Insecticide resistance monitoring of malaria vector ...dmc.gov.pk/documents/pdfs/Insecticides...

Insecticide resistance monitoring of malaria vector mosquitoes at selected sites in highly endemic districts of Pakistan.

2017 FINAL REPORT SUSCEPTIBILITY ROJECT

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ACKNOWLEGDEMENTS

IMEDVC gratefully acknowledge Dr.Asad Hafeez, Director HSA for his kind collaboration

and partnership to conduct the training session for project staff. We also appreciate the

support by DR. Abdul Baseer Achakzai, Director Malaria Control Program, who gave me

this assignment, despite considerable pressure from not so competent groups. He also

helped this project by helping in getting the WHO insecticide impregnated papers and kits

released from customs department.

We are also grateful in every possible way to DR. Asai Ardakani, WHO Representative for

Pakistan, and the WRO staff including DR. Qutubuddin Kakar, Shahzad Sardar and Ijaz

Ahmed for technical support and hope to keep up our collaboration in the future.

Acknowledgement is due to DOMC focal persons Jaipal Singh, DR. InamUllah Kakar and

Dr. Saba Faruqi for their support in the project. Many thanks go in particular to our sincere

students and colleagues including Hammad Khaleeq, Asma Sana, Saqib Razzak, Fatima Haq

and Mohammad Ali for their contribution in conducting refresher training course and data

collection in the field specially the field coordinators at Provincial level. Administrative

support by our colleagues Mr. Khurram Qureshi and Nawaz Shah is acknowledged..

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1. BACKGROUND:

1.1. MALARIA SITUATION GLOBALLY AND IN PAKISTAN

Mosquitoes are vector for number of human disease like Malaria and various viral

diseases like dengue and yellow fever1. Malaria remains one of the major endemic

diseases in the tropics due to high frequency of transmission of plasmodium species by

large number of anopheles mosquitoes2,3. Malaria control in Pakistan was started as

Malaria Control Activity in 1950. In 1961 this program became the Malaria Eradication

Program with the objective to interrupt malaria transmission with residual

insecticides, but in 1969 this program suffered technical, administrative, and financial

problems 7. Insecticide resistance in vector mosquitoes and anti-malarial drug

resistance posed major technical hurdles. The failure of the Malaria Eradication

Program led to the initiation of a five year National Malaria Control Program (MCP),

where control of vector mosquitoes was the main control strategy 4.In 2012 it was

reported that in Pakistan out of 173 million inhabitant 9% live in high transmission

districts (34 districts), with an annual parasitic index (API) ranging from 5.4 to 27.28;

20% is living in moderately endemic districts (41 districts) with API ranging from 1.04

to 4.87, and 71% living in low endemic districts with and API below 1/1000 (61districts)

The national API for the entire country was 1.69, witch classify Pakistan as a moderate

endemic country. The mapping shows that the Province of Baluchistan has the highest

parasitic index with (API 7.68) and gradually decreases in FATA (API 6.83), Sindh (API

2.92), KPK (API 2.76), Punjab (API 0.19) and AJK is the lowest with API of 0.10. In 2012

the lowest malaria incidence was confined to two provinces – Punjab and AJK with

combined population of more than 56% of total population of the country (Table 1.)

Province Highly endemic

district API > 5

Moderately endemic districts API 1-5

low endemic district API < 1

Total

Baluchistan 19 10 1 30

FATA 7 6 0 13

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Table 1. Showing District with high, medium and low level of malaria endemicity.

1.2. GLOBAL MALARIA VECTOR SITUATION:

Anopheles mosquitoes are the main vector of malaria5 as the asexual multiplication and

obligate sexual reproduction of plasmodium takes place in anopheles mosquito6. Genus

Anopheles play unequal roles in malaria transmission as some Anopheles species are not

involved in transmission of malaria because weak interrelations with humans, therefore,

taxonomic research on malaria vectors is very important7. Worldwide there are more than

3000 mosquito species8. Globally about 460 species of anopheles are known9 out of which

70 Anopheles species are capable of transmitting malaria10.

1.3. MALARIA VECTOR MOSQUITO SITUATION IN REGION AND PAKISTAN

In EMR, out of the 70 Anopheline mosquito species reported, 18 species are confirmed

vectors of malaria11. Six species, Anopheles culicifacies, An. stephensi, An. superpictus,

An. arabiensis, An. sacharovi and An. sergentii, are the most important, these 6 species

put 85% populations at risk of malaria transmission. In Pakistan 24 Anopheline

mosquito species have been reported, however only two species i.e. Anopheles

culicifacies and Anopheles stephensi are known Malaria vector

Sind 4 16 3 23

KPK 4 6 14 24

Punjab 0 3 33 36

AJK 0 0 10 10

Grand total 34 41 61 136

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Table 2. Anopheline mosquito species of Pakistan

1.4. STATUS OF INSECTICIDE USE AND RESISTANCE IN PAKISTAN

Vector control is an important part of the global malaria control strategy. Previously, use of

pesticides has been the mainstay of mosquito control in Pakistan, but unfortunately little

attention was given to application of alternative control methods and development of trained

manpower in the specialized field of medical entomology and disease vector control.

Dichlorodiphenyltrichloroethane (DDT) was used for mosquito control in the province of

Punjab from 1961 to 1975 (two cycles/year of 1-2 gm/m2), which consequently produced

strong DDT resistance in Anopheline malaria vector mosquitoes. In 1975 DDT was partially

replaced by BHC, to which resistance developed rapidly 12. In 1976 Malathion (two

cycles/year, 1 gm/m 2) was introduced for malaria vector control. At the same time,

Fenitrothion was introduced in some areas 13. After 1996, operational failure of vector

control, partly due to insecticide resistance to organophosphate insecticides and to financial

and administrative constraints as well as changes in global vector control strategy towards

selective insecticide application, influenced the MCP to shift its emphasis to selective spraying

with Pyrethroids and increased use of insecticide-treated bed nets14.

In Pakistan base-line work on insecticide resistance monitoring was carried out in 1985 as the

first large-scale field survey to map insecticide resistance status in 11 randomly selected

districts in the Punjab province. An. culicifacies was susceptible to all insecticides except DDT,

but for An. stephensi resistance to Malathion was widespread 15. During the last 40 years, not

much attention has been paid to systematically monitor the insecticide resistance status of

Anopheline mosquitoes in Pakistan. This lack of information on the resistance status of vector

mosquitoes can have serious technical and financial consequences, especially when

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Pyrethroids are used extensively for agricultural and public health purposes. Development of

undetected vector resistance to currently effective Pyrethroids can lead to uncontrollable

epidemics by vector-borne diseases 16.

Rathor et al. 2013 studied pesticide susceptibility status of anopheles mosquitoes in four

flood-affected districts of south Punjab, Pakistan and concluded that both An. stephensi and

An. culicifacies remained resistant to DDT and Malathion. Tests with three commonly used

pyrethroids, Permethrin, Lambda-cyhalothrin, and Deltamethrin detected resistance in the

majority of cases, but in a number of localities mortalities with these three Pyrethroids ranged

from 80-97% and were therefore placed under verification-required status 17. This status

indicates the presence of susceptible individuals in these populations. These results suggest

that if appropriate resistance management strategies are applied in these areas, then the

development of high levels of resistance can still be prevented or slowed. This study forms an

important evidence base for the strategic planning of vector control in the four flood-affected

districts.

Knowledge of vector/pest susceptibility to pesticides and insight into changing trends of

resistance and expected operational implications provide the evidence base to form effective

national policy and pesticide use strategies for vector-borne disease and pest control

programs. Therefore, insecticide resistance monitoring must be an integral part of disease

vector and public health pest control programs.

2. Objectives

2.1 THE MAIN OBJECTIVES OF THE STUDY ARE:

2.1.1. Capacity building of vector control staff on implementation of the insecticide

susceptibility/resistance surveillance and monitoring at Provincial level in Pakistan.

2.1.2. To assess the resistance/susceptibility status of malaria vectors against

discriminating dosage of DDT 4.0%, Malathion 5.0%, Fenitrothion 1.0%, Propoxur 0.1%,

Bendiocarb 0.1%, Permethrin 0.75%, Deltamethrin 0.05%, Lambdacyhalothrin 0.05%,

Alpha-cypermethrin, and Etofenprox 0.5%.

Note: The second objective is in line with the instructions of the WHO-GMP

informal consultations convened in 2010, 2013, December 2015 and WHO

Guidelines 18,19 recommending an expanded three-step bioassay protocol.

[Please note that Mortalities < than 98% indicate susceptibility, at mortalities

from 90% to 97%, three types of investigations are recommended:

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Step 1. Perform more tests, on the same population on wider scale, for phenotypic resistance detection. Step 2. Determine strength of phenotypic resistance detected by applying 5X & 10X dosage of Diagnostic dose. Step 3. Determine resistance mechanisms, synergistic studies, ELISA & PCR studies Please note that, for change of insecticide in use at country level, which has serious management and financial implications. For making a decision for change the above stated 3 steps (“Eco-Ento-Epidemiologica” studies (the 3EEE studies) must be completed. In addition to the above mentioned 3 step protocol, decision on resistance management require village scale operational trial , conducted in local conditions, a hut trial in a village instead of the whole village can be cost- effective and more practical. This will provide “Operationally meaningful data”, suggested as the outcome of deliberations of: WHO consultation on technical basis for coordinated action against insecticide resistance: preserving the effectiveness of modern malaria vector control, WHO Geneva, May 2010.

The present study, in view of available equipment, materials and the available

trained manpower, aimed at achieving first step of detection of phenotypic

resistance. The next two steps (step 2 and step 3 and village scale trial) shall be

the ultimate target for insecticide resistance management in a country and

covered by future studies.

2.1.3. Based on the present study results, propose recommendations on policy strategy

for rational selection, use and management of presently employed and other available or

alternative pesticides for malaria vectors in the country.

2.1.4. Promotion of regular and careful monitoring of insecticide susceptibility in vectors as the

key entomological activity where chemical insecticides are in use, maintaining systematically

updated status of susceptibility / resistance in vector mosquitoes, to ensure safe, sustainable and

rational use of insecticides.

3. RATIONALE:

Malaria is the major vector borne disease and a major public health problem in Pakistan. Despite many

years effort of malaria eradication followed by malaria control and integrated control programmes,

the disease continues to threaten millions of people. A number of major factors are known to be

responsible for persistence of malaria. Most recently, the global strategic framework for malaria

control 2016-2030 proposed emphasised, maximizing the impact of vector control, bases of local

epidemiological and entomological data 20. The IRS and long lasting insecticide treated nets are

the main tools for vector control. These chemical control methods and tools can be effective, provided

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dedicated and properly skilled and trained technical staff is employed to apply these tools with

strategic planning. Unfortunately, these skilled categories of workers are not available. Moreover, due

to excessive, indiscriminate and inappropriate use of pesticides, vector mosquitoes are becoming

resistance to various groups of insecticides and pose is a real threat to malaria control programmes.

Under constant selection pressure by insecticides, resistance in vector mosquitoes is constantly

evolving. Under such situation lack of information on insecticide resistance status, can have serious

operational implications.

Recently research on insecticide resistance of malaria vectors by Rathor et al21 in four districts of

south Punjab, Pakistan was an attempt to restart the process of generating the much needed

information on resistance status in the country. The recent work was done in a small part in one

province, while the remaining country remains unexplored and unmonitored. The above study

revealed that both An. stephensi and An. culicifacies in the tested area remained resistant to DDT and

Malathion. Tests with three commonly used Pyrethroids: Permethrin, Lambda-cyhalothrin, and

Deltamethrin detected resistance in the majority of cases, but in a number of localities mortalities with

these three Pyrethroids ranged from 80-97% and were therefore placed under verification-required

status. This status indicates the presence of susceptible individuals in these populations. These results

suggest that if appropriate resistance management strategies are applied in these areas, then the

development of high levels of resistance can still be prevented or slowed. This resistance monitoring

needs to be done in the remaining provinces. The information on vector/pest susceptibility will

provide much needed insight into changing trends of resistance.

