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International journal of Horticulture, Agriculture and Food science(IJHAF) [Vol-1, Issue-3, Sep-Oct, 2017]
https://dx.doi.org/10.22161/ijhaf.1.3.8 ISSN: 2456-8635
www.aipublications.com/ijhaf Page | 48
on the Effect of Nitrogen gastudiesSLaboratory
of the Cowpea Weevil,Mortalities) on2(N
Callosobruchus maculatus (F.)(Coleoptera :
Bruchidae ) in Iraq *
2
Ani-Maha T. AL,1
A. MohammedAldinHussam
Dept.of plant protection , College of Agriculture ,University of Baghdad , Iraq
*Part of M. Sc. Dissertation for the second author
Abstract— A labrotary studies were conducted at the
college of Agriculture , University of Baghdad to
investigate the effect of nitrogen gas (N2) at low pressure
of 160 and 180 mm mercury and a nitrogen gas
(98.5%)on mortality rates of differentdevelopmental
stages of the cowpea weevil ,Callosobruchus maculatus
(F.) from egg to adult , with different exposure periods of
1,5,6 days. Results indicated that egg , larval, pupal and
adult mortalities increasrd with increasing exposure
peroids, the complete mortality rates for all stages were
achived after 5-6 days. However, these treatment have no
significant effect on seed germination , The use of these
information in control programs for this pest , also
discussed.
Keywords— cowpea weevil, callosobruchusmaculatus
(F.) , Physical control , Nitrogen gas , Low pressure ,
Iraq .
I. INTRODUCTION
Cowpea , Vigna unguiculatis important grain legume
grownin many parts of the world. It has a high nutritanol
value of protein ranged from 17 – 34 %. As well as fat,
carbohydrates and minerals such as iron. calcium and
phosphors (Al-qaisi ,2000; shaw ,2007). Cowpea may be
infested by different insect Pests causing considerable
damage and considered to be one of the limiting factors in
cowpea production. The cowpea weevil ,C. maculatusis a
key pest of grain legumes in food stores ,it has a wide
host range including ,cowpea, chicpeas, lentils, peas and
beans(Aslam et al.2002).Infestation began in the field
when infested seeds were harvested and stored , the
insect continues to feed as a hidden infestation and
emerges as an adult causes secondary infestation and a
total destruction to seeds and decrease their
germination(Bhall et al.,2008).
Pesticides and fumigants have been extensively used all
over the world to control legume grain infestation.
However, chemical methods are being restricted globally
because toric residue and their adverse effect on
environment and human health (Bhalla et al.2008).
Different biological control methods such as,cultural
,Physical and biological were used by varies workers
(Van,2011; Ismail,2006;Trioesele et al,2015 and yamane
,2013 ).The use of controlled atmosphere vacum ,heating
,colling and sterilizing , requires relatively advanced
techniques (van,2014).
This paper was aimed to find an alternative, ecofriendly
method to insecticides and fumigants used in the present
time by using nitrogen gas for the control of this pest in
legume grain stores.
II. MATERIALS AND METHODS
This study was conducted in the insect research
laboratory at Dept., of Plant Protection, College of
Agriculture / University of Baghdad. Different stages of
Cowpea weevil , Callosobruchus maculatus (F.) were
obtained from a laboratory colony rearead on cowpea
seeds for this purpose and maintained under control
conditions of 26 ±2c˚ and humidity of 70±5% in the
incubator . Infested cowpea seeds were prepared by
relasing 1-day old adults (10 pairs for100 seeds) for a
period of 24 h for oviposition / egg laying in culture jars .
