Dezmirean H. et al.

/Scientific Papers: Animal Science and Biotechnologies, 2015, 48 (1)

Characterization of Some Breeds and Hybrids of Bombyx

mori L. Silkworms through Biological Index of Spawning
Horaiu Dezmirean1, Liviu Al. Mrghita1, Ioan Paca1, Silvia Ptruic2, Georgeta Dini3,
Anton Nagy1, Emilia Furdui1, Daniel Dezmirean1
University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca. 400372, Cluj-Napoca, Calea Mntur
3-5, Romania
2
Banat University of Agricultural Sciences and Veterinary Medicine Timioara, 300645 Timioara, Calea Aradului
119, Romania
3
University of Agronomic Science and Veterinary Medicine of Bucharest, 011464, Bucharest, Bulevardul Mrti 59,
Romania
1

Abstract
The main aim of this study was to determine the parameters of biological indexes of Bombyx mori L. spawning in
order to obtain adequate larvae for experimental breeding using MSF (Familial Sericulture Modulus). The biological
material used for experiments contained the pure breeds AB (Alb Bneasa), B1 (Bneasa 1), AC (Alb de Cislu),
AC29 (Alb chinezesc, line 29), AC29/T (Alb Chinezesc), AC/T (Alb de Cislu), RG90, IBV, S8, AJ5/F, C127, AJ17,
C108/T, AO/T and the hybrids B1xAC, ACxB1, ABxAC29, AC29xAB, ACxAB, ABxAC, ABxB1, B1xAB. For each
pure breed and hybrid the number of eggs/spawning, the number of hatching eggs, the hatching percent (%) and the
number of non-hatching eggs were determined. The analysis of these indexes was done in three consecutive years,
and the averages and spread parameters, as well as variability coefficients (V%) were calculated for each breed and
hybrid. The obtained values of biological indexes of spawning were compared with those reported in autochthonous
and foreign literature. The main conclusions of our study are that the autochthonous breeds and hybrids values are
according to the values published by other researchers and correspond for the experimental breeding.
Keywords: dispersion, eggs, prolificacy, silkworm, spawning, variability

created under the environment conditions of our

country, and the used of high quality spinning
machines for silk processing.
In the present study we establish the biological
indexes of silkworms and hybrids which are
represented by prolificacy and hatching
percentage, expressed as average of three
spawnings/breed or hybrid. From these
characteristics depends the good development of
the obtained larvae, and the result presented in this
paper represents the first step of our research
project during 2011-2013 [1, 2].
In other words, we studied here the egg stage of
silkworms, recording the parameters characteristic
for every breed and hybrid. The eggs stage is the
first one from the evolutional cycle of Bombyx
mori, and is about 300 days in univoltine species

1. Introduction
In the last period the Romanian sericulture
registered a strong decline and for this reason the
evaluation of autochthonous gene pool is
imperative. The Romanian genetic patrimony
contains 72 silkworm breeds and lines and 59
breeds of mulberry trees, some of them unique
worldwide [1].
In this moment the Romanian sericulture can be
restarted in all the country by implementation of a
new strategy. This strategy is based on silkworm
breeding using Familial Sericulture Modulus
(MSF), the used of selected biological material,
the maximum exploitation of breeds and hybrids
* Corresponding author: Daniel Dezmirean,
0264596384, ddezmirean@usamvcluj.ro

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The start of incubation process was establish every

year according to the vegetative evolution of
mulberry leaves, considering that in the day of
hatching the trees has to have 3 to 5 leaves. The
day when incubation starts is easy to establish due
to the fact that between the embryo development
and the evolution of mulberry shoots exist a
connexion, the shoots being 2 or 3 days late [3,5].
As reproduction material, the eggs of Bombyx
mori are named seed, and the eggs of one
female are called spawning [6].

(one generation/year) and about 120 days in

bivoltine ones (with two generation/year)[3].