The prevailing situation of use of insecticides i.e. wide spread use of long-lasting insecticidal nets and

indoor residual spraying may lead to high level of insecticides resistance in the vectors, which could

lead to substantial increase in malaria mortality with devastating public health consequences.

Therefore, the WHO strategy22, emphasizes on development and implementation of plans for

monitoring and managing insecticide resistance

The information generated through the proposed project for detection of insecticide resistance, in two

high endemicity Provinces, with the WHO susceptibility test kit will provide much needed evidence

base and also confirm any operational failure due to resistance in the field and provides very vital

information for the selection of a new insecticide or replacement of an insecticide in use.

4. IMPLICATIONS OF STUDY RESULTS, TO DISEASE CONTROL

PROGRAMM:

Promotion of regular and careful monitoring of insecticide susceptibility in vectors as the key

entomological activity where chemical insecticides are in use for vector control, and promotion of

rational use of pesticides by national malaria control programme, by following a strategic plan, based

on systematically updated status of susceptibility / resistance in vector mosquitoes.

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Established insecticide resistance monitoring system will help in strategically appropriate insecticide

use, prevention and reduction in resistance development in malaria vectors and consequently prevent

escalation and epidemics of malaria in Pakistan.

5. MATERIALS AND METHODS

Study DESIGN, PROTOCOL, Plan AND PROCESS:

5.1. STUDY DESIGN:

Testing of vector mosquitos with diagnostic doses of insecticide using WHO test kits involves

testing of natural population of mosquito vectors in various districts enabling us to establish

resistant/susceptible status against insecticide used in malaria vector control program. This also

allows various field populations for these characteristics. This information is essential for

strategic planning for safe and judicious application of insecticides

5.2. STUDY SITES

This research was conducted in five different malarious areas of provinces of Baluchistan,

Khyber Pakhtunkhwa (KPK), Punjab, Sindh, Federally Administered Tribal areas (FATA) as

shown in Figure 1 which annual parasite index in Pakistan.

Figure 1

The criteria for selection of sites was based on level of endemicity and availability of vector

mosquitoes at the time of vector collection and testing. Moreover selection of localities for this

purpose was carried out in consultation with malaria control program personnel’s in each

district.

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The following districts are selected for study purpose in various provinces;

Figure 2

In the province of Baluchistan tests were made in three localities in the district of Jaffarabad as

shown in the figure 2. The province has API >5. Collection/test sites:1. Goth Ghulam Sarwar/

Dera Allahyaar, 2. Goth Khaliq ali Ghandaka , 3. Haji Fateh Mhd Goth Usta Mhd. La. 28.3697

-Lo 68.35144

Khyber Pakhtunkwa

Figure 3

In province of kpk at least three localities were tested in the district of Lakki Marwat and

Bannu as shown in the figure 3, the Province has API >5. Collection/test sites: 1. Mama Khel. lat.

33.7855164 - Lo 72.1342083. 2/10/2017, 2. Serai Naurang. La. 32.825024 - Lo. 70.783653. 5/10/2017,

Jafar Abad

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3. lakki city . la.32.6045 – Lo. 70.9131. 9/10/2017

PUNJAB

Figure 4

In the province of Punjab at least three localities were tested in the district of SHEIKHU

PURA. The Province has API <5. Collection/test sites: 1. Machikai . UC. NO.53., 2. Village:

Kulkarni UC. NO Kulkarni. 3. UC. NO. 41. Marbangwab /Baag Danaiser, 13-22/10/2017

LAKKI MARWAT

SHEIKHUPURA

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SIND

Figure 5

In the province of Sindh at least three localities will be tested in the district of

Mirpur Khas, as shown in the figure 5, with API >5. Collection/test sites: 1. UC.

Yar Muhammad Ginji Khipro rd. la. 25.540490-Lo. 69.033815. 2. UC. Mir

Ghulam Hussain Talpur . la. 25.374491-Lo. 69.201705. 3. UC. Hingorono. la.

25.735317- Lo. 69.176000. 17-19/10/2017.

FATA. Figure 6

In the FATA at least four localities will be tested in the district of Khyber Agency as shown in

the figure 6. The Province has API >5. Collection/test sites: 1. UC. NO Gallani Khola. la.34.0211-Lo .71.2874. 2. Ali

Mirpur Khas

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Masjid Kalay. la 34.0243 - Lo. 71.2651. 3. Qadam . La…- Lo.. 29/9/2017-,6/10/2017, 20/10/2017..

5.3. TIMING AND DURATION OF THE STUDY:

The study was to be conducted during peak malaria vector breeding/transmission season, from

June to September 2016, however, due to delays in assigning contract and delay in release of

WHO kits, which was beyond the control of the writer, starting studies at the tail end of the

vector density period. This resulted in making certain compromises in work schedule,

However, in addition to hard work by the provincial research associates, we had to launch

additional quality control teams of IMEDVC to get reasonably enough data to draw the

susceptibility status from all 5 provinces.

5.4. STUDY POPULATION:

Natural malaria vector mosquitoes Anopheles stephensi and Anopheles culicifacies were the

study population.

5.5. PROCEDURE FOR WHO SUSCEPTIBILITY TESTS:

STEPS ADOPTED FOR CARRYING OUT ASSIGNMENT.

KHYBERR

AGENCY

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As shown in the time line given in Annex 1. the activities will be carried out as following:

Study protocol out line prepared

A capacity building activity carried out as a short training for entomologists working in various capacities in provinces.

A technical consultation will be held to brief the stakeholders

Data collected analyzed and shared with stakeholders before preparation of report.

5.6. WHO SUSCEPTIBILITY TEST KIT:

The WHO test kits were used which are simple easily usable tool for standard and

internationally comparable method for the detection of resistance in malaria vector

mosquitoes. 23,24,25,26,27,

The kit consists of Plastic tubes, with each tube fitted at one end with 16-mesh screen; slides

units, each with a screw-cap on either side, provided with a 20 mm filling hole; Insecticide-

impregnated papers; and a number of related instruments. The Details information on test kits

is given in WHO Expert Committee Report.These kits can be purchased from WHO. The kits

were provided to the project by the Director of the Malaria control programme.

5.7. SUITABLE MOSQUITO STAGES FOR TEST

Susceptibility tests using the WHO standard test kits were carried out, in field conditions, on at

least 100 adult blood-fed 2-5 days old female mosquitoes collected from field. In case, enough

numbers are not available, collection of unfed, blood fed, semi-gravid and gravid females were

tested. But their physiological status was recorded carefully.

5.8. METHODOLOGY OF INSECTICIDE RESISTANCE/SUSCEPTIBILITY TESTING IN THE FIELD:

Standard WHO adult test kits were used . Due to shortage of time only wild-caught fed

females were tested under field conditions. The mosquitoes were exposed to the discriminating

doses recommended by WHO recommended insecticide test papers were used, with a 1-h

exposure period, 10% sugar pads were provided during the 24-h holding period, after which

mortalities were calculated. Appropriate controls were run in all cases. If mortality ranged 5–20

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% in controls, corrected percentage mortalities were calculated using Abbott’s formula as

follows:

% Test mortality - % Control mortality x 100

100- % Control mortality

5.9. INTERPRETATION OF SUSCEPTIBILITY RESULTS:

The percentage of mortalities were calculated and used to establish the susceptible and resistant

status of these populations. Interpretations of the susceptibility tests was based upon the WHO

newly revised criteria which as follows:

> 98 % Mortality = Susceptible

90- 97 % Mortality =Verification required

<90 % Mortality= Resistance confirmed

Note: (.Mortality 98-100…………...Susceptible*

Mortality 90 - 97 ………..Further Verification required**

Mortality < 90 …………………. Resistance confirmed** *

(* detects phenotypic resistance, ** further studies suggested determines resistance

gene frequency and its fixation in genetic background (so called strength of

resistance), *** further studies suggested determines mechanism of resistance)

5.10. FIELD COLLECTION OF FEMALE MOSQUITOES AND CROSS CHECKING FOR SPECIES

IDENTIFICATION:

The malaria vectors (An. stephensi and An. culicifacies) were field collected and held in IC

carton cages and tested in the field or a suitable temporary established base laboratory,

having conditions similar to the field. The samples of Field collected and tested mosquitoes

were received at the Institute of Medical Entomology and Disease Vector Control (IMEDVC),

Islamabad for cross-checking the species identification.

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5.11. EQUIPMENT & SUPPLIES FOR FIELD SURVEY TEAMS:

After completion of training for susceptibility resistance tests, required materials & supplies,

for survey purposes were handed over to provincial teams for implementation of the survey.

5.12. SURVEY IMPLEMENTATION ARRANGEMENTS:

Each provincial field co-investigator was responsible for its respective provincial team which

completed its field survey within five days at three selected sites from one district of its

province. After completion of field surveys, data and samples were dispatched to the central

office, Islamabad for further species identification cross check and data analysis. .Field

specimens for cross-checking were received in special labelled vials, Fig 1. Where necessary

IMEDVC trained quality control teams were sent to provinces to improve the quality of data

generated.

Following are the lists of provincial field co-investigators:

5.13. LIST OF PROVINCIAL FIELD CO-INVESTIGATORS

5.14. DATA COLLECTION AND MANAGEMENT:

Collections were performed from three different locations of priority risk district from each province during the month of September and October. Collections were made from animal shed and agriculture fields at early morning using hand held aspirator or sweepers. After collection, specimens were brought the field laboratory set by the trained staff and malaria vector species were sorted morphologically using taxonomic keys. Females were counted and stored for further analysis.

SR.NO. NAME PROVINCE CONTACT EMAIL

1. Mr. Asif

Mehmood/Afeef

Punjab 0306-

3333136

[email protected]

2. Salahudin khan KP 0301-

8008805

[email protected]

3. Mohan Sadhu Sindh 0300-

3310600

4. Nasir Bulaidi

Samina Baluch

Balochistan 0312-

8000776

[email protected]

5. Ajmal khan FATA 0301-

8818450

[email protected]

mailto:[email protected]


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WHO slandered test kits were used to perform the experiment. From each district, at least 100 female mosquitoes were tested. Tests were performed in 3 to 4 replicates against a control for each insecticide at each locality and 24 hours readings were recorded.

The principal investigator made ensure that the study protocol was strictly adhered to and that all data are collected and recorded correctly on appropriate forms. Data analysis was carried out at IMEDVC.

5.15. STATISTICAL METHODS:DATA ANALYSIS:

Microsoft excel software program was used for data entry management. Data analysis, especially the Chi square analysis was carried out Using SPSS-16 statistical software. The final analysis showed the resistance/susceptibility status of malaria vector mosquitoes against four groups of insecticides.

6. DISSEMINATION OF RESULTS AND FURTHER ACTION BY THE MOH.

At the end of the study, the principal investigator will submit a draft report on the study and its

main outcome. This report and recommendations will be shared with the national malaria

control programme and the Ministry of Health. In the light of discussion with stake holders the

recommendations will be finalized to enable the Ministry of Health to prepare/revise the

national strategy on insecticide resistance management. The study will be presented during a

scientific meeting and will be published in scientific journals.

7. PRODUCTS / OUTPUTS AND FUTURE OUTLOOK: 1. Trained man power.

2. Establishment of status of susceptibility/resistance status in malaria vector Anopheles species against representative insecticides from four different groups.

3. Recommendation for future sustainability of insecticide susceptibility in the country in the light of survey results.

4. Recommendation for re-assessment of national strategy on efficient and sustainable resistance management and control program.

5. On the base of data generated by this national resistance/susceptibility survey formation of strategic plan for resistance management in Anopheles mosquito.

8. CAPACITY BUILDING COMPONDENT:

8.1. Short Training of Provincial Teams for susceptibility resistance testing:

Training of provincial teams, including Provincial and district entomological staff and refresher training of field Associates will be carried out for three days at central office of Institute of Medical Entomology & Disease Vector Control (IMEDVC), Islamabad. The field exercise will be

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conducted at Gujjar Khan to enhance practical skills of field staffs. Two members team were trained from each province for project implementation. Each Provincial team was guided to prepare and present a structured presentation on their provincial status on entomological work, specially related to insecticide use, evaluation and management strategy. Five participants made valuable presentations. All presented information on their malaria vector mosquito bionomics and the status of their vector control strategy. All five indicated need for revival of Post-graduate Diploma in Medical Entomology and Disease Vector Control, the program that had produce, for the first time in the country, qualified Medical Entomologists, who have been the main technical resource in fight against dengue. The presentations also revealed that only one Province had carried out their vector control needs Assessment, the remaining four has yet to carry out their Provincial Vector control needs assessment and strategy.

