Research on various kinds of fungi

Sections

I. - INTRODUCTION
II. - MATERIALS AND METHOD
III. SPECIES OF FUNGI ISOLATED FROM SAMPLES COLLECTED FROM ALL OPIUM-PURCHASING AGENCIES
IV. - REMOVAL OF ISOLATED FUNGI WITH FUNGICIDES
V. - STUDY OF THE DOMINANT CHARACTERISTICS OF FUNGI
VI. - CONCLUSIONS
VII. - LITERATURE

Details

Author: Dr. Hikmet Eltutar, Mustafa Igneciler,
Pages: 25 to 34
Creation Date: 1960/01/01

Research on various kinds of fungi

observed on Turkish opium and their removal with different sorts of fungicide

Dr. Hikmet Eltutar , chemist &
Mustafa Igneciler, agricultural engineer
Soil Products Office of Turkey, Opium Section

I. - INTRODUCTION

It was observed on some occasions that opium purchased in different regions of this country was covered with mould. We therefore proceeded to research with the purpose of determining species of fungi causing moldiness on opium and their action on the opium alkaloids.

The first part of our work on the subject was published in 1958 in the United Nations Bulletin on Narcotics, vol. X, No. 4. We then learned from The Journal of Japanese Botany, vol. 33, No. 2, February issue, pp. 24-27, that Takashi Matsushima & Haruyo Asahina of the National HygienicLaboratory, Tokyo, Japan, had been carrying out research on the same subject, results of which were shown in the above journal. These scientists succeeded in isolating a new species which they called Oospora sp., growing on Turkish opium. Thereupon, we reverted to the subject and proceeded to comparative studies on the results obtained by the Japanese scientists.

The second part of our work was reported in 1959 as" Species of fungi isolated from moldy samples of opium from ten different regions ".

During our research we did not find any Oospora sp. as determined by the Japanese scientists on Turkish opium. However, before stating definitely that no Oospora sp. growth was observed on Turkish opium, we thought that research should be first extended to the entire country. As explained hereafter, we carried out our research on 265 samples, which we personally collected, ensuring sterilization, from 75 opium purchasing agencies in Turkey.

Eleven species of fungi causing mould on opium, spreading over opium-producing areas in this country, were isolated and forwarded to Holland and England for identification. In the meantime, we proceeded to apply different fungicides with varied active elements with the purpose of removing fungi attacking opium.

We would like to express our gratitude to Dr. Haydar Okaner, Assistant General Director of the Turkish Soil Products Office, Ankara, and Mr. H?rrem Balkan, Director of the Opium Department, for their help and for making it possible for us to carry out the work on opium, this product being an important source of income for this country. We further extend our heartfelt thanks to Mr. Selman Açba, Director of the Opium Department, Istanbul, for his interest in our work and his encouragement, and to Prof. Dr. Bekir Alkan of the Ankara University and Doc. Dr. Ibrahim Karaca for their efforts and help on the isolation of fungi. Furthermore we wish to express our deep gratitude to the Commonwealth Mycological Institute, Kew, Surrey, England, and the Centraalbureau voor Schimmelcultures, Baarn, Holland, for their most valuable co-operation in the identification of fungi.

II. - MATERIALS AND METHOD

A. - Materials

The 265 samples of opium used in our tests were collected by ourselves under required conditions of sterilization, from the 75 opium-purchasing agencies of the Turkish Soil Products Office. The jars in which we placed the opium samples were previously sterilized, and their lids were closed.

TABLE 1

Origin and number of opium samples

Origin

Province

Number of samples

Dates on which Samples were drawn

Collected by

Suhut
Afyon
6
20.VII.59
M. Igneciler
Yildirimkemal
,, 4 ,, ,,
Sincanli
,, 4 ,, ,,
Emirdag
,, 6
21.VII.59
,,
Bolvadin
,, 6 ,,
.
Cay
,, 6 ,, ,,
Sultandag (Ishakli)
,, 4 ,, ,,
Afyon
,, 9
22.VII.59
,,
Aksehir
Konya
8
24.VII.59
,,
Sandikli
Afyon
4
25.VII.59
,,
Dinar
,, 4 ,, ,,
Burdur
Burdur
4 ,, ,,
Korkuteli
Antalya
6 ,, ,,
Tefenni
Burdur
5
27.VII.59
,,
Isparta
Isparta
6
28.VII.59
,,
Keciborlu
,, 6 ,, ,,
Uluborlu
,, 4 ,, ,,
Senirkent
,, 4 ,, ,,
Egridir
,, 4
29.VII.59
,,
Gelendost
,, 4 ,, ,,
S. Karaagac
,, 4 ,, ,,
Yalvac
,, 6 ,, ,,
Konya
Konya
4
31.VII.59
,,
Karaman
,, 5
3.VIII.59
,,
Ilgin
,, 4
7.VIII.59
,,
Argithan
,, 5
8.VIII.59
,,
Beysehir
,, 6
11.VIII.59
,,
Seydisehir
,, 6 ,, ,,
Bozkir
,, 5 ,, ,,
Kadinhan
,, 4
14.VIII.59
,,

