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Comparison of the Newly Introduced Rearing Methods of Cotton Stainer, Dysdercus koenigii (Hemiptera: Pyrrhocoridae) With Classical Methods.

Byline: Syed Ishfaq Ali Shah, Muhammad Rafiq, Tassawar Hussain Malik, Imran Rafi Khan, Syed Awais Sajid Shah and Zahid Hussain

Abstract

Cotton stainer, Dysdercus koenigii (Hemiptera: Pyrrhocoridae) was reared using four rearing methods including cylindrical perforated plastic bottles (CPPB), cotton seeds in soil (CSS), cotton seeds without soil (CSWS) and integrated rearing technique (IRT) at Lab. maintained temperature 282C along with 705% R.H. Data were recorded on five various parameters including number of egg batches, eggs batch,-1 shortest longevity, longest longevity and transformed %mortality. Our results based compared their mean difference among the highly significant values to the least significant values of the recorded all parameters. In IRT method, cotton stainer laid significantly higher number of egg batches (6.0) as compared to CSS (2.67), CSWS (2.33) and statistically similar to CPPB (1.33). In IRT, cotton stainer laid maximum (56.00 eggs batch-1). It was significantly higher (24.67) in CSS and statistically similar with difference of (21.67) and (3.33) CSWS and CPPB, respectively.

The shortest longevity value (2.33 days) of cotton stainer was observed in CSS method. It was significantly different from the CPPB (8.0 days) and IRT (6.34 days) while, at par with CSWS (2.34 days). Longest longevity was recorded (23.67 days) in CPPB which was significantly (5.34 days) higher in CSS, at par recorded in CSWS (2.34 days) and IRT (1.34 days). Lowest transformed %mortality of was (51.81%) achieved in CPPB. It was recorded (37.62%) and (22.81%) significantly higher in CSS and CSWS respectively. However, it was at par with IRT (8.27%). On the basis of our findings, it was concluded that cotton stainer had shown extreme love for water in all stages for development. The cylindrical perforated plastic bottles had the highest water retention capacity than the other rearing techniques and they sucked the moist seed sap in the same behavior as from the cotton bolls in field. IRT proved most suitable for cotton stainer rearing followed by CPPB.

Key words: Cotton stainer, Dysdercus koenigii, rearing methods, cylindrical perforated plastic bottles, moist cotton seed, integrated rearing technique, fecundity, biocontrol agent.

INTRODUCTION

The cotton having a share of 1.4% in GDP and 6.7% in agriculture value addition is an important source of raw material to the textile industry in Pakistan (PES, 2014). But the cotton is attacked by many chewing and sucking insects (Saeed et al., 2007) causing about 20-40% loss annually (Ahmad, 1999). Recently, cotton stainer, Dysdercus koenigii Fabricius (Hemiptera: Pyrrhocoridae), which is commonly known as red cotton bug, has resurged as a destructive pest in cotton zone of Pakistan (Ahmad and Mohammad, 1983; Ashfaq et al., 2011; Jaleel et al., 2013). Both nymphs and adults of the genus Dysdercus have strong proboscis, a needle like stylet meant for piercing and sucking of plant sap (Elzinga, 1997; Wilson et al., 2004; Shah, 2014).

Dysdercus spp. feed on emerging cotton bolls and mature cotton seeds and transmits cotton staining fungus, Nematospora gossypii that develops on immature lint and seed (Ahmad and Khan, 1980; Ahmad and Schaefer, 1987; Yasuda, 1992). Dysdercus koenigii may insert its proboscis into the soft tissues of cotton bolls and causes an abnormal out-growth (warts) on the inner surface of carpel walls and lesions on the lint (Shah, 2014). This kind of damage on the inner walls of carpels is observed when locules are removed after dissecting the bolls (Wilson et al., 2004).

A major reason for increased population of the cotton stainer is its faster egg development (Venugopal et al., 1994) and wide spread growing of transgenic cotton that have reduced insecticides application, whereas, before the introduction of transgenic varieties, cotton stainer was not even a minor pest of cotton in Pakistan (Shah, 2014). Due to large scale adoption of Bt cotton in the sub-continent sucking pest pressure especially that of D. koenigii increased very much especially during 2011 and 2012 (Ashfaq et al., 2011). Cotton stainer caused severe losses in 2011 onwards exhibiting bad opening of bolls with stained lint in major cotton growing areas of Pakistan (Shah, 2014).

