French primate disaster
Primate BSE quarantine recommendations
TSEs in primates: brief literature survey
Lemur prion sequences predicted
Prosimians: which animals are involved so far?
Earlier BSE problems at French, Australian, and Irish zoos
Further blow aimed at surviving primates
Which zoos, which primates, how many animals?
Why infect more chimps with BSE (when so many are already infected?)
Noelle Bons et al.30 Mar 99 issue of PNAS 96:4046-4051. webmaster comment:The 30 Mar 99 issue of PNAS carries an explosive French study of very high experimental standards (plus a Nobel lauriate co-author) about TSE in primates in French zoos serving BSE-tainted feed supplements through June, 1996. This is a huge scandal because it potentially affects the survival of many of the world's primate species.
It also suggests very strongly that the nvCJD epidemic will indeed be a 'plague of of biblical proportions': easy passage across many mammalian orders is seen , the doses are small, the incubation periods not long relative to lifespan, the penetrance very high in various genera of primates, the distribution of contaminated foodstuff widespread.
The primates showed contamination of the tonsils, esophagus, gastric glands, duodenum, walls of lymph and blood vessels, Peyer's patches, and spleen, in addition to dorsal and ventral root ganglian, spinal chord and brain. 14 of 26 non-experimental primates that had died for whatever reason at the Montpellier zoo had neurological signs; 5/6 autopsies were positive, indicating very high dietary penetrance in a variety of primate families. This suggests that tonsil screening in England will be very effective in identifying infected humans. The old debate about kuru, ie whether it was transmitted by cannabalism or simply by contact with the corpse may be resolved as well if gastrointestinal material was set aside.
Shockingly, the British manufacturer had continued to manufacure and distribute this speciality product, possibly to hundreds of zoos on the Continent and possibly worldwide, for 10 years after zoo ungulates and carnivores were first diagnosed with BSE in England. British zoos also apparently continue to export animals incubating disease worldwide. It is so bizarre that in the same years that Prince Phillip, as head of the World Wildlife Fund, was making strong pleas for primate conservation worldwide that these devastating shipments continued to be made.
Equally shocking, French zoos continued to use this high-risk feed despite knowning its contaminated source, even though nearly every one of the 234 species of primate is endangered in the wild, zoos are often used to sustain populations, and breeding facilities provide animals for human medical research. Only 8 French zoos and primate breeding facilities out of 89 agreed to cooperate with this study, which simply asked about primate neurological or unexplained deaths and dietary practices. A zoo in Lille acknowledged 3 primate deaths with TSE-like neurological illness. The Colorado experience with CWD shows that animal facilities can be become thoroughly contaminated and transmit disease horizontally, as well as to trace-forward herds.
It is highly questionable not to autopsy [or save samples] from valuable animals with unexplained neurological deaths fed known BSE feed, yet this is a big step up from non-cooperation. It seems that French veterinary ethics have a similar standard to those of French medicine (AIDS, hepatitis, cadaver growth hormone).
But is the problem limited to France and to primates? We all know that English zoos have been lying for years -- MAFF interfered with kudu and dogs according to the Inquiry, did it stop there: the list of species affected never made any sense given the prion sequences involved. Whose protein supplement did English zoos use for primates if not the local company's? Why would only French primates go down with TSE? (Explanation: Max the cat was bad enough, how could the UK resume beef exports if they announced Charlie the chimpanzee had BSE?)
The products are identified as "Singe 107, MP, or Marex." Singe 107 (UAR, Paris, France) was previously described as containing "products declared fit for human consumption." The present article does not identify the companies by name but says, "according to the manufacturers, this food contained various items, including gross protein, fats, corn, soya, carob bean, alfalfa, minerals, yeast, vitamis A, C, D3, and E, and cracklings (the so-called 'fifth quarter of beef' suitable for human consumption) from the same French distributor.
Will we see a list released of the customers for this special feed over the last ten years? Will we see an accounting of primate deaths at every facility in Europe? It is absolutely essential to have this information for every zoo and breeding facility worldwide.
The British manufacturer 'which distributes nutritional supplements to zoos and animal breeding facilities through a French company [and possibly others in other countries]) announced in June, 1996 that it ceased to use beef in its nutritional supplements' which the authors take to mean that they had used beef prior to 'ceasing'. On 6 July 1996, an article appeared in Lancet documenting non-experimental transmission of BSE to primates (in a rhesus monkey that had died in 1992).
It has not been clarified whether the British manufacturer or French distributor recalled existing stocks in June, 1996 or allowed warehoused material to be sold until it was all gone. Zoos may have continued to use up their inventory even after the manufacturer ceased using beef. This latter scenario would represent an uncanny similarity to the French and Canadian blood AIDS and hepatitis scandals, where known tainted stocks were used up.
In the experiment reported here, 79 primates of 11 species were feed 20-40 gm/kg body weight of these feeds in addition to their usual fruits and vegetables. For a 60 kg human, this works out to 1200-2400 gm per day [simple scaling]. There are 8.8 quarter-pound hamburgers in a kilogram. No one knows the minimum daily dose that would have made the lemurs ill, perhaps 2-4 gm/kg or for that matter 0.2-0.4 gm/kg would have sufficed -- they may have received a saturating exposure. No one knows whether a daily ration for years on end was necessary -- or just a single meal. The two Microcebus lemurs became ill after only 1-2 feedings of cattle brain.
In July 1996, Bons et al. published that a rhesus monkey and two lemurs at the Montpellier zoo had died of TSE. Here they look at an additional 20 lemurs and macaques from 3 French zoos. The zoos had to kill these lemurs anyway because they were primate hybrids, now illegal in France. Two lemurs were symptomatic, 18 were not: all were definitely positive for prion disease.
Lemurs and macaques can live for 20+ years, so what this implies is that a great many primates in French zoos are not yet symptomatic but are slowly incubating BSE-based prion disease.
We could ask whether the facilities are thoroughly contaminated (like the ones in Colorado), whether only primates are affected, whether only French zoos are affected, whether food 'fit for human consumption' ended up in the human food chain, and indeed, wonder whether there is much of a species barrier at all to humans, given that 5 other species of primate have gone down via the oral route.
Species used in study: Microcebus murinus# [Strepsirhini (prosimians) small, short-lived mouse lemur^] Eulemur fulvus mayottensis# [prosimian, lemur] Eulemur fulvus albifrons# [prosimian, lemur] Eulemur mongoz# [prosimian, lemur] Eulemus macaco# [prosimian, lemur] Lemur catta# [prosimian, (ring-tailed lemur] Varecia variegata varigata# [prosimian, ruffed lemur] Varecia variegata rubra [prosimian, ruffed lemur] Macaca mulatta#* [old world monkey] Macaca sylvanus* [old world monkey] Macaca fuscata* [old world monkey] Saimiri sciureus#* [new world monkey] ^ exposed to BSE brain orally # one or more non-experimental BSE-food exposed animal suspected or confirmed. * sequence of prion gene known. 35 other individuals pending further study. -- no prosimian prion sequence is known at this point.Previous relevent articles:
The Lancet Volume 348, Number 9019 - Saturday 6 July 1996 Noelle Bons, Nadine Mestre-Francois, Yves Charnay, Fabrizio TagliaviniSir--Although human and animal prion diseases have been experimentally transmitted to various monkey species, spontaneous occurrence of a spongiform encephalopathy in non-human primates has not been reported. We describe the development of a spongiform encephalopathy without known cause in a rhesus monkey (Macaca mulatta). The monkey was born in 1982 and was acquired from Ravensden zoo, Rushend, Northants, UK, in October, 1986, by the zoological park in Montpelier, France where it was housed in the primate colony. In Montpelier, the monkey was fed standard monkey feed including Singe 107 (UAR, Paris, France) which contains meat products declared fit for human consumption. In summer, 1991, aged 9, the previously healthy monkey became lethargic and developed mood changes, in particular, aggressiveness. The monkey became isolated from its companions and hid itself away. In June, 1992, the monkey was anaesthetised...