TABLE 1 (continued)

Origin

Province

Number of samples

Dates on which Samples were drawn

Collected by

Gümüşhaciköy
Amasya
3
27.VII.59
H. Eltutar
Merzifon
,, 4 ,, ,,
Corum
Corum
1 ,, ,,
Mecitözü
,, 4
28.VII.59
,,
Amasya
Amasya
5
28.VII.59
,,
Erbaa
Tokat
1
29.VII.59
,,
Niksar
,, 3 ,, ,,
Tokat
,, 5 ,, ,,
Zile
,, 5
30.VII.59
,,
Malatya
Malatya
4
3.VIII.59
,,
Nallihan
Ankara
5
5.VIII.59
,,
Beypazari
,, 5 ,, ,,
Geyve
Kocaeli
1
19.VIII.59
,,
Göynük
,, 6 ,, ,,
Banaz
Afyon
6
24.VIII.59
,,
Usak
Usak
7 ,, ,,
Ulubey
,, 2
25.VIII.59
,,
Sivasli
,, 3 ,, ,,
Simav
Kütahya
9
26.VIII.59
,,
Gediz
,, 6 ,, ,,
Kütahya
,, 2
27.VIII.59
,,
Tavsanli
,, 9 ,, ,,
Emet
,, 6 ,, ,,
Seyitgazi
Eskisehir
4
28.VIII.59
,,
Sivrihisar
,, 3 ,, ,,
Yunusemre
,, 4 ,, ,,
Eskisehir
,, 4 ,, ,,
Bilecik
Bilecik
2
29.VIII.59
,,
Gölpazari
,, 2 ,, ,,
Inegöl
Bursa
4
31.VIII.59
,,
Denizli
Denizli
8
6.IX.59
,,
Dazkiri
,, 3 ,, ,,
Cal
,, 6
7.IX.59
,,
Acipayam
,, 7 ,, ,,
Civril
,, 4 ,, ,,
Aydin
Aydin
2
8.IX.59
,,
Kula
Manisa
2
10.IX.59
,,
Alasehir
,, 3 ,, ,,
Salihli
,, 5 ,, ,,
Selendi
,, 3
11.IX.59
,,
Sindirgi
Balikesir
1 ,, ,,
Akhisar
Manisa
2
12.IX.59
,,
Balikesir
Balikesir
2 ,, ,,
Dursunbey
,, 5
14.IX.59
,,
Orhaneli
Bursa
4
16.IX.59
,,

Mouldy lumps were selected among opium purchased by agencies in the above districts; these lumps were cut in the middle and classified according to their appearance. About 20-30 g were taken together with their media from different lumps of opium which had the same colour and aspect, and these were placed in jars (method 1). The jars were closed and isolated with paraffin; then labels were attached to the jars with indication of the origin of the sample, the marks of the locality, the date on which the sample was drawn, and the name of the person who collected the sample.

Fungicides used in tests were supplied by the Ministry of Agriculture General Directorate of Quarantine and the Faculty of Agriculture Plant Protection Chair Laboratory.

Full size image: 18 kB

B. - Method

The medium used for growing fungi was Czapek-Agar with addition of 20% sucrose. After carefully preparing the medium and placing it in petri boxes, fungi from different samples were inoculated to each one separately. Fungi thus inoculated in petri were left to grow in an incubator at 26°C.

In order to test fungicide action on fungi, the products were evenly mixed with Czapek-Agar media with addition of 20% sucrose and placed in petris. Each species of fungi isolated from samples was separately introduced in petri boxes containing media with fungicides; these were then placed in an incubator at 26°C, and their action was tested from time to time. Products containing copper as active element were tested twice for each fungus species; the other fungicides were tested ten times.