Experiment was carried out using cowpea seeds infested
with different developmental stages of C.maculatus , As
the development occurs inside seeds , These comporised
of egg stage 1-day after oviposition , larval -19 days after
oviposition, pupal stage -22 days after oviposition and 1-
day old adults (Bhalla et al.,2008), And 10 seeds of
cowpea infested with each of the development of stage
of C.maculatusper replicate were spread in the base of
petri dishes were exposed to N2 Gas at low pressure
levels (150 and 180mm Hg) in the anaerobic bacteria
breeding container capacity of (1-2)Kg.All development
stages were exposed to N2 gas(98.5%)for different
periods of 1,5,6 days then the insect were transferred to
petri dishes (9cm)covered with muslin cloth and closed
International journal of Horticulture, Agriculture and Food science(IJHAF) [Vol-1, Issue-3, Sep-Oct, 2017]
https://dx.doi.org/10.22161/ijhaf.1.3.8 ISSN: 2456-8635
www.aipublications.com/ijhaf Page | 49
tightly with rubber bullets and then placed in the
incubator at a temperature of 26 ±2˚and a relative
humidity of 70±5% .The experiment were laid out
according to Complete Randomized Design (CRD) and 3
replicates for each treatment . Healthy cowpea seeds
were also subjected to these doses treatments as above
and to germination test . The effect of cumulative
impact of N2 gas on different developmental stages
on the percentage of mortality rates of eggs , larva ,
pupa , and adults were calculated . Data were
analayzed using SAS program (2012). All the
comparisons was considered significant when P≤ 0.05.
III. RESULTS AND DISCUSSION
Results in Tables 1 revealed that their were a
significant differences in the mortalities of egg , larva,
pupa and adults in the tretments of N2 gas under low
pressure of 150 mm Hg and control treatment .
Mortality rates of 90 , 66.66 , 73.33 and 93.33 %
respectively were achived after 24 hr from exposure
to N2 gas . Its obvious that eggs and adults were
more sensitive to this treatment more than larvae and
pupal stage during the first 24 hr . However ,
complete mortalities of developmental stages from the
egg – adult were reached after 5 – 6 days from
exposure . These results agree with that of Ismail
(2006) who found that the adult , stage of the cowpea
weevil, C. maculatus was the most sensitive followed
by the egg stage , larva , then pupa , and complete
mortalities of all these stage were obtained after 6
days from exposure to N2 gas and carbon dioxide at
low pressure (160 mm Mercury) . This results also
conside with Al –Hadidi (2002) that the adults stage
of the flower beetles , Tribolium castenum evertsand
Trogoderma granariumis the more sensitive to N2 gas
under low pressure followed by eggs , larva and pupa
respectively .
Table.1: Effects of exposure to N2 gas on the different development stages of cowpwa weevil , C.maculatus under low
pressure of (150 mm Mercury)
Results in Table (2) show a significant differences in
the mortalities of all developmemtal stages of the
cowpea weevil , C. maculatus from egg to adult in the
treatments of N2 gas and low pressure of 180 mm
mercury and the control treatments . Mortalities of
93.33 , 76.0 , 93.0 and 96.0 were obtained after 24 hr
from the exposure to N2 gas for the eggs , larva ,
pupa and adults respectively . Mortality rates were
increased for all developmental stages by the
increasing pressure from 150 mm mercury to 180 mm
mercury during the first 24 hr from the treatment .
However , Their were no significant effect was
observed from the previous experiment above when
exposure poroid increased to 5 and 6 days , as a
complete mortalities of all developmental stages were
obtained . It seems that amount of pressure is the
effecting factor which may speed the rate of
mortality than the exposure time which need a
further investigation in the future . Mbata (1994)
observed the same results that when C.subiunotatus
and C.maculatus were exposed to an atomosphere
saturated with carbon dioxide and a temperature of
32 C0
and relative humidity of 70 %
.
%Adult
mortality
% pupal
mortality
%larval
mortality
%egg mortalityExposure period
(days)
0000000000000.00Control
390993909966066300001
000000000000360660000005
0000000000000000000000006
54.554.5846714.1L.S.D
International journal of Horticulture, Agriculture and Food science(IJHAF) [Vol-1, Issue-3, Sep-Oct, 2017]
https://dx.doi.org/10.22161/ijhaf.1.3.8 ISSN: 2456-8635
www.aipublications.com/ijhaf Page | 50
Table.2: Effects of exposure to N2 gas on different developmental stages of cowpea weevil C .maculatus under low
pressure of (180mm mercury) .
