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Evidence for multiple-insecticide resistance in urban Aedes albopictus populations in southern China
Yiji Li1,2,3,
Jiabao Xu1,
Daibin Zhong4,
Hong Zhang1,
Wenqiang Yang1,
Guofa Zhou4,
Xinghua Su1,
Yang Wu1,
Kun Wu1,
Songwu Cai5,
Guiyun Yan1,4 &
Xiao-Guang Chen1
Parasites & Vectors volume 11, Article number: 4 (2018) Cite this article
Aedes albopictus (Skuse) is an invasive mosquito that has become an important vector of chikungunya, dengue and Zika viruses. In the absence of specific antiviral therapy or a vaccine, vector management is the sole method available for reducing Aedes-induced disease morbidity. Determining the resistance status of Ae. albopictus to insecticides and exploring the resistance mechanisms is essential for future vector control planning.
Aedes albopictus larvae and pupae were sampled from six sites (two sites each from urban, suburban and rural) in Guangzhou. The resistance bioassays were conducted against Bacillus thuringiensis israelensis (Bti): deltamethrin, propoxur and malathion for larvae; and deltamethrin, DDT, propoxur and malathion for adults. P450 monooxygenase (P450s), glutathione S-transferase (GSTs) and carboxylesterase (COEs) activities of adult mosquitoes were measured. Mutations at the knockdown resistance (kdr) gene were analyzed, and the association between kdr mutations and phenotypic resistance was tested.
Adult bioassays revealed varied susceptibility against DDT, deltamethrin and propoxur in the six Ae. albopictus populations. Significantly lower mortality rates were found in urban populations than suburban and rural populations. Urban mosquito populations showed resistance against DDT, deltamethrin and propoxur, while one rural population was resistant to DDT. All populations tested were susceptible to malathion. Larval bioassays results indicated that all populations of Ae. albopictus were sensitive to the larvicide Bti and malathion. Resistance to deltamethrin and propoxur was common in larval populations. The F1534S and F1534 L mutations were found to be significantly associated with deltamethrin resistance. Biochemical assays indicated elevated detoxification enzyme activities in the field mosquito populations.
Aedes albopictus populations in Guangzhou, especially in urban areas, have developed resistance to the commonly used insecticides, primarily DDT and deltamethrin. This finding calls for resistance management and developing counter measures to mitigate the spread of resistance.
Aedes albopictus (Skuse) (Diptera: Culicidae), the Asian tiger mosquito, is an important vector of dengue, chikungunya, yellow fever and Zika viruses, which have emerged as global public health threats [1,2,3,4,5]. This mosquito originated at the edges of forests and bred in natural habitats, and it was previously considered a rural vector [6]. However, Ae. albopictus has adapted well to urban environments with larvae now breeding in artificial containers and has become the most important and sometimes sole vector in urban areas [7, 8]. Aedes albopictus is the primary dengue vector in China [4, 9, 10]. Guangdong Province experienced the highest incidence of dengue in mainland China in the past 40 years [11] accounting for 90% of dengue cases in China. Several major dengue fever outbreaks have occurred in this area since 1978, and Ae. albopictus was the sole vector. Since the 1990s, more than 30,000 dengue cases were reported in Guangzhou. Most of the dengue cases are present in urban areas of Guangzhou [12].
In the absence of specific antiviral therapy or a vaccine, control of Ae. albopictus-borne diseases by vector management is the sole method available for reducing the disease burden. Adult mosquito control depends largely on insecticides. However, resistance to insecticides is rising globally [13,14,15,16,17,18]. The extensive use or non-regulated application of pesticides can hamper the efficacy of larvicide and adulticide-based control programs, as demonstrated in the vector control of Ae. aegypti [19, 20] and Culex pipiens quinquefasciatus [21]. It is reported that Ae. albopictus is resistant or incipient to the major insecticides currently or historically used for vector control across the world, i.e. DDT [16, 20, 22,23,24,25], malathion and bendiocarb [23] and pyrethroids [26, 27] such as permethrin [14, 22, 23] and deltamethrin [16, 22, 23]. Previous studies indicated that Ae. albopictus in Guangzhou was sensitive to all types of insecticides prior to 2002 [28]. Along with the rapid urbanization and recent regional economic development, insecticides were extensively and frequently used in Guangzhou city for dengue control in the past decade [29]. Recent studies have demonstrated that Ae. albopictus developed resistance against pyrethroids during the period when pyrethroids had been massively applied in Guangzhou [29,30,31].
Global surveys indicated that insecticide resistance in mosquitoes can be associated with target-site insensitivity, and/or metabolic-based detoxification. The main target site inactivity mechanisms involve (i) amino acid substitutions in the voltage gated sodium channel (VGSC) that cause a resistance phenotype to pyrethroids and DDT insecticides known as knockdown resistance (kdr) [32]; and (ii) mutations in the acetylcholine esterase sequence that lead to insensitivity of this enzyme to organophosphates [33]. Metabolic detoxification has been found to be a key resistance mechanism in Anopheles and Culex mosquitoes [34, 35]. Detoxification enzymes typically linked to insecticide resistance mainly include three major gene families, cytochrome P450 monooxygenases (P450s), carboxylesterases (COEs), and glutathione S-transferases (GSTs). So far, compared to other mosquito species of public health importance such as Anopheles spp., Culex spp. and Ae. aegypti, very little is known about the molecular or biochemical basis of resistance in Ae. albopictus. Previous studies have examined general resistance status in limited number of mosquito populations in Guangzhou city. A systematic examination of Ae. albopictus resistance status and mechanism investigation in different ecological settings would provide important information on resistance distribution and guidance on resistance management.
In this study we explored insecticide resistance of larval and adult Ae. albopictus in different settings (urban, suburban and rural) in Guangzhou. We adopted biochemical and molecular assays to identify putative resistance mechanisms in Ae. albopictus adult for target-site mutations and detoxifying enzymes up-regulation. We also investigated the insecticide application and sales in different settings in Guangzhou.
The study was conducted in dengue endemic areas in Guangzhou, Guangdong Province, China (Table 1, Fig. 1). Guangzhou, about 200 km north of Hong Kong, is the largest city in Guangdong Province and key commercial harbor in southern China, with a population of 12 million according to the 2012 census survey [36]. It is located in the sub-tropical area with an annual average temperature of 21.6 °C, and annual cumulative precipitation of about 1980 mm. We selected six sites, two each in urban, suburban and rural areas, for our study.
Table 1 Description of Aedes albopictus mosquito population collection sites from urban, suburban and rural settings in Guangzhou, China
Map of study areas and insecticide resistance status of adult Aedes albopictus in three ecological settings in Guangzhou, China
Mosquito strains and collection
Six populations of Ae. albopictus larvae were collected from different ecological settings, i.e. two each in urban, suburban, and rural, from May to October 2014 (Table 1, Fig 1). Tonghe town in Baiyun district (23°11′24″N, 113°19′48″E, 40 m above sea level, masl) and Shishu town in Yuexiu district (23°07′48″N, 113°15′0″E, 32 masl) are urban areas with a population density of > 3000 people/km2. Land usage types are primarily residential and commercial buildings as well as public services such as schools and hospitals, filled with trees and grasses. Liangtian town in Baiyun district (23°21′36″N, 113°22′12″E, 38 masl) and Xinshuikeng town in Panyu County (22°58′12″N, 113°23′24″E, 20 masl) are suburban areas with a population density of approximately 1000 people/km2, and land use includes a mixture of residential, manufacturing, and farmland. Dengcun village in Conghua County (23°30′0″N, 113°33′0″E, 27 masl) and Lanhe village in Panyu County (22°49′48″N, 113°20′24″E, 31 masl) are rural areas with a population density of < 300 people/km2, where land is primarily used for agriculture such as rice and vegetable farming, and forest.
Insecticide resistance bioassays
Larval resistance bioassays
Four insecticides of technical grade were used. Three chemical insecticides were from the Chinese Center for Disease Control and Prevention (China CDC): propoxur (95.56% pure), deltamethrin (94.62% pure) and malathion (95% pure), and one microbial larvicide: Bacillus thuringiensis israelensis (Bti) (7000 ITU/mg, Wuhan Nature's Favour Bioengineering Co., Ltd., Wuhan city, China). All insecticides except Bti (which was diluted in water) were diluted in acetone to the required dosage following WHO guidelines [37]. All bioassays were performed using Ae. albopictus collected directly from the field after species identification. A set of 25 third- and fourth-instars larvae was incubated in 99 ml of distilled water, to which 1 ml of insecticide solution at the required concentration was added. Three replicates were tested for each concentration. Five to nine concentrations, providing a range of mortalities between 10 and 95%, were used to determine LC50 values (the 50% mortality lethal concentration). Mosquitoes of the Foshan strain, which have been reared in the laboratory since 1981 without insecticide exposure, were used as a control. Larval mortality was recorded after 24 h exposure. Control bioassays were conducted by adding 1 ml of acetone to 99 ml of distilled water. Temperature and relative humidity were maintained at 27 ± 2 °C and 80–90%, respectively, in an incubator with a 16:8 h light:dark period.
Adult resistance bioassays
For each strain, five batches of 20 non-blood-fed females (3–5 day-old; n = 100) reared from field-collected immature Ae. albopictus were subjected to insecticide susceptibility test against 0.05% deltamethrin, malathion (0.8%), propoxur (0.1%) and DDT (4%) following the standard WHO tube test protocol [38]. Briefly, we defined resistant mosquitoes as mosquitoes that survived 24 h after the end of the bioassay, and susceptible mosquitoes as the mosquitoes that were knocked down during the 60 min exposure time or that died within the 24 h recovery period. Mosquitoes were considered knocked down if they were unable to fly when they were mechanically stimulated. [39] Mosquitoes of the Foshan strain, which have been reared in the laboratory since 1981 without insecticide exposure, were used as a control (hereafter referred to as laboratory susceptible strain). WHO test and control papers were supplied by China CDC, except for deltamethrin which was supplied by the School of Biological Sciences, Universiti Sains Malaysia (11800 Minden, Penang, Malaysia). The knockdown time (KDT) of females was reported every 10 min during the 60 min exposure period. Mortality was scored after the 24 h recovery period. After the bioassay, one leg of each mosquito was removed and stored individually in 95% alcohol for subsequent DNA analysis.
Metabolic enzyme activity assays
Three metabolic enzymes, P450s, GSTs, and COEs, were analyzed on single individuals from 3 to 5 day-old F0 females without insecticide exposure, and on the laboratory susceptible strain following previously published protocols [40]. Briefly, mean absorbance values for each tested mosquito and enzyme were converted into enzyme activity and standardized based on the total protein amount. Total protein was measured for each mosquito using the method of Bradford [40]. All measurements were done in duplicate. COE activity was measured following the method of Hosokawa & Satoh [41]. Spontaneous hydrolysis was used as a blank control. COE activity was calculated as μmol of p-nitrophenol formed per min per mg protein, using the formula: Δabsorbance/min – Δblank/min) × 1.0/16.4 × 0.05 × protein (mg/ml). An absorption coefficient of 16,400 M was used [42]. For each mosquito population and each insecticide, 30 female adult mosquitoes were tested.
DNA extraction and kdr mutations detection
Genomic DNA was extracted from individual mosquitoes using Fast Tissue-to-PCR Kit (Sigma-Aldrich, Missouri, USA) following the product protocol. Extracted DNA was stored at 4 °C or used immediately for PCR. All survivors and 40 randomly selected (when available) dead specimens after DDT and deltamethrin bioassay exposure were genotyped at the VGSC gene to detect mutations within domains II, III and IV, by direct sequencing of PCR products that contained the specific domains following previously published protocols [43]. Sequences were aligned and analyzed with Sequencher 5.0 (Gene Codes, Ann Harbor, Michigan, USA).
Insecticide usage and sales survey
An insecticide sales and usage survey was conducted in May 2014 with a usage questionnaire from individual residents, the community, and agriculture, as well as insecticide sales from the shops. Insecticides usage included brand name, component content and frequency of application for agricultural and/or public health. At each site, residential insecticide usage surveys were administered to 80 households, 20 households for agriculture application, 8 communities for adult and larvae mosquito control, and 10 shops were administered insecticide sales surveys.
Mosquito resistance status was classified based on the 2013 WHO standard [39]: resistant if mortality is less than 90%, probable resistant if mortality between 90 and 97%, and susceptible if mortality > 97%. Larval median lethal concentration (LC50) and adult 50% knockdown times (KDT50) were estimated with the log-probit model [44]. For the same insecticide, among-site difference in LC50 was tested by pair-wise comparison of the regression slopes of the Probit model. Intensity of resistance was measured using resistant ratio (RR50) defined by (LC50 of field population)/(LC50 of susceptible population) for insecticide concentration, or by (KDT50 of field population)/(KDT50 of susceptible population) for knockdown time. Generalized linear model (GLiM) was used to examine whether adult mortality in the WHO standard tube bioassay differ significantly among localities. Association between kdr mutations and resistance phenotype was examined using Fisher's exact test, and odds ratio was calculated for each kdr allele. Statistical comparisons of detoxification enzyme levels between the laboratory susceptible strain and the field populations were assessed with the Student's t-test. In the case that multiple comparisons were conducted, the significance level was adjusted accordingly.