TABLE 2

Names of fungicides used in tests, names of manufacturers and active elements

Product

Manufacturer

Percentage of active element

Copramat
Ciba
Ziran, 27.25
   
Copper, 32.84
Cu- Sul
Neptune
Copper, 10.50
   
Sulphur, 19.35
Brestan
Hoechst
Triphenyl-sincacetate, 20
Hortexan
Agro-Merck
Lindane, 20
   
Phenylqueck-silveracetate, 3.20
Formalin
Merck
Formaldehyde

Full size image: 200 kB

As for the determination of the dominant characteristics of fungi species with regard to each other, we used petri boxes of 15-18 cm diameter. We poured Czapek-Agar media with 20% sucrose in these, and inoculated isolated fungi at determined distances (method 2).

Growth of the above in an incubator at 26°C was studied. This test was repeated three times.

III. SPECIES OF FUNGI ISOLATED FROM SAMPLES COLLECTED FROM ALL OPIUM-PURCHASING AGENCIES

As explained in the "Materials" section, a laboratory number was given to each sample of mouldy opium with regard to its origin. Fungi drawn from mouldy parts of the above opium were placed in 20% sucrose-added Czapek-Agar medium, and kept in an incubator at 26°C. It was observed that these fungi appeared visibly 3 days after having been planted. Later on, they were taken out of the incubator and left at room temperature (18°C). A week later, their characteristic colours and shapes could be observed. These fungi were then classified according to their colour and shape.

In order to obtain pure cultures from fungi which were believed to be different from the colour and shape standpoint, isolation was repeated several times, and then these were forwarded to England and Holland for identification.

Fungi species growing on Turkish opium

Aspergillus niger van Tieghem

Aspergillus repens Corda Sacc. (Cda) de Bary

Aspergillus flavus Link

Aspergillus wentii Wehmer

Aspergillus ostianus Wehmer

Fusarium solani (Mart.) App. et emend. Sny. et Hans

Fusarium exysporum Schlect

Penicillium granulatum Bain

Penicillium expansum Link

Penicillium cyclopium Westling

Penicillium stoloniferum Thom

Fungi indicated by letters A, B, C, D, E, F and G in table 3 are the following:

A: Aspergillus niger van Tieghem.

B: Aspergillus repens (Corda) Sacc. (Cda) de Bary.

C: Penicillium expansum Link.

Penicillium cyclopium Westling.

Penicillium stoloniferum Thom.

Penicillium granulatum Bain.

D: Aspergillus flavus Link.

E: Fusarium solani (Mart.) App. et Wr. emend. Sny. et Hans. Fusarium oxysporum Schlect.

F: Aspergillus wentii Wehmer.

G: Aspergillus ostianus Wehmer.

As will be noted from table 3, three different kinds of fungus causing mouldiness on opium and spreading over production areas have been found. These are Aspergillus, Penicillium and Fusarium. Among them, the following species were determined:

  1. Aspergillus: Asp. niger van Tieghem; Asp. repens Corda Sacc. (Cda) de Bary; Asp. flavus Link; Asp. wentii Wehmer; Asp. ostianus Wehmer.

  2. Penicillium: P. granulatum Bain; P. expansum, Link; P. Cyclopium Westling; P. stoloniferum Thom.

  3. Fusarium: F. oxysporum Schlect.; F. solani (Mart.) App. et Wr. emend. Sny. et Hans.

On 265 samples used in tests, Aspergillus was found on 240, Penicillium on 165 and Fusarium on 16. With regard to origin, Aspergillus was found everywhere, Penicillium was fond in 71 districts excluding Çorum, Geyve, Kadinhan and Kütahya; as for Fusarium, it was found in only 12 districts, namely: Yildirimkemal, Sincanli, Çay, Ilgin, Argithan, Egirdir, S. Karaagac, Yalvac, Simav, Gediz, Zile and Denizli.

The spread of fungi species through opium-producing areas is as follows:

 
Number of districts
Aspergillus niger
73
Aspergillus repens
55
Aspergillus flavus
27
Aspergillus wentii
12
Aspergillus ostanius
5
Fusarium solani and F. oxysporum
11
4 kinds of Penicillium (as mentioned above)
71

In conclusion, it was observed that Aspergillus niger, Penicillium granulatum, Penicillium expansum, Penicillium cyclopium and Penicillium stoloniferum are widely spread over the Turkish opium-producing areas, whereas Aspergillus wentii, Aspergillus ostianus, Fusarium solani and Fusarium oxysporum are found in small quantities, and that ecological and climatic conditions have no relation with the development of such fungi.