The Lancet Volume 348, Number 9035 - Saturday 26 October 1996 R M RIdley, H F BakerSir--In a report to the European Union Commission, Prof C Weissmann of Zurich University has recommended that experiments be set up to investigate oral transmission of BSE to primates to see whether human beings are at risk from eating beef products.1 Bovine spongiform encepholopathy (BSE) has already been transmitted to primates by intracerebral injection of infected brain tissue homogenate so the "species barrier" between cattle and primates is not absolute. BSE has also been transmitted by feeding infected brain to mice so clearly transmission of this agent by the oral route is possible. During the BSE epidemic not only were cattle fed on contaminated, rendered material, but large numbers of other species in zoos, farms, and domestic ownership were also fed on pelleted food containing rendered material. A few animals of several species of antelope and cat but not monkeys became affected despite these monkeys being fed for many years on pellets which contained up to four times the amount of the implicated, rendered meat and bone meal that was included in cattle pellets. Thus a "species barrier" of some sort exists between cattle and primates. But will Weissmann be able to quantify this in a way which will have public health implications? In 10 years' time he may be able to determine the smallest amount of raw cow brain necessary to cause disease if fed to perhaps a few dozen monkeys. But what will this tell us about the maximum quantity of bovine-derived material (from which specified offals have been removed) which can be consumed without causing illness when the sample size could be up to 50 million people in the UK? As every drug company knows, extensive tests in clinical trials in large numbers of people cannot ensure that a new drug, when available on general prescription, will not cause serious illness in a handful of patients such that the drug has to be withdrawn. At best Weissmann's experiments will be irrelevant; at worse they will provide a degree of reassurance which is unwarranted. There is nothing that can be done to reduce any risk to which people have been exposed in the past, and there is now no alternative but to do the best we can to eliminate BSE entirely.
Lancet letter R M Ridley, H F Baker, C P WindleSir--We have kept a large breeding colony of common marmosets (Callithrix jacchus) for use in neuropsychological research for almost 20 years. This primate species develops spongiform encephalopathy with an incubation period of 3.5 or 4 years after intracerebral injection of scrapie-affected or bovine spongiform encephalopathy (BSE)-affected brain homogenate. The majority of marmosets in the colony are used for experimental purposes and are killed when under 3 years of age, but the breeding animals are aged 3´10 years and a small number of animals have been kept for up to 15 years in an investigation of ageing. All animals are fed a daily diet of egg sandwiches, fruit, and a proprietary brand of UK-produced New World monkey pellets. According to the manufacturers these pellets contained a 20% inclusion of ruminant-derived meat-meal protein until April 1996 (cattle-feed supplements before 1988 contained about 4% of meat and bone meal). From 1985 onwards, ruminant-derived feed probably contained increasing amounts of BSE agent from affected cattle and although the Ruminant Feed Ban of 1988 prohibited the feeding of this material to ruminants, its use in other animal feeds was legal until April 1996. We estimate that more than 100 marmosets born in our colony between 1980 and 1990 lived for more than 5 years and were exposed to ruminant-derived protein in their diet for their entire life. Each adult marmoset (weight 350 g) was fed about 2 g meat meal per day. [Note French primates were fed 10-20x of this level for longer periods. -- webmaster]
It seems likely, therefore, that the mature monkeys in our colony were exposed to significant amounts of infective agent for more than 5 years and in some cases for up to 10 years. With the exception of those animals which were injected intracerebrally with infected brain in our primary transmission studies, no animal in our colony has ever developed spongiform encephalopathy. [A cite is needed here to detailed published methods of ascertainment. The sensitivity of these methods would have to be reviewed in the light of the French results. -- webmaster] As part of general husbandry all animals are monitored daily for signs of ill-health and any with severe illness or persistent poor condition are killed.
31 Mar 99 webmaster[Open letter sent to directors of primate facilities worldwide]
"The research article by Noelle Bons et al. in the 30 Mar 99 issue of PNAS 96:4046-4051 and the accompaning NY Times story raise grave questions about risks to primates worldwide from transmissible spongiform encephalopathies.
A British manufacturer of monkey chows distributed BSE-contaminated feed to zoos and primate research facilities in at least two countries for the decade ending in June, 1996. This prion disease easily crosses the species barrier to prosimians, new world monkeys, old world monkeys, as well as great apes, including humans, across France (and surely England as well). Other mammalian orders are also affected. Contaminated facilities may have shipped animals internationally as well as reintroduced them to the wild.
In light of 30 years of experience with captive-animal transmission of prion disease in ungulates, these animals are a hazard to their cage mates and to care-providers (through shedding of agent) and have the potential to irrevocably contaminate primate housing facilities and veterinary equipment. These animals are completely inappropriate for use in medical or behavioral research or in breeding colonies. Captive animals reintroduced to the wild may establish the infection in wild populations. (CWD has reached 6% level in wild populations in contact with released animals.)
This family of diseases is not diagnosable in living primates, has no treatment, and is invariably fatal (provided the animal does not die of something else first). Animals may be highly infectious for years prior to displaying symptoms.
These facts call for:
(1) an immediate worldwide quarantine in trafficking of primates until the scope of the epidemic is clarified,
(2) obligatory IHC neuro- and lymphoreticular autopsy on all primates worldwide that die or are euthanized for any reason,
(3) immediate cessation of reintroduction programs to the wild and destruction of all reintroductions from known contaminated facilities to the extent practicable.
(4) open disclosure on the Internet by every primate holding facility worldwide of their exact source of animal protein supplements (eg, rendered cattle byproducts from UK), displays of neurological symptoms in living primates, deaths of primates from whatever reason, results of autopsy, and current availability of tissue samples to qualified researchers, all in detail back to 1985.
(5) obligatory and permanent government closure of non-cooperating institutions, including incineration of carcasses.
These requirements may seem onerous but they are basically identical to those already adopted by various US states and regional animal health associations in the context of a related prion disease, CWD. Indeed, it may already be too late for many primate colonies.
High-risk primates should be isolated. These are defined as:
-- any primate exposed to "Singe 107, MP, or Marex" brand monkey chows or similar UK or French products, 1984-1999
-- any cage mate, sibling, or offspring of a primate ever exposed to these chows,
-- any primate exposed to facilities or veterinary instruments connected with a suspected or subsequently diagnosed primate ,
-- any primate with a trace-back history to a suspect or confirmed colony.
Be aware that prion diseases involve many political and economic ramifications. Affected countries can implement significant mis-information and dis-information campaigns to avoid stigma and economic losses. Research can be stiffled to prevent embarassing histories of cover-up. 81 of 89 French facilities are not cooperating with the epidemic investigation at this time. Overall, it is very analgous to AIDS. (Note prion disease are not viral.)
Prion science is too complex to summarize here. A very large and active Internet resource is maintained at www.mad-cow.org."