2. Materials and methods

The biological material used during three
experimental years comes from the autochthonous
genetic pool of Bombyx mori L. species. A part of
it is from S.C. Sericarom Bneasa (Bucharest),
and its stock in USAMV Cluj-Napoca, containing
over 60 breeds and hybrids with different descent.
For this study we used the pure breeds: AB (Alb
Bneasa), B1 (Bneasa 1), AC (Alb de Cislu),
AC29 (Alb chinezesc linia 29), AC29/T (Alb
Chinezesc), AC/T (Alb de Cislu), RG90, IBV, S8,
AJ5/F, C127, AJ17, C108/T, and AO/T.
The simple hybrids analysed during the
experimental years were: B1xAC, ACxB1,
ABxAC29, AC29xAB, ACxAB, ABxAC, ABxB1,
and B1xAB. A large description of these breed and
hybrids are done by Dezmirean et al (2010) [1].
The incubation represents the period of time
necessary for the complete development of the
embryo from egg; the eggs are kept under optimal
condition of temperature, humidity, light and
ventilation. If these conditions are respected a
high hatching percentage is obtained.
The room where the incubation was made had a
south exposure, was thermo-isolated and it was
provided by an adequate system of heating,
humidification and ventilation. For the constant
registration of environmental conditions a thermohydrometer was used.
The incubation process was realized using the
gradual ascending temperature method; this method
is the most recommended because it respects the
physiological processes which take place during the
incubation period. Briefly, this method comprise in
removal of the eggs from freezer, their maintenance
for three days at 150C, then the ascending of
temperature to 18-200C for two days; the next four
days the temperature is maintained at 23-240C, and
then until the end of incubation process, 25-260C are
assured. On the whole incubation process the natural
light was assured, and in the moment of eggs
bleaching (when the embryo consume the pigment
batch of the egg), the artificial light was used. After
this moment the light was suspended for 36 hours in
order to obtain a high and grouped hatching
percentage, and then the normal light was assured
again [3, 4].

3. Results and discussion

Generally, the egg had 1.5/1 mm, the size being a
breed character; its size determine the number of
eggs in one gram ( 1300-2000 eggs/g). The
number of spawning eggs laid by a female
depends by the breed and it is approximately 670
in autochthonous breeds, smaller in Chinese
breeds and bigger in Russian ones [4, 7].
The colour of fresh eggs is yellow, and after a few
days change to gray, grey, and lavender, black or
red, according to the breed. The colour of
Romanian eggs is dark gray in AB and B1 breeds
and grey-greenish in AC and AC29 [8].
The colour of chorion has different shading, too
(white, yellow, green or grey). The eggs and
chorions colour represent a breed character and it
has an important role in selection and
improvement of silkworms.
The incubation method described above is used
for obtaining a high hatching percentage, and
especially grouped in a short period of time. This
fact has an important applicative use, due to the
fact that a good grouping of hatching gives a good
subsequent development of experimental works.
In our experiments (three consecutive years) the
period of incubation process was on average 13
days. The hatching was about 3 days, and the
highest hatching percentage was obtained in the
second day. The larvae obtained in this day were
kept for the next experimental procedures, in order
to maintain the homogeneity of experimental lots.
Table 1 attest that the smaller average number of
laid eggs was registered in IBV breed (462
eggs/spawning), and the biggest one in AB (627
eggs/spawning). These values are according to
those obtained by other researchers. Trag (1992),
reports in parental breeds a number between 520
and 630 eggs/spawning [9]; Rao et al. (1990)

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regarding the biological indexes obtained in IBV

breed demonstrate that not always a high number
of spawning eggs means a high number of
obtained larvae.
The average values of hatching percentage in
AC29 breed is superior of that reported by Benea,
which in 2006 [12] registered 90.71% in the same
breed. In AC breed the average value was 96.55%,
this value is a little higher from that obtained by
Paca (2004) which registered for the same breed
95.74% [14].

between 415 and 458 eggs/spawning [10];