IV. - REMOVAL OF ISOLATED FUNGI WITH FUNGICIDES

Into 100 cm 3 of the medium used in tests, we added 1 g of each fungicide containing a different active element with 3 g of Formalin, and we prepared different kinds of medium with this. After placing the media in petri containers, we inoculated isolate fungus species in each one separately, then left it to grow in an incubator at 26°C together with a blank (same species in media free of fungicide).

As will be noted from table 4, some products had no action on fungi, whilst others showed conclusive and lasting action.

TABLE 3

Species of fungi isolated from samples collected from all opium-purchasing agencies

Origin

District No.

Laboratory No.

Species of isolated fungi

Origin

District No.

Laboratory No.

Species of isolated fungi

Salihli
2 1
A
         
Amasya
2 49
A
 
C
     
  3 2  
B
          3 50
A
 
C
   
F
  1 3
A
 
C
        1 51  
No growth
       
  4 4  
B
C
        4 52
A
B
     
F
                    5 53
A
   
C
   
Seyitgazi
1 5
A
 
C
                       
  3 6      
D
   
Dinar
1 54    
C
     
  2 7    
C
D
     
C/1
55
A
B
       
  4 8      
D
      2 56
A
 
C
     
                   
C/A
57
A
         
Acipayam
1 9
A
       
F
                 
  3 10
A
B
     
F
                 
  2 11
A
 
C
     
Ccrum
1 58
A
         
  4 12  
B
                         
                 
Geyve
1 59
A
         
Eskisehir
1 13
A
                           
  3 14
A
                           
  2 15
A
 
C
     
Korkuteli
1 60
A
 
C
     
  4 16  
B
         
C/1
61
A
 
C
D
   
                    2 62    
C
     
                   
C/2
63
A
 
C
     
Akhisar
2 17
A
 
C
                       
 
C/2
18
A
B
C
D
   
Bilecik
2 64
A
 
C
     
  1 19    
C
        1 65
A
 
C
     
 
C/1
20
A
   
D
                     
Sultandag
2 21
A
       
F
Tefenni
2 66    
C
     
 
C/2
22          
F
 
C/2
67
A
B
C
     
  1 23
A
B
C
        1 68    
C
     
 
C/1
24  
B
         
C/1
69  
No growth
       
Yildirimkemal
2 25
A
 
C
     
Erbaa
1 70    
C
D
   
 
C/1
26
A
 
C
                       
  1 27
A
 
C
                       
 
C/2
28
A
B
C
 
E
 
Karaman
1 71
A
         
                   
C/1
72
A
B
C
     
Suhut
2 29
A
 
C
        2 73
A
 
C
D
   
 
C/2
30
A
B
         
C/2
74
A
 
C
     
  1 31    
C
                       
 
C/1
32
A
B
                         
                 
Argithan
1 75
A
 
C
     
                   
C/1
76
A
 
C
 
E
 
Simav
1 33        
E
F
  2 77
A
 
C
D
   
  3 34
A
 
C
D
 
F
 
C/2
78
A
 
C
     
  2 35
A
   
D
                     
  4 36
A
       
F
                 
                 
S.Karaagac
2 79
A
 
C
     
Ilgin
2 37
A
   
D
E
   
C/2
80  
B
       
 
C/2
38
A
 
C
        1 81
A
 
C
 
E
 
  1 39
A
 
C
D
     
C/1
82
A
B
   
E
 
 
C/1
40
A
 
C
D
E
                   
                 
Yalvac
1 83    
C
D
   
Seydisehir
1 41
A
 
C
D
     
C/1
84  
No growth
       
 
C/1
42
A
 
C
        2 85  
B
C
     
  2 43
A
           
C/2
86
A
 
C
 
E
 
 
C/2
44  
B
 
D
                     
Orhaneli
1 45  
B
C
     
Konya
1 87    
C
     
  3 46  
B
         
C/2
88  
B
       
  2 47    
C
        2 89
A
 
C
     
  4 48  
B
         
C/1
90  
B
       

TABLE 3 (continued)

Origin

District No.

