Br. vet .
1. (1990) . 146, 175
PATHOLOGICAL CHANGES IN MALE GENITALIA OF CATTLE INFECTED WITH TRYPANOSOMA VIVAX AND
TR YPANOSOMA CONGOLENSE
V . O . SEKONI*, C . O . NJOKUt, J . KUMI-DIAKAt and D . I . SARORt *National Animal Production Research Institute, Ahmadu Bello University, P.M. B. 1096, Shika, .aria, ,Nigeria; tFaculty of Veterinary Medicine, Ahmadu Bello University, .aria, Nigeria
SUMMARY
Samples for histological studies were taken from the genitalia of 14 bulls (five infected with Trypanosoma vivax, five with T. congolense and four uninfected control animals), slaughtered 12, 22 or 30 weeks post-infection . Infection with Y58 strain of T. vivax and strain 2295 of T. congolense caused various grades of lesions in the male reproductive organs, especially the testes and epididymides . T. congolense produced more severe degenerative changes than T. vivax . It is concluded that in long-standing cases, the result of trypanosome infection is either serious infertility or even sterility .
INTRODUCTION
Testicular degeneration has been reported in sheep and goats infected with T. vivax (Isoun & Anosa, 1974 ; Anosa & Isoun, 1980) and in sheep, rabbit and mice infected with T brucei (Ikede, 1979 ; Ikede & Akpavie, 1982 ; Anosa & Kaneko, 1984) . In contrast there is little information on the pathological effects of trypanosomiasis in cattle apart from preliminary studies by Isoun, Akpokodje & Anosa (1975) who reported testicular damage in a White Fulani (Bunaji) bull infected with T. vivax. Genital lesions can lead to deterioration of seminal characteristics and eventually to cessation of spermatogenesis when the testes themselves are involved (Lagerlof, 1936 ; Rao, 1971 ; Kumi-Diaka, 1981) . Bulls infected with T vivax or T. congolense have elevated reaction, deteriorated semen characteristics and in some instances a cessation of semen production (Sekoni et al., 1988) . The aim of the present investigation was to conduct a detailed study of the genital pathology of bulls infected with T. vivax or T. congolense .
MATERIALS AND METHODS
Fourteen normal healthy Zebu bulls aged between 3 and 5 years were used . The semen characteristics were indicative of normal genital function . The bulls were housed individually in fly-proof accommodation throughout the investigation . They were fed on concentrates, fresh pasture, hay, silage, salt licks and water ad libitum.
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BRITISH VETERINARY JOURNAL, 146, 2
The bulls were placed into two groups of five animals and a third group of four . The three groups were •kept in the same building but in different rooms, completely partitioned from each other to prevent mixed infections . Groups 1 and 2 comprising five bulls each were infected with T. vivax and T. congolense respectively and the four animals in group 3 served as controls . The Y58 strain of T vivax and strain 2295 of T congolense, both isolated from natural infections in cattle in Northern Nigeria, were used to infect the bulls . The stabilates were first inoculated into mice and then transferred into goats . Two ml of blood containing approximately 2X10' trypanosomes from the infected goats were inoculated i .v . into each bull. Parameters monitored twice weekly post-infection were : rectal temperatures, packed cell volume, haemoglobin and total plasma proteins . Measurement of scrotal circumference, palpation of the scrotal contents and internal genitalia (via rectum) was carried out weekly . Histopathological studies
Samples for histopathological studies were taken from the genitalia of all 14 bulls . Thirteen of the bulls were slaughtered at 12, 22 or 30 weeks post-infection ; the remaining animal died 8 weeks after infection with T congolense. The samples were taken from the testes, epididymides, seminal vesicle, prostate gland, ampullae, pelvic urethra and penis following gross examination of the organs . Tissues taken at necropsy were fixed in Bouin's solution for 72 hours and processed according to standard histopathological techniques . They were stained with haematoxylin and eosin (H & E), periodic acid schiff (PAS) and Van Gieson (VG) . The presence and conditions of the geminal epithelia and Sertoli cells were studied microscopically . The interstitial tissues were examined and the occurrence of lesions and their frequency recorded . Degrees of testicular degeneration were scored as shown in Table I . Sections from the epididymides and accessory glands were similarly studied and lesions scored as in Table I .