Some work on the bionomics and life cycle of D. koenigii has been reported in India (Kamble, 1971) but information on these aspects of the pest was insufficient under environmental conditions of Pakistan. Insecticides and bio-control agents play important role and were normally more effective on early weaker growth stages of insect pests. Environmentally safe plant protection is a major challenge for sustainable cotton production because, producing high yield products had always been an important part of farming. Chemical control is an essential and sometimes unavoidable component to achieve high yield but, it is important to know and understand the drawbacks of continued insecticide applications. About 90% of the farmers use chemical insecticides (Prayogo et al., 2005). It is estimated that in Pakistan, farmers spend US$300 million on pesticides annually, of which more than 80% is being used on cotton, especially for bollworms (Rao, 2007).

However, we could minimize the indiscriminate use of insecticides with alternate, environmentally safe control measures technique (Rafiq et al., 2014).

For basic and applied studies like screening of most suitable insecticides, insecticides monitoring on regular basis against insect pests, promotion of biological control of predators and parasitoids, all needs its mass culture comprising different developmental stages. Since large scale rearing of D. koenigii was practically difficult therefore, we conducted experiments on different rearing techniques under laboratory conditions to find out the most suitable, easiest and economical rearing method for sufficient rearing of the pest.

MATERIALS AND METHODS

Mass culturing in the Lab.

Adults of cotton stainer, Dysdercus koenigii were collected during November, 2013 from cotton fields of Multan District and were kept at biological control laboratory. Plastic made general mass rearing cages measuring 30x18x28 inches (LxWxD) and having three aerations holes (4x6 inches each) covered with fine mesh were used to rear cotton stainer. About one inch layer of field soil with minute quantity of sand was spread on floor of cages to provide natural substrate. Insecticide free delinted seed of Bt cotton variety CIM-599 in a plastic petri-dishes with filter paper discs underneath were offered as a food along with fresh reproductive plant parts taken from field and green house. Lab. temperature was maintained at 282C along with 705% R.H. and 11:13 hours (L:D) to get eggs of the pest.

Fifth instar nymphs were collected from the bulk and kept in separate general mass rearing cages for further development. Upon maturity to adult stage, ten adult pairs randomly selected for each replication while each treatment was replicated three times.

Rearing techniques

Selected cotton stainer pairs were released in the plastic experimental rearing cages measuring 15x9x15 inches (L x W x D) having two aerations holes (3x3 inches each) covered with fine mesh. A substratum used was one inch thick layer of field-soil mixed with minute quantity of sand and spread on the floor of cages. The following treatments were installed in respective cages after releases to evaluate mean number of egg batches, eggs batch,-1 shortest longevity, longest longevity and % mortality for comparison of different classical methods with new introduced rearing technique.

Cylindrical perforated plastic bottles (CPPB)

Nine CPPB (three for each replication) of three inches length with 1.90 inches diameter while, twenty- four holes drilled to each plastic bottle homogeneously all around and stuffed with wet surgical cotton wool. Delinted cotton seeds were inserted randomly in twelve holes while rests were left empty for moisture purpose (Fig. 1). Each perforated bottle was placed inverted in plastic petri-dishes (2 inches diameter). Few dried cotton leaves were kept near each perforated bottle like (Fig. 4) for egg laying purpose. Daily 10ml water was evenly sprayed twice daily (morning and evening hours) over the perforated bottles and soil of the cages including the dried leaves. Seeds were replaced with three days intervals.

Cotton seed in soil (CSS)

Nine plastic petri-dishes were used each having a filter paper disc inside with field soil above and 12 moist delinted cotton seeds on the top (Fig. 2). Each experimental cage had three petri-dishes whereas, no dried leaves were provided and daily 10 ml water was evenly sprayed twice a day (morning and evening hours) over the petri-dishes and soil of the cages. Seeds were replaced with three days intervals.