8.2. Following areas were included in the training program

Mosquito collection techniques from field

Vector control strategies

vector bionomics and taxonomy from 5 Provinces

Classification of pesticides used in public

Susceptibility test procedure GIS Mapping Statistics and Criteria for establishing susceptibility status.

A copy of report on the short training is attached to this report.

9. RESULTS AND DISCUSSION Anopheles stephensi and Anopheles culicifacies the two malaria vectors in Pakistan when exposed to various diagnostic doses of three pyrethroids (Deltamethrin, Lambdacyhalothrin and Permethrin), one organophosphate (Malathion), one chlorinated hydrocarbon (DDT) and one Carbamate (Bendiocarb) showed a range of mortalities in various three localities of five Provinces each. The data generated from tree sites each from Baluchistan, FATA, Sind, Khyber Pakhtukhwa and Punjab was recorded in the field in a systematic manner in specially designed tables. The tables included information on the vector bionomics, total tested, mortality percentage at 24 hours and corrected mortality with Abbott formula if necessary and record on controls. The status of susceptibility was assessed according to the latest WHO guidelines, (see section 5.9 in the main text of the report).The Annex I includes120 tables showing detailed information on results of the tests made. Each table shows results of tests on one species against one insecticide, carried out at three sites in one district. In this way test results were obtained on at least four insecticides: Deltamethrin, Lambdacyhalothrin, Malathion and DDT representing four groups of insecticides (Chlorinated hydrocarbons, Organophosphates, Carbamates and Pyrethroids). In some cases tests were also made on Bendiocarb and Permethrin.

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These 120 basic data tables were color coded for susceptibility status of each species against each insecticide: Green for susceptible (SS), yellow for Verification Required status (VR) and red indicated resistance (RR). For the purpose of ease of analysis through a wider picture in one place the 120 tables were summarized in 10 tables given in Annex II: A1 and A1.1 for Baluchistan, A2 and A2.1 for SIND, A3 and A3.1 for Punjab, A4 and 4.1 for Khyber Pakhtoonkhwa and A5 and A5.1 for FATA. In order to bring the wider picture in one view The 10 tables were further summarized in final two tables given in Annex III: B1 giving comparative description of susceptibility status in 5 Provinces against four insecticides for Anopheles stephensi and B2 for Anopheles culicifacies. It was observed that in: Baluchistan An. Stephensi against Deltamethrin in one locality had RR status but 2 localities showed increased mortalities but remained within VR status. For Lambdacyhalothrin all three localities showed RR status. For Malathion one locality showed resistance but in 2 localities showed increased mortalities but remained within VR status. For DDT complete resistance was observed. An. culicifacies against Deltamethrin one locality showed resistance and 2 localities showed VR status. For Lambdacyhalothrin, Malathion and DDT all localities had RR status. FATA Both An. stephensi and An. culicifacies had the same status: all susceptible against Deltamethrin and Permethrin but resistant to both Malathion and DDT. Sind An.stephensi against Deltamethrin was in the range of VR in all localities, RR in all localities to Lambdacyhalothrin and DDT, and VR status to Bendiocarb and Malathion. An. culicifacies had VR status in all localities for Deltamethrin, Bendiocarb and Malathion. It was resistant to both Lambdacyhalothrin and DDT. Punjab An. stephensi against Deltamethrin had SS status in one locality and in 2 localities slight lowered mortalities put it in VR status. However, all localities showed RR status to Permethrin, Malathion and DDT. An. culicifacies against Deltamethrin had RR status in 2 localities, but slightly increased mortalities in one locality placed it in VR status. For Lambdacyhalothrin it was resistant in one locality but in 2 localities slightly increased mortalities observed placed it in VR status. with Permethrin the VR status was observed in all localities. For Malathion and DDT the status was RR in all localities. KPK An. stephensi against Deltamethrin was susceptible in all localities. With Malathion RR status was observed in 2 localities but slight increased mortalities in one locality placed it in VR status. For Bendiocarb and DDT overall resistant status was observed in all localities. An. culicifacies was Susceptible to Deltamethrin in all localities and and resistant to Bendiocarb, Malathion and DDT. The chi-square test, using the SPSS programme were perform to find out, weather there is any significance or Non- significance differences present in the corrected moralities of the female mosquitoes that not survive the diagnostic dose after 24 hour of holding period in different

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localities of the districts. The chi- square test indicates that there was non-significant difference in mortalities of mosquitoes in different localities of district.

10. DISCUSSION AND CONSLUSIONS Malaria in Pakistan still persists with various degrees of intensities in different Provinces. The Plasmodium falciparum dominates the northern part of the country with high mortality and morbidity. The country has in late nineteen seventies, faced malaria control failure due to undetected resistance to DDT and Organophosphates in use, in between some carbamates were used. Presently the malaria control is heavily depending on Pyrethroids for vector control, it may be the use of long lasting nets or the IRS. It is a matter of great concern that unregulated mixtures of various groups insecticides are in use for public health or agriculture in the country. Under these circumstances judicious use of pesticides, supported by appropriate insecticide resistance monitoring in this country assumes top priority, especially for the pyrethroids which are also use in agriculture in large quantities.

In this study tests were intentionally made on Malathion and DDT, to see if discontinuation of use of the two insecticides has resulted in reversal of resistance that had reached high level in nineties. The results indicate that in all Provinces both An. stephensi and An. Culicifacies are still resistant to DDT and Malathion in the most Provinces. The results show a general susceptibility to Deltamethrin and Permethrin, but resistance Lambdacyhalothrin is apparent in some Provinces. However in a number of districts verification required status against pyrethroids indicates presence of susceptible population and that if appropriate resistance management strategies are applied, development of high level of resistance can be prevented or at least slowed down. The results of these studies form important evidence base for strategic planning of vector control. According to the WHO the susceptibility tests have limitations22, and on the basis of the susceptibility status determined by the WHO kits and insecticide impregnated papers, change of insecticide being used in the country cannot be decided. However, in the light of data obtained from Provinces where higher % age of mortalities reaching near to susceptibility level were observed with the Pyrethroid Deltamethrin, it can be recommended that at present for all practical purposes the use of Deltamethrin (already in use) can be continued in all four Provinces and FATA 11. RECOMMENDED RESISTANCE MANAGEMENT STRATEGIC PLAN ACTIVITIES 11.1. Future resistance monitoring Activities

20

Establish a country wide system for regular monitoring of susceptibility level in malaria vector

mosquitoes, using WHO kits with discriminating/diagnostic doses and consider this activity as the

key entomological activity of malaria control programme.

Malaria control Programme needs to implement expanded three-step bioassay protocol18,19

consisting of:

Step 1. Perform more tests, on the same population on wider scale, for phenotypic resistance detection. Step 2. Determine strength of phenotypic resistance detected by applying 5X & 10X dosage of Diagnostic dose. Step 3. Determine resistance mechanisms, synergistic studies, ELISA & PCR studies. In addition to the above mentioned 3 step protocol “Eco-Ento-Epidemiological” studies (the

3EEE studies) for a decision on resistance management require village scale operational trial ,

conducted in local conditions. If possible, a hut trial in a village instead of the whole village

can be cost-effective and more practical. This will provide more operationally meaningful

data28

Research based actions for resistance management

Based on susceptibility monitoring a review of patterns of resistance, cross-resistance and possible

resistance reversal in vectors shall be carried out for resistance management technical actions.

In case of control failure, based on assessment of vectors resistance / susceptibility , search for

potential alternative insecticides on scientific basis.

Identify patterns of resistance mechanisms of resistance and in malaria vectors, based on that

evidence, propose policy & strategy for vector / pest Resistance management (Guidelines of WHO

2016, Synergist studies).

Investigate if wide spread use of long-lasting insecticidal nets and indoor residual spraying may lead

to high level of insecticides resistance in the vectors, which could lead to substantial increase in

malaria mortality with devastating public health consequences.

11.2. Capacity building for resistance monitoring

In view increasing problems of vector-borne diseases in the country, It is essential that

technically qualified medical Entomologists are trained through appropriate Post-graduate

diploma or degree programs.

11.3. RESISTANCE MANAGEMENT STRATEGIES ON APPLIED FIELD TECHNIQUES

(Rotations, mosaic, combination and mixtures applications)

Carry out applied field research and apply the outcome results in the appropriate resistance

management. Strategies: such as the use of: Rotation of insecticides where different types of

21

insecticides are used in rotation in order to slow down resistance development, combination of

insecticide where two or more insecticide exposure with different mode of action are used to reduce

survival possibilities. In case of use of mosaic pattern of application of different insecticides in adjacent

areas are applied in a pattern, expecting that insects surviving insecticide in one area may migrate to

the other area and may not survive the second type of insecticide. Further operational research is

required to establish the applicability and effectiveness of mosaics and combination strategies for

malaria control. In case of use of mixture of insecticide two or more insecticides with independent

resistance mechanisms, are applied together. This principle is similar to that of artemisinin-based

combination therapy in malaria treatment.

11.4. Establish inter-sectorial steering committee

Establish strong inter-sectorial steering committee for insecticide resistance management

decision making and provide the authorities with recommendations on operational policy

recommendations in the context of integrated vector management (IVM). The committee shall

draw its members from the following stakeholders:

Ministry of health (Chair), National Vector and malaria control Programme (Secretariat), Ministry of

Agriculture: irrigation and pesticide council, Ministry of Environment, Ministry of Industry, Council for

Environment and Natural Resources, Research and academic institutions, Other relevant local experts

(e.g. experts on insecticide resistance) and WHO country office staff invited to discuss specific topics.

The Committee shall have regular quarterly meetings to discuss and decide on vector control

interventions.

12. REFERENCES CITED

1. Roosendaal J.A. Vector Control- Methods for use by individual and communities World Health Organization, Geneva.1997 2. Appawu M.A., Baffoe-Wilmot A., Afari E.A., Dunyo S., Koram K.A., Nkrumah F.K. Malaria vector studies in two ecological zones in Southern Ghana. African Entomology.2001; 9, 1:59-65. 3. Awolola T.S., Okwa O, Hunt R.H., Ogunrinade A.F., Coetzee M., Dynamics of the malaria vector population in costal Lagos, southwestern Nigeria. Annals of Tropical Medicine and Parasitology. 2002; 96,1:75-82. 4. World Health Organization. Global malaria control and elimination: report of a technical review. 2008. http://apps.who.int/iris/bitstream/10665/43903/1/9789241596756_eng.pdf 5. World Health Organization. Global malaria program. Indoor residual spraying: use of indoor residual spraying for scaling up global malaria control and elimination. 2006:1. http://apps.who.int/iris/bitstream/10665/69386/1/WHO_HTM_MAL_2006.1112_eng.pdf

6. World health Organization. Roll Back Malaria in Eastern Mediterranean Region. Achievements, challenges and the way forward. 2004:5. (WHO-EM/MAL/305/E).

http://apps.who.int/iris/bitstream/10665/43903/1/9789241596756_eng.pdf

http://apps.who.int/iris/bitstream/10665/69386/1/WHO_HTM_MAL_2006.1112_eng.pdf

22

7. World health Organization. Roll Back Malaria in Eastern Mediterranean Region. Achievements, challenges and the way forward. 2004:5. (WHO-EM/MAL/305/E).

8. Sylvie C, Frank R, Souleymane D, et al. an insight into immunogenic salivary protein of Anopheles gambiae in African children. Malaria J. 2007; 6:75. 9. Rathor HR. The role of vectors in emerging and re-emerging diseases in Eastern Mediterranean Region.Eastern Mediterranean Health J. 1996; 2:61-67. 10. Aslam K. the mosquitoes of Pakistan. A checklist.MosqSyst Newsletter.1971; 3:147-159. 11. Rathor HR, Toqir G. Malathion resistance in Anopheles stephensi Liston in Lahore, Pakistan. Mosquitoes News 1980; 40:526-531 12. Rathor HR, Sheikh ML, Toqir G, et al. preliminary survey of insecticide susceptibility in Anopheline population of selected index localities of Faisalabad District of Punjab Province. Pakistan J Med Res. 1983; 22:25-32.