Table I Grading of genital degeneration Epididymides and Grade
Testes
accessory glands
Severe +++
Germinal epithelial layers reduced to one or two with predominantly Sertoli cells . Sixty (60)% plus tubules affected
Lesions observed in 60% or more tubules
Moderate ++
Germinal epithelial layers reduced to 3 with at least 50% of tubules affected
Lesions observed in at least 50% of tubules
Mild +
Most tubules normal with slight reduction in the germinal epithelial layers
Most tubules are normal
T. VIVAX AND T. CONCOLENSE IN BULLS
1 77
Estimation of epididymal sperm reserve At least 50 epididymal tubules per slide were observed at X100 for determining spermatozoa content. The epididymal sperm reserve was then expressed as a percentage of the number of tubules with normal sperm reserve out of the total examined .
RESULTS All the bulls infected with T. vivax and T. congolense developed chronic trypanosomiasis and were all positive for trypanosomes within 8 days of infection . Clinical signs which were more pronounced in the T congolense infected bulls included fluctuating pyrexia (105-106°F), rapid weight loss, lethargy, weakness, dull and rough hair coats, pale mucous membranes, and enlarged superficial lymph nodes ; anaemia and hydropericardium occurred in some animals at post mortem . The packed cell volume, haemoglobin, and total plasma protein of the infected bulls decreased and differed significantly from control bulls .
Gross findings The male genitalia of the infected and control bulls appeared normal .
Histopathological findings Control bulls. The testes, epididymides, epididymal sperm reserve (100%) seminal vesicles, prostate glands, ampullae, urethra, penis, glans penis and prepuce were normal . T . vivax infected bulls. One animal had mild and one moderate testicular degeneration with severe testicular degeneration in the remaining three . In one animal, the infection had destroyed all the cellular structures within the interstitial tissues and seminiferous tubules . Two animals had mild and three had moderate epididymal degeneration . All the infected bulls had significant decreases in their epididymal sperm reserve ranging between 20 and 50% . Three animals had mild and four had moderate degeneration of the seminal vesicles and prostate glands . None of the bulls had lesions in the Cowper's glands, while only one animal had moderate degeneration of the ampullae . No lesions were present in the urethra, penis, or glans penis of any of the bulls . T. congolense infected bulls . Testicular degeneration was more severe in the bulls infected with T. congolense than in those infected with T vivax. One animal had moderate and four had severe degeneration . In three animals, all the cellular structures within the interstitial tissues and seminiferous tubules were destroyed . Seminiferous tubules were also collapsed in the testes of the three animals (Fig . 1) . Similarly, epididymal degeneration was more marked in the T congolense infected animals (Fig . 2) . Two animals had severe, one moderate and two had mild epididymal degeneration . All the infected animals had marked depletion of the epididymal sperm reserves which were zero % in two, 20% in one and 50% in two bulls . Degenerative changes were also more marked in the seminal vesicles of the T congolense infected bulls . Two animals had severe and three had mild degeneration of the seminal vesicles . Degenerative changes which were mild in three animals and severe in one animal were recorded in the prostate glands of the T congolense infected animals . Degeneration of the
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BRITISH VETERINARY JOURNAL, 146, 2
Fig. 1 . Severe testicular degeneration in a White Fulani bull which died 8 weeks post-infection with T. congolense. Note collapsed tubules . (H & E X200) .
Fig. 2 . Severe epididymal degeneration in a White Fulani bull which died 8 weeks post-infection with T. congolense (H & E X400) .
T VIVAX AND T. CONGOLENSE IN BULLS
1 79
prostate glands was also more marked in the T congolense infected bulls in which the only severe case was recorded . There were no lesions in the Cowper's glands of any of the infected animals . Only one infected bull had mild degeneration of the ampullae . No lesions were present in the urethra, penis, glans penis and prepuce of any of the bulls .