Cotton seed without soil (CSWS)

This set included nine plastic petri-dishes (three for each replication) filled with moist cotton wool having a filter paper disc above and 12 delinted cotton seeds on the top (Fig. 3). No dried leaves were provided and daily 10 ml water was evenly sprayed over the petri-dishes and soil of the cages twice a day. Seeds were replaced with three days intervals.

Integrative rearing technique (IRT)

In this treatment, cotton stainers were reared in combination of all above mentioned three treatments and were termed as Integrated Rearing Technique (IRT).

Each experimental cage had a single perforated bottle, petri-dishes with and without soil (Fig. 4). Dried leaves were provided near the perforated bottle and daily 10ml water was evenly sprayed over perforated bottle, petri- dishes, soil of the cages and dried cotton leaves. Seeds were replaced with three days intervals.

Statistical analysis

Experiment was planned and laid out using Completely Randomized Design (CRD). Data were recorded on five parameters including mean number of egg batches, mean eggs batch mean shortest longevity, mean longest longevity, and mean %mortality.

The %mortality was calculated by dividing the number of dead cotton stainers per replicate with total number of releases per replicate. Data collected on afore mentioned parameters were analyzed using Analysis of Variance ANOVA with Tukey's HSD (Honestly Significant Difference) post hoc test. The %mortality before the analysis was arcsine transformed in degrees using the following expression presented by Gomez and Gomez (1984).

(Equation)

However, before transformation all the 100 values were replaced by using the following formula,

(Equation)

where n is the number of insects released, whereas in our case n=20.

RESULTS

Table I shows comparison of egg batches and number of eggs/batch laid, longevity and mortality of cotton stainer using different rearing techniques.

Egg batches laid

In the IRT method, the cotton stainer laid (6.00) significantly higher mean number of egg batches as compared to CSS (2.67), without soil (2.33) and statistically similar to CPPB (1.33) (Table I). The coefficient of variation (CV) for the data on mean number of egg batches was 16.01% and P = 0.0069 while, in the CPPB method they laid statistically similar mean number of egg batches (1.34) as compared to CSS method and CSWS (1.0 egg batch).

Number of eggs batch-1 laid

In IRT method, cotton stainer laid 56.00 mean number of eggs batch-1. It was significantly higher 24.67 mean number of eggs batch-1 than in CSS and statistically similar with difference of 21.67 and 3.33 cotton CSWS and CPPB methods respectively (Table I), having C.V. = 20.21% and P = 0.0186. In perforated bottle method they laid 52.67 eggs batch-1 with difference of 21.34 and 18.34 respectively from CSS and CSWS methods (statistically at par).

Longevity

The cotton stainer's mean shortest longevity 2.33 days was recorded in CSS method. A total of 10 pairs of the cotton stainer were released in each replication of the four rearing methods. The mean shortest longevity value of cotton stainer in CSS method was significantly different from the CPPB (8.0 days) and IRT (6.34 days) (Table I) having C.V.=37.16% and P = 0.0132; while the value was statistically at par with CSWS (2.34 days). The CPPB was statistically similar to IRT and CSWS method respectively with a difference of 1.66 and 5.66 mean numbers of days.

Longest longevity of cotton stainer was recorded 23.67 days in CPPB. It was significantly 5.34 days higher in CSS method while, statistically at par recorded in CSWS 2.34 days and IRT methods 1.34 days (Table I), having C.V.=5.39% and P = 0.0028. In IRT longest longevity of cotton stainer was 22.33 days reported. It was significantly 4.00 days higher from the treatment of CSS while statistically at par 1.00 days with CSWS method.

Mortality (%) of cotton stainer

Lowest transformed percent mortality of cotton stainer was 51.81 achieved in CPPB method. It was recorded 37.62% and 22.81% significantly higher in CSS and CSWS methods respectively than the perforated bottles (Table I), having C.V.= 10.16% and P = 0.0008. However, it was statistically at par with IRT method (8.27%). In IRT, cotton stainer transformed percent mortality was 60.08% recorded which is significantly 29.35% lower from cotton seed in soil 29.35% while, statistically similar 14.54% in CSWS treatment.