Using the resistance ratio of 2.0 as the threshold value for declaring resistance for mosquito larvae, all mosquito populations were susceptible to Bti with RR50 ranging from 0.39 to 1.06, and susceptible to malathion with RR50 ranging from 0.74 to 1.94 (Table 2). All six populations were resistant against deltamethrin, with LC50 ranging 0.011 to 0.038 mg/l and RR50 ranging from 11 to 38 (Table 2). The urban population exhibited the highest LC50 in testing against deltamethrin and propoxur.
Table 2 Resistance bioassay results of larval Aedes albopictus in urban, suburban and rural settings in Guangzhou, China. Sites in the same column connected by different letters represent a significant difference in resistance levels at P < 0.05
Using the 90% mortality rate resistance threshold value designated by the WHO [45], two urban and one rural Ae. albopictus populations were resistant against DDT. Populations from urban areas had the lowest mortality rates against DDT (75–80%). Only one urban population showed resistance to deltamethrin, and this population was also resistant to propoxur (Additional file 1: Table S1). All six populations were susceptible to malathion. Moreover, KDT50 of field Ae. albopictus populations were longer than those of control population when exposed to DDT, deltamethrin, and malathion, as indicated by KRR50 > 1.0 for field populations from all sites (Additional file 1: Table S1).
Metabolic enzyme activities and association with resistance
Comparison of the detoxification enzyme activity among the field populations and the laboratory susceptible strain without insecticide exposure found that P450 levels were significantly higher in adults from SPX and SBL (t(39) = 1.87, P = 0.034; t(33) = 5.26, P < 0.001) (Fig. 2). GST levels were significantly higher in four populations, and COE levels were significantly higher in one population (t(39) = 2.11, P = 0.021) (Fig. 2) compared to the laboratory susceptible strain.
Activity of detoxification enzymes in field collected Aedes albopictus adult mosquitoes from Guangzhou, China in comparison to the laboratory susceptible strain. Error bar is the standard error of the mean. An asterisk (*) indicates a significant difference between the population and the laboratory susceptible strain at P < 0.05. Abbreviations: P450, monooxygenases (P450s); COEs, carboxylesterase; GSTs, glutathione-S transferases; S, laboratory susceptible strain
kdr genotyping
Sequences of domains II (480 bp), III (2347 bp) and IV (280 bp) of the VGSC gene were obtained from a total of 111 resistant or susceptible mosquitoes after deltamethrin resistance bioassay and 305 individuals after DDT resistance bioassay. All mutations in codons 989, 1011 and 1016 within domains II or IV were synonymous (codon nomenclature based on Musca domestica VGSC gene according to the accepted kdr codon nomenclature method). Genotyping of mosquitoes after deltamethrin bioassay was done in UBT and UYS populations due to the very small number of resistant individuals in the other four populations. At codon 1534, polymorphisms were detected in both populations with two mutated codons, F1534S and F1534 L. Both kdr mutations F1534S and F1534 L conferred protection against deltamethrin, with odds ratios of 33.6 and 9.3 for F1534S (P < 0.001), and odds ratios of 15.7 and 19.8 for F1534 L (P < 0.05) in the UBT and UYS populations, respectively (Table 3). In populations that underwent DDT resistant bioassay, mutated codons at the VGSC gene was detected in all populations except one rural population (RPL). However, kdr mutations were not significantly associated with mosquito resistance to DDT after a significance level adjustment for multiple comparisons (Table 4). No kdr mutation was detected in the laboratory susceptible populations.
Table 3 Association between mutations at codon 1534 of the voltage-gated sodium channel gene and phenotypic resistance to deltamethrin in two Aedes albopictus populations from Guangzhou, China
Table 4 Genotyping results of the voltage-gated sodium channel gene at 1534 codon and association with resistance to DDT in five Aedes albopictus populations in Guangzhou, China. The significance threshold is P < 0.01 after Bonferroni correction for multiple testing
Insecticide use and sales survey
Field surveys revealed that biological insecticides, pyrethroids, organophosphates and carbamates (except organochlorine) were used for agricultural and public health pest control in the study areas (Table 5). Overall, diverse types of insecticides are being used in the study sites. The pyrethroids-based insecticides were more commonly used than organophosphates. Pyrethrins (mainly cypermethrin and beta-cypermethrin) were the insecticides most frequently used in Guangzhou for community level adult Ae. albopictus control. However, the frequency of pyrethrin applications was higher in suburban and rural areas (5–7 times/week) than in urban areas (0–1 time/week). Pyrethrin and organophosphorus were the main insecticides used in suburban areas for agricultural purposes with a frequency of 1–2 times/week. In rural areas, organophosphorus and carbamates were the main insecticides used in agriculture with a frequency of 12 times/month.
Table 5 Survey of insecticide types and usage in three study settings in Guangzhou, China
Organophosphorus (mainly temephos, mevinphos and fenthion) and biological insecticides (mainly Bti) were used for Ae. albopictus larval control but only in urban areas with a frequency of 1–2 times/month. No insecticide/larvicide was used for larval control in suburban or rural areas (Table 5).
Guangzhou is a dengue epidemic area and has been experiencing frequent outbreaks of dengue fever in China over the past 40 years. Also, it is prone to Zika virus outbreaks due to the presence of recently imported Zika cases in the area [46]. Thus, it is necessary to address the insecticide resistance problem of Ae. albopictus since insecticides were popularly used for mosquito control in the past 40 years. The present study is by far the most comprehensive research into insecticide resistance in Ae. albopictus mosquitoes from different ecological settings in Guangzhou. Two non-synonymous mutations at position 1534 of kdr gene domain III were identified with significant associations to deltamethrin resistance. Additionally, biochemical assays indicated that the three classes of detoxification enzymes may play a role in insecticide resistance in adult mosquitoes. Furthermore, insecticide usage surveys indicated a diverse use of insecticides in the study areas.
Our study area (Guangzhou) is in the subtropical region with a suitable climate for Ae. albopictus development and reproduction. Since the infamous and most deadly 2014 dengue epidemic in Guangdong Province, the city of Guangzhou has intensified vector control programs primarily through more frequent insecticide sprays [47]. In the present study, we illustrated that insecticide usage varied in different ecological settings (urban, suburban, and rural). Insecticide spray in urban areas was more frequent and intense than in suburban and rural areas. Adult Ae. albopictus populations from urban areas were more resistant to deltamethrin, DDT and propoxur than populations from rural areas, while no resistance to malathion was detected in the populations examined. Larval Ae. albopictus populations from urban areas were also more resistant to deltamethrin than populations from suburban and rural areas, whereas, all populations were susceptible to Bti and malathion.
According to the insecticide use survey, we observed frequent deltamethrin insecticide applications in the community, which coincided with elevated deltamethrin resistance. Usage of pyrethroids in urban areas was more frequent than in suburban and rural areas. DDT was used extensively in the 1960s in China for agricultural pest control, but was banned in the 1980s [15, 18]. Since the 1980s, especially in recent years, pyrethroids were massively used to control Ae. albopictus in China [29, 48]. The large scale mosquito control program within urban areas likely contributed to the increasing selection pressure on insecticide resistant Ae. albopictus. Wide usage of insecticide treated nets (ITNs) and long-lasting insecticidal nets (LLINs) in African countries has been linked to the rapid development of pyrethroids resistance in malaria vector mosquitoes over the past decade [49, 50]. The present study suggests that pyrethroid resistance is emerging in Ae. albopictus in our study area, and it is important to develop an appropriate insecticide resistance management plan. Meanwhile, there is an urgency to adopt alternative effective vector control methods that are not reliant on chemical insecticide such as odor-baited traps, larval resource reduction [20] and biological control [7], as well as new chemical insecticides [30].
Among all the populations from Guangzhou tested for kdr mutations, two mutations (F1534S and F1534 L) were detected, and these mutations were positively associated with pyrethroid resistance. This result is consistent with previous studies which found that F1534S mutation was correlated with the deltamethrin resistance [51,52,53]. Thus, monitoring the kdr mutation frequency may aid the surveillance of pyrethroid resistance in Ae. albopictus. In addition, we found significantly higher P450, GST and COE enzyme activities in the field mosquitoes. Literature has reported potential role of GST in DDT resistance [24, 54, 55] and P450s in pyrethroid resistance in mosquitoes, but the precise role of these detoxification enzymes in Ae. albopictus insecticide resistance needs further study.
An interesting finding from this study is the revealed patchy distribution of insecticide resistant Ae. albopictus. Within the urban and rural areas, mosquito populations differed considerably in resistance. One implication of this finding is that we need to monitor the insecticide resistance status in local mosquito populations, and develop efficient mosquito control strategies that take the patchy distribution of resistance into consideration. Currently, biological insecticides such as Bti are not frequently applied for mosquito control in China and no resistance has been detected, thus they can be considered as alternative insecticides for vector control.
Our findings urgently call for timely surveillance of insecticide resistance as well as attention to the roles of metabolic detoxification enzymes and kdr mutations in insecticide resistant Ae. albopictus. The threat of dengue outbreak calls for an intensified and effective vector control program. Appropriate insecticide resistance management and additional vector control tools that are not reliant on synthetic insecticides are urgently needed to reduce dengue transmission.
Bti :
Bacillus thuringiensis israelensis
COEs:
Carboxylesterase
DDT:
Dichloro-diphenyl-trichloroethane
GSTs:
kdr:
P450s:
P450 monooxygenase
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Chen H, Li K, Wang X, Yang X, Lin Y, Cai F, et al. First identification of kdr allele F1534S in VGSC gene and its association with resistance to pyrethroid insecticides in Aedes albopictus populations from Haikou City, Hainan Island, China. Infect Dis Poverty. 2016;5:31.
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We thank Lijie Yao, Xueli Cai, Tong Liu, Zhuanzhuan Liu, Guiquan Lai, Qiankun Chen, Hehai Huang and Yuhang Huang for their assistance with the larval collection and insecticide usage survey.
This study was supported by the grants from the National Key R&D Program of China (2016YFC1200500, 2017YFC1201000), the National Nature Science Foundation of China (81371845, 81420108024, 81528013), the Natural Science Foundation of Guangdong Province (2014A030312016, 2016A020251001), the Guangzhou Synergy Innovation Key Program for Health (201508020263, 201605030010) and the National Institutes of Health, USA (R01 AI083202 and D43 TW009527).
All the important information regarding the article is available in the main text.
Department of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, China
Yiji Li
, Jiabao Xu
, Hong Zhang
, Wenqiang Yang
, Xinghua Su
, Yang Wu
, Kun Wu
, Guiyun Yan
& Xiao-Guang Chen
Key Laboratory of Translation Medicine Tropical Diseases of Ministry of Education, Hainan Medical University, Haikou, Hainan, China
Department of Pathogen Biology, Hainan Medical University, Haikou, Hainan, China
Program in Public Health, School of Medicine, University of California, Irvine, USA
Daibin Zhong
, Guofa Zhou
& Guiyun Yan
Department of Vector Control, Centers for Disease Control and Prevention of Guangdong Province, Guangzhou, China
Songwu Cai
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YL, GY and XC, conceived the study and coordinated its implementation. YL, DZ, XC and GY participated in the experimental design. YL, JX, HZ, WQ, XS, YW, KW, and SC performed the experiments and drafted the manuscript which was critically revised by JX, GZ and XC. All authors read and approved the final manuscript.
Correspondence to Guiyun Yan or Xiao-Guang Chen.
No specific permits were required for mosquito field specimens' collection, which were performed in urban, suburban and rural areas of Guangzhou, China. This study did not involve endangered or protected species.
Knockdown time (KDT) and mortality rate of Aedes albopictus populations from urban, suburban and rural settings in Guangzhou, China using the standard WHO tube susceptibility bioassay against four insecticides. (XLSX 12 kb)
Li, Y., Xu, J., Zhong, D. et al. Evidence for multiple-insecticide resistance in urban Aedes albopictus populations in southern China. Parasites Vectors 11, 4 (2018) doi:10.1186/s13071-017-2581-y
Accepted: 10 December 2017
Aedes albopictus
Biochemical assay