DISCUSSION The results showed that trypanosomiasis had a devastating effect on the male reproductive and accessory reproductive organs . The pathogenic effects of T congolense were more severe than those of T vivax in contrast to observations in West Africa where the latter is more pathogenic (Leeflang, 1978 ; Agu, 1983 ; Anosa, 1983) . The most severe degenerative changes were observed in the testes and epididymides . The decrease or absence of epididymal sperm reserves in infected bulls was most probably the result of a decrease or cessation of gonadal sperm production due to impairment of spermatogenesis (Isoun & Anosa, 1974 ; Anosa & Isoun, 1980) . The pathogenesis of testicular degeneration in trypanosomiasis is not well understood . It is possible that the intermittent pyrexia that accompanies the numerous parasitaemic peaks following infection could be partly responsible (Anosa & Isoun, 1980) . Pyrexia is known to affect spermatogenesis adversely (Lagerlof, 1936 ; Gustafsson, 1966 ; Johnson, Gomes & Vandemark, 1970 ; Arthur, 1975) . However, other factors could also be involved such as the capacity of pathogenic trypanosomes to cause extensive, generalized tissue and organ damage . Other mechanisms involved might be anoxia caused by anaemia, immunological factors, production of biologically active and toxic substances by the trypanosomes, physical swelling and disruption of tissue architecture and increased vascular permeability (Boreham & Cox, 1978 ; Tizard et al., 1978 ; Morrison, Murray & McIntyre, 1981) . The practical significance of the degenerative changes caused by trypanosomiasis on the male reproductive organs is the adverse effects on fertility brought about by disruption in testosterone production, spermatogenesis, and semen characteristics . It is highly likely that the net result of severe degenerative changes in the reproductive organs caused by trypanosomiasis is either serious infertility or even sterility in long-standing cases .
REFERENCES Acu, W . E .
(1983) . ACIAR Proceedings 4, 70 . ANOSA, V . O . (1983) . .Zentralblatt für Veterinärmedizin 30, 717 . ANOSA, V . O . & ISOUN, T . T . (1980) . ,Journal of the Nigerian Veterinary Association 3, 102 . ANOSA, V . O . & KANEKO, J . J . (1984) . Veterinary Pathology 21, 238 . ARTHUR, G . H . (1975) . Veterinary Reproduction and Obstetrics, 4th edn . London : Baillière Tindall . BOREHAM, P . F . L . & Cox, F. E . G . (1978) . Proceedings of workshop held at Nairobi, Kenya, 20-23 November. GUSrAFSSON, B . (1966) . Acta Veterinaria Scandinavica 7, supplement 17 . IKEDE, B . O . (1979) . Research in Veterinary Science 26, 145 . IKEDE, B . O . & AKPAVIE, S . O . (1982) . Research in Veterinary Science 32, 374 . ISOUN, T . T . & ANOSA, V. O . (1974) . Tropenmedizin und Parasitologie25, 469 . IsouN, T . T ., AKPOKODJE, J . U . & ANOSA, V . O. (1975) . Journal of the Nigerian Veterinary Medical Association 4, 2 .
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BRITISH VETERINARY JOURNAL, 146, 2
JOHNSON, A . D ., GoMES, W . R. & VANDEMARK, N . L . (1970) . In The Testis, Vol . 1, Ch . 4, ed . G . M . H . Waites . New York and London : Academic Press . KUMI- DIAKA, J . (1981) . PhD thesis, A . B . U ., Zaria, Nigeria . LAGERLOF, N . (1936). Veterinary Record 48, 1159 . LEEFLANG, P. (1978) . PhD thesis, University of Utrecht . MORRISON, I ., MURRAY, M . & MCINTYRE, I . (1981) . FAO/SIDA follow up seminar on Veterinary Pathology, Faculty of Veterinary Medicine, University of Nairobi, 17-27 September . RAO, A . R . (1971) . PhD thesis, Royal Veterinary College, Stockholm, Sweden . SEKONI, V . O ., KUMI-DIAKA, J ., SAROR, D . I . & NJOKU, C . O . (1988) . British Veterinary Journal144, 388 . TIZARD, I . A ., NIELSON, K ., SEED, J . R . & HALL, J . E . (1978) . Microbiology Review 42, 661 . (Accepted for publication 21 April 1989)
ANNOUNCEMENT COMISA Comisa, the Consultation Mondiale de l'Industrie de la Santé Animale (the Representative Body of the Worldwide Animal Health Industry) was officially launched on 23 October 1989 in Washington D .C . Its broad objectives include : the worldwide promotion of science and development of animal health products establishing and developing formal relations between the animal health industry and international bodies dealing with animal health affairs developing and procuring common agreement of policies to be adopted by the animal health industry in its dealings with public international organizations, both governmental and non-governmental . The Secretary of the new body with headquarters in Brussels is Dr . David Miller . Tel : 44 276 698292 .