DISCUSSION

Lab. temperature and humidity Muthupandi et al. (2014) reared and maintained D. cingulatus adults under lab. conditions of 3110C, 755% R.H. and 11:13 hours (L:D). Jaleel et al. (2013) reared and maintained adults of D. koenigii at 282C, 705% R.H. and 11:13 hours (L:D). Varma and Patel (2012) reared D. koenigii at an average room temperature of 25.507.36C and average relative humidity of 55.1921.36%. Sahayaraj and Jeeva (2012) maintained D. cingulatus in the laboratory at 282C and 705% R.H. Sahayaraj and Ilyaraja (2008) were collected D. cingulatus adults and nymphs from cotton fields and were maintained under laboratory conditions 272C, 705% R.H. and 11:13 (L:D).

Similarly, we maintained our lab temperature at 282C, R.H. at 705% and 11:13 hours (L:D). Cotton stainers are reported to be sensitive to changes in temperature and humidity (Jaleel et al., 2013). It was re- confirmed and keenly observed in rearing process that cotton strainers are very much sensitive to changes in temperature and humidity. Very few of them were survived for egg laying in general mass rearing cages when brought a bulk of cotton stainer adults from field to lab. Fluctuations in the temperature and humidity adversely affected its population. Low mortality of cotton stainer was observed in mass reared from eggs laid of field collected population of cotton stainer in general mass rearing cages. Our results revealed that highest number of eggs and egg batches were found in the IRT Mass rearing of cotton stainer in the lab.

Table I.- Eggs batch,-1 shortest longevity, longest longevity and %mortality of different rearing techniques.

###Rearing methods

Parameters###Overall MSE

###CPPB###CSS###CSWS###IRT

Egg batches###4.670.33ab###3.330.33b###3.670.33b###6.000.58a###4.420.36

Eggs batche-1###52.675.55ab###31.334.67b###34.333.28ab###56.006.66a###43.583.95

Longevity shortest###10.331.45a###2.331.20b###4.670.88ab###8.671.86a###6.51.12

Longest###23.670.67a###18.330.88b###21.330.67ab###22.330.33a###21.420.66

Mortality (%)###51.812.63c###89.430.00a###74.627.41ab###60.081.92bc###68.984.66

Table II.- Eggs batch,-1 shortest longevity, longest longevity and %mortality of different rearing techniques.

###Rearing methods

Parameters

###CPPB###CSS###CSWS###IRT

Egg batches###45###34###34###57

Eggs batch-1###4463###2440###2440###4769

Shortest longevity###813###04###36###511

Longest longevity###2325###1720###1720###2223

Mortality (%)###47.8756.79###89.4389.43###89.4389.43###56.7963.44

Muthupandi et al. (2014) reared D. cingulatus in plastic containers of 20 inches diameter on moist Gossypium arboretum L. seed. Jaleel et al. (2013) reared D. Koenigii in plastic chamber 4x4 inches on soaked fuzzy cottonseeds of MNH-886 (Bt). The plastic chambers were half filled with sterilized soil as natural medium for oviposition. Filter paper was placed on the soil to maintain moderate moisture level in the plastic chamber. Nymphs were collected from the field and confined to plastic jars with filter paper on the bottom and mouth closed with muslin cloth. Fresh seeds of okra were provided at two days interval until adult emergence (Verma et al., 2013). Sahayaraj and Jeeva (2012) reared D. cingulatus on water soaked cotton seeds and fresh cotton leaves in plastic container.

Sontakke et al. (2013) were reared D. cingulatus on moist cotton seed in glass bottles covered its mouth with muslin cloth and rubber bands. Sahayaraj and Ilyaraja (2008) reared D. cingulatus in plastic containers 20x10x15 inches on soaked cotton seeds.