An outstandingly beautiful single male guppy, no matter how many trophies he may come home with, does not make a champion guppy strain… or a champion guppy breeder. You can be justifiably proud of your win in the single classes because the competition is brisk, but the secret of true champions in the guppy world is a strain of guppies that is capable of producing a number of matching males that are as close to carbon copies as possible.
In the single classes it doesn't matter whether a fish is from a true strain or from a random outcross. All that matters is that the guppy be a superb individual specimen. The quality of a strain becomes progressively more important in the matched male classes. Random outcrosses may produce beautiful individuals, but rarely will they produce two beautiful individuals that match in every characteristic well enough to win in the two-matched-male classes (known as Tank Classes), much less five outstanding mirror images needed to win in the Breeder Class. Strains that have been developed to the point where they produce consistently uniform males of high quality are valuable and highly sought after. The time and effort and money that has gone into the development of such strains make them well worth the initial investment.
If you were lucky enough, and smart enough, to locate and purchase breeding stock from one of the best strains available, and have paid attention to the raising of the offspring, even as a newcomer in the hobby you could already have guppies in your tanks capable of winning championships or helping you pay for your hobby. Even if these are the first show guppies you have ever raised, you can already be well ahead of more experienced guppy breeders who might have skimped on the purchase of their breeding stock and have had to struggle along wasting time and effort in trying to improve inferior stock. Naturally, this depends to a large degree on whether your care and conditions have allowed the fish to blossom to their full potential.
In past articles we have discussed how to select those individuals that are of show quality. Now we'll take a look at matching. To find out just how well your guppy males really match each other, try picking out your five best males and putting them together in a separate container where you can see them from all sides. At first glance they may all look pretty much alike, but when you start comparing them feature for feature, as the judges of matched competitions do in a show, you can begin to appreciate the value of a good true-breeding strain. Are the caudals all exactly the same shape and size… or does one have an indentation here or an extended ray there, or perhaps one has a rounded corner or concave trailing edge? Is the color and pattern of the tail the same in each fish, or is one a little brighter color, another have a few streaks of darker color or a different iridescence where the tail leaves the peduncle? Are the dorsals the same width? Length? Shape? Color? Are the bodies all the same size and shape, or is one smaller or larger; what about the one that has a bulkier pot belly or a narrower peduncle? Are the body markings the same down to the various splashes or dots of color? Do the fish all carry their finnage equally well? This may all sound like nit picking, but I have seen shows where the presence or absence of one tiny black dot on the body meant the difference between first and second place.
It should also be noted here that fish with multi, or bicolor patterns in the finnage are apt to be less uniform than fish with solid colored caudals. Even in the purest of strains the patterned tails show more variation because of the added factor to the matching… pattern as well as color. When matching multis or bicolors for competition, select first for colors that match as closely as possible and then look for the closest pattern match from among those showing the desired color.
When you figure the odds of getting five trophy caliber males in the first place and then add in the odds of having them all matched in every characteristic you can begin to realize why the quality of a strain is the single most important thing in guppy raising. NO amount of super food or super care is going to give your guppies the genetic makeup needed for uniformity and show-quality characteristics.
When selecting guppies for show competition, most experienced exhibitors will select their matched male entries first, knowing that the combination of top quality and good matching is harder to attain than finding a single outstanding male. Winning in matched male classes is not only more prestigious but, deservedly, earns the breeder many more points toward the annual championship awards.
If, after evaluating your stock as to both the number of show quality individuals and the good matching of the males, your strain does not seem to measure up, don't automatically assume you were "taken" by the breeder from whom you purchased your strain. That is possible, of course, but it's usually not the case if you bought from a reputable breeder It is a known fact that guppies react differently to different environmental factors. For example, guppies of the same strain may be one color in acid water and quite a different shade in alkaline water. Lights, temperatures, foods, water, etc., can all have visible effects on physical appearances. Unfortunately, when you acquired the guppies from the breeder, you could not hope to also acquire his identical set up and handling techniques. It sometimes takes several generations of working with a new strain to discover the conditions that will make it perform best for you, so don't be too hasty to blame the breeder. I once obtained breeding stock of a champion blue delta strain from one of the top breeders. In the first generation most of the males showed up with yellow and blue variegated tails… they weren't even very pretty. Knowing the breeder, I stuck with it and amazing as it may sound, after two generations had been raised in my tanks, the color settled down to a solid green, No, I never could never get them to turn blue, but I did end up with a very lovely solid green show strain. It had to have been the effect of something in my environment, because when my "greens" were passed onto breeders in other areas, they sometimes became pure blue again.
The time to be most suspicious about not having obtained high quality stock is if the first generation of young show a wide assortment of colors and characteristics. Different conditions and handling won't make helter-skelter differences in a good strain. if environment makes any changes to the appearances, they will Usually be quite uniform differences if the strain is true breeding to begin with.
Throughout the discussions of evaluating show fish I have given tile points allotted to each characteristic as set forth in the IFGA standards, because its important for a breeder or exhibitor to learn the weight given to each trait in the final balance. I highly recommend practicing the actual pointing of your fish until these values become automatic. If you feel that you have learned them all well enough to successfully pick out your "most apt to win" show fish, test yourself with the following exercise: Visually select what you consider to bc your top three fish and place them in first, second and third position. Now, go back and, with pencil and paper, completely and honestly point each characteristic of each fish. If the point totals place the three fish in the same order as your original visual selection, you have mastered the point system well enough to feel confident you are not leaving your best fish at home when you enter a show.
From this point on you can safely use the same short cut method of selecting your best fish as is used by show judges, who cannot possibly point every fish in a class. Instead of going through the arithmetic of pointing each feature and adding up the total, judges who thoroughly know the standards and the values placed on each characteristic mentally start each fish with 100 points and deduct for any short comings the fish may have. Although this system usually comes out amazingly close to actually pointing the fish, judges still point fish completely in close contests and you will probably come out ahead by doing the same when in doubt.
A renowned aquarist, Midge Hill has been keeping and breeding guppies for 23 years and is a member of the Guppy Hall of Fame. Active in the showing and judging of guppies throughout the United States from the time guppy competition first started in California. Midge has won numerous trophies, including an IFGA Championship for the Albino Class. The author was one of the founding members of the Pan Pacific Guppy Association in 1965 and served in many capacities for that association, including the office of president. She also originated its publication, "Guppy Roundtable," which she edited for six years while serving as Guppy Judging Chairman and Seminar Instructor. Currently a lecturer, fish photographer, artist and author of numerous articles on guppies and related subjects. Midge is also active in the International Fancy Guppy Association where she has served as a member of the IFGA Judging Board since 1974. In addition, she edited the IFGA Bulletin for three years. Midge Hill is professionally associated with the aquarium hobby as one of the founders of Golden State Aquatics, Inc. and also as a Contributing Editor for Freshwater and Marine Aquarium Magazine.
This entry was posted in Guppy Blog - Guppies corner and tagged GUPPY GAMBITS BY MIDGE HILL, Guppy Show Classes, Show Guppies, show guppy. Bookmark the permalink.