Results in Table (3) indicated that their were no
significant differences in the percentage of cowpea
seeds germination between the cowpea seed exposed
to N2 gas from (1 – 6 ) days and the control
treatment under low pressure of 150 and 180 mm
mercury
Oyebanji et al., (2015) found that using the atmospheric
pressur, Ahigh seed germination rate up to (88.2%)
during the storage and noticed that the longer the seed
storage period, the lower the germination rate.
Table.3: Effects of N2 gas on the germination of cowpea seeds
REFERENCES
[1] Al- Hadidi , I . K 2002 . The life of beetles
Tribolium castaneum (Coleoptera: Tenebrionidae)
(Herbst) and Trogoderma granarium Everts
(Coleopteran: Dermestidae) in some local wheat
products and Its sensitivity to low pressure, carbon
dioxide and nitrogen, Master Thesis, College of
Education, University of Mosul,Iraq.68pp0
[2] AL –qaisi , M . D . Mahdi damad (2000). Future
prospects for legumes production and their role in
food security. Journal of Agriculture and
Development in the Arab World,3:50-550
[3] Aslam, M. ; K. A. Khan and M. Z. H. Bajwa 2002.
Potency of Some Spices Against Callosobruchus
chinensis (L.). J. Biol. Sci. 2: 449-4520
[4] Bhalla, S.; K.Gupta; B.Lal; M.L. Kapur and R.K.
Khetarpal . 2008. Efficacy of various non-chemical
methods against pulse, Callosobruchusmaculatus
Fab. ENDURE International Conference
2008 Diversifying crop protection, 12-15 October.
La Grande-Motte, France – Oral presentations – p. 4
[5] Ismail , A . Y 2006 . Effect of low pressure ,
Nitrogen an carbon Dioxide Gases on the mortlity of
the cowpea Bruchid , Callosobrochus maculatus (F.)
(Bruchidae : Coleoptera) . Arab J . of prot . 24 , : 28
– 31 0
[6] Mbata, C., C. Reichnuth and T. Ofuya. 1994.
Comporative toxicity of carbon dioxide to two
Callosobruchus species Pages 120-22. In:
Proceedings of the 6th International Workshop
Conference on Stored Product Protection. E.
Highley, E.J. Wright, H.J. Banks and B.R. Champ
(Editors), Canberra, Australia, 17-23 April , 1994
CAB International, Wallingfored, Oxon0
[7] Oyebanji, A .O; Olagbaju, A.R; Zaka, K.O ;
Ilesanmi, F.F and M. F.Olorunfemi .2015. Quality
and acceptability of inert-atmosphere-metal-silo
stored paddy rice as food and planting material.
Journal of Agricultural and Crop Research 3 (1), 11-
200
[8] Shaw, Monica. (2007).100 Most Protein Rich
Vegetarian Foods .SmarterFitter Blog.28 October0
%adult
mortality
% pupal
mortality
%larval
mortality
%egg mortalityExposure period
(days)
0000000000000000Control
360663909936066390991
0000000000000000000000005
0000000000000000000000006
54.554.554.554.5L.S.D
% of cowpea seed germinationPeriod (Day)Tretment (Bar)
301-6Control
390
5 399
396
8411L.S.D
390
11 399
0006
84.7L.S.D
International journal of Horticulture, Agriculture and Food science(IJHAF) [Vol-1, Issue-3, Sep-Oct, 2017]
https://dx.doi.org/10.22161/ijhaf.1.3.8 ISSN: 2456-8635
www.aipublications.com/ijhaf Page | 51
[9] Triroesele , B . ; K . Thomas and s. seketeme . 2015
control of cowpea weevil, Callosobruchusmaculatus
(F.) (Coleoptera : Bruchidae) , using natural plant
products , Insects , 6 : 77 – 840
[10]Van, H . 2011. Biological methods of Bruchid
control in Tropics: a review in: curvent topics in
Tropical Entomology, 12 (1-2-3): 87 – 1020
[11]Yamane, T. 2013. Biorational control methods for
protection of stored grain legumes against Bruchid
beetles, Agricultural Sciences, 4 (12) 5 pages.