13. Rathor HR, Toqir G. Malathion resistance in Anopheles stephensi Listton in Lahore, Pakistan. Mosquitoes News 1980; 40:526-531.

14. Rathor HR, Toqir G, Risen WK. status of insecticide resistance in Anopheline mosquitoes of Punjab, Pakistan. Southeast Asian J Trop Med Pub Health. 1980; 11:332-340.

15. Rathor HR, Toqir G, Rashid S, et al. Insecticide resistance in Anopheline mosquitoes of Punjab province, Pakistan. Pak J Zool. 1985; 17:35-49.

16. Tiwari S, Susanta KG, Vijay PO, et al. reduced susceptibility to selected Pyrethroids in urban malaria vector Anopheles stephensi: a case study in Bangalore city India. Malaria J. 2010; 9:179.

17. Rathor HR, Nadeem G, Khan IA. Pesticide susceptibility status of Anopheles mosquitoes in four flood-affected districts of South Punjab, Pakistan. Vector-Borne and Zoonotic Diseases. 2013 Jan 1; 13(1):60-6.

18. World Health Organization . Test procedure for Insecticide resistance monitoring in malaria vector mosquitoes. 2013. [email protected] http://www.who.int/malaria

19. World Health Organization ( second edition). Test procedure for Insecticide resistance monitoring in malaria vector mosquitoes. 2016.

20. World Health Organization. Global Technical Strategy for Malaria 2016-2030. Geneva. 2012 (http://apps.who.int/iris/bitstream/10665/176712/1/9789241564991_eng.pdf.

21 Rathor HR, Nadeem G, Khan IA. Pesticide susceptibility status of Anopheles mosquitoes in four flood-affected districts of South Punjab, Pakistan. Vector-Borne and Zoonotic Diseases. 2013 Jan 1; 13(1):60-6.


http://www.who.int/malaria


23

22. World Health Organization. Global plan for insecticide resistance management in malaria vectors. Geneva. 2012 (http://apps.who.int/iris/bitstream/10665/44846/1/9789241564472_eng.pdf

23 World Health Organization. Global plan for insecticide resistance management in malaria vectors. Geneva. 2012 (http://apps.who.int/iris/bitstream/10665/44846/1/9789241564472_eng.pdf

24 World Health Organization, 2010. The technical basis for coordinated action against insecticide resistance: Preserving the effectiveness of modern malaria vector control. Meeting report Global Malaria Programme, WHO HEADQUARTERS, GENEVA, 4–6 May 2010.

25. World Health Organization. Report of the WHO informal consultations test procedures for insecticide resistance monitoring in Malaria Vectors: Bioefficacy and persistence of insecticides on the treated surface. 1998 (WHO/CDS/CPC/MAL/98.12).

26. World Health Organization. Supplies for Monitoring Insecticide Resistance in Disease Vectors Procedures and Conditions. 2005 (WHO/MAL/95.1073 and WHO/CTD/VBC/95.998).

27 World Health Organization . Test procedure for Insecticide resistance monitoring in malaria vector mosquitoes. 2013. [email protected] http://www.who.int/malaria

28. WHO consultation on technical basis for coordinated action against insecticide resistance: preserving the effectiveness of modern malaria vector control, WHO Geneva, May 2010.




http://www.who.int/malaria

24

Table.1 site 1. Baluchistan Species. An. stephensi Tested with. Deltamethrin. 0.05% District. Jaffarabad Baluchistan UC. NO. Ghulam Sarwar Goth. Dera Allahyaar GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 13/10/2017

Replicate

Total Tested Females

Dead after 24 hours

Corrected Mortality

Status

Total tested in control

Dead after 24 hours

% Mortality

1 22 19

2 25 22

3 26 24

4 27 25

Total 100 90 % 90% V.R 50 2 4%

Table.2 site 1. Baluchistan Species. An. stephensi Tested with. Lambdacyhalothrin. 0.05% District. Jaffarabad Baluchistan UC. NO. Ghulam Sarwar Goth. Dera Allahyaar GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 13/10/2017 .

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 9

2 26 11

3 24 7

4 25 7

Total 100 34 34% RR 35 0 0.0%

Table.3 site 1. Baluchistan Species. An. stephensi Tested with. DDT .4% District. Jaffarabad Baluchistan UC. NO. Ghulam Sarwar Goth. Dera Allahyaar GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 13/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 6

2 27 8

3 26 4

4 25 7

Total 103 25 24.27%

RR 40 2 5%

Table.4 site 1. Baluchistan Species. An. stephensi Tested with. Malathion. 5% District. Jaffarabad Baluchistan UC. NO. Ghulam Sarwar Goth. Dera Allahyaar GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 13/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 22

2 25 23

3 24 21

4 26 23

Total

100 89 89 % RR 48 2 4.16%

Basic Data Tables (24 tables for each Province), a total of 120 tables. ANNEX I

25

Table.5 site 1. Baluchistan Species. An.culicifacies Tested with. Deltamethrin. 0.05% . District. Jaffarabad Baluchistan UC. NO. Ghulam Sarwar Goth. Dera Allahyaar GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 13/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 26 25

2 26 24

3 24 22

4 24 23

Total 100 94 94 % V.R 25 1 4%

Table.6 site 1. Baluchistan Species. An.culicifacies Tested with. Lambdacyhalothrin. 0.05% District. Jaffarabad Baluchistan UC. NO. Ghulam Sarwar Goth. Dera Allahyaar GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 13/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 12

2 25 11

3 26 13

4 24 10

Total 100 46 46% RR 15 0 0.0%

Table.7 site 1. Baluchistan Species. An.culicifacies Tested with. DDT 4% . District. Jaffarabad Baluchistan UC. NO. Ghulam Sarwar Goth. Dera Allahyaar GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 13/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 27 10

2 25 9

3 26 11

4 24 8

Total 102 38 37.25% RR 22 1 4.54%

Table.8 site 1. Baluchistan Species. An.culicifacies Tested with. Malathion 5% . District. Jaffarabad Baluchistan UC. NO. Ghulam Sarwar Goth. Dera Allahyaar GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 13/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 21

2 25 20

3 25 23

4 25 20

Total 100 84 84% RR 23 1 4.34%

26

Table.5 site 2. Baluchistan Species. An.culicifacies Tested with. Deltamethrin. 0.05% . District. Jaffarabad Baluchistan UC. NO. Goth Khaliq ali Ghandaka GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 26 23

2 26 22

3 24 21

4 24 22

Total 100 88 88 % RR 25 1 4%

Table.6 site 2. Baluchistan Species. An.culicifacies Tested with. Lambdacyhalothrin. 0.05% District. Jaffarabad Baluchistan UC. NO. Goth Khaliq ali Ghandaka GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 14

2 26 13

3 24 11

4 25 10

Total 100 48 48% RR 15 0 0.0%

Table.7 site 2. Baluchistan Species. An.culicifacies Tested with. DDT 4% . District. Jaffarabad Baluchistan UC. NO. Goth Khaliq ali Ghandaka GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 13

2 25 12

3 25 10

4 25 9

Total 100 44 44% RR 22 1 4.54%

Table.8 site 2. Baluchistan Species. An.culicifacies Tested with. Malathion 5% . District. Jaffarabad Baluchistan UC. NO. Goth Khaliq ali Ghandaka GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 27 21

2 25 20

3 26 19

4 24 22

Total 102 82 80.39 % RR 20 1 5%

27

Table.1 site 3. Baluchistan Species. An. stephensi Tested with. Deltamethrin. 0.05% District. Jaffarabad Baluchistan UC. NO. Haji Fateh Mhd Goth Usta Mhd GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 24/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 24 22

2 24 24

3 26 24

4 27 25

Total 101 95 94.05%

V.R 50 1 2%

Table.2 site 3. Baluchistan Species. An. stephensi Tested with. Lambdacyhalothrin. 0.05% District. Jaffarabad Baluchistan UC. NO. Haji Fateh Mhd Goth Usta Mhd GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 24/10/2017.

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 20

2 25 15

3 25 8

4 25 9

Total

100 52 52% RR 53 2 3.77%

Table.3 site 3. Baluchistan Species. An. stephensi Tested with. DDT .4% District. Jaffarabad Baluchistan UC. NO. Haji Fateh Mhd Goth Usta Mhd GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 24/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 8

2 24 9

3 26 7

4 25 7

Total 100 31 31% RR 50 2

4%

Table.4 site 3. Baluchistan Species. An. stephensi Tested with. Malathion. 5% District. Jaffarabad Baluchistan UC. NO. Haji Fateh Mhd Goth Usta Mhd GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 24/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 27 24

2 29 25

3 22 20

4 22 19

Total

100 88 88% RR 54 2 3.70%

28

Table.5 site 3. Baluchistan Species. An.culicifacies Tested with. Deltamethrin. 0.05% . District. Jaffarabad Baluchistan UC. NO. Haji Fateh Mhd Goth Usta Mhd GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 24/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 24

2 25 22

3 25 23

4 25 23

Total 100 92 92% V.R 24 1 4.16%

Table.6 site 3. Baluchistan Species. An.culicifacies Tested with. Lambdacyhalothrin. 0.05% District. Jaffarabad Baluchistan UC. NO. Haji Fateh Mhd Goth Usta Mhd GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 24/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 24 10

2 27 13

3 23 11

4 26 11

Total 100 45 45% RR 20 0 0.0%

Table.7 site 3. Baluchistan Species. An.culicifacies Tested with. DDT 4% . District. Jaffarabad Baluchistan UC. NO. Haji Fateh Mhd Goth Usta Mhd GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 24/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 12

2 27 13

3 23 11

4 26 9

Total 101 45 44.55 % RR 24 1 4.16%

Table.8 site 3. Baluchistan Species. An.culicifacies Tested with. Malathion 5% . District. Jaffarabad Baluchistan UC. NO. Haji Fateh Mhd Goth Usta Mhd GR Coordinates: latitude. 28.3697 Longitude. : 68.35144 24/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 24 19

2 25 20

3 27 22

4 24 18

Total 100 79 79 % RR 20 0 0.0%

29

Table.1 site 1. Fata Species. An. stephensi Tested with. Deltamethrin. 0.05% District. District: Khyber. Ag. UC. NO Gallani Khola GR Coordinates: latitude.34.0211 Longitude. : 71.2874 29/9/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 20

2 22 22

3 22 21

4 20 20

5 20 20

Total 104 103 99.0 % SS 41 1 2.4%

Table.2 site 1. Fata Species. An. stephensi Tested with. Permethrin District. District: Khyber. Ag. UC. NO Gallani Khola GR Coordinates: latitude.34.0211 Longitude. : 71.2874 29/9/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 26 26

2 27 27

3 25 25

4 28 28

Total

106 106 100 % SS 25 2 8%

Table.3 site 1. Fata Species. An. stephensi Tested with. DDT .4% District. District: Khyber. Ag. UC. NO Gallani Khola GR Coordinates: latitude.34.0211 Longitude. : 71.2874 29/9/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 27 10

2 26 7

3 26 7

4 27 9

Total 106 33

34.90 % RR 20

0 0%

Table.4 site 1. Fata Species. An. stephensi Tested with. Malathion. 5% District. District: Khyber. Ag. UC. NO Gallani Khola GR Coordinates: latitude.34.0211 Longitude. : 71.2874 29/9/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 24 11

2 24 10

3 23 7

4 25 12

5 25 10

Total

111 50 40.47 % RR 26 2 7.6

30

Table.5 site 1. Fata Species. An.culicifacies Tested with. Deltamethrin. 0.05% . District. District: Khyber. Ag. UC. NO Gallani Khola GR Coordinates: latitude.34.0211 Longitude. : 71.2874 29/9/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 25

2 26 26

3 27 27

4 25 25

5

Total 103 102 99.00 % SS 22 1 4.5%

Table.6 site 1. Fata Species. An.culicifacies Tested with. Permethrin District. District: Khyber. Ag. UC. NO Gallani Khola GR Coordinates: latitude.34.0211 Longitude. : 71.2874 29/9/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 24