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text:
Op Ed December 2, 2020
The National Infrastructure Strategy – better, greener, and faster?
By Genevieve Redgrave - World Infrastructure Journal
"Better, greener, faster" is the motto of the long-awaited National Infrastructure Strategy published last week alongside the Chancellor's latest Comprehensive Spending Review. Over two years on from when the National Infrastructure Commission (NIC) first made its recommendations, the Government finally published its response, setting out its vision for the future of the UK's infrastructure.
With broad-sweeping aims of "levelling up" and not-too-distant goals of net-zero and full-fibre connectivity, does this report have the right strategy?
A united kingdom?
The term regional powerhouse has long haunted the political landscape, with grand claims of improving economies seen as left behind. Yet, with a seemingly growing north-south divide and the capital's leading role in the planned economic recovery, it would not be too obtuse to suggest the powerhouse remains firmly in London.
This Infrastructure Strategy, however, aims to change that by "making cities the engines of growth and revitalising towns. " This includes increasing rural network connectivity to 95 per cent, creating jobs in the most deprived areas through the Free Ports Programme, revitalising 100 town centres through the Towns Fund and creating green growth clusters in traditionally industrial areas. The specifics of these areas are yet to be known but could follow on from the industrial hubs of renewable energy that have significantly improved local economies across the UK.
National infrastructure improvements rely heavily on levelling up the connections between communities yet travel outside of London remains woefully inadequate. The regular criticisms of costly and inefficient train networks, infrequent bus services and dilapidated roads that most of the nation face are not going away just yet. However, with greater investment, the out-of-date infrastructure underpinning these services can travel into the modern world.
The Government has set out £4.2 billion for intra-city transport outside of London, an integrated rail plan to improve trains in the Midlands and the North, £27 billion for strategic roads, and the Union Connectivity Review which aims to increase the physical links of the four nations of the UK.
These travel routes may also be useful for the 22,000 civil servants who are to be moved out of London and the south-east by 2030 - although it is still unclear which departments will be leaving and exactly where they will be heading. This is part of a wider devolution settlement in the strategy.
The highly anticipated UK Infrastructure Bank will also be based in the North of England and will look to co-invest with the private sector on infrastructure projects. While the full terms and regulations of this institution will be set out in 2021, it seems the financer will be able to lend to local authorities for key projects and provide them with advice.
This could prove a useful resource for cash-strapped councils, but it remains questionable if many will simply not want to take the risk of borrowing when their finances have already been hit so hard by the pandemic. A levelling-up plan based on local authorities borrowing is very unlikely to lead to the rapid revolution of infrastructure that so many areas need to meet the demands of the future.
A greener future?
Following on from the announcement that petrol and diesel vehicles will no longer be sold from 2030, the discussion on electric and renewable infrastructure has been supercharged. Decarbonising the economy and adapting to the threats of climate change are rapidly rising on the agenda ahead of COP26 next year in Glasgow and show no signs of slowing down.
The strategy sets out "significant investment" in offshore wind and modern ports which will be helped by the world's largest windfarm coming to the shores of Yorkshire. This investment comes alongside £525 million in nuclear, £1.3 billion in electric charging infrastructure, £1 billion for carbon capture and the large-scale rollout of existing heat decarbonisation technology. For those 336,000 properties also at risk from climate-related threats such as extreme flooding or cliff erosion, £5.2 billion is set to be invested by 2027 for their protection.
Sustainability is also at the heart of new housing developments moving forward with large-scale reform of environmental regulations taking place. This aims to create a quicker and simpler framework for assessing environmental impact to ensure the environment is protected at every stage.
Simultaneously, the delivery of housing is set to be "accelerated and improved" by transforming the construction sector to make it "more productive, more sustainable and more internationally competitive. " The strategy sets out the aim of accelerating investment and skills to ensure the necessary infrastructure is delivered faster. However, this is at odds with the decision to place more emphasis on "quality design and better monitoring and evaluation" which many have suggested will simply increase delays on projects. It can only be hoped that "better, greener, faster" is not a pick-and-mix for developments.
The industry response
The long-awaited National Infrastructure Strategy came as a monumental moment for those key stakeholders in the infrastructure sector.
The Institution of Civil Engineers called for "more clarity on how the Government intends to deliver on UK net-zero commitments" but added that "the publication of the NIS should still be considered as an important moment for the infrastructure sector. "The most prominent reaction was that of the National Infrastructure Commission (NIC) whose recommendations were the catalyst for the report. Sir John Armitt, Chair of the NIC said:
"Publication of the UK's first-ever National Infrastructure Strategy is an important moment, given the vital role infrastructure can play in supporting economic recovery and growth across the whole of the UK, as well as tackling climate change. We are pleased to see the Government's strategy responds closely to our own independent assessment of the country's infrastructure needs and how to address them.
"We are particularly pleased that government has accepted the Commission's proposal for an infrastructure investment bank, to help catalyse private investment.
"Alongside announcements in the Spending Review, the Strategy represents a solid down payment on a long-term fiscal commitment to infrastructure.
"As government acknowledges, there are currently some missing pieces of the jigsaw when it comes to energy policy. The 10 point plan set out by the Prime Minister last week set out serious commitments in areas such as wind power, decarbonising heating, developing hydrogen and carbon capture and storage. We look forward to seeing detail on the delivery plans for achieving these targets in forthcoming announcements.
"We welcome reform of the Treasury's Green Book to assess investments against policy objectives. The proposed focus on investment in local infrastructure is welcome, as are intra-city budgets for mayors. However, to achieve the aim of levelling up, more long-term funding and control for cities is necessary to bring transformational urban public transport projects to places outside London.
"The Commission will monitor the Government's progress on delivery and very much hope this strategy marks the beginning of a renewed focus on long term infrastructure policy around which industry and investors can plan with confidence. "
In a period of such uncertainty, the popular phrase "building back better" has become all-encompassing, failing to offer any real clarity about how exactly this can actually be achieved.
While far from comprehensive, the National Infrastructure Strategy does offer a vision for what this can look like, and large scale and long terms aims are a step in leading the UK's infrastructure forwards. If this Strategy is harnessed and put into practice, it just might be possible that the nation can actually build back better, greener and faster.
#wijnews #infrastructure #genevieveredgrave #nis #coronavirus