[12]SAS. 2012. Statical Analysis System, User,s Guide .
Statistical. Version 9.1th ed . SAS. Inc. Cary. N.C
USA.

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  • 1. International journal of Horticulture, Agriculture and Food science(IJHAF) [Vol-1, Issue-3, Sep-Oct, 2017] https://dx.doi.org/10.22161/ijhaf.1.3.8 ISSN: 2456-8635 www.aipublications.com/ijhaf Page | 48 on the Effect of Nitrogen gastudiesSLaboratory of the Cowpea Weevil,Mortalities) on2(N Callosobruchus maculatus (F.)(Coleoptera : Bruchidae ) in Iraq * 2 Ani-Maha T. AL,1 A. MohammedAldinHussam Dept.of plant protection , College of Agriculture ,University of Baghdad , Iraq *Part of M. Sc. Dissertation for the second author Abstract— A labrotary studies were conducted at the college of Agriculture , University of Baghdad to investigate the effect of nitrogen gas (N2) at low pressure of 160 and 180 mm mercury and a nitrogen gas (98.5%)on mortality rates of differentdevelopmental stages of the cowpea weevil ,Callosobruchus maculatus (F.) from egg to adult , with different exposure periods of 1,5,6 days. Results indicated that egg , larval, pupal and adult mortalities increasrd with increasing exposure peroids, the complete mortality rates for all stages were achived after 5-6 days. However, these treatment have no significant effect on seed germination , The use of these information in control programs for this pest , also discussed. Keywords— cowpea weevil, callosobruchusmaculatus (F.) , Physical control , Nitrogen gas , Low pressure , Iraq . I. INTRODUCTION Cowpea , Vigna unguiculatis important grain legume grownin many parts of the world. It has a high nutritanol value of protein ranged from 17 – 34 %. As well as fat, carbohydrates and minerals such as iron. calcium and phosphors (Al-qaisi ,2000; shaw ,2007). Cowpea may be infested by different insect Pests causing considerable damage and considered to be one of the limiting factors in cowpea production. The cowpea weevil ,C. maculatusis a key pest of grain legumes in food stores ,it has a wide host range including ,cowpea, chicpeas, lentils, peas and beans(Aslam et al.2002).Infestation began in the field when infested seeds were harvested and stored , the insect continues to feed as a hidden infestation and emerges as an adult causes secondary infestation and a total destruction to seeds and decrease their germination(Bhall et al.,2008). Pesticides and fumigants have been extensively used all over the world to control legume grain infestation. However, chemical methods are being restricted globally because toric residue and their adverse effect on environment and human health (Bhalla et al.2008). Different biological control methods such as,cultural ,Physical and biological were used by varies workers (Van,2011; Ismail,2006;Trioesele et al,2015 and yamane ,2013 ).The use of controlled atmosphere vacum ,heating ,colling and sterilizing , requires relatively advanced techniques (van,2014). This paper was aimed to find an alternative, ecofriendly method to insecticides and fumigants used in the present time by using nitrogen gas for the control of this pest in legume grain stores. II. MATERIALS AND METHODS This study was conducted in the insect research laboratory at Dept., of Plant Protection, College of Agriculture / University of Baghdad. Different stages of Cowpea weevil , Callosobruchus maculatus (F.) were obtained from a laboratory colony rearead on cowpea seeds for this purpose and maintained under control conditions of 26 ±2c˚ and humidity of 70±5% in the incubator . Infested cowpea seeds were prepared by relasing 1-day old adults (10 pairs for100 seeds) for a period of 24 h for oviposition / egg laying in culture jars . Experiment was carried out using cowpea seeds infested with different developmental stages of C.maculatus , As the development occurs inside seeds , These comporised of egg stage 1-day after oviposition , larval -19 days after oviposition, pupal stage -22 days after oviposition and 1- day old adults (Bhalla et al.,2008), And 10 seeds of cowpea infested with each of the development of stage of C.maculatusper replicate were spread in the base of petri dishes were exposed to N2 Gas at low pressure levels (150 and 180mm Hg) in the anaerobic bacteria breeding container capacity of (1-2)Kg.All development stages were exposed to N2 gas(98.5%)for different periods of 1,5,6 days then the insect were transferred to petri dishes (9cm)covered with muslin cloth and closed