1. (1990) . 146, 175
PATHOLOGICAL CHANGES IN MALE GENITALIA OF CATTLE INFECTED WITH TRYPANOSOMA VIVAX AND
TR YPANOSOMA CONGOLENSE
V . O . SEKONI*, C . O . NJOKUt, J . KUMI-DIAKAt and D . I . SARORt *National Animal Production Research Institute, Ahmadu Bello University, P.M. B. 1096, Shika, .aria, ,Nigeria; tFaculty of Veterinary Medicine, Ahmadu Bello University, .aria, Nigeria
SUMMARY
Samples for histological studies were taken from the genitalia of 14 bulls (five infected with Trypanosoma vivax, five with T. congolense and four uninfected control animals), slaughtered 12, 22 or 30 weeks post-infection . Infection with Y58 strain of T. vivax and strain 2295 of T. congolense caused various grades of lesions in the male reproductive organs, especially the testes and epididymides . T. congolense produced more severe degenerative changes than T. vivax . It is concluded that in long-standing cases, the result of trypanosome infection is either serious infertility or even sterility .
INTRODUCTION
Testicular degeneration has been reported in sheep and goats infected with T. vivax (Isoun & Anosa, 1974 ; Anosa & Isoun, 1980) and in sheep, rabbit and mice infected with T brucei (Ikede, 1979 ; Ikede & Akpavie, 1982 ; Anosa & Kaneko, 1984) . In contrast there is little information on the pathological effects of trypanosomiasis in cattle apart from preliminary studies by Isoun, Akpokodje & Anosa (1975) who reported testicular damage in a White Fulani (Bunaji) bull infected with T. vivax. Genital lesions can lead to deterioration of seminal characteristics and eventually to cessation of spermatogenesis when the testes themselves are involved (Lagerlof, 1936 ; Rao, 1971 ; Kumi-Diaka, 1981) . Bulls infected with T vivax or T. congolense have elevated reaction, deteriorated semen characteristics and in some instances a cessation of semen production (Sekoni et al., 1988) . The aim of the present investigation was to conduct a detailed study of the genital pathology of bulls infected with T. vivax or T. congolense .
MATERIALS AND METHODS
Fourteen normal healthy Zebu bulls aged between 3 and 5 years were used . The semen characteristics were indicative of normal genital function . The bulls were housed individually in fly-proof accommodation throughout the investigation . They were fed on concentrates, fresh pasture, hay, silage, salt licks and water ad libitum.
176
BRITISH VETERINARY JOURNAL, 146, 2
The bulls were placed into two groups of five animals and a third group of four . The three groups were •kept in the same building but in different rooms, completely partitioned from each other to prevent mixed infections . Groups 1 and 2 comprising five bulls each were infected with T. vivax and T. congolense respectively and the four animals in group 3 served as controls . The Y58 strain of T vivax and strain 2295 of T congolense, both isolated from natural infections in cattle in Northern Nigeria, were used to infect the bulls . The stabilates were first inoculated into mice and then transferred into goats . Two ml of blood containing approximately 2X10' trypanosomes from the infected goats were inoculated i .v . into each bull. Parameters monitored twice weekly post-infection were : rectal temperatures, packed cell volume, haemoglobin and total plasma proteins . Measurement of scrotal circumference, palpation of the scrotal contents and internal genitalia (via rectum) was carried out weekly . Histopathological studies
Samples for histopathological studies were taken from the genitalia of all 14 bulls . Thirteen of the bulls were slaughtered at 12, 22 or 30 weeks post-infection ; the remaining animal died 8 weeks after infection with T congolense. The samples were taken from the testes, epididymides, seminal vesicle, prostate gland, ampullae, pelvic urethra and penis following gross examination of the organs . Tissues taken at necropsy were fixed in Bouin's solution for 72 hours and processed according to standard histopathological techniques . They were stained with haematoxylin and eosin (H & E), periodic acid schiff (PAS) and Van Gieson (VG) . The presence and conditions of the geminal epithelia and Sertoli cells were studied microscopically . The interstitial tissues were examined and the occurrence of lesions and their frequency recorded . Degrees of testicular degeneration were scored as shown in Table I . Sections from the epididymides and accessory glands were similarly studied and lesions scored as in Table I .