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News Coronavirus COVID vaccine passports no 'magic bullet' for reopening Australia's international borders
6:00am, May 22, 2021 Updated: 6:51pm, May 22
COVID vaccine passports no 'magic bullet' for reopening Australia's international borders
COVID vaccine passports are not a 'magic bullet', IATA says. Photo: Supplied
Cait Kelly Reporter
The federal government is in talks with airlines for a global coronavirus vaccine passport, but a leading epidemiologist has warned Australia needs to inoculate at least 85 per cent of the population against COVID-19 before international borders reopen.
This comes as Prime Minister Scott Morrison's idea to develop a vaccine app for interstate travel has been slammed by some health experts, who have called on the government to help fix the hotel quarantine system instead.
The government is currently in talks with the International Air Transport Association, a trade association made up of 290 of the world's airlines, to use their vaccine passport system to unlock quarantine-free travel.
Vinoop Goel, IATA's regional director for airports and external relations in Asia Pacific, told The New Daily talks with the Australian government were "ongoing".
"We are working with a number of airlines on trials of the IATA Travel Pass, including over 30 airlines that have announced publicly. Qantas and Air New Zealand are participating in the pilots," Mr Goel said.
"Singapore, Panama and Estonia have announced that they will accept the IATA Travel Pass for entry."
The Morrison government is eyeing Singapore and some Pacific Island countries as possible destinations with which Australia could open a travel bubble.
IATA's vaccine passport no 'magic bullet'
The IATA Travel Pass consists of a global registry of health requirements, a global registry of testing/vaccination centres, a lab app to securely share test and vaccination certificates with passengers, and a contactless travel app that enables passengers to create a digital passport and manage their travel documentation online.
Digital "green pass" vaccination certificates are already being embraced around the world, including by Israel, the EU, and the UK, where international travellers will be asked to put in their COVID vaccine status through the NHS app.
Mr Goel said IATA's travel pass wouldn't be a "magic bullet" as governments also need to develop plans for getting citizens vaccinated and opening borders.
"The IATA travel pass alone is not the magic bullet for restarting travel if governments do not remove restrictions such as quarantine," he said.
"Hence the priority is still for governments to establish a roadmap towards reopening borders without quarantine and relaxing travel restrictions, using testing and vaccination, once it is safe to do so, and to work with the industry on the roadmap so as to allow airlines to plan ahead and provide the necessary connectivity."
PM flags COVID passports for interstate travel
Mr Morrison has flagged the potential use of COVID passports for interstate travel in Australia, but Marylouise McLaws, an infectious diseases expert at the University of New South Wales and member of the World Health Organisation's COVID-19 response team, said the plan was "unnecessary".
"I don't think we need it. It may well have been mentioned to encourage people to get vaccinated," Professor McLaws said.
"There are many other ways to get people vaccinated, rather than excluding people from travel."
The best thing we can do is fix the faults in our hotel quarantine system, she said.
"We have no circulating virus, the only risk we have is inadequacies of quarantine facilities," she said.
"If the authorities took on best practice nationally and every hotel quarantine was practised at that level, we should not need a vaccine passport."
Australia's path to reopening borders
Calls for a plan for reopening Australia's international borders have been growing.
This week, Virgin Australia chief executive Jayne Hrdlicka was forced to clarify her comments that the Australian border cannot remain shut forever, and that "some people may die" as a result of reopening.
"If I had time again I would use different words to make the same point," Ms Hrdlicka told reporters in Brisbane.
But she wasn't the only one trying to press the federal government for a plan to open up – the New South Wales government also called for a clear timetable.
Many questions remain about how Australia will get there – including how many of us need to be vaccinated before we can open up.
The target we should be aiming for
Australia needs to vaccinate at least 85 per cent of the population before reopening, Professor McLaws said.
If you take the midpoint for efficacy of both the AstraZeneca and Pfizer vaccines, then look at it in conjunction with the transmissibility level (which needs to be higher because of mutant strains), Professor McLaws said Australia needs to vaccinate 85 per cent of the population at a minimum.
"You want as many people to get vaccinated," Professor McLaws said.
"I would go for 100 per cent. You won't get it, but we can't aim for 80 per cent, it's not high enough."
Right now, the most important factor for getting the borders open is to get people aged between 20 and 59 vaccinated, because they are typically more social and will "ring-fence" older Australians, Professor McLaws said said.
"I wouldn't be contemplating [opening the borderl] until you have the majority of that group vaccinated," she said.
"We are relying on them, not just for their own health, they are going to be our saviours. The heroes."
Australian Politics Coronavirus vaccine COVID-19
Celine Dion cancels world tour leg over health problems
Half-a-million vaccinations per week soon: PM urges Australians to get jab