Medline survey 31 Mar 99 webmaster
Paste the parenthetic expression into the text box at Medline to recover abstracts. Some false hits occur. 233 articles: ((((scrapie[All Fields] OR kuru[All Fields]) OR Creutzf*[All Fields]) OR prion[All Fields]) AND primate[All Fields]) 77 articles: ((((scrapie[All Fields] OR kuru[All Fields]) OR Creutzf*[All Fields]) OR prion[All Fields]) AND chimpanzee[All Fields]) 30 articles: ((((scrapie[All Fields] OR kuru[All Fields]) OR Creutzf*[All Fields]) OR prion[All Fields]) AND squirrel monkey[All Fields]) 9 articles: ((((scrapie[All Fields] OR kuru[All Fields]) OR Creutzf*[All Fields]) OR prion[All Fields]) AND rhesus[All Fields]) 45 articles: ((((scrapie[All Fields] OR kuru[All Fields]) OR Creutzf*[All Fields]) OR prion[All Fields]) AND macaca[All Fields]) 01 articles: ((((scrapie[All Fields] OR kuru[All Fields]) OR Creutzf*[All Fields]) OR prion[All Fields]) AND prosimian[All Fields]) ((((scrapie[All Fields] OR kuru[All Fields]) OR Creutzf*[All Fields]) OR prion[All Fields]) AND prosimian[All Fields])A prelimary search for primate species affected shows:
PNAS 1999 96:4046-4051 Microcebus murinus, Eulemur fulvus mayottensis, Eulemur fulvus albifrons, Eulemur mongoz, Eulemus macaco, Lemur catta, Varecia variegata varigata C R Acad Sci III 1997 Dec;320(12):971-9 Eulemur fulvus mayottensis C R Acad Sci III 1996 Aug;319(8):733-6 Macaca mulatta Lancet 1996 Jul 6;348(9019):55 rhesus monkey Nature 1996 Jun 27;381(6585):743-4 macaques Vet Rec 1993 Apr 17;132(16):403-6 common marmosets (Callithrix jacchus) Dev Biol Stand 1993;80:9-13 chimpanzee and squirrel monkey. other new-world primates, Ital J Neurol Sci 1983 Apr;4(1):61-4 monkeys Acta Neuropathol (Berl) 1983;61(3-4):300-4 Macaca nemestrina Acta Neuropathol (Berl) 1981;53(4):337-41 Rhesus monkey. spider monkey, marmoset, squirrel monkey Rev Neurol (Paris) 1981;137(12):785-805 squirrel monkey spider monkey J Infect Dis 1980 Aug;142(2):205-8 Saimiri sciureus Electroencephalogr Clin Neurophysiol 1978 Nov;45(5):611-20 rhesus monkey Epilepsia 1978 Jun;19(3):257-64 Macaca mulatta Primates Med 1978;10:254-60 marmoset monkeys (Saguinus sp.) Brain 1976 Dec;99(4):637-58 squirrel monkeys J Neuropathol Exp Neurol 1976 Nov-Dec;35(6):593-605 primates. Eight animals Immunopathologie du systeme nerveux 1976 Aug 23;: monkeys (Saimiri sciureus) Immunopathologie du systeme nerveux 1976 Aug 23;: chimpanzees J Med Primatol 1976;5(4):205-9 gibbon and sooty mangabey pigtailed macaque. table lists all the species with transmission by 1976 Brain 1975 Dec;98(4):595-612 spider monkey Rev Electroencephalogr Neurophysiol Clin 1975 Oct-Dec;5(4):335-43 chimpanzee Proc Soc Exp Biol Med 1975 Jul;149(3):723-4 stumptail macaque (Macaca arctoides) Adv Neurol 1975;10:341-6 primates Adv Neurol 1975;10:319-39 primates Intervirology 1975-76;6(3):150-5 patas monkey (Erythrocebus patas) squirrel monkey Adv Neurol 1975;10:291-317 primates Intervirology 1974;2(1):14-9 marmoset monkeys Science 1973 Oct 5;182(107):67-8 primates Biomedicine 1973 Jul;18(4):328-35 chimpanzee to chimpanzee Nature 1972 Dec 8;240(5380):351 rhesus monkey (Macaca mulatta) 8.5 years after inoculation. Nature 1972 Mar 10;236(5341):73-4 cynomolgus monkey (Macaca fascicularis) Nature 1971 Apr 30;230(5296):588-91 new world monkeys J Neuropathol Exp Neurol 1969 Jul;28(3):353-70 chimpanzees and spider monkeys Science 1968 Nov 8;162(854):693-4 spider monkey (Ateles geoffreyi)
31 Mar 99 webmasterNo lemur (prosimian) prion sequences have been determined and none are in the works. This is unfortunate in terms of understanding species barrier implications of BSE transmission to these primates. However, H. Schatzl et al. determined a great many primate prion sequences some years back and recently posted additional rodent (outgroup) sequences for cotton rats and gerbil.
It is fairly simple to predict the sequence of prosimian prion protein at the time of divergence from the primate lineage using a method introduced earlier on this site for prion promoters:. Outgroup arbitration is a topologically driven consensus technique that extracts additional information from alignments. It requires sequences from 3 or more species clamped to a definitely known topology of divergence. In the case of the prosimian prion (rodent, (lemur, (NW monkey, (OW monkey, great ape)))) is the topology.
The geometry of the tree is an additional consideration. Here, the geometry is favorable as branch lengths are approximately equal. Had the (prosimian, NW monkey) divergence been too recent relative to the rate of mutation, there would be few opportunities to apply the technique; if the divergence had been too early, the tree is essentially a trichotomy.
| Sequence Feature Analysis by Outgroup Arbitration | ||||
| Distribution of Feature | Allocating events to primate tree | |||
| rodent | lemur | monkey | Interpretation | Outgroup arbitration works because two rare events are far less likely than one rare event -- a small number squared vs a small number. The method is much stronger than simple consensus or maximal parsimony inference but does require knowing the topology of the relevent tree. Mechanisms such as slippage tandem repeats must also be discounted as they are not at all rare.
The idea of outgroup arbitration is that the outgroup can resolve feature ambiguity in the ingroup when it can cast a deciding vote. In nested outgroup arbitration, a restored sequence (here, the ancestral rodent) can then serve as a noise-filtered outgroup to as the process is repeated. The table below lists all combinations of scenarios (either gaps or point mutations; may include sequencing errors). Outgroup arbitration is only applicable to half of these. The other cases must be deferred to a more distant outgroup. |
| + | + | + | none | |
| + | + | - | monkey event | |
| + | _ | + | lemur event | |
| - | + | + | deferred | |
| - | - | - | none | |
| - | - | + | monkey event | |
| - | + | - | lemur event | |
| + | - | - | deferred | |
The ancestral prosimian prion sequence by this method is then given by (lower case represents outgroup arbitration):
MANLGyWMLVLFVATWSDLGLCKKRPKPGGWNTGGSRYPGQGSPGGNRYPPQGGGGWGQP c HGGGWGQPHGGGWGQPHGGGWGQPHGGGWGQGGGTHNQWnKPSKPKTnMKHMAGAAAAGA hs VVGGLGGYMLGSAMSRPLIHFGNDYEDRYYRENMYRYPNQVYYRPVDQYSNQNNFVHDCV NITIKQHTVTTTTKGENFTETDVKMMERVVEQMCITQYErESQAYYQRGSSMVLFSSPPV k ILLISFLIFLIVG 253 aaAligning this with human prion, we find 6 conservative differences in the mature region and 1 in the signal region (97% agreement); lemur and cattle differ by 11 positions in the mature region:
>lemur MANLGYWMLVLFVATWSDLGLCKKRPKPGGWNTGGSRYPGQGSPGGNRYPPQGGGGWGQP HGGGWGQPHGGGWGQPHGGGWGQPHGGGWGQGGGTHNQWNKPSKPKTNMKHMAGAAAAGA VVGGLGGYMLGSAMSRPLIHFGNDYEDRYYRENMYRYPNQVYYRPVDQYSNQNNFVHDCV NITIKQHTVTTTTKGENFTETDVKMMERVVEQMCITQYERESQAYYQRGSSMVLFSSPPV ILLISFLIFLIVG >cattle MVKSHIGSWILVLFVAMWSDVGLCKKRPKPGGGWNTGGSRYPGQGSPGGNRYPPQGGGGW GQPHGGGWGQPHGGGWGQPHGGGWGQPHGGGGWGQGGSHSQWNKPSKPKTNMKHVAGAAA AGAVVGGLGGYMLGSAMSRPLIHFGNDYEDRYYRENMHRYPNQVYYRPVDQYSNQNNFVH DCVNITVKEHTVTTTTKGENFTETDIKMMERVVEQMCITQYQRESQAYYQRGASVILFSS PPVILLISFLIFLIVG >human MANLGCWMLVLFVATWSDLGLCKKRPKPGGWNTGGSRYPGQGSPGGNRYPPQGGGGWGQP HGGGWGQPHGGGWGQPHGGGWGQPHGGGWGQGGGTHSQWNKPSKPKTNMKHMAGAAAAGA VVGGLGGYMLGSAMSRPIIHFGSDYEDRYYRENMHRYPNQVYYRPMDEYSNQNNFVHDCV NITIKQHTVTTTTKGENFTETDVKMMERVVEQMCITQYERESQAYYQRGSSMVLFSSPPV ILLISFLIFLIVGThis method obviously does not extend lemur prion sequence evolution to the present era, nor distinguish different prosimians. These would involve an additional 4-5 residues change most likely at the more variable residue positions (given below). Although species barriers and incubation periods cannot be reliably predicted from sequence (or 3D structure) alone, the differences among primates including prosimians are generally quite minor.