Table I Grading of genital degeneration Epididymides and Grade
Testes
accessory glands
Severe +++
Germinal epithelial layers reduced to one or two with predominantly Sertoli cells . Sixty (60)% plus tubules affected
Lesions observed in 60% or more tubules
Moderate ++
Germinal epithelial layers reduced to 3 with at least 50% of tubules affected
Lesions observed in at least 50% of tubules
Mild +
Most tubules normal with slight reduction in the germinal epithelial layers
Most tubules are normal
T. VIVAX AND T. CONCOLENSE IN BULLS
1 77
Estimation of epididymal sperm reserve At least 50 epididymal tubules per slide were observed at X100 for determining spermatozoa content. The epididymal sperm reserve was then expressed as a percentage of the number of tubules with normal sperm reserve out of the total examined .
RESULTS All the bulls infected with T. vivax and T. congolense developed chronic trypanosomiasis and were all positive for trypanosomes within 8 days of infection . Clinical signs which were more pronounced in the T congolense infected bulls included fluctuating pyrexia (105-106°F), rapid weight loss, lethargy, weakness, dull and rough hair coats, pale mucous membranes, and enlarged superficial lymph nodes ; anaemia and hydropericardium occurred in some animals at post mortem . The packed cell volume, haemoglobin, and total plasma protein of the infected bulls decreased and differed significantly from control bulls .
Gross findings The male genitalia of the infected and control bulls appeared normal .
Histopathological findings Control bulls. The testes, epididymides, epididymal sperm reserve (100%) seminal vesicles, prostate glands, ampullae, urethra, penis, glans penis and prepuce were normal . T . vivax infected bulls. One animal had mild and one moderate testicular degeneration with severe testicular degeneration in the remaining three . In one animal, the infection had destroyed all the cellular structures within the interstitial tissues and seminiferous tubules . Two animals had mild and three had moderate epididymal degeneration . All the infected bulls had significant decreases in their epididymal sperm reserve ranging between 20 and 50% . Three animals had mild and four had moderate degeneration of the seminal vesicles and prostate glands . None of the bulls had lesions in the Cowper's glands, while only one animal had moderate degeneration of the ampullae . No lesions were present in the urethra, penis, or glans penis of any of the bulls . T. congolense infected bulls . Testicular degeneration was more severe in the bulls infected with T. congolense than in those infected with T vivax. One animal had moderate and four had severe degeneration . In three animals, all the cellular structures within the interstitial tissues and seminiferous tubules were destroyed . Seminiferous tubules were also collapsed in the testes of the three animals (Fig . 1) . Similarly, epididymal degeneration was more marked in the T congolense infected animals (Fig . 2) . Two animals had severe, one moderate and two had mild epididymal degeneration . All the infected animals had marked depletion of the epididymal sperm reserves which were zero % in two, 20% in one and 50% in two bulls . Degenerative changes were also more marked in the seminal vesicles of the T congolense infected bulls . Two animals had severe and three had mild degeneration of the seminal vesicles . Degenerative changes which were mild in three animals and severe in one animal were recorded in the prostate glands of the T congolense infected animals . Degeneration of the
1 78
BRITISH VETERINARY JOURNAL, 146, 2
Fig. 1 . Severe testicular degeneration in a White Fulani bull which died 8 weeks post-infection with T. congolense. Note collapsed tubules . (H & E X200) .
Fig. 2 . Severe epididymal degeneration in a White Fulani bull which died 8 weeks post-infection with T. congolense (H & E X400) .
T VIVAX AND T. CONGOLENSE IN BULLS
1 79
prostate glands was also more marked in the T congolense infected bulls in which the only severe case was recorded . There were no lesions in the Cowper's glands of any of the infected animals . Only one infected bull had mild degeneration of the ampullae . No lesions were present in the urethra, penis, glans penis and prepuce of any of the bulls .
