer LMPB1 yielded 21 RAPD profiles, primer LMPB4 yielded 19 profiles, and primer HLWL74 yielded 26 profiles. A total of 55 composite profiles were identified by combining the profiles derived from the three primers. Source A yielded 50 composite RAPD profiles, whereas source B yielded only ten composite profiles. In addition, 27 of the 55 composite profiles were derived from individual isolates and RAPD types 11 and 18 included 49 and 27 isolates respectively. Fish from retail source A clearly harbored far more RAPD types than did source B. The results clearly indicated that two major retail sources in close geographic proximity can vary considerably with respect to the incidence and numbers of L. monocytogenes present on the fish tissue. It was not possible to determine whether the processors furnishing fish to retail outlet A or the supermarket itself was responsible for the notably higher incidence and numbers of L. monocytogenes on fish from retail source A compared to fish from retail source B

Enhanced toxicity of Bacillus thuringiensis subspecies kurstaki and aizawai to black cutworm larvae (Lepidoptera: Noctuidae) with Bacillus sp. NFD2 and Pseudomonas sp. FNFD1

Bacillus thuringiensis subspecies kurstaki and aizawai are important control agents for lepidopteran pests. Bioassays were designed to test B. t. kurstaki and aizawai against second- and-fourth instar black cutworm larvae with and without Bacillus sp. NFD2 and Pseudomonas sp. FNFD1 bacteria. B. thuringiensis subsp. aizawai (XenTari) was more toxic to both second- and fourth-instar black cutworm, Agrolis ipsilon (Hufnagel) (Lepidoptera: Noctuidae), larvae than B. t. kurstaki (DiPel) at 7 d after treatment (DAT). When DiPel was combined with NFD2 or FNFD1 versus second instars, the LC 50s were 5.0X and 4.7X lower, respectively, than with DiPel alone. DiPel combined with both NFD2 and FNFD1 versus second instars resulted in an LC50 value 7.7X lower than with DiPel alone. When XenTari was combined with NFD2 or FNFD1 versus second instars, the LC50s were 5.2X and 3.8X lower, respectively, than with XenTari alone. XenTari combined with both NFD2 and FNFD1 versus second instars resulted in an LC50 9.7X lower than with XenTari alone. When DiPel was combined with NFD2 or FNFD1 versus fourth instars, the LC50s were 4.4X and 3.4X lower, respectively, than with DiPel alone. DiPel combined with both NFD2 and FNFD1 versus fourth instars resulted in an LC50 5.0X lower than with DiPel alone. When XenTari was combined with NFD2 or FNFD1 versus fourth instars, the LC50s were 5.7X and 3.3X lower, respectively, than with XenTari alone. XenTari combined with both NFD2 and FNFD1 versus fourth instars resulted in an LC50 6.7X lower than with XenTari alone. © 2011 Entomological Society of America

Enhanced toxicity of Bacillus thuringiensis japonensis strain Buibui toxin to oriental beetle and northern masked chafer (Coleoptera: Scarabaeidae) larvae with Bacillus sp. NFD2

Bacillus thuringiensis japonensis strain Buibui (Btj) has the potential to be an important control agent for pest scarabs. Bioassays using autoclaved and nonautoclaved soil showed there were always lower LC50 values associated with nonautoclaved soil. We identified five other bacteria found in the hemolymph of insects killed by Btj and used them in bioassays to see whether we could enhance the control achieved with Btj alone. One bacterium, designated NFD2 and later identified as a Bacillus sp., showed the greatest enhancement of Btj in preliminary experiments and was used in bioassays with Btj versus oriental beetle, Anomala orientalis (Waterhouse), and northern masked chafer, Cyclocephala borealis Arrow (Coleoptera: Scarabaeidae), larvae. This bacterium alone was nontoxic to grubs in bioassays. A combination of this bacterium with Btj in nonautoclaved soil resulted in a significantly lower LC50 value (0.23 g toxin per g soil) from all other treatments for A. orientalis with one exception; the LC50 where NFD2 was added back into autoclaved soil (0.29 g toxin per g soil). A combination of this bacterium with Btj in nonautoclaved soil resulted in a significantly lower LC50 value (48.29 g toxin per g soil) from all other treatments for C. borealis with the exception of the treatment where Bacillus sp. NFD2 was added back to autoclaved soil (96.87 g toxin per g soil) with Btj. This research shows that other soil bacteria can be used to enhance the toxicity of Btj and possibly other Bts. © 2010 Entomological Society of America