Coccinellidae beetles (Coleoptera) fauna of district Layyah (Punjab), Pakistan

Beetles belonging to family Coccinellidae are predators of various sucking insect pests, some of them are phytophageous. A lot of work for the exploration of these beetles has been done in Pakistan. Layyah being a multi crop area was selected as study area for the exploration of Coccinellidae beetles. A survey was conducted for the collection of Coccinellidae adult specimens from the territory of Layyah district of southern Punjab, Pakistan, during 2018-19. A total 568 adult specimens were collected from seven localities. Out of which, ladybird beetles belonging to seven genera, with ten species namely; Coccinella septempunctata Linnaeus, 1758, Brumoides suturalis (Fabricius, 1798), Henosepilachna vigintioctopunctata (Fabricius, 1775, Henosepilachna elaterii (Rossi, 1794), Hippodamia variegata (Goeze, 1777), Scymnus (Pullus) quadrillum Motschulsky, 1858, Scymnus (Pullus) posticalis Sicard, 1913, Scymnus (Scymnus) nubilus Mulsant, 1850, Micraspis allardi (Mulsant, 1866), Pharoscymnus flexibilis (Mulsant, 1853), have been recorded with their prey and host plants for the first time from the surveyed area. The coccinellids distribution is also given and map was prepared by using ArcGIS map tool. Explored predatory beetles may be used as biocontrol agents in the region for the management of various sucking insect pests of various crops, fruits vegetables etc.


Introduction
The voracious and non-specialist predator, beetles belong to the family Coccinellidae, are the beneficial insects for having ample impacts on diverse insect pest species like aphids, mites, scale insects, whiteflies (Obrycki and Kring, 1998). The predatory behavior of the Coccinellidae varies within the species as they have a positive impact by feeding on the insect pests and intraguild cannibalism included in their negative feeding behavior (Lucas, 2012). Most of the species are economically important predators, while some are also plant or being mycetophagous (Sutherland and Parrella, 2009). So, the predatory behavior of the Coccinellids includes different transitions throughout different trophic levels which covers primary carnivores to herbivorous insect. Similarly, the female Coccinellids are being proved stronger predators than the male beetles (Chowdhury et al., 2008). Most of the Coccinellidae species adults are with bright and shining color patterns with patches or spots against the contrasting backgrounds. Similarly, these color patterns have been found defensive as signs of warning or danger or distasteful for their bird predators (Moreton, 1969). The farmers usually spray pesticides to kill pests extensively which kills both the pests and their natural enemies. The indiscriminate use of chemicals may affect lady bird beetles searching behavior, feeding behavior, mating behavior, number of eggs, and overall life span (Alinejad et al., 2014;Afza et al., 2020). However, ladybird beetles have great potential to survive the lethal effects of applied chemicals which makes them preferable for any integrated pest management (Mughal et al., 2017) According to the latest classifications, Coccinellinae Latreille, 1807 and Microweiseinae Leng, 1920, are now subfamilies of Coccinellidae family (Slipinski, 2007;Seago et al., 2011). Moreover, it is reclassified to superfamily Coccinelloidea (Coleoptera: Polyphaga: Cucujiformia) (Robertson et al., 2015). Overall, more than 6000 species with 360 genera have been reported worldwide. Additionally, more than 300 species have been reported from Indo-Pak subcontinent and more than 71 species specifically from Pakistan. (Fleming, 2000;Irshad, 2001;Boopathi et al., 2020). Most recently and locally, Irshad (2001) 71 species, Rafi et al. (2005) 75 species, Khan et al. (2007) 12 species, Ali et al. (2014aAli et al. ( , 2014bAli et al. ( , 2015 29 species, Ahmed et al. (2017) 9 species, Iqbal et al., (2017, 2019a, 2019b, 2020) 5 species, described predatory ladybird beetles with their associated prey insect pests from different locations of provinces in Pakistan. However, till now there is still deficiency of information about this economically important biological control agent's family from Pakistan. The current study was intended to explore the ambiguous species riches and distribution of Coccinellidae family from the district Layyah of Punjab, Pakistan.

Collection and identification
The ladybird beetle specimens were collected in the morning (8:00-10:00 am) and afternoon (2:00 to 4:00 pm) from different intended areas of district Layyah as shown in figure 1. The ladybird beetle's distribution map is prepared by using ArcGIS map tool. The adult ladybird beetles were collected through hand-picking and net sweep method and killed by using cyanide insect killing glass bottle (Iqbal et al., 2017). The killed specimens were brought to the insect taxonomy laboratory of Fareed Biodiversity Conservation Centre, Department of Agricultural Engineering, Khwaja Fareed University of Engineering and Information Technology, Rahim Yar Khan, Punjab, Pakistan, and preserved in wooden boxes. Each specimen was pinned and mounted on small triangular plastic card tips with the labelling information of their locality, date of collection, host plants etc. Before placing specimens, the wooden boxes were treated with Coopex Powder (®) and naphthalene tablets were placed to deter the other insects feeding on preserved insects. The morphological and taxonomic characters of ladybird beetles were studied on the basis of available published literature and checklists by Poorani, (2002), Rafi et al. (2005), Ali et al. (2015Ali et al. ( , 2018, Ashfaque et al. (2015) and other website links like NBAII, (2009) under the microscope ZMS 2000 compound microscope. However, Photographs of the adult specimens were taken using Nikon Digital camera (SMZ 1500), which was attached to the stereo microscope and photographs were processed by using Helicon focus 6.7.1 and Adobe Photoshop CS 6.0.

General comments
Coccinella septempunctata found similar with the given diagnostic characters description by Ali et al. (2018) and first time reported from Layyah district.

General comments
Micraspis allardi found similar with given diagnostic characters description by Ali et al. (2018) and first time reported from Layyah district.

General comments
Pharoscymnus flexibilis found similar with given diagnostic characters description by Ali et al. (2018) and first time reported from Layyah district.

Discussion
The current surveys were anticipated to explore the indigenous Coccinellidae fauna from southern Punjab district Layyah. A total of 10 species belonging to 7 genera with their prey, associated host plants and distribution were recorded. Previously Poorani (2002) from India, Ali et al. (2015Ali et al. ( , 2018 from Sindh province, Ashfaque et al. (2015) from Northern area of Pakistan, Ahmed et al. (2017) from Sargodha, Gilani (1976) from Faisalabad and Rafi et al. (2005) from different localities of Pakistan reported these species. Other than these species, Shah (1985) reported about sixteen ladybird species with their distribution and pests host plants from Peshawar. Hussain et al. (2018) described four species from rainfed and irrigated localities of Gujrat. Din (2002) reported similar species from Chitral and Rafi et al. (2005)  The persuasion for higher crops production has been directed towards intensive farming systems. That, however, become the reason for high pest populations followed by high intensity pesticides usage and other pest control techniques. The current study will be supportive in mass production of ladybird beetles against pests, reduction in number of insecticide applications and ultimately helping in reduction of environmental pollution.

Conclusion
The current study results contribute to a baseline to study ladybird beetle's ecology, biology, phytophagy, and predatory behavior against different pests. The current reported species of Coccinellidae family were diversified and abundant. These species may be utilized as potential biological control agents after evaluating their predatory potentials. Finally, it leads the researchers towards the conservatory biological control strategies for the endemic plant protection programs.