The newest versions of Microsoft's Office productivity suite are an interesting departure from the Office of old. There are several versions and editions of Office, and each one offers something a little bit different. There's the well-known local desktop version, Office 2016, and the relative newcomer, cloud-based Office 365. The apps in each are comparable, but Office 365 offers more flexibility with its access-from-anywhere model.
Companies and IT administrators need to know what they're getting from the different Office editions -- and smaller shops are not exclusively limited to the Office for Business versions. Businesses must also consider whether to deliver the mobile or desktop version, and they might even find it's time to switch to the cloud-based subscription suite. One bright spot in this migration process is that Microsoft has added security features to Office 2016, so moving off an older version could improve existing security measures. This handbook outlines everything businesses need to know about choosing which Office to deliver to users.


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Thomas the Tank Engine Character 'Oliver' may return for the line's 150th anniversary year in 2022
South Devon Railway is hopeful that family favourite Thomas the Tank Engine™ character 'Oliver' may return to steam again at the SDR for the line's 150th anniversary year in 2022
Two weeks ago saw the 'Family' theme week in the fourth of the six-week long, nationwide 'Love Your Railway' campaign from July 26 to September 6, and in which the South Devon Railway (SDR) has been actively involved very successfully.
So, mention of the overhaul of one of the classic steam loco characters from the late Rev. Wilbert Awdry's now world-famous Thomas the Tank Engine™ series of children's books is fitting as the engine is hopefully getting closer to completion of a 15-year overhaul this year at the SDR.
The overhaul should now speed up thanks to a recent grant of £5,000 from the Veronica Awdry Charitable Trust towards new boiler stays for the final restoration of former GWR Collett designed auto-tank 0-4-2 No. 1420, AKA 'Oliver' in the Thomas the Tank Engine™ children's books.
Veronica Awdry is the daughter of the late Rev.W Awdry and her charitable trust helpfully distributes some royalty funds regularly for appropriate heritage railway projects, and which has been vital in the Covid 19 period with limited income and big losses suffered on all heritage railways.
Veronica Awdry comments: "Steam loco No. 1420 is a truly iconic engine for the South Devon Railway because members of the class were seen in use on the quintessential GWR branch line from the 1930's right up until its closure as a freight railway in 1962, and then under the Dart Valley and South Devon Railway's 52-year operation of the scenic seven-mile line from 1969 to date. Featuring as 'Oliver' in the Thomas series of books, I know he was one of my late father's favourite characters.
"And, with the SDR celebrating its 150th anniversary year in 2022 having first opened in 1872, there is a concerted effort underway to complete the overhaul. SDR supporters are hoping to see 'Oliver' finished in time for service next year, although quite a lot of work is still needed. The popular engine is owned by the SDR Association and it last ran on the SDR in 2006 when it was stopped for a protracted overhaul mainly due to a leaking boiler foundation ring and other faults."
Last year, a further £22,180 grant from the National Lottery Heritage Fund (NLHF) was made towards moving the overhaul of No.1420 forward to cover the manufacture of the remaining, vital firebox boiler plates and a new foundation ring, which was critical to enable the SDR to re-build the new boiler using its own skilled staff and to keep the loco restoration timeline on track to maintain future operations.
The earlier NLHF grant was made possible by money raised by National Lottery players, and was a considerable boost to the SDR which sadly was simply unable to run any trains last year.
Although steam loco No. 1420, AKA 'Oliver' is nearing completion in the SDR's Buckfastleigh workshops with the wheels and frames having already been restored, plus considerable boiler work, there is still a long list of work to be completed and funds raised to pay for it so further specific donations towards finishing the tasks would be warmly welcomed by the SDR.
The outstanding works include fitting a new boiler barrel, foundation ring, tube plate and completion of a roughly assembled firebox which is being drilled for assembly with new stays. And, although the chassis is now back on its wheels, none of the valves, pistons and motion are fitted and nothing has been done yet to overhaul these parts. Boiler fittings now in use on other SDR locos will need to be found, and there is the small issue of the loco requiring new side tanks which will take some making as they are riveted, but completely different designs left and right due to the reverser.
The South Devon Railway is the oldest heritage railway in the West Country having re-opened in 1969, and usually welcomes up to 100,000 visitors every year. Last year, it carried only 1,750 passengers to March and incurred losses of some £2 million, offset by Government and Council grants and an 'SOS' fund-raising campaign which raised almost £1.4 million.
It is a registered charity and has a small number of some 26 paid staff and over 600 volunteers who usually run the railway every day from March to October, plus at Christmas and New Year too.
The SDR has an extensive, well-equipped workshop which repairs the line's own locomotives as well as undertaking complex, contract engineering jobs for other heritage railways and owners, plus main-line Train Operating Companies too.
As a result, it is well placed to complete the largely new boiler for GWR loco No.1420 and help bring the popular steam engine back to life again soon, hopefully in 2022.
The SDR has a number of other 'Family' related elements around the railway's footprint, including a large play area, miniature railway around the 12-acre site and a Riverside walk at Buckfastleigh, along with the Dartmoor Otters and Buckfast Butterfly centre, plus the Totnes Rare Breeds Farm, and Pixie Falls en-route near Totnes. It's a family friendly attraction wherever you look.
Thomas the Tank Engine is back for two days at South Devon Railway on 24-26th September 2021.
Read more and book tickets here
Devon's Top Attractions
NEW Bear Experience at Wildwood Escot!
Christmas in Devon – Let's Get Festive!!