2 25 25

3 25 25

4 25 24

5

Total 100 98 98.0 % SS 25 0 0%

Table.7 site 1. Fata Species. An.culicifacies Tested with. DDT 4% . District. District: Khyber. Ag. UC. NO Gallani Khola GR Coordinates: latitude.34.0211 Longitude. : 71.2874 29/9/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 24 13

2 25 12

3 26 13

4 22 10

5 20 10

Total 117 58 49. 5 % RR 25 0 0%

Table.8 site 1. Fata Species. An.culicifacies Tested with. Malathion 5% . District. District: Khyber. Ag. UC. NO Gallani Khola GR Coordinates: latitude.34.0211 Longitude. : 71.2874 29/9/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 12

2 26 13

3 26 13

4 26 13

5

Total 103 51 49.5 % RR 23 0 0%

31

Table.1 site 2. Fata Species. An. stephensi Tested with. Deltamethrin. 0.05% District: Khyber. Ag. Name. Ali Masjid Kalay GR Coordinates: latitude. 34.0243 Longitude. 71.2651 6/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 25

2 26 26

3 25 25

4 27 27

Total 102 102 100 % SS 25 0 0%

Table.2 site 2. Fata Species. An. stephensi Tested with. Permethrin District: Khyber. Ag. Name. Ali Masjid Kalay GR Coordinates: latitude. 34.0243 Longitude. 71.2651 6/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 27 27

2 27 27

3 26 26

4 25 25

Total

105 105 100 % SS 24 0 0%

Table.3 site 2. Fata Species. An. stephensi Tested with. DDT .4% District: Khyber. Ag. Name. Ali Masjid Kalay GR Coordinates: latitude. 34.0243 Longitude. 71.2651 6/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 26 13

2 25 12

3 24 12

4 25 13

Total 100 50 50.0 % RR 20 0 0%

Table.4 site 2. Fata Species. An. stephensi Tested with. Malathion. 5% District: Khyber. Ag. Name. Ali Masjid Kalay GR Coordinates: latitude. 34.0243 Longitude. 71.2651 6/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 12

2 26 13

3 28 14

4 27 10

Total 105 47 44.7 % RR 20 0 0%

32

Table.5 site 2. Fata Species. An.culicifacies Tested with. Deltamethrin. 0.05% . District: Khyber. Ag. Name. Ali Masjid Kalay GR Coordinates: latitude. 34.0243 Longitude. 71.2651 6/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 25

2 25 24

3 26 26

4 27 27

Total 103 102 99.0 % SS 25 0 0%

Table.6 site 2. Fata Species. An.culicifacies Tested with. Permethrin District: Khyber. Ag. Name. Ali Masjid Kalay GR Coordinates: latitude. 34.0243 Longitude. 71.2651 6/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 26 26

2 27 26

3 26 26

4 25 25

Total 104 103 99.0 % SS 25 0 0%

Table.7 site 2. Fata Species. An.culicifacies Tested with. DDT 4% . District: Khyber. Ag. Name. Ali Masjid Kalay GR Coordinates: latitude. 34.0243 Longitude. 71.2651 6/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 26 14

2 25 13

3 25 12

4 26 14

Total 102 53 51.9 % RR 15 0 0%

Table.8 site 2. Fata Species. An.culicifacies Tested with. Malathion 5% . District: Khyber. Ag. Name. Ali Masjid Kalay GR Coordinates: latitude. 34.0243 Longitude. 71.2651 6/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 12

2 26 13

3 27 14

4 27 9

Total 105 48 45.7 % RR 20 0 0%

33

Table.1 site 3. Fata Species. An. stephensi Tested with. Deltamethrin. 0.05% District. Khyber Ag. UC. NO…. Name. Qadam GR Coordinates: latitude. Longitude. : 20/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 26 25

2 28 28

3 27 27

4 27 27

Total 108 107 99.0 % SS 25 1 4%

Table.2 site 3. Fata Species. An. stephensi Tested with. Permethrin District. Khyber Ag. UC. NO…. Name. Qadam GR Coordinates: latitude. Longitude. : 20/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 24

2 25 24

3 25 25

4 25 25

Total

100 98 98.0 % SS 23 0 0%

Table.3 site 3. Fata Species. An. stephensi Tested with. DDT .4% District. Khyber Ag. UC. NO…. Name. Qadam GR Coordinates: latitude. Longitude. : 20/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 26 16

2 24 15

3 26 16

4 26 15

Total 102 62 60.7 %

RR 20 0

0%

Table.4 site 3. Fata Species. An. stephensi Tested with. Malathion. 5% District. Khyber Ag. UC. NO…. Name. Qadam GR Coordinates: latitude. Longitude. : 20/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 23 12

2 22 12

3 25 12

4 25 12

5 25 12

Total 120 57 47.5 % RR 25 1 4%

34

Table.5 site 3. Fata Species. An.culicifacies Tested with. Deltamethrin. 0.05% . District. Khyber Ag. UC. NO…. Name. Qadam GR Coordinates: latitude. Longitude. : 20/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 24 24

2 25 25

3 25 25

4 26 25

Total 100 99 99.0 % SS 18 0 0%

Table.6 site 3. Fata Species. An.culicifacies Tested with. Permethrin District. Khyber Ag. UC. NO…. Name. Qadam GR Coordinates: latitude. Longitude. : 20/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 23 22

2 26 26

3 27 26

4 25 25

Total 101 99 98.0 % SS 25 1 4%

Table.7 site 3. Fata Species. An.culicifacies Tested with. DDT 4% . District. Khyber Ag. UC. NO…. Name. Qadam GR Coordinates: latitude. Longitude. : 20/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 23 12

2 27 13

3 25 18

4 27 13

Total 102 56 49.32 % RR 17 2 11.0%

Table.8 site 3. Fata Species. An.culicifacies Tested with. Malathion 5% . District. Khyber Ag. UC. NO…. Name. Qadam GR Coordinates: latitude. Longitude. : 20/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 10

2 26 16

3 27 13

4 28 13

Total 108 52 48.1 % SS 20 0 0%

35

Table.1 site 1. KPK Species. An. stephensi Tested with. Deltamethrin. 0.05% District: KPK….. UC. NO…. Name. ……mama khel GR Coordinates: latitude. 33.7855164 Longitude. : 72.1342083 2/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 24 24

2 26 25

3 23 23

4 27 26

Total 100 98 98% SS 45 1 2.22%

Table.2 site 1. KPK Species. An. stephensi Tested with. Bendiocarb District: KPK….. UC. NO…. Name. ……mama khel GR Coordinates: latitude. 33.7855164 Longitude. : 72.1342083 2/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 19

2 24 18

3 26 23

4 25 20

Total

100 80 80% RR 43 2 4.65%

Table.3 site 1. KPK Species. An. stephensi Tested with. Malathion. 5% District: KPK….. UC. NO…. Name. ……mama khel GR Coordinates: latitude. 33.7855164 Longitude. : 72.1342083 2/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 17

2 24 18

3 23 21

4 25 18

5 10 5

Total 102 79 77.45 RR 38 1 2.6%

Table.4 site 1. KPK Species. An. stephensi Tested with. DDT .4% District: KPK….. UC. NO…. Name. ……mama khel GR Coordinates: latitude. 33.7855164 Longitude. : 72.1342083 2/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 13

2 20 15

3 18 10

4 18 13

5 25 16

Total 101 67 66.33% RR 33 1 3.03%

36

Table.5 site 1. KPK Species. An.culicifacies Tested with. Deltamethrin. 0.05% . District: KPK….. UC. NO…. Name. ……mama khel GR Coordinates: latitude. 33.7855164 Longitude. : 72.1342083 2/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 20

2 25 25

3 26 25

4 20 20

5 10 10

Total 101 100 99% SS 25 1 4%

Table.6 site 1. KPK Species. An.culicifacies Tested with. Bendiocarb District: KPK….. UC. NO…. Name. ……mama khel GR Coordinates: latitude. 33.7855164 Longitude. : 72.1342083 2/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 21 15

2 21 16

3 20 15

4 20 16

5 20 16

Total 102 78 76.47% RR 25 1 4%

Table.8 site 1. KPK Species. An.culicifacies Tested with.. DDT 4% District: KPK….. UC. NO…. Name. ……mama khel GR Coordinates: latitude. 33.7855164 Longitude. : 72.1342083 2/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 10

2 25 8

3 25 11

4 25 12

Total 100 41 35.8% RR 25 2 8%

Table.7 site 1. KPK Species. An.culicifacies Tested with. Malathion 5% District: KPK….. UC. NO…. Name. ……mama khel GR Coordinates: latitude. 33.7855164 Longitude. : 72.1342083 2/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 16

2 26 13

3 26 14

4 26 13

Total 103 56 54.36% RR 26 1 3.84%

37

Table.1 site 2. KPK Species. An. stephensi Tested with. Deltamethrin. 0.05% District: UC. NO…. Name. … Serai Naurang… GR Coordinates: latitude. 32.825024 Longitude. 70.783653 5/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 20

2 25 25

3 20 20

4 20 20

5 20 19

Total 105 104 98.01% SS 45 2 4.44%

Table.2 site 2. KPK Species. An. stephensi Tested with. Bendiocarb District: UC. NO…. Name. … Serai Naurang… GR Coordinates: latitude. 32.825024 Longitude. 70.783653 5/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 16

2 20 16

3 20 17

4 23 16

5 20 16

Total

103 81 78.6% RR 33 1 3.03%

Table.3 site 2. KPK Species. An. stephensi Tested with. Malathion. 5% District: UC. NO…. Name. … Serai Naurang… GR Coordinates: latitude. 32.825024 Longitude. 70.783653 5/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 24

2 20 18

3 20 15

4 25 20

5 10 4

Total 100 81 81% RR 33 1 3.03%

Table.4 site 2. KPK Species. An. stephensi Tested with. DDT .4% District: UC. NO…. Name. … Serai Naurang… GR Coordinates: latitude. 32.825024 Longitude. 70.783653 5/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 12

2 20 14

3 18 19

4 21 11

5 23 11

Total 102 67 65.68% RR 25 1 4%

38

Table.5 site 2. KPK Species. An.culicifacies Tested with. Deltamethrin. 0.05% . District: UC. NO…. Name. … Serai Naurang… GR Coordinates: latitude. 32.825024 Longitude. 70.783653 5/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 20

2 20 20

3 20 22

4 20 20

5 24 20

Total 104 102 98.07% SS 25 1 4%

Table.6 site 2. KPK Species. An.culicifacies Tested with. Bendiocarb District: UC. NO…. Name. … Serai Naurang… GR Coordinates: latitude. 32.825024 Longitude. 70.783653 5/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 10

2 25 12

3 25 10

4 25 12

Total

100

44

44%

RR

25

0

0%

Table.7 site 2. KPK Species. An.culicifacies Tested with. Malathion 5% District: UC. NO…. Name. … Serai Naurang… GR Coordinates: latitude. 32.825024 Longitude. 70.783653 5/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 16

2 20 20

3 25 20

4 25 20

5 15 10

Total 105 86 81.90% RR 25 1 3.03%

Table.8 site 2. KPK Species. An.culicifacies Tested with.. DDT 4% District: UC. NO…. Name. … Serai Naurang… GR Coordinates: latitude. 32.825024 Longitude. 70.783653 5/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 12

2 25 13

3 25 12

4 25 13

Total

100 50 50%

RR 25

1

4%

39

Table.1 site 3. KPK Species. An. stephensi Tested with. Deltamethrin. 0.05% District: UC. NO…. Name. … lakki city … GR Coordinates: latitude.32.6045 Longitude. 70.9131 9/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 20

2 20 20

3 20 20

4 20 20

5 20 20

Total 100 100 100% SS 35 1 2.85%

Table.2 site 3. KPK Species. An. stephensi Tested with. Bendiocarb District: UC. NO…. Name. … lakki city … GR Coordinates: latitude.32.6045 Longitude. 70.9131 9/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 23 20