Vassileva et al. (2004) between 506 and 560
eggs/spawning [11]; Benea (2006) between 505
and 660 eggs/spawning [12]; Furdui (2011)
reports an average number of eggs/spawning of
533, the smaller value was registered in IBV breed
(456 eggs/spawning), and the highest one (533
eggs/spawning) in AC/T [13].
The average hatching percentage of breeds was
96%, the smaller one was in AC29 breed (94.03%),
and the biggest one in IBV (97.40%). The results

Breed
AB
AC
B1
AC29
AC29/T
AC/T
RG90
IBV
S8
AJ5/F
C127
AJ17
C108/T
AO/T
Breeds
average

Simple hybrid
ABxAC
ACxAB
B1xAC
ACxB1
ABxB1
B1xAB
ABxAC29
AC29xAB
Hybrids
average

Table 1. Prolificacy and hatching percentage of studied silkworm breeds

Number of eggs /
Number of hatching
Hatching percentage
Number of non-hatching
hatching
eggs
(%)
eggs
627
597
95.27
30
609
588
96.55
21
608
581
95.61
27
530
499
94.03
32
524
500
95.36
24
574
550
95.88
24
560
537
95.90
23
462
450
97.40
12
517
501
96.84
16
502
481
95.88
21
464
442
95.26
22
480
461
95.91
20
509
486
95.61
22
477
454
95.24
23
532

509

96

23

Table 2. Prolificacy and hatching percentage of studied silkworm hybrids

Number of
Number of hatching Hatching percentage Number of non-hatching
eggs/hatching
eggs
(%)
eggs
623
602
96.52
21
587
552
93.44
35
618
587
94.74
62
610
576
94.35
67
622
592
95.08
31
597
573
96
25
587
559
95.28
28
595
567
95.19
29
605

576

259

95

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Table 3. Averages and dispersion parameters calculated for hatching percent at pure breeds and hybrids
Breeds

SD

V%

Hybrids

SD

V%

AB
AC
B1
AC29
AC29/T
AC/T
RG90
IBV
S8
AJ5/F
C127
AJ17
C108/T
AO/T

95.27
96.55
95.61
94.03
95.36
95.88
95.90
97.40
96.84
95.88
95.26
95.91
95.61
95.24

1.76
1.23
2.55
2.95
3.49
1.18
0.37
1.58
0.97
0.52
1.56
2.01
2.36
1.51

1.84
1.27
2.67
3.14
3.66
1.23
0.39
1.62
1.01
0.55
1.64
2.09
2.47
1.59

ABxAC
ACxAB
B1xAC
ACxB1
ABxB1
B1xAB
ABxAC29
AC29xAB

96.52
93.99
94.74
94.35
95.08
95.88
95.28
95.19

2.28
3.06
2.74
1.53
0.93
1.24
1.60
1.96

2.36
3.26
2.89
1.62
0.98
1.29
1.68
2.06

The variability coefficients (V%) were in all cases

under 15%, demonstrating that the average of
breed samples were representative, and the
samples were homogeneous. In the case of hybrids
the smallest variability coefficient (V%) was
registered for ABxB1 hybrid (0.98%), and the
bigger for ACxAB (3.26%), in this case the
samples being homogeneous, too.
The hatching period was about 3 days; the larvae
hatched in the second day were selected for the
next experimental rearing.

The biological indexes of the eight studied hybrids

are presented in Table 2.
The average number laid by the hybrids was 605
eggs/spawning, with a hatching percent of 95%.
These values are similar to those obtained by
Matei (1993) [15], which reported 93.6% in
parental breeds and 96.3% in simple hybrids and
to those reported by Furdui (2011), which reports
an average value of prolificacy of 95.43% [13].
The smaller number of eggs/spawning and
hatching percentage was determined in ACxAB
hybrid (587 eggs/spawning and 93.44%).
The biggest eggs/spawning was determined in
ABxAC (623), and its hatching percentage was
96.52%, which represented the maximum value
registered in the studied hybrids. For these two
hybrids, Paca (2004) reported a hatching
percentage of 93.41% for ACxAB and 96.66% for
ABxAC [14].
The coefficient of linear regression (R2) calculated
for hatching percentage and the number of
eggs/spawning was 0.07 for parental breeds and
0.02 for hybrids, values that demonstrate that
these two characters are not powerfully influenced
one by other. The same situation was
demonstrated by Furdui (2011), too [13].
The averages and dispersion parameters for
hatching percentage of breeds and hybrids are
presented in Table 3. The smaller variability
coefficient was calculated for RG90 breed
(0.39%), and the biggest for AC29/T (3.66); in
these breeds the standard deviations were lower,
too.