We may further exploit the plethora of prion sequence data to predict (in a probabalistic manner) sequences of individual prosimians by a method that might be called double cladistic fourier filtration. Simply put, residues at different positions along any polypeptide chain have estimatable characteristic rates of change. A position averaging a PAM on a 10 million year time scale is an unsuitable (plesiomorphic) character for primate orders or families (ie, the fourier coefficient is mismatched) but might be quite servicable in distinguishing microcebus from galago. For lemuridae vs cheirogaleidae, a convolution vis-a-vis residues of 30 my time scales makes more sense.
In other words, just adapt the band pass of fourier filtration to the taxonomic level. Weightings must couple band pass filtration with the geometric topology of the tree as estimated from the fossil record or molecular clocks. The method offers no information whatsoever about which changes occur in which lineages.
As a practical matter, anyone can read off results of cladistic fourier filtration from an alignment of prion sequences simply by keeping one eye on the divergences and their dates, discounting synapomorphy. Taking the prosimian node as the point of departure for middle and high pass filteration, we can define the residues of use at the respective taxonomic levels as:
MANLGyWMLVLFVATWSDLGLCKKRPKPGGWNTGGSRYPGQGSPGGNRYPPQGGGGWGQP c HGGGWGQPHGGGWGQPHGGGWGQPHGGGWGQGGGTHNQWnKPSKPKTnMKHMAGAAAAGA h s VVGGLGGYMLGSAMSRPLIHFGNDYEDRYYRENMYRYPNQVYYRPVDQYSNQNNFVHDCV NITIKQHTVTTTTKGENFTETDVKMMERVVEQMCITQYErESQAYYQRGSSMVLFSSPPV k ILLISFLIFLIVG MANLGYWmLvLFVATWSDlGLCKKRPKPGGWNTGGSRYPGQGSPGGNRYPPQGGGGWGQP l/i a v HGGGWGQPHGGGWGQPHGGGWGQPHGGGWGQGGGTHNQWNKPsKPKTNMKHMAGAAAAGA n VVGGLGGYMLGSAMSRPLIHFGNDYEDRYYRENMyRYPNQVYYRPVDQYSNQNNFVHDCV h/n NITIKQHTVTTTTKGENFTETDVKMMERVVEQMCiTQYERESqAYYQRGSSMvLFSSPPV v e i ILLISFLIFLIVG MANLGyWMLVLFVAtWSDLGLCKKRPKPGGWNTGGSRYPGQGSPGGNRYPPQGGGGWGQP c m HGGGWGQPHGGGWGQPHGGGWGQPHGGGWGQGGgtHnQWnKPSKPKTnMKHmAGAAAAGA - s s/g h/g s v VVGGLGGYMLGSAMSRPlIHFGnDYEDRYYRENMYRYPNQVYYRPVDQYsNQNnFVHDCV i/m s n s/t NITIKQHTVTTTTKGENFTETDvKmMERVVEQMCITQYerESQAYYQRGSSmVLFSSPPV i/m i q k a/v ILLISFLIFLIVGOf course we cannot really determine prion sequences from our armchairs -- we can focus a telescope in Chile over the Internet but we cannot yet ask the robotic sequenator to lift the primate tray from the freezer and PCR the primer in our text box (a small Visa card charge applies). However, the goal here is not to predict specific sequences, but to identify their surprises -- the residual changes, those not given above (hence double filtration). For it is exactly here, not in the drift, that evolutionary progression in normal function is concentrated.
31 May 99 webmaster GenBank Taxonomy
Strepsirhini(prosimians)
Cheirogaleidae(dwarf and mouse lemurs)
Cheirogaleus
Cheirogaleus major
Cheirogaleus medius(fat-tailed dwarf lemur)
Microcebus
Microcebus murinus
Mirza
Mirza coquereli
Daubentoniidae(aye-ayes)
Daubentonia
Daubentonia madagascariensis(aye-aye)
Galagonidae(galagos)
Galago
Galago alleni
Galago demidovii
Galago moholi
Galago senegalensis(northern lesser bushbaby)
Galago sp.(bush baby)
Otolemur(greater bushbabies)
Otolemur crassicaudatus(thick-tailed bush baby)
Otolemur garnetti
Indridae
Indri
Indri indri
Propithecus(sifakas)
Propithecus diadema
Propithecus tattersalli(Tattersall's sifaka)
Propithecus verreauxi(white sifaka)
Propithecus sp.(sifaka)
Lemuridae(lemurs)
Eulemur
Eulemur coronatus
Eulemur fulvus
Eulemur macaco
Eulemur mongoz
Eulemur rubriventer
Hapalemur
Hapalemur griseus
Lemur
Lemur catta(ring-tailed lemur)
Lemur sp.
Varecia
Varecia variegata(ruffed lemur)
Loridae(lorises)
Loris
Loris tardigradus(slender loris)
Nycticebus
Nycticebus coucang(slow loris)
Perodicticus
Perodicticus potto(potto)
Megaladapidae
Lepilemur
Lepilemur dorsalis
Lepilemur mustelinus(weasel lemur)
Lepilemur ruficaudatus
Lepilemur septentrionalis
12 Apr 99 webmaster The Denver, Toronto, Wyoming zoos, and 4 US research facilities have been affected by CWD.The full text of Tuesday's PNAS primate-BSE article is becoming broadly available, but as the article is not lucidly written, it is worth going over it carefully to see exactly what it says, who the authors are, the earlier history of TSEs in French and other zoos, and what the implication might be (with parallels to the chronology of an older prion disease in wildlife and zoos, chronic wasting disease, as summarized earlier.
-- From 'Spongiform encephalopathy in an imported cheetah in France' [Vet Rec 1997 Sep 13;141(11):270-1 ] and the MAFF site, we learn that the French cheetah was born in GB, as were Australian and Irish cheetahs. Five cheetahs have died in England, 8 altogether. These deaths say nothing about contaminated feed in French, Australian, or Irish zoos, but rather document that English zoos have exported non-primates already incubating BSE to other zoos. The cheetah of course is critically endangered.
-- C R Acad Sci III 1997 Dec;320(12):971-9 N Bons et al. Submitted 26 May 1997, accepted 7 Nov 97.
This mainly addresses 3 individual lemurs [Eulemur fulvus mayottensis, 1500 gm], #481, 474, and 584. The first originated in Madagascar in 1974 and had no connection to England other than monkey chow Singe 107. It was euthanized in Dec 92 at the age of 18 with confirmed TSE. #474 was born at the Montpellier zoo in June 1988 and was euthanized in Dec 92 at the age of 4.5 with confirmed TSE. Its alimentary positivity establishes that the monkey chow was still infectious post-June 1988.
#584 was born at the Montpellier zoo in June 95 and was euthanized in Sept 96 at the age of 15 months because of a wasting disease ("weight loss for unkown reasons") but lacked TSE (figure 17 shows negative jejunum transverse section). Despite the formalin, this animal might still be tested for intra-cerebral infectivity. Although the animal was very young, it might be argued that the monkey chow was no longer significantly infectious after June of 1995. (It was reformulated a year later.) It is not clear whether #584 was a cagemate of the affected two (no horizontal transmission?). The PNAS paper puzzlingly lists #584 as suffering from neurological signs and blindness.
These animals showed up at the university because of an arrangement with the zoo director, Marcel Gallet, to furnish any primate dying from or affected by a chronic neurological disorder. The remains were stored in formalin. JY Robert at Besancon, F Haclewyn at Lille, and Y Rumpler at Strasbourg also cooperated on behalf of their respective zoos in the PNAS study.
No explanation is offered for the 60 months that passed between the deaths of the 2 lemurs and the publication of the paper. Numbers are given for total lemurs at the zoo in the PNAS article.