2 24 22

3 22 21

4 21 20

5 14 10

Total

104 93 89.42% RR 33 1 3.03%

Table.3 site 3. KPK Species. An. stephensi Tested with. Malathion. 5% District: UC. NO…. Name. … lakki city … GR Coordinates: latitude.32.6045 Longitude. 70.9131 9/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 19

2 19 17

3 21 20

4 20 20

5 21 18

Total 101 94 92.28% V.R 30 3 10%

Table.4 site 3. KPK Species. An. stephensi Tested with. DDT .4% District: UC. NO…. Name. … lakki city … GR Coordinates: latitude.32.6045 Longitude. 70.9131 9/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 12

2 20 18

3 20 14

4 20 15

5 13 10

Total 103 69 63.72 RR 33 3 9.09%

40

Table.5 site 3. KPK Species. An.culicifacies Tested with. Deltamethrin. 0.05% . District: UC. NO…. Name. … lakki city … GR Coordinates: latitude.32.6045 Longitude. 70.9131 9/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 25

2 25 25

3 26 26

4 26 26

Total 102 102 100%

SS 25 0 0%

Table.6 site 3. KPK Species. An.culicifacies Tested with. Bendiocarb District: UC. NO…. Name. … lakki city … GR Coordinates: latitude.32.6045 Longitude. 70.9131 9/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 15

2 25 20

3 20 15

4 25 15

5 10 5

Total 100 70 70% RR 20 0 0%

Table.7 site 3. KPK Species. An.culicifacies Tested with. Malathion 5% District: UC. NO…. Name. … lakki city … GR Coordinates: latitude.32.6045 Longitude. 70.9131 9/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 15

2 20 20

3 20 20

4 20 15

5 20 10

Total 105 80 76.19% RR 25 1 4%

Table.8 site 3. KPK Species. An.culicifacies Tested with.. DDT 4% District: UC. NO…. Name. … lakki city … GR Coordinates: latitude.32.6045 Longitude. 70.9131 9/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 22 11

2 20 11

3 20 11

4 25 11

5 15 8

Total

102 52 49.01% RR 26 1 3.84%

41

Table.1 site 1. PUNJAB Species. An. stephensi Tested with. Deltamethrin. 0.05% District: Punjab UC. NO. 53 Name Village Machikai GR Coordinates: latitude. 15/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 24 20

2 24 22

3 24 21

4 23 21

5 15 13

Total 110 97 86.95% RR 48 4 8%

Table.2 site 1. PUNJAB Species. An. stephensi Tested with. Lambdacyhalothrin. 0.05% . District: Punjab UC. NO. 53 Name Village Machikai GR Coordinates: latitude. 15/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 22

2 25 23

3 25 22

4 25 23

Total 100 90 89% RR 50 4 8%

Table.3 site 1. PUNJAB Species. An. stephensi Tested with. Permethrin.0.75% District: Punjab UC. NO. 53 Name Village Machikai GR Coordinates: latitude. 15/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 20

2 25 22

3 25 21

4 25 23

Total 100 86 84% RR 48 4 8%

Table.4 site 1. PUNJAB Species. An. stephensi Tested with. Malathion. 5% District: Punjab UC. NO. 53 Name Village Machikai GR Coordinates: latitude. 15/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 20

2 25 20

3 25 21

4 25 19

Total 100 80 77.14 RR 48 6 12.5%

42

Table.5 site 1. PUNJAB Species. An.Stephensi Tested with. DDT. 4% District: Punjab UC. NO. 53 Name Village Machikai GR Coordinates: latitude. 15/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 26 12

2 24 11

3 25 12

4 25 11

Total 100 46 40% RR 48 5 10%

Table.6 site 1. PUNJAB Species. An.culicifacies Tested with. Deltamethrin. 0.05% . District: Punjab UC. NO. 53 Name Village Machikai GR Coordinates: latitude. 13/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 24 22

2 25 21

3 26 23

4 25 22

Total 100 88 86.95% RR 48 4 8%

Table.7 site 1. PUNJAB Species. An.culicifacies Tested with. Lambdacyhalothrin. 0.05% District: Punjab UC. NO. 53 Name Village Machikai GR Coordinates: latitude. 13/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 21

2 25 23

3 25 24

4 25 22

Total 100 90 89.36% RR 50 3 6%

Table.8 site 1. PUNJAB Species. An.culicifacies Tested with. Permethrin 0.75% District: Punjab UC. NO. 53 Name Village Machikai GR Coordinates: latitude. 13/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 24

2 25 22

3 26 25

4 24 22

Total 100 93 92% V.R 48 4 8%

43

Table.9 site 1. PUNJAB Species. An.culicifacies Tested with. . Malathion .5% . District: Punjab UC. NO. 53 Name Village Machikai GR Coordinates: latitude. 22/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 23

2 24 21

3 26 24

4 25 22

Total 100 90 89% RR 48 4 8%

Table.10 site 1. PUNJAB Species. An.culicifacies Tested with. DDT 4% . District: Punjab UC. NO. 53 Name Village Machikai GR Coordinates: latitude. 22/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 10

2 25 13

3 25 11

4 25 12

Total 100 46 40% RR 48 5 10%

Table.1 site 2. PUNJAB Species. An. stephensi Tested with. Deltamethrin. 0.05% Village Kulkarni UC. NO Kulkarni GR Coordinates: latitude. 15/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 18

2 22 19

3 20 18

4 18 16

5 20 18

Total 100 89 89% RR 40 1 2.5%

Table.2 site 2. PUNJAB Species. An. stephensi Tested with. Lambdacyhalothrin. 0.05% . Village Kulkarni UC. NO Kulkarni GR Coordinates: latitude. 16/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 19

2 21 20

3 22 19

4 23 19

5 20 18

Total 106 95 87% RR 40 6 15%

44

Table.3 site 2. PUNJAB Species. An. stephensi Tested with. Permethrin.0.75% Village Kulkarni UC. NO Kulkarni GR Coordinates: latitude. 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 22

2 24 21

3 26 23

4 25 22

Total 100 88 86% RR 49 4 8%

Table.4 site 2. PUNJAB Species. An. stephensi Tested with. Malathion. 5% Village Kulkarni UC. NO Kulkarni GR Coordinates: latitude. 18/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 19

2 25 20

3 25 19

4 25 21

Total 100 79 77% RR 49 4 8%

Table.5 site 2. PUNJAB Species. An.Stephensi Tested with. DDT. 4% Village Kulkarni UC. NO Kulkarni GR Coordinates: latitude. 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 14

2 24 10

3 26 14

4 25 13

Total 100 51 51% RR 49 2 4%

Table.6 site 2. PUNJAB Species. An.culicifacies Tested with. Deltamethrin. 0.05% . Village Kulkarni UC. NO Kulkarni GR Coordinates: latitude. 12/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 18

2 21 19

3 21 20

4 20 18

5 18 15

Total 100 90 88% RR 40 4 10%

45

Table.7 site 2. PUNJAB Species. An.culicifacies Tested with. Lambdacyhalothrin. 0.05% Village Kulkarni UC. NO Kulkarni GR Coordinates: latitude. 10/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 24

2 25 24

3 25 22

4 25 22

Total 100 92 92% V.R 42 2 4.76%

Table.8 site 2. PUNJAB Species. An.culicifacies Tested with. Permethrin 0.75% Village Kulkarni UC. NO Kulkarni GR Coordinates: latitude. 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 23

2 25 23

3 24 22

4 26 24

Total 100 92 92% V.R 49 2 4%

Table.9 site 2. PUNJAB Species. An.culicifacies Tested with. . Malathion .5% . Village Kulkarni UC. NO Kulkarni GR Coordinates: latitude. 21/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 21

2 25 22

3 25 22

4 25 22

Total 100 87 87% RR 50 2 4%

Table.10 site 2. PUNJAB Species. An.culicifacies Tested with. DDT 4% . Village Kulkarni UC. NO Kulkarni GR Coordinates: latitude. 22/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 26 12

2 24 11

3 25 12

4 25 13

Total 100 48 48% RR 48 2 4%

46

Table.1 site 3. PUNJAB Species. An. stephensi Tested with. Deltamethrin. 0.05% UC. NO. 41 Mar bangwab Village Baag Danaiser 2 GR Coordinates: latitude. 15/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 23

2 25 23

3 25 23

4 25 23

Total 100 92 92% V.R 40 1 2.5%

Table.2 site 3. PUNJAB Species. An. stephensi Tested with. Lambdacyhalothrin. 0.05% . UC. NO. 41 Mar bangwab Village Baag Danaiser 2 GR Coordinates: latitude. 16/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 22

2 26 23

3 25 22

4 24 22

Total 100 89 89% RR 45 2 4%

Table.4 site 3. PUNJAB Species. An. stephensi Tested with. Malathion. 5% UC. NO. 41 Mar bangwab Village Baag Danaiser 2 GR Coordinates: latitude. 18/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 19

2 25 18

3 25 18

4 25 18

Total 100 73 73% RR 48 1 2%

Table.3 site 3. PUNJAB Species. An. stephensi Tested with. Permethrin.0.75% UC. NO. 41 Mar bangwab Village Baag Danaiser 2 GR Coordinates: latitude. 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 24 21

2 24 21

3 26 23

4 26 22

Total 100 87 87% RR 48 2 4%

47

Table.5 site 3. PUNJAB Species. An.Stephensi Tested with. DDT. 4% UC. NO. 41 Mar bangwab Village Baag Danaiser 2 GR Coordinates: latitude. 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 10

2 20 10

3 20 9

4 20 11

5 20 10

Total 100 50 50% RR 50 2 4%

Table.6 site 3. PUNJAB Species. An.culicifacies Tested with. Deltamethrin. 0.05% . UC. NO. 41 Mar bangwab Village Baag Danaiser 2 GR Coordinates: latitude. 12/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 23

2 25 23

3 25 24

4 25 22

Total 100 92 92% V.R 40 1 2.5%

48

Table.7 site 3. PUNJAB Species. An.culicifacies Tested with. Lambdacyhalothrin. 0.05% UC. NO. 41 Mar bangwab Village Baag Danaiser 2 GR Coordinates: latitude. 10/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 24 22

2 24 21

3 26 23

4 26 24

Total 100 90 89% RR 47 4 8%

Table.8 site 3. PUNJAB Species. An.culicifacies Tested with. Permethrin 0.75% UC. NO. 41 Mar bangwab Village Baag Danaiser 2 GR Coordinates: latitude. 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 23

2 24 21

3 26 24

4 25 24

Total 100 92 92% V.R 48 2 4%

Table.9 site 3. PUNJAB Species. An.culicifacies Tested with. . Malathion .5% . UC. NO. 41 Mar bangwab Village Baag Danaiser 2 GR Coordinates: latitude. 21/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 22

2 25 21

3 25 20

4 25 22

Total 100 85 85% RR 50 2 4%

Table.10 site 3. PUNJAB Species. An.culicifacies Tested with. DDT 4% . UC. NO. 41 Mar bangwab Village Baag Danaiser 2 GR Coordinates: latitude. 22/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 26 14

2 24 12

3 25 11

4 25 13

Total 100 50 50% RR 50 2 4%

49

Table.1 site 1. Sindh Species. An. stephensi Tested with. Deltamethrin. 0.05% District: Mirpur khas UC. Yar Muhammad Ginji Khipro rd GR Coordinates: latitude. 25.540490 Longitude. 69.033815 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 20

2 20 20

3 24 22

4 22 21

5 25 23

Total 111 106 94.28% V.R 24 3 12.5%

Table.2 site 1. Sindh Species. An. stephensi Tested with. Lambdacyhalothrin. 0.05% . District: Mirpur khas UC. Yar Muhammad Ginji Khipro rd GR Coordinates: latitude. 25.540490 Longitude. 69.033815 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 14

2 20 15

3 23 16

4 25 15

5 24 14

Total 112 74 61.36 % RR 25 3 12%

Table.3 site 1. Sindh Species. An. stephensi Tested with. Malathion. 5% District: Mirpur khas UC. Yar Muhammad Ginji Khipro rd GR Coordinates: latitude. 25.540490 Longitude. 69.033815 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 20