Laboratory No.

Species of isolated fungi

Origin

District No.

Laboratory No.

Species of isolated fungi

Senirkent
1 91
A
B
C
     
Emet
2 133
A
B
C
   
F
 
C/1
92
A
B
          3 134
A
 
C
   
F
  2 93    
C
        1 135
A
B
C
   
F
 
C/2
94
A
B
          4 136
A
       
F
                 
Gediz
2 137
A
B
C
     
Gelendost
1 95
A
 
C
        3 138
A
   
D
   
 
C/1
96
A
 
C
        1 139
A
     
E
 
  2 97    
C
        4 140  
B
       
 
C/2
98
A
B
C
                       
                 
Sivasli
1 141
A
         
Uluborlu
1 99
A
B
          3 142
A
 
C
   
G
 
C/1
100
A
B
C
        2 143
A
 
C
     
  2 101  
B
C
                       
 
C/2
102
A
B
C
     
Inegöl
1 144
A
 
C
     
                    3 145
A
 
C
     
Keciborlu
1 103
A
B
C
     
Inegöl
2 146
A
B
C
     
 
C/1
104
A
 
C
        4 147
A
         
  2 105    
C
                       
 
C/2
106
A
 
C
                       
                 
Beysehir
1 148
A
B
C
     
                   
C/1
149
A
B
C
     
Mecitözü
2 107    
C
        2 150
A
 
C
     
  4 108
A
 
C
       
C/2
151
A
       
F
  1 109    
C
                       
  3 110  
B
       
Tavasanli
2 152
A
 
C
     
                    3 153
A
B
       
                    1 154
A
 
C
     
Sincanli
2 111
A
 
C
D
E
                   
 
C/2
112
A
 
C
D
   
Kula
2 155
A
 
C
     
  1 113
A
 
C
D
      1 156
A
 
C
     
 
C/1
114
A
 
C
 
E
                   
                 
Aksehir
1 157
A
B
C
     
Nallihan
2 115    
C
   
F
  3 158
A
 
C
     
  3 116
A
       
F
  2 159
A
B
C
     
  1 117
A
       
F
                 
  4 118
A
       
F
Isparta
1 160
A
B
C
     
                   
C/1
161  
B
       
                    2 162
A
 
C
     
Niksar
2 119
A
 
C
       
C/2
163
A
 
C
     
  3 120  
B
                         
  1 121
A
 
C
                       
                 
Gümüshaciköy
2 164
A
 
C
     
                    3 165      
D
   
                    1 166    
C
     
Dazkiri
1 122
A
 
C
                       
  3 123
A
B
                         
  2 124
A
B
C
     
Selendi
2 167
A
 
C
     
                    3 168
A
 
C
D
   
                    1 169
A
 
C
     
Beypazari
1 125
A
 
B
C
D
                   
  3 126
A
 
B
     
Emirdag
2 170
A
 
C
   
F
  2 127
A
           
C/1
171
A
B
     
F
  4 128
A
            1 172
A
 
C
   
F
                   
C/2
173
A
       
F
Merzifon
2 129
A
 
C
                       
  3 130
A
B
C
D
   
Zile
1 174  
B
C
D
   
  1 131
A
 
C
        4 175
A
     
E
 
  4 132
A
B
          2 176  
B
       
                    3 177
A
B
       

TABLE 3 (conclusion)

Origin

District No.

Laboratory No.

Species of isolated fungi

Origin

District No.

Laboratory No.

