Molecular entomology

The genus Heissiella Péricart, 1984, previously known from India and Bhutan, is newly recorded for China, with the species Heissiella sinica sp. nov. described as new. Dorsal habitus photograph and illustrations of characters of the species are provided, together with a key to species of Heissiella.

In India honey bees like Apis cerana and Apis mellifera are reared for commercial purpose. The other bees like rock bee (Apis dorsata), little bee (Apis florea) and dammer bee (Trigona sp) are not domesticated because of their ecophysiology. However, the kani tribes living in Karayar area of Kalakad-Mundanthurai Tiger Reserve area (KMTR) of Western Ghats have developed a novel method of rearing the dammer bees. The objective of the study is to understand the traditional skill of Kani tribes to rear the stingless bee (Trigona sp.) which was not used for regular bee keeping. The present study also gives importance to evolve commercial strategies to get honey (medicinal honey) from Trigona sp. The findings of the present study will help to preserve the art of rearing stingless bees and to introduce modern techniques to harvest pure, clean and uncontaminated honey by suitable modifications in the indigenous practice of stingless bee rearing. The honey produced by dammer bees is a panacea for much human illness. The Kani tribes have developed a bamboo pole bee hive to keep this stingless bee belonged to the species Trigona irredipensis. The dammer bees build an interesting type of comb made of propolis, mud and about 600-700gm honey is collected per year per hive. Because of rich medicinal value the honey collected from this beehive is sold at a rate of Rs.1000/Kg.

Spodoptera exigua (Lepidoptera: Noctuidea) is a polyphagous pest that attacks many
important agricultural crops. Identifying an insect specimen is a crucial step in entomology.
This study demonstrated a molecular method to identify the species of pest and its
parasitoid in the case of a lack of a morphological identification key. To facilitate the
identification of these pest-parasitoid species, a DNA barcoding approach was used for
accurate and time-consuming identification based on nucleotide sequencing analysis of
the mitochondrial Cytochrome Oxidase subunit I (COI) gene. DNA barcoding sequences
successfully identified both pest and parasitoid species by comparing barcode sequences
to the GenBank database. This study provided evidence of Pediobius sp. as a parasitoid
of S. exigua in Malaysia.

In this study, a first record on interaction of insect with fern species has been presented from Malaysia. Molecular identification of cytochrome oxidase I gene (COI) has revealed the Herpetogramma platycapna (Meyrick) is a gregarious pest of Angiopteris evecta (G. Forst.) Hoffm. The pest species also has been reconfirmed based on morphological characters.

Sequences from the DNA barcode region of the mitochondrial COI gene are an effective tool for specimen identification and for the discovery of new species. The Barcode of Life Data Systems (BOLD) (www.boldsystems.org) currently hosts 4.5 million records from animals which have been assigned to more than 490,000 different Barcode Index Numbers (BINs), which serve as a proxy for species. Because a fourth of these BINs derive from Lepi-doptera, BOLD has a strong capability to both identify specimens in this order and to support studies of faunal overlap. DNA barcode sequences were obtained from 4503 moths from 329 sites across Pakistan, specimens that represented 981 BINs from 52 families. Among 379 species with a Linnaean name assignment, all were represented by a single BIN excepting five species that showed a BIN split. Less than half (44%

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In  the  Indian  subcontinent,Visceral  Leishmaniasis  (VL)  is  transmitted  through  the  established  vector  Phlebotomus argentipes.  There  are  several  reports  on  development  of  DDT  resistance,  the  insecticide  of  choice  to  P.  argentipes,  based on  bio-assay  test.  In  this  study,  metabolic  based  mechanisms  for  DDT  resistance  were  investigated  in  P.  argentipes  to facilitate  the  development  of  novel  strategy  to  control  resistance  development  in  the  vector.  Wild  sand  flies  collected  from Baizalpur  (District-Sonepur),  Chandi  (District-Nalanda)  and  Khusroopur  (District-Patna)  of  Bihar,  India  were  reared  for  F1 progeny  out  of  which  one  sub  sample  was  stored  at  -800C  for  biochemical  assay  while  the  other  was  used  for  insecticide bio-assay  test.  Biochemical  assay  was  conducted  for  detoxifying  enzymes  (Glutathione  S-transferase  (GST), Acetylcholinestrase  (AChE),  Cytochrome  p450  (Cyt  p450)  and  non-  specific  esterase  (ETS).  The  total  protein  pooled  from elevated  GST  samples  were  also  separated  on  one  dimension  gel  electrophoresis.  P.  argentipes  population  from  Baizalpur, Chandi  and  Khusroopur  villages  were  found  highly  resistant  to  DDT.  F1  progeny  of  P.  argentipes  showed  elevated  GST  in 60%  population.  There  were  no  elevation  in  AChE,  Cyt  p450  and  ETS  activity.  Furthermore,  the  GST  elevation  was  also observed  in  12%  SDS-PAGE  at  around  42  kDa  and  was  confirmed  through  western  blotting.  This  study  provides preliminary  biochemical  evidence  of  GST  enzyme  elevation  in  DDT  resistant  Sand  fly  population,  which  probably  resulted from  Malaria  and  Kala-azar  control  activities  in  the  area.