4. Conclusions
For phenotype characterization of autochthonous
silkworm breeds and hybrids experimental
growing were done in three consecutive years.
The
measurements
achieved
and
data
interpretation permitted us to state the following
conclusions regarding the biological indexes of
spawning.
The prolificacy was express by the number of
eggs laid by a female and by the hatching
percentage. The number of eggs fluctuated
between 462 eggs/spawning in IBV breed and 627
eggs/spawning in AB. For hybrids the smaller
number of eggs/spawning was registered in
ACxAB hybrid (587), and the biggest at ABxAC
(623).
The hatching percentage fluctuated between
94.03% in AC29 breed and 97.40% in IBV.
93.44%. The smallest hatching percent was

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Hibrids of Cocoon Colour Sex-Limited Breed of

Silkworm, Bombyx mori L. under different
environments of temperature, J. Ent. Res. Soc., 2008,
10, pp.1-12.
6. Paca, I., Mrghita, A.L., Morar, R., Pusta, D.,
Dezmirean, D., Cmpean, A., Macri, A., Furdui, E.,
Orioan, R., Bagita, C., Research concerning the
correlation between the characteristics of raw and dry
cocoon at some mulberry silkworm breeds (Bombyx
mori L.), Bulletin of USAMV Cluj-Napoca, Veterinary
Medicine, 2010, 67:1, pp. 202-209.
7. Matei A., Oprescu A., Paca I., Doli M., Dezmirean
D., Sudiul manifestrii heterozisului la unele caractere
cantitative la Bombyx mori. Anual Session of Sci.
Comm. Probleme actuale i de perspectiv n
zootehnie, Iai, 2002, 63.
8. Lazr, ., O. C. Vornicu, Bazele biologice i tehnice
ale produciei sericicole, Ed. Ion Ionescu de la Brad,
Iai, 2013.
9. Trag, A. R., Evolution of high yielding bivoltine
silkworm (Bombyx mori L.) Genotypes, Sericologia
1992, 32:2, pp. 321-324.
10. Rao, S.G., Sahai, V., Combining ability and
heterosis studies in bivoltine strains of silkworm,
Bombyx mori L., U.P.J. Zoolegy 9, 1990, pp. 150-164.
11. Vassileva, Y., Tzenov, P., Braslavski, M.,
Poluciavane na hibridi ot tipa partenoclon po poroda,
Jivotnovodni nauki, XLI, 2004, pp. 42-44.
12. Benea, M., PhD Thesis. USAMV Cluj-Napoca,
2006.
13. nFurdui, E.M., PhD Thesis. USAMV Cluj-Napoca,
2011.
14. Paca I., PhD Thesis. USAMV Cluj-Napoca, 2004.
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diferitelor sisteme de hibridare la Bombyx mori L.,
Hungarian Symposium Musayev E. NDT and E Int.,
1993,
38,
pp.
59-68.

registered in ACxAB hybrid (93.44%) and

96.52% was the biggest one in reciprocal crossing.
The coefficient of linear regressions (R2)
calculated for hatching percentage and the number
of eggs/spawning was 0.07 for pure breeds and
0.02 for hybrids, demonstrating that the two
characters are not powerful interrelated.
The variability coefficients (V%) calculated for
the number of laid eggs and hatching percentage
were in all cases under 15%, so the samples
averages of studied breeds were representative and
the analyzed samples were homogenous.
Acknowledgments
The authors are grateful to S.C. Sericarom Bneasa
(Bucharest) Research Unit, for providing the
biological material.

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