Biodiversity of Culicidae Mosquitoes in District Bagh, Azad Jammu and Kashmir

Muhammad Shahbaz1,*, Nosheen Farooq1, Abu ul Hassan Faiz1, Arshad Javid2, Irfan Baboo3, Misbah Shoukat1 and Muhammad Aslam Khan4 1Department f Zoology, Women University, Azad Jammu and Kashmir, Bagh 2Department of Wildlife and Ecology,University of Veterinary and Animal Sciences, Lahore 3Department of Zoology, Cholistan University of Veterinary and Animal Sciences, Bahwalpur 4Department of Human Genetics and Molecular Biology, University of Health Sciences, Lahore Article Information Received 01 October 2019 Revised 11 December 2019 Accepted 03 January 2020 Available online 29 June 2020


B ecause of its geographical location and ecology
Pakistan is one of the hotspots for mosquito-vectorial diseases (Chan et al., 1995;Stark and Schoneberg, 2012), that's why as early as 1971, mosquito biodiversity was initiated in Pakistan (AslamKhan, 1971(AslamKhan, , 1972. Due to the latest occurrences of dengue (Shakoor et al., 2012), it has created greatest attention in mosquito study in Pakistan (Mukhtar et al., 2011;Ilahi and Suleman, 2013;Rasheed et al., 2013).
Earlier in 1969, from the Changa Manga National Forest, AslamKhan and Salman (1969) studied the bionomics of mosquitoes and described 29 species of mosquitoes, many of which were uncommon, rare and reported for the first time from Pakistan. In Pakistan, the first ever effort to describe the Culicidae fauna was done by Aslamkhan (1971Aslamkhan ( , 1972 who recorded 134 species of mosquitoes from Pakistan, of which 91 species from West Pakistan and 89 from East Pakistan (now Bangladesh). From 1934 to 1971, one species of Anopheles and three species of Culex were included in the mosquito fauna of Pakistan (AslamKhan, 1971). Later, AslamKhan (1972) documented 16 endemic species of mosquito from Pakistan.
More than 3500 species of mosquitoes have been documented, which belong to 42 genera and divided into three subfamilies such as Culicinae, Anophelinae and Toxorhynchitinae (Knight and Stone, 1977).
Currently the family Culicidae is divided into two subfamilies, 113 genera, 11 tribes and 3526 species (Harbach, 2007). Nearly 3523 species have been documented globally in 111 genera from different regions up till now (Harbach, 2012). The genus Anopheles has 7 subgenera and 460 species. Culex has 763 species belonging to 26 subgenera. The genus Aedes has 927 species, which belong to 70 subgenera. Most members of the family Culicidae are public health importance (Wilkerson et al., 2015;Freitas et al., 2015).
The purpose of this study was to enhance the knowledge of the culicids.

Materials and methods
The present study was carried out at District Bagh ( Supplementary Fig. S1). The biodiversity of mosquitoes of the study area was never documented before.
The current study on the culicidae was accomplished in the district of Bagh, Azad Jammu and Kashmir from May, 2017 to October, 2017. Azad Kashmir is located at latitude 33° to 36° and longitude 73° to 75° and covers an area of about 13,297 square kilometers. This state of O n l i n e

F i r s t A r t i c l e
Azad Kashmir is the western part of Himalayan range. The geography of the study area is commonly mountainous and woody with fertile valleys and small grasslands. Its climate is sub-tropical highlands form. Average extreme temperature of district Bagh varies from 20°C to 32°C while the average lowest temperature varies from 4°C to 7°C. Average yearly rain fall ranges from 1000 to 2000 millimeter. The general elevation above sea level, ranges from south to north from 360 meters to 6325 meters, respectively (Hussain, 2013). The mosquitoes of the study area were explored, from May, 2017 to October, 2017 in the morning and evening. The priority of collection was given to the city of Bagh and surroundings areas due to estimated advanced definite abundance in these regions while urban and agricultural zones were also measured.
The mosquitoes were collected from human dwelling (indoor) and adjoining of human dwelling (outdoor) using pyrethrum spray technique as defined by WHO (1992) in the morning between 6 am to 8 am and 6 pm to 9 pm. Apart from this, catches in outdoor shelters like gardens, nurseries and wild vegetation was also made in day time. The mosquitoes were collected at outdoor using mouth aspirator and torch light and the collected mosquitoes were identified by using the keys (Christophers, 1933;Nagpal and Sharma, 1995;Smart, 2003).
The mosquitoes were killed with the help of ethyl acetate vapours and then mounted on piece of thick paper maintained by entomological pin and nail polish. These specimens were preserved in collection boxes comprising naphthalene balls.
The Shannon diversity index (H) was used to characterize species diversity at eight study sites.

Results and discussion
A total of 2895 specimens of mosquitoes were collected from the study area. A total of eleven species were identified (Table I).
Data show eight areas and eleven species of which the most abundant species was Armigeres subalbatus and minimum species was Aedes aegypti. Culex pipiens was present in all 8 areas of district Bagh. A total of 431 specimens of Culex pipiens were collected, greatest number was 143 from the Hodda Bari and least number was 25 which was from Kiayat. The total number of Culex vishnui was 244, the greatest number was 56 from the Hodda Bari and minimum number was 16 from the Kiayat. It was not present in Kotayra. The total number of Culex pseudovishnui was 231, the miximum number was 47 from Hodda Bari and minimum number was 20  (Table I).
The data revealed that Culex pipiens was present in in all the eight sites, Culex vishnui was present in all the sites except Kotayra. Culex pseudovishnui was present in all the sites except Ghaziabad. Culex fatigan was present in Kotayra, Hodda Bari, Ghaziabad and Dhirkot and it was absent in Reyara, Noman Pora, Norgala and Kiayat. Culex barraudi was present in all eight sites. Culex fuscocephala was present in all sites except Reyara and Hodda Bari. Culex epidesmis and Anopheles barrianensis was present in all sites. Aedes aegyti was present in Kotayra, Hodda Bari, Kiayat and Dhirkot. Aedes micropterus was present in all sites except Reyara. Armigeres subalbatus was present in all sites.
The number of male specimen from the eight areas of district Bagh, AJK. Total 1414 species of female mosquitoes were collected from this district. The number of female Culex pipiens (220), Culex vishnui (109), Culex pseudovishnui (126), Culex fatigan (59), Culex barraudi (92), Culex fuscocephala (105), Culex epidesmis (117), Anopheles barrianensis (143), Aedes aegypti (27), Aedes micropterus (103) and Armigerus subalbatus (313). Maximum percentage was Armigeres subalbatus (22.13) percent and minimum percentage was Aedes aegypti (1.90%). This study shows that the percentage of male Culex pipiens was lesser than the female, the percentage of male Culex vishnui was greater than the female, the percentage of male Culex pseudovishnui was less than the female, the percentage of male Culex fatigan was less than the female, the percentage of male Culex barraudi was greater than the female, the percentage of male Culex fuscocephala was almost equal to the female, the percentage of male Culexepidesmis was greater than the female, the percentage of male Anopheles barrienensis was less than the female, the percentage of male Aedes aegypti was greater than the female, the percentage of male Aedes micropterus was less than the female, and the percentage of male Armigeres subalbatus was greater than the female.

Supplementary material
There is supplementary material associated with this article. Access the material online at: https://dx.doi. org/10.17582/journal.pjz/20191001191052

Statement of conflict of interest
The authors declare no conflict of interest.