Chloroquine resistance in P. falciparum is conferred by mutations in the pfcrt gene with K76T mutation being definitive in as far as chloroquine resistance is concerned. The prevalence of point mutations at codons 74, 75 and 76 of P. falciparum of the pfcrt gene was determined by dot�blot/probe hybridization analysis in 121 samples collected in May 2009 twelve years after chloroquine cessation in Kenya. We found out that 80.17% of the field isolates from Mbita harbor the T76 mutation while 19.83% have the wild type allele K76 [ T test, P= 0.108 (2005 versus 2009)]. Majority of the isolates had MNT haplotype at codons 74, 75 and 76. Conclusion: The study showed a significant expansion of the pfcrt 76K allele and thus expansion of parasite populations that are sensitive to chloroquine.

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 LC 50 values associated with nonautoclaved soil. We identiÞed Þve 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 identiÞed 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, Cyclo-cephala 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 signiÞcantly lower LC 50 value (0.23 g toxin per g soil) from all other treatments for A. orientalis with one exception; the LC 50 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 signiÞcantly lower LC 50 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.

The genetic variability of the coffee berry borer (Hypothenemus hampei) in Colombia was evaluated using the amplified fragment length polymorphism (AFLP) technique. Sixty H. hampei DNA samples from 16 coffee producing areas throughout Colombia were analyzed. We performed AFLP´s with these DNA samples using the two most polymorphic selective primers from a previous investigation. The results revealed very low genetic variation in the Colombian samples but enough polymorphism to perform a phylogenetic analysis. The findings indicated that there were multiple H. hampei introductions into Colombia probably from Peru, Ecuador, and Brazil. There was a main line of H. hampei present in every coffeeproducing area in Colombia that spread throughout the country, perhaps due to coffee pickers moving within coffee regions and transporting infested coffee berries or adult H. hampei females. We also found that a line of H. hampei present in Costa Rica matched the fingerprint profile of this main line in Colombia, thus raising the possibility that the line of H. hampei in Costa Rica originated from Colombia. We designed sequence tagged sites STS markers from AFLP polymorphisms to answer some issues related to the biology and ecology of H. hampei. The design of co-dominant markers using Genome Walking and Single Stranded Conformational Polymorphism SSCP will be also of enormous relevance for a better understanding of the biology and ecology of H. hampei.

Sequences from the DNA barcode region of the mitochondrial COI gene are an effective tool for specimen identification and for the discovery of new species. The Barcode of Life Data Systems (BOLD) (www.boldsystems.org) currently hosts 4.5 million records from animals which have been assigned to more than 490,000 different Barcode Index Numbers (BINs), which serve as a proxy for species. Because a fourth of these BINs derive from Lepidoptera, BOLD has a strong capability to both identify specimens in this order and to support studies of faunal overlap. DNA barcode sequences were obtained from 4503 moths from 329 sites across Pakistan, specimens that represented 981 BINs from 52 families. Among 379 species with a Linnaean name assignment, all were represented by a single BIN excepting five species that showed a BIN split. Less than half (44%) of the 981 BINs had counterparts in other countries; the remaining BINs were unique to Pakistan. Another 218 BINs of Lepidoptera from Pakistan were coupled with the 981 from this study before being compared with all 116,768 BINs for this order. As expected, faunal overlap was highest with India (21%), Sri Lanka (21%), United Arab Emirates (20%) and with other Asian nations (2.1%), but it was very low with other continents including Africa (0.6%), Europe (1.3%), Australia (0.6%), Oceania (1.0%), North America (0.1%), and South America (0.1%). This study indicates the way in which DNA barcoding facilitates measures of faunal overlap even when taxa have not been assigned to a Linnean species.

The relative residual toxicity of selected insecticides against whitefly was assessed in the Entomology laboratory at College of Horticulture, Mudigere during the year 2014-2015. The treatments viz., imidachloprid (15.70 to 22.97%), triazophos (10.43 to 21.97%), thiamethoxam (20.77 to 37.97%), cyantraniliprole (20.03 to 34.37%), azadirhactin (10.40 to 18.27%), fipronil (6.60 to 29.10%) and buprofezin (7.93 to 17.00%) showed high level of residual toxicity up to sixth day after the treatment. From eighth day after treatment, toxicity started decreasing. However, in treatments acephate and emamectin benzoate, residual toxicity started decreasing from sixth day itself. Although toxicity of spinosad (15.23%) was noticed to be high at two days after treatment, its toxicity started decreasing from fourth day itself. At sixth day after treatment, the order of residual toxicity was thiamethoxam > cyantraniliprole > fipronil > imidachloprid > triazophos > azadirhactin > bup...

The established vector for Kala-azar (VL) Phlebotomus argentipes is developing tolerance/resistance against the common insecticide (DDT). However, the insecticidal effect of wild plant i.e. Clerodendron infortunatum leaf (Hexane extract) was found effective in killing 63% sand flies that can be enhanced with formulation of lead molecules. Hence, its toxicity evaluation to humans is pre-requisite. Percentage Hemolytic Inhibition activity of the extract was observed with IC50 value of 25.80 mg/L, 18.68 mg/L & 15.90 mg/L at 450 nm, 490 nm & 655 nm wavelengths respectively. The extract was found non-toxic and safer to use as a new insecticide against Sand fly. © 2014 Elixir All rights reserved ARTICLE INF O Artic le hi story: Received: 25 July 2014; Received in revised form: 29 September 2014; Accepted: 24 October 2014;