2 20 20

3 23 22

4 25 24

5 24 23

Total 112 109 96.59 % V.R 25 3 12%

Table.4 site 1. Sindh Species. An. stephensi Tested with. DDT 4% District: Mirpur khas UC. Yar Muhammad Ginji Khipro rd GR Coordinates: latitude. 25.540490 Longitude. 69.033815 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 9

2 20 7

3 23 8

4 25 9

5 22 7

Total 110 40 30.43 % RR 24 2 8%

50

Table.5 site 1. Sindh Species. An.Stephensi Tested with. Bendiocarb 0.1% District: Mirpur khas UC. Yar Muhammad Ginji Khipro rd GR Coordinates: latitude. 25.540490 Longitude. 69.033815 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 19

2 20 20

3 20 20

4 22 21

5 21 19

Total 103 99 95.65 % V.R 25 2 8%

Table.6 site 1. Sindh Species. An.culicifacies Tested with. Deltamethrin. 0.05% . District: Mirpur khas UC. Yar Muhammad Ginji Khipro rd GR Coordinates: latitude. 25.540490 Longitude. 69.033815 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 22 21

2 22 21

3 24 23

4 22 21

5 20 19

Total 110 105 94.28 V.R 24 3 12.5%

Table.7 site 1. Sindh Species. An.culicifacies Tested with. Lambdacyhalothrin. 0.05% District: Mirpur khas UC. Yar Muhammad Ginji Khipro rd GR Coordinates: latitude. 25.540490 Longitude. 69.033815 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 14

2 24 15

3 25 17

4 22 13

5 24 13

Total 115 72 55.29% RR 20 3 15%

Table.8 site 1. Sindh Species. An.culicifacies Tested with. Malathion. 5% District: Mirpur khas UC. Yar Muhammad Ginji Khipro rd GR Coordinates: latitude. 25.540490 Longitude. 69.033815 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 24 23

2 25 24

3 22 21

4 20 19

5 23 22

Total 114 109 94.31% V.R 25 3 12%

51

Table.9 site 1. Sindh Species. An.culicifacies Tested with. . DDT 4% . District: Mirpur khas UC. Yar Muhammad Ginji Khipro rd GR Coordinates: latitude. 25.540490 Longitude. 69.033815 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 10

2 22 11

3 20 9

4 25 13

5 22 11

Total 109 54 43.33% RR 20 2 10%

Table.10 site 1. Sindh Species. An.culicifacies Tested with. Bendiocarb 0.1% District: Mirpur khas UC. Yar Muhammad Ginji Khipro rd GR Coordinates: latitude. 25.540490 Longitude. 69.033815 17/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 24

2 22 21

3 20 19

4 20 20

5 20 19

Total 107 103 95% 25 3 12%

Table.1 site 2. Sindh Species. An. stephensi Tested with. Deltamethrin. 0.05% District: Mirpur khas UC. Mir Ghulam Hussain talpur GR Coordinates: latitude. 25.374491 Longitude. 69.201705 18/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 19

2 20 19

3 19 19

4 20 19

5 22 20

Total 101 96 94.31 % V.R 25 3 12%

Table.2 site 2. Sindh Species. An. stephensi Tested with. Lambdacyhalothrin. 0.05% . District: Mirpur khas UC. Mir Ghulam Hussain talpur GR Coordinates: latitude. 25.374491 Longitude. 69.201705 18/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 10

2 20 11

3 21 11

4 22 12

5 20 10

Total 103 54 44% RR 23 3 13%

52

Table.3 site 2. Sindh Species. An. stephensi Tested with. Malathion. 5% District: Mirpur khas UC. Mir Ghulam Hussain talpur GR Coordinates: latitude. 25.374491 Longitude. 69.201705 18/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 20

2 20 20

3 20 20

4 21 20

5 21 20

Total 102 100 98% SS 22 3 13.63%

Table.4 site 2. Sindh Species. An. stephensi Tested with. DDT 4% District: Mirpur khas UC. Mir Ghulam Hussain talpur GR Coordinates: latitude. 25.374491 Longitude. 69.201705 18/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 3

2 20 3

3 21 4

4 22 4

5 23 3

Total 106 15 14 % RR 25 1 4%

Table.5 site 2. Sindh Species. An.Stephensi Tested with. Bendiocarb 0.1% District: Mirpur khas UC. Mir Ghulam Hussain talpur GR Coordinates: latitude. 25.374491 Longitude. 69.201705 18/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 20

2 19 18

3 21 20

4 22 21

5 23 22

Total 105 101 95% V.R 24 2 8%

Table.6 site 2. Sindh Species. An.culicifacies Tested with. Deltamethrin. 0.05% . District: Mirpur khas UC. Mir Ghulam Hussain talpur GR Coordinates: latitude. 25.374491 Longitude. 69.201705 18/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 19

2 22 21

3 24 23

4 21 19

5 21 20

Total 108 102 93% V.R 25 3 12%

53

Table.7 site 2. Sindh Species. An.culicifacies Tested with. Lambdacyhalothrin. 0.05% District: Mirpur khas UC. Mir Ghulam Hussain talpur GR Coordinates: latitude. 25.374491 Longitude. 69.201705 18/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 12

2 19 11

3 20 12

4 21 13

5 23 14

Total 103 62 54.28% RR 24 3 12.5%

Table.8 site 2. Sindh Species. An.culicifacies Tested with. Malathion. 5% District: Mirpur khas UC. Mir Ghulam Hussain talpur GR Coordinates: latitude. 25.374491 Longitude. 69.201705 18/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 20

2 20 20

3 21 20

4 22 21

5 20 19

Total 103 100 96% V.R 25 3 12%

Table.9 site 2. Sindh Species. An.culicifacies Tested with. . DDT 4% . District: Mirpur khas UC. Mir Ghulam Hussain talpur GR Coordinates: latitude. 25.374491 Longitude. 69.201705 18/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 10

2 22 11

3 21 11

4 23 12

5 20 11

Total 106 55 46% RR 25 2 8%

Table.10 site 2. Sindh Species. An.culicifacies Tested with. Bendiocarb 0.1% District: Mirpur khas UC. Mir Ghulam Hussain talpur GR Coordinates: latitude. 25.374491 Longitude. 69.201705 18/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 24 23

2 23 22

3 21 20

4 20 19

5 22 21

Total 109 105 95.47% V.R 26 3 11.53%

54

Table.1 site 3. Sindh Species. An. stephensi Tested with. Deltamethrin. 0.05% District: Mirpur khas UC. Hingorono GR Coordinates: latitude. 25.735317 Longitude. 69.176000 19/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 19

2 20 20

3 21 20

4 22 20

5 20 18

Total 103 97 93% V.R 23 2 8%

Table.2 site 3. Sindh Species. An. stephensi Tested with. Lambdacyhalothrin. 0.05% . District: Mirpur khas UC. Hingorono GR Coordinates: latitude. 25.735317 Longitude. 69.176000 19/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 21 12

2 21 11

3 20 13

4 22 12

5 23 11

Total 105 59 56 % RR 25 1 4%

Table.3 site 3. Sindh Species. An. stephensi Tested with. Malathion. 5% District: Mirpur khas UC. Hingorono GR Coordinates: latitude. 25.735317 Longitude. 69.176000 19/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 20

2 20 19

3 20 20

4 20 20

5 21 19

Total 101 98 96% V.R 25 3 12%

Table.4 site 3. Sindh Species. An. stephensi Tested with. DDT 4% District: Mirpur khas UC. Hingorono GR Coordinates: latitude. 25.735317 Longitude. 69.176000 19/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 21 12

2 20 13

3 25 10

4 23 11

5 22 12

Total 111 58 44% RR 21 3 14%

55

Table.5 site 3. Sindh Species. An.Stephensi Tested with. Bendiocarb 0.1% District: Mirpur khas UC. Hingorono GR Coordinates: latitude. 25.735317 Longitude. 69.176000 19/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 21 20

2 20 20

3 21 20

4 20 19

5 20 18

Total 102 97 94% V.R 24 3 12.5%

Table.6 site 3. Sindh Species. An.culicifacies Tested with. Deltamethrin. 0.05% . District: Mirpur khas UC. Hingorono GR Coordinates: latitude. 25.735317 Longitude. 69.176000 19/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 20

2 22 21

3 19 18

4 20 20

5 22 20

Total 103 99 96 % V.R 23 1 4%

Table.7 site 3. Sindh Species. An.culicifacies Tested with. Lambdacyhalothrin. 0.05% District: Mirpur khas UC. Hingorono GR Coordinates: latitude. 25.735317 Longitude. 69.176000 19/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 15

2 20 14

3 21 15

4 20 13

5 20 15

Total 101 72 68% RR 25 2 8%

Table.8 site 3. Sindh Species. An.culicifacies Tested with. Malathion. 5% District: Mirpur khas UC. Hingorono GR Coordinates: latitude. 25.735317 Longitude. 69.176000 19/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 25 24

2 22 21

3 23 22

4 25 24

5 20 20

Total 115 111 95% V.R 25 2 8%

56

Table.9 site 3. Sindh Species. An.culicifacies Tested with. . DDT 4% . District: Mirpur khas UC. Hingorono GR Coordinates: latitude. 25.735317 Longitude. 69.176000 19/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 15

2 20 14

3 23 13

4 20 11

5 22 13

Total 105 66 62 % RR 21 1 4%

Table.10 site 3. Sindh Species. An.culicifacies Tested with. Bendiocarb 0.1% District: Mirpur khas UC. Hingorono GR Coordinates: latitude. 25.735317 Longitude. 69.176000 19/10/2017

Replicate


Dead after 24 hours

Corrected Mortality

Status


Dead after 24 hours

% Mortality

1 20 20

2 20 19

3 20 20

4 20 20

5 22 20

Total 102 99 96% V.R 25 3 12%

57

Summary Table A 1: Province and species against Insecticides. Province: Baluchistan Specie: An.Stephensi Annex II

Deltamethrin Lambdacyhalothrin Permethrin DDT Malathion

Localities

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

test

ed

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

Mo

rtal

ity

%

Stat

us**

No

. of

fem

ales

test

ed

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

Ghulam Sarwar Goth- Jaffarabad

100 90% V.R 100 34% RR 103 24.27% RR 100 89% RR

Goth Khaliq ali Ghandaka- Jaffarabad 100 89% RR 100 34% RR 101 26.73% RR 100 89% RR

Haji Fateh Mhd Goth-Jaffarabad 101 94.05% V.R 100 52% RR 100 31% RR 100 88% RR

Summary table A1.1: Province and species against Insecticides Province: Baluchistan Specie: An.culicifacies


Localities

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

test

ed

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

Ghulam Sarwar Goth- Jaffarabad

100 94% V.R 100 46% RR 102 37.25% RR 100 84% VR

Goth Khaliq ali Ghandaka- Jaffarabad 100 88% RR 100 48% RR 100 44% RR 102 80.39% RR

Haji Fateh Mhd Goth-Jaffarabad 100 92% V.R 100 45% RR 101 44.55% RR 100 79% RR

58

Summary table A2: Province and species against Insecticides Province: Sindh Specie: An.Stephensi Deltamethrin Lambdacyhalothrin Bendiocarb DDT Malathion

Localities

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

test

ed

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

Mo

rtal

ity

%

Stat

us**

No

. of

fem

ales

test

ed

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

Yar Muhammad Ginji

Khipro rd-Mirpur

Khas 111 94.28% V.R 112 61.36% RR 103 95.77 V.R 110 30.43% RR 112 96.59% V.R

Mir Ghulam Hussain

talpur-Mirpur Khas 101 94.36% V.R 103 44% RR 105 95% V.R 106 14% RR 102 98.79% SS

Hingorono-

MirpurKhas 103 93% V.R 105 56% RR 102 94% V.R 111 44% RR 101 96% V.R

Summary table A2.1 Province and species against Insecticides Province: Sindh Specie: An.culicifacies

Deltamethrin Lambdacyhalothrin Bendiocarb DDT Malathion

Localities

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

test

ed

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

test

ed

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

Yar Muhammad Ginji

Khipro rd-Mirpur Khas 110 94.28% V.R 115 55.29 % RR 107 95% V.R 109 43.33% RR 114 94.31% VR