The monkey chow is described as 'aliments industriel contenant des farines de viand de betail' containing 'element carnes suspects' or 'ailments industriels renfermant des proteines animales.' Betail simply means cattle, farine means flour, viande is meat. The PNAS article described these as 'cracklings' (rendered meat products). Cracklings in Europe refer to the cooked meat fraction produced during rendering, probably pelleted or otherwise formed products. N Bons called this product 'cretons de boeuf' in a Nature interview.
-- C R Acad Sci III 1996 Aug;319(8):733-6 N Bons et al. Submitted 8 Jul 96, accepted 11 Jul 96. An announcement of this article appeared as a letter in the July 1996 issue of Lancet.
This concerns 1 numbered and 3 unnumbered rhesus monkeys (Macaca mulatta) all acquired from the Ravensden zoo, UK by the zoological park in Montpelier. Monkeys #455 and #2 were acquired in Oct 86; monkey #3 in 1988.
Monkey #4 was born in 1982 at the same UK zoo but no date of acquisition is given. It was still healthy as of July 1996 and had a baby. This monkey has disappeared from table 1 of the PNAS article (unless it is among the M. sylvatus or M. fuscata).
Monkey #455 was euthanized in June 1992 at age 10 and was confirmed with TSE (and AD). This was the first zoo primate with confirmed nvCJDp as it preceded the two lemurs confirmed in Dec 1992. All remains except the brain were incinerated, ruling out comparable study of the gastrointestinal tract. Monkeys #2 and #3 were killed in 1991 and 1993 at 11 years of age with the same clinical neurological symptoms but were incinerated completely. In the PNAS article, #2 is said to have died of pneumonia, inconsistent with this article.
These monkeys were all fed Singe 107 monkey chow. They may have been exposed to contaminated feed both in the UK and Montpellier, especially #3.
No explanation is offered for the 49 months that passed between the death of macaque #455 and the publication of the paper. No reason is given for the total incineration of #3 which died with neurological signs a year after #455 and 2 lemurs had been confirmed. Numbers are given for total macaques at the zoo in the PNAS article.
-- The PNAS paper, submitted 21 Dec 98, has no further data on the 5 dead squirrel monkeys, one of which died with neurological signs in 1990. These animals originated at 'Frejus' which is presumbably the Parc Zoologique de Frejus on the Cote d'Azur.
A broad range of dates for dates of death of primates with neurological symptoms, with most in the early 1990's. However, table 1 does not include the 18 lemurs from Besancon and Strasbourg apparently all still living in 1998 and all positive. The 3 primate deaths with neurological signs at Lille were in Jan 96. This says that the primate epidemic is still going strong and that the penetrance may be very high indeed (supported separately by Table 1). Stopping the bad monkey chow in 1996 will hardly stop nvCJDp as these animals can live for 25 years or more.
It is really a shame that so much time elapsed between the first primates dying in 1989-92, the first reports in 1996, and the first attempt at a survey in 1999. A lot of animals would have been exported to other countries in this time frame. Zoos are surprisingly unconcerned about the origins and deaths of their primates; a great many suspicious deaths must simply have resulted in incineration with no autopsy or samples saved. This raises very serious questions whether these zoos have the professional competence and integrity to hold primates.
-- From Lancet 1996 Jul 6;348(9019):55 we learn of a spongiform encephalopathy in a rhesus monkey (Macaca mulatta born in 1982 and acquired from Ravensden zoo, Rushend, Northants, UK, in October, 1986, by the zoological park in Montpelier, France where it was housed in the primate colony and fed standard monkey feed including Singe 107 (UAR, Paris, France). In summer, 1991, aged 9, the monkey came down with symptoms later confirmed as TSE. This may have been one of the three rhesus monkeys mentioned above, the other two may have originated from Ravensden zoo as well or been cage mates or offspring. This case is the fourth animal exported from English zoos proven to have subsequently died of prion disease. Note that this animal, as well as the French cheetah, was exposed to contaminated feed both in England and in France.
-- From PNAS Vol. 96, Issue 7, 4046-4051, March 30, 1999 we learn of 20Ýlemurs of 3 genera fed UK-exported beef protein dietary supplements at three different primate facilities (Montpellier, BesanÁon zoo, and Strasbourg breeding facility). The 2 hybrids at the Montpellier zoo were already symptomatic and confirmed by prion IHC etc.
The 6 from BesanÁon zoo and 12 from Strasbourg (18 lemurs) were asymptomatic but all well along in confirmed TSE neuropathology. (Healthy primates would scarcely be sacrificed as controls, but because of a new French law forbidding primate hybrids, these had to be euthanized anyway. For this reason, lemurs were the only samples available of healthy animals from other zoos. The data does not imply in any way that lemurs were disproportionately affected among primate species.)
The origin of confirmed non-hybrid lemurs housed at the Montpellier zoo include Madagascar (#481, 586), Montpellier (#474), and Paris (#476). An additional 3 lemurs at the Montpellier zoo but originating from Mulhouse died with neurological signs but are not yet studied in Table 1. Apparently an additional 2 Montpellier lemurs are among the 20 confirmed hybrids. These cases show some animals have acquired nvCJDp from French zoo food along and not as a result of having been exposed in England and exported.
Mulhouse sounds English but is actually a town in eastern France; the Parc Zoologique et Botanique Tel : 03 89 31 85 10 - Fax 03 89 31 85 26, web site showing showing a yawning cheetah andmany publications in primate cytogenetics.
Ominously, the Mulhouse zoo maintain the international studbook on Hylobates concolor, the crested gibbon.
The zoo at Lille reported 3 primate deaths in Jan 96 in primates after neurological illnesses similar to those seen in the Montpellier primates; no tissue was saved, their origin and species are not reported. Eight other French zoos with primates "denied any suspicious or neurological deaths"; 81 other French zoos did not respond. A complete accounting of all primates in all French zoos is urgently required.
The primate article in question is:
PNAS Vol. 96, Issue 7, 4046-4051, March 30, 1999 full text for subscribers, single articles not purchasable.N–elle Bons, Nadine Mestre-Frances, Patrick Belli, FranÁoise Cathala, D. Carleton Gajdusek, and Paul Brown The work was funded by the Region Languedoc-Roussillon, the Ministry of National Education, Research and Technology, and the Mediterranean Association for the Study on Cerebral Aging.
The first author is a neurobiologist at the Ecole Pratique des Hautes Etudes, Laboratoire de Neuromorphologie Fonctionnelle, UniversitÈ Montpellier II, Montpellier,ÝFrance with 46 publications at Medline, 3 others of which concern TSE in primates [abstracts below] and 14 of which concern other amyloidoses in primates. Contact information is email: ephemcb@crit.univ-montp2.fr, Tel: (33) 04.67.52.40.08, Fax:(33) 04.67.63.33.27. [fluent in English]
The second author, Nadine Mestre-Frances, co-authored the group's 4 articles on primate TSE and 7 other papers on primate amyloidoses, and is apparently a post-doc in the lab the first author.
The third author, FranÁoise Cathala, has 60 publication on scrapie and familial CJD dating back to 1971, including 4 earlier concerning primates. In 1985, she dscribed a case of CJD in a 19-year old girl [Eur J Epidemiol 1985 Mar;1(1):42-7. In 1981], later 'transmission to primates of a CJD-like disease from two familial cases of Alzheimer disease' is discussed [Neurology 1981 Mar;31(3):323-5].
The fourth author, Patrick Belli, is affiliated with the Centre National d'Etudes Veterinaires et Alimentaires, Pathologie Bovine, Lyon,ÝFrance. He has 5 earlier TSE papers, including 2 on BSE, 2 on scrapie, 1 on the French zoo cheetah. The CNEVA Web site exists but is dysfunctional.
Gajdusek and Brown have well-known histories of research in this area.
Abstracts of earlier work on prion disease in French primates and zoo animals:
Baron-T, Belli-P Madec-J-Y Moutou-F Vitaud-C Savey-M. Vet Rec 1997 Sep 13;141(11):270-1 Spongiform encephalopathy iy in an imported cheetah in France.