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redpajama_set_name: RedPajamaGithub


RCW 46.83.030: Deposit, control of funds—Support.
All funds appropriated by the city, town, or county to the operation of the traffic school shall be deposited with the city, town, or county treasurer and shall be administered by the governing body of the city, town, or county. The governing bodies of every city, town, or county participating in the operation of traffic schools are authorized to make such appropriations by ordinance or resolution, as the case may be, as they shall determine for the establishment and operation of traffic schools, and they are further authorized to accept and expend gifts, donations, and any other money from any source, private or public, given for the purpose of said schools.


The new consultancy will offer everything from strategic communications to advocacy advertising to government relations.
Indent Communications and Torch Agency have come together to form One Persuasion, a Toronto-based consultancy focused on public strategy, advertising, advocacy and research.
Through the merger, Indent Communications' Daniel Charron and Dan Robertson join Torch's Hamish Marshall, Travis Freeman and Brendan Jones. Charron and Robertson have retained their titles as partners of creative and strategy, respectively, while Marshall has been appointed research partner, Freeman as digital partner and Jones as creative partner. The One team has also brought on Terrance Oakey to lead its government relations practice.
The fully bilingual agency will be headquartered in Toronto and have additional offices in Ottawa and Vancouver, due to the nature of its business plan, Daniel Charron explained in an email to strategy.
Indent Communications and Torch decided to merge in order to be able to offer more services and expertise to their clients, he said.
One hopes to attract national clients through its broad range of services, including strategic communications, advocacy advertising, research, branding, political consulting, issues management, digital marketing and government relations.


The following command removes the skip activation flag from all logical volmes that are thin snapshots.
The following example shows a more complex selection criteria statement. In this example, all logical volumes are tagged with "mytag" if they have a role of origin and are also named lvol or the logical volume size is more than 5g.


I've pushed myself harder and harder with every novel I've written, both in going deeper with my thoughts and trying to get purer in the craft.
Yes, the better I've gotten as a writer (not just my assessment—below are the average Goodreads ratings), the more my book sales have plummeted. From once being considered a major mass market pull by my Indian publishers, I'm just a touch above the midlist. Reliable but not blockbuster. Relevant but dispensable.
If I've learnt anything in my corporate career, it's that a product earns mass success when it meets the unmet needs of the masses. Every time I've created an advertising campaign that works or launched a successful new product in my job, I'd done my homework to know my consumers inside-out and created something that satisfies an unmet, often unarticulated need for them.
I've never done that for my writing.
I haven't kept the consumer in mind for my books. On the contrary, I've written to answer questions that have tormented me—the meaning of success in Johnny Gone Down, the nature of suffering in The Seeker/The Yoga of Max's Discontent—and enveloped them in the same kind of pacy narratives that interest me as a reader.
It doesn't work, at-least from a mass sales perspective.
The less mainstream the questions I've asked myself have gotten, the less my books have sold. To sell a million copies, you need to touch a million hearts. To touch a million hearts, you need to know their hopes, their dreams, their aspirations, their burning pains.
I can guess. People are consumed by conflicts of love, the drama of relationships, the dreams of upward mobility.
That's why The Seeker didn't sell a million copies.
It didn't deserve to. I hadn't done my consumer work.
But curiously enough, I'd write it all over again. And for my next book, I'm likely going to skip the consumer research again. Because each book I've written has opened windows to new worlds for me, helped me find meaning where I felt none existed, and transformed me in immeasurable ways. And in answering my own questions, I hope I've touched a few lives along the way.
The one decision you need to make as a writer.
Are you creating a product for people or are you trying to deepen your own soul?
In the former, you'll end up selling a lot of copies, garner adoring audiences, and become someone whose voice is heard and respected above the din of individual opinion.
In the latter, you'll lose out on all of these but you'll gain something else. A growing profundity, the quiet joy of an unadulterated pursuit in a noisy world, and the enthusiasm of boundless personal curiosity as you seek deeper, more meaningful experiences with each creation.
Both have their place. That's why I don't understand the vitriolic directed at a James Patterson in the US or a Chetan Bhagat in India. They know their exact consumers and are creating the exact products that meet their consumers' needs. Much to admire there–even if that's not a choice you decide to make.
My one advice for a debut novelist.
I'm a hypocrite. For all my talk about writing being a personal journey, I love publishing a novel. My editors at both Harper and Random House have challenged my stories to help me grow tremendously as a person. Readers' emails and messages never fail to lift my heart. I get excited when I hit the bestseller lists. All in all, I like everything that comes with being a "traditionally" published novelist.
If I'd started with a novel as personal as The Seeker, I'd probably have none of those.
I got lucky. Unknowingly, Keep off the Grass satisfied the growing market demand for campus stories while being a personal passion project since I wanted to write a trippy, road-trip adventure, being a personal fan of the genre.
Be more deliberate than me to get a major publishing deal though. You don't have to be a sell-out. Just figure out the intersection of your personal passion with a market need by looking at what genres sell, what people are talking about, and where the zeitgeist is going. That's the recipe for a winning story you'll also dig writing.
Of course, if you have that one idea that grabs you such that it doesn't let go, then forget all this above. Just go for it. From Fifty Shades to Amish, each year there's at breakout that defies all analyses. Here's wishing it's you!
P.S: What drives your writing? I'd love to hear from you in the comments below.
Insightful as always Karan. Think this will help me resolve my current decision making.. Have you moved to Mumbai already..
How you? Hope you doing good. Your avid reader her again.
Well written article, much needed.
Needed this article at this time.
Karan I am trying to consider self publish my book by January 2017.
I never got any reply from major publishing companies, for the proposal and Outline manuscript I have sent.
I have a strong sense in my mind, that I have written something out of my heart, which people would love to read , leave them a feeling and marvelous impact after reading the last word in the last page of my book.
Karan could you give me some advice or check list or "How I did it?" tips for self publishing.
Chaitanya, I'd love to, man, but I've never self-published so I don't know if my advice is any use. Good luck, man.
Please clarify. Want to write which sells more. Is this decision determined by outside activities? When will you change your natural alignment and for what ,you will change.
I'm not sure I understand the question, Prabhu.
My apologies man, do send me a paypal so I can refund your purchase.
I just loved your book The Seeker. There were many points in the book that lent itself to deep reflection. I would actually, love to write such a book but I know I can't. I love reading and writing romance and I don't feel authentic when I do something else. I spent the last two years trying to write a thriller and it didn't work out because I don't like reading thrillers!
It's sometimes embarrassing for a woman with growing kids like me to tell people that I like reading and writing romances but I've realised that it's what I love and I can't help it. I have to be true to myself.
Romance is terrific for a woman with growing kids And thank you for liking The Seeker, that means a lot.
Both these books 40000 copies in total, which is much lesser than yours in spite of being better books.
But I am a fan, not of your writing, but of the fact that you are not a sell out. It would have been so much easier for you to keep coming out with different versions of Keep off the Grass, but you didnt. For that courageous decision, you have my respect. Keep writing.
Thanks man, I'd love to read The Underdog. Where can I find it?
Thank you Bhanupriya, you and I have a lot in common–I'd love to read your book.
Hi Karan.. The Seeker might not have sold million copies, dont worry about that. You are not a commercial movie producer who selects scripts for the mass. How many people really think about their purpose in life? How many of us would actually think of leaving this materialistic world for the sake of enlightenment? Not many… so you should be feeling awesome that your heart could conjure such deep meaningful thoughts. This book certainly changed my heart.. though I practise Buddhism, I was not able to get any direction in my life. After reading The Seeker I was a changed person. I am going re read it as many times till I can get to the bottom of every of word, every sentence in the book.
I have never tries writing, if I ever do that it has to be something that touches my heart. I do like the idea of knowing what people would want to read.. but that wouldnt be creativity anymore. That would be just a product. Writing like any form of Art has to be your own creation.. your individual feelings and thoughts. Thank you Karan. Keep following your heart.
Very, very kind of you, Sonia, that truly means a lot. I appreciate how deeply you've understood the book.
Thanks for taking the trouble to discern and write about this and then share it!
I can relate to this completely, although my context wasnt writing novels, but facilitating experiential growth-promoting workshops. Same downward spiral from 1995 to 2013… eventually stopped even attemptiing to oragnise them in the last 3 years.
Yes, but. There are 2 thoughts i am trying to hang on to. First, I don't have anyway to quickly access information on this, but isn't there the long term value of a book to consider? Not the blockbuster value? Not the fad? I'm thinking of 'Siddhartha' and 'Search in Secret India.' I'd love to know what Hesse and Brunton would have to say about it.
Also, what would be the effect of your reasoning about writing for your perceptions of the needs and desires of others eventually do to your own sense of who you are? It seems to me that there are at least 2 kinds of artists and you need to figure out which one to be. It's possible to do both–but maybe rare? I'm thinking of 2 southwestern artists I have always loved, Amado Pena and R.C. Gorman. They both produced tons of quick pieces and, with printmaking, made tons of money–and they caught a lot of criticism for it. But both continued with very serious work, I think. Maybe Pena has gone deeper now. I haven't kept up. But you already figured that out.
Anyway, I wait with anticipation and affection to see what you will do next.
No offense, but Keep off the Grass was the best. Seeker was ok. Haven't read Jonny, so can't say. It not my place to suggest, but since you're good with adventure-type backgrounds, how about a simple adventure story the kind Michael Crichton wrote. Or something like Indiana Jones.
Hi Karan! I keep on coming to this blog of yours. I like the angle you provide. Actually, it is very difficult to delve into ourselves for everything we want to do, I mean doing everything for a reason and then, you have this world where vanity I guess plays a pretty important role. I have just finished writing a book and I am self publishing it, actually vanity publishing it. I just love to write, I don't like to market it or promote my book. Now when the book is complete, it has absolutely no value to me. Even while I am honestly writing this comment, I am not sure whether I am trying to promote my book at the same time. What's your take to all this "creating the right balance" jargon?
I have been a voracious reader who has been struggling to read with a struggling motherhood…I loved what you wrote. It is matter of fact and non objective. I really do not care about genres…I like writers. I like your work. Keep it up.
Yes its true we have to choose what we want all the time, not just in writing, in everything. From the most trivial, what should I eat today to the most profound,what do I want to do with my life. Once you make choices the path is laid out. We, however, do not know where it will lead us. I am no one to suggest anyone anything but I believe Karan,you would be more at peace with yourself if you remain true to yourself. I rest my case.
Well said, Shalu, thanks–and I can well understand the paucity of time to read/write with parenthood. Thank you for sharing.
Hi Karan, I love your writing and also this has been a productive blog. Thank you for sharing a detailed insight.