Species of isolated fungi

Bolvadin
2 171
A
   
D
     
Alasehir
1 224
A
 
C
     
 
C/2
179
A
 
C
D
        2 225
A
B
       
  1 180
A
B
                           
 
C/1
181
A
 
C
D
   
G
                 
                   
Civril
1 226
A
B
C
     
                      2 227
A
B
C
     
Göynök
2 182  
B
C
          3 228
A
B
C
     
  3 183
No growth
              4 229
A
B
C
     
  1 184    
C
                         
  4 185
A
B
                           
                   
Yunusemre
2 230
A
 
C
     
                      4 231  
B
       
Cay
2 186
A
B
C
   
F
    1 232    
C
     
 
C/2
187
A
B
C
   
F
    3 233
A
B
C
     
  1 188
A
 
C
                         
 
C/1
189
A
     
E
F
                   
Tokat
1 190
A
B
         
Sivrihisar
1 234    
C
     
  3 191  
B
            3 235
A
 
C
   
G
  2 192
A
B
C
          2 236
A
 
C
     
  4 193
A
   
D
   
G
                 
                   
Ulubey
1 237
A
B
C
     
Malatya
1 194  
B
C
          2 238
A
         
  3 195
A
   
D
                       
  2 196      
D
     
Sandikli
1 239
A
   
D
   
  4 197
A
   
D
       
C/1
240
A
 
C
     
                      2 241
A
         
Burdur
1 198
A
 
C
         
C/2
242    
C
D
   
 
C/1
199  
B
C
                         
  2 200
A
           
Bozkir
2 243
A
         
 
C/2
201
No growth
             
C/2
244
A
 
C
     
                      1 245
A
B
C
     
                     
C/2
246
A
 
C
   
G
Egirdir
2 202    
C
                         
 
C/2
203
A
 
C
                         
  1 204
A
 
C
       
Aydin
2 247
A
         
 
C/1
205        
E
      1 248
A
 
C
     
Banaz
1 206
A
 
C
       
Cal
1 249
A
         
  3 207
A
B
C
          3 250
A
B
       
  2 208
A
 
C
          2 251
A
 
C
     
  4 209
A
 
C
          4 252
A
 
C
     
Balikesir
1 210
A
B
C
       
Denizli.
1 253
A
 
C
 
E
 
  2 211    
C
          3 254
A
 
C
D
   
                      2 255
A
         
                      4 256
A
B
C
 
E
 
Dursunbey
2 212
A
 
C
                         
  4 213
A
       
F
                   
  1 214
A
B
C
       
Gölpazari
1 257
A
B
C
     
  3 215
A
 
C
D
        2 258
No growth
         
Afyon
1 216
A
B
C
       
Kadinhan
1 259      
D
   
 
C/1
217
A
 
C
         
C/1
260
A
   
D
   
  2 218
A
 
C
          2 261
A
         
 
C/2
219
A
       
F
   
C/2
262
A
   
D
   
Usak
1 220
A
B
C
       
Kütahya
1 263
A
         
  3 221
A
   
D
        2 264
A
B
       
  2 222
A
B
C
                         
  4 223  
B
         
Sindirgi
1 265
A
 
C
D
   

TABLE 4

Number of days elapsed before growth of fungi in artificial media with fungicide

 

Number of days

Name of product

Fungus mark

1

2

3

4

5

6

7

8

...

24

25

26

27

28

...

60

Hortexan, 1 g
A
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
B
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
C
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
D
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
E
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
F
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
G
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
Blank
 
-
-
-
+
+
+
+
+
 
+
+
+
+
+
 
+
                                   
Cu-Sul, 1 g
A
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
 
B
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
 
C
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
 
D
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
 
E
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
 
F
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
 
G
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
Blank
 
-
-
-
+
+
+
+
+
 
+
+
+
+
+
 
+
                                   
Coprantol, 1 g
A
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
 
B
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
 
C
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
 
D
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
 
E
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
 
F
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
 
G
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
Blank
 
-
-
-
+
+
+
+
+
 
+
+
+
+
+
 
+
                                   
Copromat, 1 g
A
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
 
B
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
 
C
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
 
D
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
 
E
-
-
-
-
+
+
+
+
 
+
+
+
+
   
+
 
F
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
 
G
-
-
-
-
+
+
+
+
 
+
+
+
+
+
 
+
Blank
 
-
-
-
+
+
+
+
+
 
+
+
+
+
+
 
+
                                   
Brestan, 1 g
A
-
-
-
-
-
-
-
-
 
-
-
+
+
+
 
+
 
B
-
-
-
-
-
-
-
-
 
-
+
+
+
+
 
+
 
C
-
-
-
-
-
-
-
-
 
-
+
+
+
+
 
+
 
D
-
-
-
-
-
-
-
-
 
+
+
+
+
+
 
+
 
E
-
-
-
-
-
-
-
+
 
+
+
+
+
+
 
+
 
F
-
-
-
-
-
-
-
+
 
+
+
+
+
+
 
+
 
G
-
-
-
-
-
-
-
-
 
-
+
+
+
+
 
+
Blank
 
-
-
-
+
+
+
+
+
 
+
+
+
+
+
 
+
                                   
Formalin, 3 g
A
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
B
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
C
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
D
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
E
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
F
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
G
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
Blank
 
-
-
+
+
+
+
+
+
 
+
+
+
+
+
 
+

The sign (-) in the above table signifies that there was no growth of fungus. The sign (+) signifies that growth had begun. The capital letters stand for fungus species as indicated in table 3. As will be noted from the table, blanks developed visibly 3 days after being planted. All fungi in media with Cu-Sul, Coprantol and Copromat which contain copper began to grow on the fifth day following the date on which they were planted.