Mir Ghulam Hussain

talpur-Mirpur Khas 108 93% V.R 103 54.28% RR 109 95.47% V.R 106 46% RR 103 96% V.R

Hingorono-MirpurKhas 103 96% V.R 101 68% RR 102 96% V.R 105 62% RR 115 95% V.R

59

Summary table A3.1 Province and species against Insecticides Province: Punjab Specie: An.culicifacies


Localities

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

Mo

rtal

ity

%

Stat

us**

No

. of

fem

ales

test

ed

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

Machikai-Sheikupura

100 86.95% RR 100 89.36 % R.R 100 92% V.R 100 40% RR 100 89% RR

Kurlkai-Sheikupura

100 88% RR 100 92% V.R 100 92% V.R 100 48% RR 100 87 RR

Baag Danaiser2-Sheikhupura

100 92% V.R 100 89% RR 100 92% V.R 100 50% RR 100 85% RR

Summary table A3. Province and species against Insecticides Province: Punjab Specie: An.Stephensi


Localities

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

test

ed

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

test

ed

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

test

ed

Co

rrec

ted

M

ort

alit

y %

Stat

us**

Machikai-Sheikupura

110 86.95% RR 100 89% RR 100 84% RR 100 40% RR 100 77.14% RR

Kurlkai-Sheikupura

100 89% RR 106 87% RR 100 86% RR 100 51% RR 100 77% RR

Baag Danaiser2-Sheikhupura

100 92% V.R 100 89% RR 100 87% RR 100 50% RR 100 73% RR

60

Summary table A4: Province and species against Insecticides Province: Khyber Pakhtunkhwa Specie: An.stephensi

Deltamethrin Bendiocarb Permethrin DDT Malathion

Localities

No

. of

fem

ales

test

ed

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

test

ed

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

Mo

rtal

ity

%

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

Mamakhel-

KPK 100 98% SS 100 80% RR 101 66.33% RR 102 77.45% RR

Serai Narang 105 98.01% SS 103 78.6% RR 102 65.68% RR 100 81% RR

Lucki city 100 100% SS 104 89.42% RR 103 63.72% RR 101 92.28% V.R

Summary table A4.1: Province and species against Insecticides Province: Khyber Pakhtunkhwa Specie: An.culicifacies


Localities

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

test

ed

Co

rrec

ted

Mo

rtal

ity

%

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

Mamakhel-

KPK 101 99% SS 102 76.47% RR 100 35.8% RR 103 54.36% RR

Serai Narang 104 98.07% SS 100 44% RR 100 50% RR 105 81.90% RR

Lucki city 102 100% SS 100 70% RR 102 49.01% RR 105 76.19% RR

61

Summary table A5: Province and species against Insecticides Province: FATA Specie: An.stephensi


Localities

No

. of

fem

ales

te

sted

Co

rrec

ted

Mo

rtal

ity

%

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

Mo

rtal

ity

%

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

Mo

rtal

ity

%

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

Mo

rtal

ity

%

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

Mo

rtal

ity

%

Stat

us**

Gallani Khola-

Kyber Ag 104 99% SS 106 100% SS 106 34.90% RR 111

40.47%

RR

Ali Masjid Kalay-

khyber Ag 102 100% SS 105 100% SS 100 50% RR 105

44.7%

RR

Qadam-Khyber Ag 108 99% SS 100 98% SS 102 60.7% RR 120 47.5

% RR

Summary table A5.1: Province and species against Insecticides Province: FATA Specie: An.culicifacies


Localities

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

Mo

rtal

ity

%

Stat

us**

No

. of

fem

ales

test

ed

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

No

. of

fem

ales

te

sted

Co

rrec

ted

M

ort

alit

y %

Stat

us**

Gallani Khola-Kyber

Ag 103 99% SS 100 98% SS 117 49.5% RR 103

49.5%

RR

Ali Masjid Kalay-

khyber Ag 103 99% SS 104 99% SS 102 51.9% RR 105

45.7%

RR

Qadam-Khyber Ag 100 99% SS 101 98% SS 102 49.32% RR 108 48.1

% RR

62

Final Summary Table B 1: 5 Provinces against 4 insecticides for Specie: An.Stephensi ANNEX III

Deltamethrin Lambdacyhalothrin Permethrin Bendiocarb DDT Malathion

Districts Localities

To

tal

tes

ted

Co

rrec

ted

Mo

rta

lity

%

Sta

tus*

*

Total

teste

d

Co

rrec

ted

Mo

rta

lity

%

Sta

tus*

*

To

tal

test

ed

Co

rrec

ted

Mo

rta

lity

%

Sta

tus*

*

Tota

l

teste

d Co

rrec

ted

Mo

rta

lity

%

Sta

tus*

*

To

tal

test

ed

Co

rrec

ted

Mo

rta

lity

%

Sta

tus*

*

Punjab

Machikai-Sheikupura 11

0

86.95

% V.R 100 89% RR 100 84% RR

100 40% RR 100

77.14

% RR

Kurlkai-Sheikupura 10

0 89% V.R 106 87% RR 100 86% RR

100 51% RR 100 77% RR

Baag Danaiser2-

Sheikhupura

10

0 92% SS 100 89% RR 100 87% RR

100 50% RR 100 73% RR

Chi square χ2 =3.00 df=2

p=0.223

χ2 =3.00 df=1

p=0.083

χ2 =3.00 df=2

p=0.223

χ2 =3.00 df=2

p=0.223

χ2 =3.00 df=2

p=0.223

Sindh:

Mirpur

Khas

Yar Muhammad

Ginji Khipro rd-

11

1

94.28

% V.R 112

61.36

% RR 103

93.65

% V.R 110

30.43

% RR 112

96.59

% V.R

Mir Ghulam Hussain

talpur-

10

1

94.31

% V.R 103 44% RR 105 95% V.R 106 14% RR 102

97.38

% V.R

Hingorono- 10

3 93% V.R 105 56% RR 102 94% V.R 111 44% RR 101 96% V.R


p=0.199

χ2 =6.00 df=4

p=0.199

χ2 =6.00 df=4

p=0.199

χ2 =6.00 df=4

p=0.199

χ2 =6.00 df=4

p=0.199

Baluchista

n:

Jafarabad

Ghulam Sarwar

Goth-

10

0 90% V.R 100 34% RR

103

24.27

% RR 100 98% SS

Goth Khaliq ali

Ghandaka

10

0 89% RR 100 34% RR

101

26.73

% RR 100 89% RR

Haji Fateh Mhd

Goth-

10

1

94.05

% V.R 100 52% RR

100 31% RR 100 88% RR


p=0.223

χ2 =0.75 df=1

p=0.386

χ2 =6.00 df=4

p=0.199

χ2 =3.00 df=2

p=0.223

Khyber

Pakhtunk

hwa

Mamakhel 100 98% SS

100 80% RR 101 66.33

% RR 102

77.45

% RR

Serai Narang 105 98.01

% SS

103 78.6% RR 102

65.68

% RR 100 81% RR

Lucki city 100 100% SS

104 89.42

% RR 103

63.72

% RR 101

92.28

% SS

Chi Square χ2 =3.00 df=2 χ2 =6.00 df=4 χ2 =6.00 df=4 χ2 =6.00 df=4

63

p=0.223 p=0.199 p=0.199 p=0.199

FATA:

Khyber

Agency

Gallani Khola 104 99% SS

106 100% SS

106 34.90

% RR 111 45% RR

Ali Masjid Kalay-

khyber ag 102 100% SS

105 100% SS

100 50% RR 105 44.7% RR

Qadam- 108 99% SS 100 98% SS 102 60.7% RR 120 47.5% RR


.p=0.223

χ2 =3.00 df=2

p=0.223

χ2 =6.00 df=4

p=0.199

χ2 =6.00 df=4

p=0.199

64

Final Summary Table B2: 5 Provinces against 4 insecticides for Specie: An.culicifacies

Deltamethrin Lambdacyhalothrin Permethrin Bendiocarb DDT Malathion D

istr

ict

Lo

cali

ties

To

tal

test

ed

Corrected

Mortality

% Sta

tus*

*

To

tal

test

ed

Correcte

d

Mortalit

y %

Sta

tus*

*

To

tal

test

ed

Corre

cted

Morta

lity %

Sta

tus*

*

To

tal

test

ed

Corr

ected

mort

ality

%

Sta

tus

To

tal

test

ed

Correct

ed

Mortali

ty %

Sta

tus*

*

To

tal

test

ed

Correcte

d

Mortalit

y %

Sta

tus*

*

Punjab

Machikai-Sheikupura 100 86.95% RR 100 90% V.R 100 92% V.R 10

0 40% RR 100 89% RR

Kurlkai-Sheikupura 100 88% RR 100 92% V.R 100 92% V.R 10

0 48% RR 100 87 RR

Baag Danaiser2-Sheikhupura 100 92% V.R 100 89% RR 100 92% V.R 10

0 50% RR 100 85% RR

Chi square χ2 =6.00 df=4 p=0.199 χ2 =6.00 df=4 p=0.199 χ2 =6.00 df=4 p=0.199 χ2 =6.00 df=4 p=0.199

Sindh:

MirpurKhas

Yar Muhammad Ginji Khipro

rd 110 94.28% V.R 115 41.17% RR

10

7 95%

V.

R

10

9 43.33% RR 114 94.31% VR

Mir Ghulam Hussain talpur- 108 93% V.R 103 54.28% RR 10

9 95.47%

V.

R

10

6 46% RR 103 96% V.R

Hingorono-MirpurKhas 103 96% V.R 101 68% RR 10

2 96%

V.

R

10

5 62% RR 115 95% V.R

Chi square χ2 =6.00 df=4 p=0.199 χ2 =6.00 df=4 p=0.199 χ2 =6.00 df=4

p=0.199

χ2 =6.00 df=4 p=0.199 χ2 =6.00 df=4 p=0.199

Baluchistan

Ghulam Sarwar Goth-

Jaffarabad 100 94% V.R 100 46% RR

1

0

0

37.25% RR 100 48% RR

Goth Khaliq ali Ghandaka-

Jaffarabad 100 88% RR 100 48% RR

1

0

0

44% RR 102 80.39% RR

Haji Fateh Mhd Goth-

Jaffarabad 100 92% V.R 100 45% RR

1

0

1

44.55% RR 100 79% RR


Khyber

Pakhtunkhw

a

Mamakhel-KpK 101 99% SS 10

2

76.47

%

R

R

1

0

0

35.8% RR 103 56% RR

Serai Narang 104 98.07% SS 10

0 44%

R

R

1

0

0

50% RR 105 81.90% RR

Lucki city 102 100% SS 10

0 70%

R

R

1

0

2

49.01% RR 105 76.19% RR

Chi Square χ2 =6.00 df=4 p=0.199 χ2 =6.00 df=4

p=0.199

χ2 =6.00 df=4 p=0.199 χ2 =6.00 df=4 p=0.199

FATA Gallani Khola-Kyber ag 103 99% SS 100 98% S

S

1

1

7

49.5% RR 103 49.5% RR

65

Ali Masjid Kalay-khyber ag 103 99% SS 104 99% S

S

1

0

2

51.9% RR 105 45.7% RR

Qadam-Khyber ag 100 99% SS 101 98% S

S

1

0

2

49.32% RR 108 48.1% RR


66

Fig,1

67

PICTURE 2. Refresher training workshop organized by Institute of medical entomology and disease vector control

(IMEDVC) Islamabad

68

PICTURE 3. REFRESHER TRAINING WORKSHOP ORGANIZED BY INSTITUTE OF MEDICAL

ENTOMOLOGY AND DISEASE VECTOR CONTROL (IMEDVC) ISLAMABAD

70

PICTURE 4. REFRESHER TRAINING WORKSHOP ORGANIZED BY INSTITUTE OF MEDICAL ENTOMOLOGY AND

DISEASE VECTOR CONTROL (IMEDVC) ISLAMABAD

71

PICTURE 5. FIELD WORK FOR SUSCEPTIBILITY TESTING IN 5 PROVINCES.

SIND

BALUCHISTAN

72

KPK

PUNJAB

73

FATA

Insecticide resistance monitoring of malaria vector ...dmc.gov.pk/documents/pdfs/Insecticides...

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