DISCUSSION The results showed that trypanosomiasis had a devastating effect on the male reproductive and accessory reproductive organs . The pathogenic effects of T congolense were more severe than those of T vivax in contrast to observations in West Africa where the latter is more pathogenic (Leeflang, 1978 ; Agu, 1983 ; Anosa, 1983) . The most severe degenerative changes were observed in the testes and epididymides . The decrease or absence of epididymal sperm reserves in infected bulls was most probably the result of a decrease or cessation of gonadal sperm production due to impairment of spermatogenesis (Isoun & Anosa, 1974 ; Anosa & Isoun, 1980) . The pathogenesis of testicular degeneration in trypanosomiasis is not well understood . It is possible that the intermittent pyrexia that accompanies the numerous parasitaemic peaks following infection could be partly responsible (Anosa & Isoun, 1980) . Pyrexia is known to affect spermatogenesis adversely (Lagerlof, 1936 ; Gustafsson, 1966 ; Johnson, Gomes & Vandemark, 1970 ; Arthur, 1975) . However, other factors could also be involved such as the capacity of pathogenic trypanosomes to cause extensive, generalized tissue and organ damage . Other mechanisms involved might be anoxia caused by anaemia, immunological factors, production of biologically active and toxic substances by the trypanosomes, physical swelling and disruption of tissue architecture and increased vascular permeability (Boreham & Cox, 1978 ; Tizard et al., 1978 ; Morrison, Murray & McIntyre, 1981) . The practical significance of the degenerative changes caused by trypanosomiasis on the male reproductive organs is the adverse effects on fertility brought about by disruption in testosterone production, spermatogenesis, and semen characteristics . It is highly likely that the net result of severe degenerative changes in the reproductive organs caused by trypanosomiasis is either serious infertility or even sterility in long-standing cases .
REFERENCES Acu, W . E .
(1983) . ACIAR Proceedings 4, 70 . ANOSA, V . O . (1983) . .Zentralblatt für Veterinärmedizin 30, 717 . ANOSA, V . O . & ISOUN, T . T . (1980) . ,Journal of the Nigerian Veterinary Association 3, 102 . ANOSA, V . O . & KANEKO, J . J . (1984) . Veterinary Pathology 21, 238 . ARTHUR, G . H . (1975) . Veterinary Reproduction and Obstetrics, 4th edn . London : Baillière Tindall . BOREHAM, P . F . L . & Cox, F. E . G . (1978) . Proceedings of workshop held at Nairobi, Kenya, 20-23 November. GUSrAFSSON, B . (1966) . Acta Veterinaria Scandinavica 7, supplement 17 . IKEDE, B . O . (1979) . Research in Veterinary Science 26, 145 . IKEDE, B . O . & AKPAVIE, S . O . (1982) . Research in Veterinary Science 32, 374 . ISOUN, T . T . & ANOSA, V. O . (1974) . Tropenmedizin und Parasitologie25, 469 . IsouN, T . T ., AKPOKODJE, J . U . & ANOSA, V . O. (1975) . Journal of the Nigerian Veterinary Medical Association 4, 2 .
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BRITISH VETERINARY JOURNAL, 146, 2
JOHNSON, A . D ., GoMES, W . R. & VANDEMARK, N . L . (1970) . In The Testis, Vol . 1, Ch . 4, ed . G . M . H . Waites . New York and London : Academic Press . KUMI- DIAKA, J . (1981) . PhD thesis, A . B . U ., Zaria, Nigeria . LAGERLOF, N . (1936). Veterinary Record 48, 1159 . LEEFLANG, P. (1978) . PhD thesis, University of Utrecht . MORRISON, I ., MURRAY, M . & MCINTYRE, I . (1981) . FAO/SIDA follow up seminar on Veterinary Pathology, Faculty of Veterinary Medicine, University of Nairobi, 17-27 September . RAO, A . R . (1971) . PhD thesis, Royal Veterinary College, Stockholm, Sweden . SEKONI, V . O ., KUMI-DIAKA, J ., SAROR, D . I . & NJOKU, C . O . (1988) . British Veterinary Journal144, 388 . TIZARD, I . A ., NIELSON, K ., SEED, J . R . & HALL, J . E . (1978) . Microbiology Review 42, 661 . (Accepted for publication 21 April 1989)
ANNOUNCEMENT COMISA Comisa, the Consultation Mondiale de l'Industrie de la Santé Animale (the Representative Body of the Worldwide Animal Health Industry) was officially launched on 23 October 1989 in Washington D .C . Its broad objectives include : the worldwide promotion of science and development of animal health products establishing and developing formal relations between the animal health industry and international bodies dealing with animal health affairs developing and procuring common agreement of policies to be adopted by the animal health industry in its dealings with public international organizations, both governmental and non-governmental . The Secretary of the new body with headquarters in Brussels is Dr . David Miller . Tel : 44 276 698292 .