C R Acad Sci III 1997 Dec;320(12):971-9 Bons N, Mestre-Frances N, Guiraud I, Charnay YWe report on two animals of a non-human primate species Eulemur fulvus mayottensis, housed in the local zoo and fed over a number of years with a food containing cattle meat, that developed serious neurological symptoms associated with prion immunoreactivity in brain and various viscera. Microscopy of the brains showed neuronal vacuolation with patchy/perivacuolar immunolabelling with an abnormal isoform of prion protein (IR-PrP), an important characteristic of spongiform encephalopathy. For the first time, we report the presence in the same severely ill animals of IR-PrP in the gastrointestinal tract, detected by immunocytochemistry with mono- and polyclonal antibodies directed against various parts of the PrP.
Strong PrP labelling was observed in the epithelial cells lining the pharyngeal and gastrointestinal lumen. The tonsils and the walls of the lymph and blood vessels below the intestinal epithelium were also labelled. There were no such immunoreactions in healthy lemurians killed as controls, i.e. a younger congener of the same species housed under the same conditions, and others belonging to the smaller species Microcebus murinus, reared in the laboratory and never fed on commercial food products containing cattle meat. These results demonstrate a strong PrP accumulation in the brain, the gastrointestinal tract and underlying lymphoreticular structures in these primates living in a zoological park and suffering from a spongiform encephalopathy.
C R Acad Sci III 1996 Aug;319(8):733-6 Bons N, Mestre-Frances N, Charnay Y, Salmona M, Tagliavini FThis paper reports the occurrence of a spongiform encephalopathy (SE) in young adult monkeys housed in a zoological park. Three rhesus monkeys (Macaca mulatta) acquired from the same zoo and maintained on feed containing animal protein, developed a progressive neurological disorder with behavioural abnormalities and physical deterioration and died at the age of 10-year-old. Neuropathological examination of one of these animals revealed a spongiform encephalopathy similar to that observed in monkeys following experimental transmission of Creutzfeldt-Jakob disease (CJD). In particular, several brain regions exhibited vacuolation of nerve cell bodies and processes accompanied by astrogliosis. Immunohistochemical analysis showed prion protein (PrP) immunoreactivity at the periphery of vacuolated neurons. The spontaneous occurrence of a SE in these young monkeys might be related to consumption of protein of animal origin.
Lancet 1996 Jul 6;348(9019):55 Bons N, Mestre-Frances N, Charnay Y, Tagliavini F [no abstract available, free full text at Lancet]Sir--Although human and animal prion diseases have been experimentally transmitted to various monkey species, spontaneous occurrence of a spongiform encephalopathy in non-human primates has not been reported. We describe the development of a spongiform encephalopathy without known cause in a rhesus monkey (Macaca mulatta).
The monkey was born in 1982 and was acquired from Ravensden zoo, Rushend, Northants, UK, in October, 1986, by the zoological park in Montpelier, France where it was housed in the primate colony. In Montpelier, the monkey was fed standard monkey feed including Singe 107 (UAR, Paris, France) which contains meat products declared fit for human consumption. In summer, 1991, aged 9, the previously healthy monkey became lethargic and developed mood changes, in particular, aggressiveness. The monkey became isolated from its companions and hid itself away. In June, 1992, the monkey was anaesthetised with pentobarbital and perfused intracardially with saline followed by 4% paraformaldehyde.
The brain was dissected into 5 mm coronal blocks, which were embedded in paraplast. 6 µm serial sections were stained with haematoxylin and eosin and thioflavine S for amyloid, or immunolabelled with antibodies: monoclonal antibody to a synthetic peptide homologous to residues 106-126 of human prion protein (PrP)...
There were conspicuous spongiform changes and gliosis in the grey matter, particularly in the cerebral cortex where the deep layers were mostly involved. A number of neurons in the affected areas contained large vacuoles; these were single or multiple and distended the soma to produced ballooned cells with a marrow rim of cytoplasm. Immunohistochemistry showed that vacuolated neurons and distended nerve-cell processes were surrounded by PrP-immunoreactive material (figure). The severe vacuolation of neurons observed in this rhesus monkey is similar to the vacuolation seen in squirrel monkeys after inoculation with brain material of patients with Creutzfeldt-Jakob disease (CJD).1 Immunological labelling of neurons in this monkey also resembled those seen in the brains of patients with CJD. Immunostaining showed anti-þ protein immunoreactive deposits in the neuropil of the cerebral cortex.
As far as we are aware, this is the first reported case of spontaneously developed spongiform encephalopathy in a monkey. The feeding of this monkey with animal protein raises the possibility of cross-species transmission of the disease through contaminated foodstuff, similar to that suggested for bovine spongiform encephalopathy.
1 Zlotnik I, Grant DP, Dayan AD, Earl CJ. Transmission of Creutzfeldt-Jakob from a man to squirrel monkey. Lancet 1974; 24: 435-38.
11 Apr 99 webmasterThis pair of news stories in the 8 Apr 99 Nature is yet another bizarre twist in the primate saga, researchers wanting to deliberately infect more primates. The zoos and breeding facilities are already full of them -- one cannot usefully give a second experimental dose of BSE to an animal already incubating the disease. And the controls? (Note the English have already funded an ominous "Refurbishment of the MRC Primate Facility Dr A F Dixon, Cambridge" on their BSE projects page, suggesting large scale experimental infections are intended.)
The first order of business is mandatory autopsy of all captive primates worldwide that die for any reason, not to capture and infect the few remaining wild ones and put them in contaminated facilities. Biopsies are quite feasible as well.
Absurdly, England is said in the story to have opposed primate experimentation on ethical grounds, this after deliberately exposing primates worldwide for a decade with BSE to make a few quid selling monkey chow. With Prince Phillip head of the World Wildlife Fund primate conservation and Jane Goodall taking the high road with the chimps, the British government itself took the low road. Collateral damage to Europeans or the world's primates were written off as insignificant compared to rebuilding piddling beef exports.
Think for a minute about selecting 1 primate species out of 234 at random (French law against hybrids) and finding that 20 of 20 are dying of BSE. What does that say about probable levels of infectivity in the the other 233 species? Why should the mess be limited to primates? -- I haven't seen any negative autopsies coming out of zoos for the other species.
How many ongoing unrelated biomedical research projects in Europe are unknowingly using infected primates and what are the implications of terminating those experiments?
I share the widely expressed sentiment about the stupidity of many of the proposed experiments -- it is too late to worry about anatomical minuteae of the route of infection or the early role of the immune system. No one is planning to eat TSE-contaminated material again. The nvCJD exposure peaked ten years ago. Why autopsy a primate when you can autopsy people?
It is too late to intervene therapeutically at intermediary steps; besides, we have no idea how and by the time we do it will be even more too late. And this is true for secondary infections as well (oral route studies have undemonstrated relevence to injection nor to second passage within a species). Look at the human tonsil collection to see how the epidemic is shaping up. Use transgenic mice with human prions.
If a case can be made for these primate experiments, nobody has made it yet.
Yes it would have been nice to have done western blot strain-typing, but this is only a convenient distraction for people who cannot or will not face up to reality. These primates did not get prion plaques in their brains by eating grubs and berries in a Madagascar rain forest, any more than the zoo kudus did in 1986 from eating a bale of hay. Primates across France and England (minimally) were fed BSE-tainted feed for over a decade -- little will be learned in the end from pedantic concerns over western blots.
The scientific reaction so far has been irresponsible in the extreme: quietly burn the dead primates without autopsy, hope that nvCJDp does not transmit horizontally (unlike scrapie and chronic wasting disease), and exploit every opportunity to demand more money to pursue personal agendas. If there was any scientific leadership in Europe, the first priorities would be in forcing an investigation of this endangered species debacle and getting on with therapeutics. BSE itself is largely of historical interest -- we do not have the luxury or leisure to study it.