Through our foods, we are helping to unlock the potential of communities around the world. Our commitment to responsibly sourcing our 10 priority ingredients by 2020 is a key way we are doing so. This work starts on the farm, supporting the people and communities who grow our ingredients. That's why, as part of Breakfasts for Better Days™, we've pledged to improve the livelihoods of 500,000 farmers by advancing climate-smart agriculture practices that increase productivity and climate resiliency, while using fewer natural resources. Within this commitment, we also recognize the important role smallholder farmers and women play in the global agriculture community. As a result, we've already provided this support to nearly 20,000 smallholder farmers in our direct supply chain and have similarly helped more than 10,000 women farmers. To date, we've engaged nearly 300,000 farmers toward our 2025 goal.


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A Bold Peace, a film by Matthew Eddy and Michael Dreiling
A Bold Peace
a film by Matthew Eddy and Michael Dreiling (2017, 89 mins)
Bullfrog Films Distribution
In his famous "Cross of Iron" speech in 1953, President Eisenhower critiqued the military-industrial complex while asking, "Is there no other way the world may live?" In Costa Rica today, we glimpse another way to live.
In 1948, Costa Rica dismantled their military establishment and intentionally cultivated security relationships with other nations through treaties, international laws, and international organizations. Free of the burden of military spending, they used the financial savings to invest in their people, creating strong public institutions including public higher education and universal health care. In short, Costa Ricans created a society committed to peace, solidarity, and international law. They have survived with safety and relative prosperity for nearly 70 years without a standing army.
A BOLD PEACE details the events which shook the country to its foundations, culminating in the 1948 civil war and the decision to abolish the military. Over the decades, the Costa Rican model has survived several serious crises, but the current threats may be the most formidable of all.
Art for Social Change and Cultural Awakening. An Anthropology of Residence in Taiwan
CFP – Beauty and the Beast: photography, body and sexual discourse in the Middle East and Central Eurasia
The Return, a film by Kelly Duane de la Vega, Katie Galloway
How to Let Go of the World, a film by Josh Fox
Celling Your Soul, a film by Joni Siani


when nobody's waiting by your side ?
Lay-la, you got me on my knees.
Lay-la, I'm begging darlin' please.
Lay-la, darlin' won't you ease my worried mind.
and your old man won't let you down.
I turned the whole world upside down.
and tell me all my love's invain.


Murdered wife and sexually assaulted sister-in-law - motivated by sexual passion for sister-in-law.
Strangled 21y female after leaving nightclub late at night and buried in sandpit - assaulted and robbed two other girls who had left night club.
Strangled wife during argument at home.
Strangulation of 47y male friend - offenders and victim under influence of alcohol and marijuana following funeral earlier that day for offender's brother - angered by victim's continued reference to deceased brother - perceived lack of sensitivity to aboriginal culture - various acts of degradation performed on body after death and dumped in nearby bush.
Suffocated wife and two infant children and dumped bodies in bushland - highly charged emotional state at time of killing.
Brain injury at least affected judgment.
Female - mistreated, abused and assaulted 2y son over long period of time – murdered victim by asphyxiation although actual method of killing unclear –wrapped body in plastic, placed in suitcase and threw in nearby pond – spontaneous offence giving in to anger and influenced by several factors and pressures – had declined to give up victim to custody of mother or DOCS despite asking for help – aware victim about to be returned to lawful guardian - slightly below mid-range.
Female – strangled and stabbed female victim and stabbed in abdomen – victim had entered relationship with offender's close male friend, with whom offender had lived and shared some sexual involvement – feeling isolated - some degree of planning.
Depression – anxiety – had to terminate pregnancy - good rehabilitation prospects – studying and employed – good character.


Collaborative Divorce centers on you, the client.
It's about what you want and need, not about court action. So, first, you pledge to be collaborative. The key difference between Collaborative Law Practice and conventional divorce is the pledge to reach an agreement without going to court. You and your spouse keep control of the decisions yourselves, rather than giving that control to a judge. In order to accomplish that, all of the parties consent in writing to be a part of a respectful process that leads to an out-of-court resolution. With Collaborative Law Practice, the goal is to develop effective relationships, solve problems jointly, and prevent a court battle.
Next, we open lines of communication. Even under the best of circumstances, communication can be strained as a relationship is ending. Yet keeping the lines of communication open is essential for reaching an agreement. Collaborative Law Practice provides for face-to-face meetings with you, your spouse and your respective family law attorney and other advisers as needed. These sessions are intended to produce an honest exchange of information and expression of needs and expectations. When the issues are openly discussed, problem solving can be direct and solutions-oriented.
Then, we reach an agreement everyone can live with. If you've reached a decision about divorce, you may be reassured to know that Collaborative Family Law Practice focuses on attaining resolution. The Collaborative Model is designed to protect the interests of children and help you and your spouse move forward with new lives. Throughout, the Collaborative Process emphasizes cooperating to find solutions. And instead of being an endless airing of grievances, Collaborative Law Practice encourages spouses to reach a workable settlement by building on areas on mutual agreement.
You get support when you need it the most.
Divorce usually involves many considerations, from concern for children to financial questions to property matters. Along with handling the overall emotional impact, managing by yourself can be difficult. For that reason, Collaborative Law Practice offers you a supportive approach. You and I, as your family law specialist, work as a team. Other professionals including mental health professionals, child specialists, and financial consultants can become part of your team, helping find constructive solutions to deal with divorce's wide-ranging issues and establish goals for the future.
It focuses on the future.
Divorce ends a marriage but need not sever family ties or relationships. Especially when children are involved, lifelong responsibilities remain. By preserving respect and encouraging cooperation, Collaborative Law Practice helps parents and children keep family bonds while embracing new lives. Divorce will always remain a significant life event. In the face of it, Collaborative Law Practice can lead you and your family to a compassionate ending, and a healthy new beginning.