  • 2. International journal of Horticulture, Agriculture and Food science(IJHAF) [Vol-1, Issue-3, Sep-Oct, 2017] https://dx.doi.org/10.22161/ijhaf.1.3.8 ISSN: 2456-8635 www.aipublications.com/ijhaf Page | 49 tightly with rubber bullets and then placed in the incubator at a temperature of 26 ±2˚and a relative humidity of 70±5% .The experiment were laid out according to Complete Randomized Design (CRD) and 3 replicates for each treatment . Healthy cowpea seeds were also subjected to these doses treatments as above and to germination test . The effect of cumulative impact of N2 gas on different developmental stages on the percentage of mortality rates of eggs , larva , pupa , and adults were calculated . Data were analayzed using SAS program (2012). All the comparisons was considered significant when P≤ 0.05. III. RESULTS AND DISCUSSION Results in Tables 1 revealed that their were a significant differences in the mortalities of egg , larva, pupa and adults in the tretments of N2 gas under low pressure of 150 mm Hg and control treatment . Mortality rates of 90 , 66.66 , 73.33 and 93.33 % respectively were achived after 24 hr from exposure to N2 gas . Its obvious that eggs and adults were more sensitive to this treatment more than larvae and pupal stage during the first 24 hr . However , complete mortalities of developmental stages from the egg – adult were reached after 5 – 6 days from exposure . These results agree with that of Ismail (2006) who found that the adult , stage of the cowpea weevil, C. maculatus was the most sensitive followed by the egg stage , larva , then pupa , and complete mortalities of all these stage were obtained after 6 days from exposure to N2 gas and carbon dioxide at low pressure (160 mm Mercury) . This results also conside with Al –Hadidi (2002) that the adults stage of the flower beetles , Tribolium castenum evertsand Trogoderma granariumis the more sensitive to N2 gas under low pressure followed by eggs , larva and pupa respectively . Table.1: Effects of exposure to N2 gas on the different development stages of cowpwa weevil , C.maculatus under low pressure of (150 mm Mercury) Results in Table (2) show a significant differences in the mortalities of all developmemtal stages of the cowpea weevil , C. maculatus from egg to adult in the treatments of N2 gas and low pressure of 180 mm mercury and the control treatments . Mortalities of 93.33 , 76.0 , 93.0 and 96.0 were obtained after 24 hr from the exposure to N2 gas for the eggs , larva , pupa and adults respectively . Mortality rates were increased for all developmental stages by the increasing pressure from 150 mm mercury to 180 mm mercury during the first 24 hr from the treatment . However , Their were no significant effect was observed from the previous experiment above when exposure poroid increased to 5 and 6 days , as a complete mortalities of all developmental stages were obtained . It seems that amount of pressure is the effecting factor which may speed the rate of mortality than the exposure time which need a further investigation in the future . Mbata (1994) observed the same results that when C.subiunotatus and C.maculatus were exposed to an atomosphere saturated with carbon dioxide and a temperature of 32 C0 and relative humidity of 70 % . %Adult mortality % pupal mortality %larval mortality %egg mortalityExposure period (days) 0000000000000.00Control 390993909966066300001 000000000000360660000005 0000000000000000000000006 54.554.5846714.1L.S.D