I think most biologists will say, clean up your mess first before talking about infecting more animals. Additionally there will be strong opposition from the animal rights groups, the general public, the conservation community, and the governments (the latter for bad reasons of course). A host of new questions will be raised about zoos, their levels of deceit, and care providers who remained silent as their animals were dying. -- webmaster opinion.
Nature 398, 449 (1999) 8 April 1999 Declan Butler[PARIS] A study published last week that appears to show that primates are easily infected orally with bovine spongiform encephalopathy (BSE) may, if confirmed, have implications for the understanding of the new variant of Creutzfeldt-Jakob disease (vCJD) in humans.
The study could also provide a better and faster animal research model than those currently available. But researchers are angry that more extensive primate experiments, recommended to the European Commission in 1996 by one of its expert committees, have not been supported (see below). This reluctance, argue some scientists, stemmed from opposition in a number of member states to research using primates.
Two young Microcebus murinus lemurs, fed 0.5 g of cattle brain infected with the agent that causes BSE and autopsied five months later, were found to have prion infections, despite showing no outward clinical signs of the disease. Three control animals showed no signs of infection.
The group also autopsied two symptomatic and 18 apparently healthy large monkeys and lemurs from French zoos that had eaten animal feed produced in the United Kingdom. All were found to be infected.
The work was published in the 30 March issue of the Proceedings of the National Academy of Sciences (PNAS) by a Franco-American team including NoÎlle Bons of the University II of Montpellier, Paul Brown of the US National Institute of Neurological Disorders and Stroke in Bethesda, Maryland, and D. Carleton Gajdusek, the 1976 Nobel prize winner for his work on prions, now at the Institut Alfred Fessard near Paris.
Charles Weissman, from the University of Z¸rich, in Switzerland, points out that a major caveat in the study is that infection was assessed by immunostaining of proteinase-resistant prion protein, rather than by Western blotting of abnormal prion protein. Fixed brain samples which cannot be tested using a blot were taken at the time, explains Bons, adding that ongoing experiments will use frozen samples and Western blotting.
At the same time, Weissmann -- who chaired the committee that advocated more primate studies in 1996 -- says that he hopes the new findings may encourage more interest in these. The findings, if confirmed, could have "very worrisome implications" for humans, he adds, as they suggest that primates closely related to us may be highly susceptible to oral transmission of BSE.
Dominique Dormont, whose group demonstrated the first transmission of BSE to a macaque (see Nature 381, 743; 1996), points out that Microcebus murinus may be a valuable new animal model. This tiny species weighs just 100 grammes and has a short lifespan of eight to ten years, making it much more suitable for research than macaques and other larger, longer-lived primates.
According to Bons, another lemur from another experiment has fallen ill with symptoms of spongiform encephalopathy one year after being fed contaminated cattle brain. The short time to incubation and illness suggest the species would be ideal for research purposes, she says, warning at the same time against the dangers of extrapolating results directly to humans.
In the experimental PNAS study, infection was found throughout the tonsils and peripheral tissues, and confirms an oral route of transmission of BSE, says Dormont. The agent appears to pass through the digestive tract, across the spleen, and up the spinal cord into the brain, says Brown.
Bons argues that the results also suggest that nervous degeneration and wide peripheral infection occurs early in the incubation phase, raising the prospect that in humans asymptomatic individuals could be carrying high levels of infections -- which might have implications for secondary transmission. "We know nothing of the kinetics of nvCJD in humans [where living individuals cannot be autopsied]," says Brown, arguing that primate studies could therefore yield new information on this.
The dose of brain fed to the lemurs is equivalent to a 70 kg man eating a dose of 500 g. The larger experiments needed to establish the minimum dose needed for infection could not be carried out, however. This is "unfortunate," says Weissmann, pointing out that his committee called in 1996 for precisely such experiments, as well as ones on maternal transmission.
Bons is also studying two baby lemurs born from infected parents. She says she is collecting samples to establish whether and how maternal transmission occurs.
The PNAS paper contains another potentially controversial element. According to Bons, the the only meat in the zoo primates' food was British 'cretons de boeuf', a meat residue approved for human consumption. A fuller inquiry into the source of infection is needed, she says.
Nature 398, 449 (1999)[PARIS] Dominique Dormont, a prion expert who is head of the French government's spongiform encephalopathy advisory committee, is -- like many BSE/CJD researchers -- dismayed at what he claims are the years lost because of an apparent lack of political support for work using primates.
Dormont says that a proposal for experiments on macaques, submitted in 1996, was not approved because some states -- including the United Kingdom -- opposed it on animal-welfare grounds.
Large-scale primate experiments can only be carried out at a continental level, says Dormont, because of the high costs and logistics. He says that some countries felt such experiments would be a waste of money, as, by the time their results came through, the results of human exposure to BSE would already be beginning to appear.
Paul Brown, of the US National Institute of Neurological Disorders and Stroke, one of the authors of the PNAS paper (see above), adds that he advocated similar experiments in the United States at roughly the same time. But he claims that government safety restrictions on BSE and nvCJD materials have all but halted research.
"I can understand that they don't want every Tom, Dick and Harry working on these agents," says Brown. "But the regulations are blocking all research in the handful of labs who have forty years experience handling these agents."
NoÎlle Bons, from the University II of Montpellier, another author of the paper, wrote to Edith Cresson, the then European commissioner for research, in 1998, presenting her with the preliminary results of the PNAS paper. She also complained that she had heard nothing regarding a research proposal on BSE she had submitted the previous year. Bons claims that, despite meeting a member of Cresson's staff, she remains in the dark as to support for her project.
2 Apr 99 webmasterIt is quite feasible to construct a worldwide database of which zoos hold how many of which species of primate, from information on the Web. I did this using Dec 1998 data for 5,988 captive animals representing 8 of the 56 genera of primates. Of these 665 are at 23 British and 555 at 15 French zoos. This extrapolates roughly to 42,000 captive primates of 53 genera with 8,500 in the UK and France (this would turn up additional zoos). At the bottom is the data for the rest of Europe.
How many TSE deaths these zoos have had, how many are silently incubating the disease, and whether BSE in exported primates transmits horizontally as efficiently as CWD are the questions of the hour. A close reading of the PNAS study gives support to a truly catastrophic scenario.