We reared D. koenigii in plastic cages measuring 30x18x28 inches (LxWxD) having three aeration holes (4x6 inches each) covered with fine mesh on delinted moist cotton seed and fresh reproductive parts of cotton plant in a general mass rearing; while for experimental purpose, they were reared in 15x9x15 inches (LxWxD) having two aeration holes (3x3 inches each) covered with fine mesh called the plastic experimental rearing cages. About one inch ticked layer of field-soil was spread over the cage floor with minute quantity of raw sand in both the general mass and experimental rearing cages. In plastic experimental rearing cages (Figs. 1, 2, 3) moist seed of CIM-599 (Bt) were provided in different mediums. Cotton seed used for stock feed is an important alternate source of food for cotton strainers (Wilson et al., 2008). On the basis of above mentioned results, we therefore recommend that the most efficient commercial mass rearing methods for D. koenigii are the IRT and CPPB.

Fecundity

Dysdercus cingulatus lays eggs singly or in small, loose clusters on the bottom of petri-dish (Verma et al., 2013). D. koenigii laid eggs singly or in clusters in sand, soil, or on plants (Jaleel et al., 2013). In our findings, D. koenigii preferred to lay eggs in the moist soil under plants debris or a narrow hide moist place. Based upon our results, we will strongly recommended that for commercial rearing of D. koenigii moist soil and plant debris for cotton stainer egg laying medium are the best one. A single female may lay 10-50 eggs, scattered on the provided substrate (Ahmad and Mohammad, 1983). Fecundity of female ranged between 55-56 eggs per female (Verma et al., 2013). D. koenigii laid eggs varied from 39-183 (Jaleel et al., 2013). The fecundity of red cotton bug was 115 eggs per female (Verma and Patel 2012).

Likewise, we found in our study the ranges of eggs per batch as 12-105, 19-91, 13-77 and 18-62 with respective average of 55.0, 52.0, 31.0 and 34.0 eggs per batch respectively in IRT, CPPB, CSS and CSWS rearing methods,. We collected eggs mostly from the moist soil and under moist dried leaves in the rearing cages.

Longest and shortest longevity

Survival period of D. koenigii adults was 20.856.12 days (Jaleel et al., 2013). D. cingulatus longevity of adults was 20-24 with an average of 21.61.81 days (Varma et al., 2013). The average longevity of D. koenigii was 22.331.44 days. Whereas, the cotton stainer adults survived for 25, 23, 22 and 20 days, respectively in the CPPB, the IRT, the CSWS and the CSS rearing methods.

Mode of damage

Cotton stainer had been declared as one of the most destructive cotton pest in other parts of the world (Sprenkel, 2000). Most of its nymphal stages as well as its adults feed on the seeds of cotton bolls. Shah (2014) in his study had observed very keenly its mode of damage. Both nymphs and adults of this insect have a very strong proboscis. They insert and used proboscis up and down and sucked the sap seeds from inside the cotton boll. We found as cotton strainers efficiently responded in the perforated bottles method because they inserted proboscis with up and down movement in the holes of perforated bottles and sucked sufficient sap from the seed in seed inserted holes and moisture from wool of empty holes of the perforated bottles.

CONCLUSION

On the basis of above mentioned findings, it was concluded that cotton stainers extremely love moisture (mesophylic). The perforated bottles had the highest moisture retention capacity than the other rearing techniques and cotton stainer suck the cell sap from the moist cotton seed in the same behavior when it insert its proboscis into cotton boll in the field, reached to the seed, sucked the sap from the seed with frequent upward and downward moment of the proboscis. It means that provision of same feeding behavior can greatly matter in the nourishment of insect especially the cotton stainer.

We therefore, highly recommend the IRT and CPPB methods for rearing of the cotton stainers particularly for commercial rearing of the insect on biological control aspects.

ACKNOWLEDGMENT

The principal author is extremely thankful to Dr. Kohno Katsuyuki (Entomologist) NARO Institute of Vegetable and Tea Sciences, Japan for reviewing this manuscript and providing constructive and valuable comments to improve the results and discussion. We also appreciated field staff of CCRI, Multan i.e., Abdul Mateen, Muhammad Aslam, Muhammad Akbar and Javed for their constant support in collection and rearing of cotton stainer during the whole course of experiment.

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Author:Shah, Syed Ishfag Ali; Rafig, Muhammad; Malik, Tassawar Hussain; Khan, Imran Rafi; Sajid, Syed Awais
Publication:Pakistan Journal of Zoology
Article Type:Report
Date:Jun 30, 2016
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