  • 3. International journal of Horticulture, Agriculture and Food science(IJHAF) [Vol-1, Issue-3, Sep-Oct, 2017] https://dx.doi.org/10.22161/ijhaf.1.3.8 ISSN: 2456-8635 www.aipublications.com/ijhaf Page | 50 Table.2: Effects of exposure to N2 gas on different developmental stages of cowpea weevil C .maculatus under low pressure of (180mm mercury) . Results in Table (3) indicated that their were no significant differences in the percentage of cowpea seeds germination between the cowpea seed exposed to N2 gas from (1 – 6 ) days and the control treatment under low pressure of 150 and 180 mm mercury Oyebanji et al., (2015) found that using the atmospheric pressur, Ahigh seed germination rate up to (88.2%) during the storage and noticed that the longer the seed storage period, the lower the germination rate. Table.3: Effects of N2 gas on the germination of cowpea seeds REFERENCES [1] Al- Hadidi , I . K 2002 . The life of beetles Tribolium castaneum (Coleoptera: Tenebrionidae) (Herbst) and Trogoderma granarium Everts (Coleopteran: Dermestidae) in some local wheat products and Its sensitivity to low pressure, carbon dioxide and nitrogen, Master Thesis, College of Education, University of Mosul,Iraq.68pp0 [2] AL –qaisi , M . D . Mahdi damad (2000). Future prospects for legumes production and their role in food security. Journal of Agriculture and Development in the Arab World,3:50-550 [3] Aslam, M. ; K. A. Khan and M. Z. H. Bajwa 2002. Potency of Some Spices Against Callosobruchus chinensis (L.). J. Biol. Sci. 2: 449-4520 [4] Bhalla, S.; K.Gupta; B.Lal; M.L. Kapur and R.K. Khetarpal . 2008. Efficacy of various non-chemical methods against pulse, Callosobruchusmaculatus Fab. ENDURE International Conference 2008 Diversifying crop protection, 12-15 October. La Grande-Motte, France – Oral presentations – p. 4 [5] Ismail , A . Y 2006 . Effect of low pressure , Nitrogen an carbon Dioxide Gases on the mortlity of the cowpea Bruchid , Callosobrochus maculatus (F.) (Bruchidae : Coleoptera) . Arab J . of prot . 24 , : 28 – 31 0 [6] Mbata, C., C. Reichnuth and T. Ofuya. 1994. Comporative toxicity of carbon dioxide to two Callosobruchus species Pages 120-22. In: Proceedings of the 6th International Workshop Conference on Stored Product Protection. E. Highley, E.J. Wright, H.J. Banks and B.R. Champ (Editors), Canberra, Australia, 17-23 April , 1994 CAB International, Wallingfored, Oxon0 [7] Oyebanji, A .O; Olagbaju, A.R; Zaka, K.O ; Ilesanmi, F.F and M. F.Olorunfemi .2015. Quality and acceptability of inert-atmosphere-metal-silo stored paddy rice as food and planting material. Journal of Agricultural and Crop Research 3 (1), 11- 200 [8] Shaw, Monica. (2007).100 Most Protein Rich Vegetarian Foods .SmarterFitter Blog.28 October0 %adult mortality % pupal mortality %larval mortality %egg mortalityExposure period (days) 0000000000000000Control 360663909936066390991 0000000000000000000000005 0000000000000000000000006 54.554.554.554.5L.S.D % of cowpea seed germinationPeriod (Day)Tretment (Bar) 301-6Control 390 5 399 396 8411L.S.D 390 11 399 0006 84.7L.S.D
  • 4. International journal of Horticulture, Agriculture and Food science(IJHAF) [Vol-1, Issue-3, Sep-Oct, 2017] https://dx.doi.org/10.22161/ijhaf.1.3.8 ISSN: 2456-8635 www.aipublications.com/ijhaf Page | 51 [9] Triroesele , B . ; K . Thomas and s. seketeme . 2015 control of cowpea weevil, Callosobruchusmaculatus (F.) (Coleoptera : Bruchidae) , using natural plant products , Insects , 6 : 77 – 840 [10]Van, H . 2011. Biological methods of Bruchid control in Tropics: a review in: curvent topics in Tropical Entomology, 12 (1-2-3): 87 – 1020 [11]Yamane, T. 2013. Biorational control methods for protection of stored grain legumes against Bruchid beetles, Agricultural Sciences, 4 (12) 5 pages. [12]SAS. 2012. Statical Analysis System, User,s Guide . Statistical. Version 9.1th ed . SAS. Inc. Cary. N.C USA.