France 114 Mulhouse 67 La Palmyr 54 Romagne 50 Fontaine 50 Beauval 42 Lille Zo 35 Besancon 34 Peaugres 31 Paris Zoo 15 Thoiry 14 Lisieux Z 13 Amiens 13 La Plaine 12 Paris Jp 8 Touroparc 3 Obterre UK 94 Twycross 83 Banham 52 Colchestr 48 London Rp 47 Jersey 37 Marwell 37 Edinburgh 33 Chester 30 Chard 26 Dudley 24 Burford 24 Blackpool 22 Paignton 20 Bristol 15 Southport 15 Battersea 13 Bekesbrne 11 Chesingtn 9 So Lakes 8 Lympne 8 Alfriston 5 Colwynbay 4 Whipsnade
12......Fr......Allenopithecus......Paris Jp 5......Fr......Allenopithecus......Mulhouse 2......Fr......Allenopithecus......Lille Zo 8......Uk......Allenopithecus......Edinburgh 3......Uk......Allenopithecus......Twycross 2......Uk......Allenopithecus......Paignton 25......Uk......Alouatta......Twycross 8......Uk......Alouatta......Lympne 5......Uk......Alouatta......Bristol 4......Uk......Alouatta......Banham 2......Uk......Alouatta......Paignton 1......Uk......Alouatta......Bekesbrne 11......Fr......Ateles......Fontaine 4......Fr......Ateles......Mulhouse 3......Fr......Ateles......Beauval 3......Fr......Ateles......Touroparc 2......Fr......Ateles......Amiens 2......Fr......Ateles......Fontaine 2......Fr......Ateles......Obterre 1......Fr......Ateles......Fontaine 15......Uk......Ateles......Colchestr 12......Uk......Ateles......Burford 10......Uk......Ateles......Banham 8......Uk......Ateles......Chester 8......Uk......Ateles......Twycross 7......Uk......Ateles......Banham 7......Uk......Ateles......Twycross 4......Uk......Ateles......Bristol 4......Uk......Ateles......London Rp 3......Uk......Ateles......Paignton 3......Uk......Ateles......Twycross 2......Uk......Ateles......Blackpool 2......Uk......Ateles......Burford 2......Uk......Ateles......Colwynbay 2......Uk......Ateles......Dudley 2......Uk......Ateles......Twycross 1......Uk......Ateles......Blackpool 1......Uk......Ateles......Bristol 1......Uk......Ateles......Colwynbay 1......Uk......Ateles......Southport 1......Uk......Ateles......Southport 1......Uk......Ateles......Twycross 1......Uk......Cacajao......Twycross 11......Fr......Callimico......Beauval 4......Fr......Callimico......Mulhouse 2......Fr......Callimico......La Palmyr 1......Fr......Callimico......Lille Zo 8......Uk......Callimico......Jersey 8......Uk......Callimico......London Rp 8......Uk......Callimico......Marwell 5......Uk......Callimico......Paignton 4......Uk......Callimico......Alfriston 4......Uk......Callimico......Chard 4......Uk......Callimico......Edinburgh 3......Uk......Callimico......Colchestr 3......Uk......Callimico......Twycross 2......Uk......Callimico......Dudley 1......Uk......Callimico......Colwynbay 1......Uk......Callimico......Southport 23......Fr......Callithrix......La Palmyr 17......Fr......Callithrix......Mulhouse 17......Fr......Callithrix......Peaugres 12......Fr......Callithrix......La Palmyr 11......Fr......Callithrix......Besancon 10......Fr......Callithrix......Beauval 10......Fr......Callithrix......Lille Zo 8......Fr......Callithrix......Beauval 5......Fr......Callithrix......Fontaine 4......Fr......Callithrix......Fontaine 4......Fr......Callithrix......La Palmyr 4......Fr......Callithrix......La Palmyr 4......Fr......Callithrix......La Palmyr 3......Fr......Callithrix......Mulhouse 3......Fr......Callithrix......Peaugres 3......Fr......Callithrix......Romagne 2......Fr......Callithrix......Lille Zo 2......Fr......Callithrix......Lisieux Z 2......Fr......Callithrix......Romagne 1......Fr......Callithrix......Lille Zo 1......Fr......Callithrix......Mulhouse 14......Uk......Callithrix......Banham 13......Uk......Callithrix......Edinburgh 13......Uk......Callithrix......London Rp 13......Uk......Callithrix......Twycross 12......Uk......Callithrix......Twycross 11......Uk......Callithrix......Banham 10......Uk......Callithrix......London Rp 9......Uk......Callithrix......Blackpool 9......Uk......Callithrix......Jersey 9......Uk......Callithrix......Jersey 8......Uk......Callithrix......Colchestr 7......Uk......Callithrix......Colchestr 7......Uk......Callithrix......Edinburgh 6......Uk......Callithrix......Battersea 6......Uk......Callithrix......Bristol 4......Uk......Callithrix......London Rp 4......Uk......Callithrix......Twycross 3......Uk......Callithrix......Blackpool 3......Uk......Callithrix......Edinburgh 3......Uk......Callithrix......Marwell 3......Uk......Callithrix......Paignton 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5 De Callithrix Magdeburg 26 De Callithrix Magdeburg 19 De Callithrix Berlinzoo 16 De Callithrix Halle 14 De Callithrix Frankfurt 13 De Callithrix Frankfurt 13 De Callithrix Koln 11 De Callithrix Augsburg 10 De Callithrix Berlinzoo 9 De Callithrix Augsburg 7 De Callithrix Duisburg 7 De Callithrix Heidelbrg 7 De Callithrix Koln 7 De Callithrix Hannover 6 De Callithrix Krefeld 6 De Callithrix Munster 6 De Callithrix Berlinzoo 5 De Callithrix Heidelbrg 5 De Callithrix Magdeburg 4 De Callithrix Munich 4 De Callithrix Duisburg 3 De Callithrix Leipzig 3 De Callithrix Munich 3 De Callithrix Saarbruck 3 De Callithrix Stuttgart 3 De Callithrix Heidelbrg 2 De Callithrix Magdeburg 2 De Callithrix Stuttgart 2 De Callithrix Magdeburg 1 De Cebus Berlinzoo 1 De Cebus Karlsruhe 3 De Cebus Magdeburg 2 De Cebus Neuwied 4 De Cebus Rostock 1 De Cebus Rostock 0 De Eulemur Berlinzoo 37 De Eulemur Koln 11 De Eulemur Koln 11 De Eulemur Wuppertal 10 De Eulemur Koln 9 De Eulemur Saarbruck 9 De Eulemur Koln 8 De 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Callithrix Helsinki 1 Fi Allenopithecus Szeged 1 Hu Callithrix Budapest 2 Hu Cebus Budapest 42 Hu Varecia Budapest 4 Hu Ateles Belfast 6 Ir Ateles Belfast 4 Ir Ateles Fota 4 Ir Ateles Dublin 2 Ir Ateles Dublin 2 Ir Ateles Belfast 1 Ir Ateles Belfast 1 Ir Ateles Dublin 1 Ir Callimico Belfast 11 Ir Callimico Dublin 2 Ir Callithrix Belfast 18 Ir Callithrix Belfast 7 Ir Callithrix Belfast 2 Ir Cebus Dublin 1 Ir Eulemur Fota 3 Ir Eulemur Fota 2 Ir Varecia Belfast 2 Ir Varecia Belfast 9 Ir Varecia Dublin 2 Ir Varecia Fota 2 Ir Varecia Fota 4 Ir Ateles Puntaverd 1 It Callithrix Bussoleng 2 It Callithrix Roma 2 It Cebus Bussoleng 3 It Cebus Puntaverd 3 It Cebus Roma 1 It Eulemur Bussoleng 6 It Eulemur Bussoleng 3 It Eulemur Bussoleng 2 It Eulemur Bussoleng 2 It Eulemur Roma 2 It Varecia Bussoleng 2 It Varecia Bussoleng 9 It Varecia Bussoleng 9 It Alouatta Apeldoorn 3 Ne Ateles Rotterdam 9 Ne Ateles Apeldoorn 7 Ne Ateles Antwerp 6 Ne Ateles Amsterdam 4 Ne Ateles Apeldoorn 2 Ne Ateles Emmen 2 Ne 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Varecia Singapore 2 Sp Varecia Singapore 5 Sp Callithrix Moscow 12 Su Callithrix Rostov 2 Su Callithrix Moscow 1 Su Cebus Moscow 2 Su Cebus St Peters 6 Su Cebus St Peters 4 Su Eulemur St Peters 7 Su Eulemur Moscow 3 Su Varecia Moscow 2 Su Varecia St Peters 1 Su Varecia St Peters 3 Su Varecia St Peters 3 Su Ateles Basel 9 Sw Ateles Basel 5 Sw Callimico Irchel 154 Sw Callimico Bern 4 Sw Callimico Zurich 3 Sw Callithrix Zurich 9 Sw Callithrix Zurich 7 Sw Callithrix Zurich 1 Sw Cebus Zurich 8 Sw Varecia Basel 2 Sw Varecia Total Zurich 9 Sw Callithrix Kolmarden 12 Swe Callithrix Eskilstun 7 Swe Callithrix Boras 3 Swe Callithrix Furuvik 2 Swe Callithrix Furuvik 1 Swe Cebus Furuvik 8 Swe
12 Apr 99 webmasterThere are 3,070 captive chimps worldwide (687 in Europe: 123 UK, 39 France) of which a good many would have been fed mad monkey chow for years or been cagemates of such. HIV, hepatitis C, and toxicology testing -- how does one interpret neurological side effects in a chimp with nvCJDp?
Note the English have already funded an ominous "Refurbishment of the MRC Primate Facility Dr A F Dixon, Cambridge" on their BSE projects page, suggesting large scale experimental primate infections are intended.
Like the French, they are refusing to account for their zoos.
(A very gross account of what the USA did with its hundreds of chimps at Holloman Air Force Base is also online.)
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