In media with Brestan, Aspergillus wentii, Fusarium appeared on the 8th day; Aspergillus flavus the 24th day; Aspergillus repens, Aspergillus ostianus and all Penicillium species appeared on the 25th day; Aspergillus niger developed 33on the 26th day. No species planted in media containing Hortexan and Formalin developed within 60 days. Cu-Sul, Coprantol, Copramat and Brestan had no complete action on fungi and we therefore had to be content with these tests.

As for Hortexan and Formalin, these products showed conclusive and lasting action, and research was continued using the above fungicides in order to determine their minimum action dosage. The results are shown in table 5.

TABLE 5

Minimum doses of fungicides showing conclusive action

 

Number of days

Name of product

Fungus mark

1

2

3

4

5

6

7

8

...

24

25

26

27

28

...

60

Hortexan, 0.500 g
A
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
B
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
C
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
D
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
E
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
F
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
G
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
Blank
 
-
-
-
+
+
+
+
+
 
+
+
+
+
+
 
+
                                   
Hortexan, 0.250 g
A
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
B
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
C
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
D
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
E
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
F
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
G
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
Blank
 
-
-
-
+
+
+
+
+
 
+
+
+
+
+
 
+
                                   
Formalin, 2 g
A
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
B
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
C
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
D
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
E
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
F
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
G
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
Blank
 
-
-
-
+
+
+
+
+
 
+
+
+
+
+
 
+
                                   
Formalin, 1 g
A
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
B
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
C
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
D
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
E
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
F
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
 
G
-
-
-
-
-
-
-
-
 
-
-
-
-
-
 
-
Blank
 
-
-
-
+
+
+
+
+
 
+
+
+
+
+
 
+

It will be observed from the above table that Hortexan and Formalin showed conclusive action even with minimum doses. On basis of these results obtained in artificial media, tests will be carried out on natural media (opium).

V. - STUDY OF THE DOMINANT CHARACTERISTICS OF FUNGI

Research was carried out with the purpose of determining whether fungi causing mouldiness on opium showed any predominance over each other under equal conditions regarding medium, temperature and culture.

In tests carried out on the subject, as described in the "Materials" section of this report, 20% sucrose added Czapek-Agar was placed in large petri containers of 15-18 cm diameter, then each species of fungi was planted at equal distances using a fine needle. Fungi thus inoculated developed in three days at 25°C and colonies covered entirely the petri surface within a week. No marked difference was observed on the spreading area of fungi in the petri containers.

VI. - CONCLUSIONS

  1. - Fungi species causing mouldiness on opium throughout opium-producing regions of Turkey are the following:

Aspergillus niger van Tieghem

Aspergillus repens Corda Sacc. (Cda) de Bary

Aspergillus flavus Link

Aspergillus wentii Wehmer

Aspergillus ostianus Wehmer

Fusarium solani (Mart.) App. et Wr. emend. Sny. et Hans.

Fusarium oxysporum Schlect.

Penicillium granulation Bain

Penicillium expansum Link

Penicillium Cyclopium Westling

Penicillium stoloniferum Thom

  1. - Spread of the above species in this country is shown in table 4.

  2. - Fungicides which gave the best results for the removal of fungi were Hortexan and Formalin.

VII. - LITERATURE

  1. Thom, Charles & Raper, Kenneth B.: 1945, A Manual of the Aspergilli, The Williams and Wilkins Company, Baltimore, 373.

  2. Centraalbureau voor Schimmelcultures, 1947 , List of Cultures, Baarn, Holland, 145.

  3. Takashi Matsushima, Haruryo Asahina: 1958, Oospora Sp. isolated from moldy Opium, The Journal of Japanese Botany, vol. 33, No. 2 (February), pp. 24-27.

  4. Eltutar, H. & Igneciler, M.: Bulletin on Narcotics, vol. X, No. 4, October-December 1958, pp. 12-16.