NOTES FROM 1989 APV WORKSHOP August 1989 TB reactors in "non-infected animals": Many people are getting positive TB reactors ... believed not to be a true positive. Most probably are Type III (Arthus) reaction. Particularly a problem in Orangs, Patas, Cebus, and baboons. Has been seen in rhesus, aotus and cyno also. Have looked at TB media, various TB tuberculin products (at .1 and .2 ml ID) and saline controls. Often seen as a reaction that occurs within 15 minutes of injection and persist for many days. SIV in Indian Rhesus: Antigenemia seen 14 days post infection, but can persist to result in future related death. Can result in carrier state with recurrent antigenemia. The disease is seen as a decline in helper T cells. Generally, animals < 1 month old when infected die early; animals 1 mo to 1 year have a long slow decline in health; and animal > 1 year old live a long time (chronic carriers?). SIV in Chinese Rhesus: Antigenemia last 12 - 13 days (not as long). Compared to the Indian animal, Chinese Rhesus die earlier and more often. There are fewer chronic carriers than Indian Rhesus. SIV pathology: Multifocal white lesions of small intestine (mycotic). Syncytial Giant Cell Pneumonia (Retroviral pneumonia). Increased antigenemia = Decreased antibodies; as antibodies decline, antigenemia increases SIV Therapy: Not much works. Tang for rehydration, nasogastric feeding. What accounts for the differences seen in SIV response? 1) Genetic variability (Ind. vs Chin.) to susceptibility 2) Individual variability to susceptibility 3) Virus variability DISCUSSION NOTES ON SIV: 1. Delta is looking at 'developing' a Chinese strain of SIV to study the disease in that strain of animal. 2. So far there has not been any Chinese macaque found infected with SIV at purchase. 3. HIV2 will infect Chinese macaques but will not cause disease. COMPARISON OF INDIAN (IM) AND CHINESE MACAQUES (CM): CM is more aggressive. Hybrid vigor does work! CM cross IM = greater aggressiveness (in any colony this animal will become the alpha male). LD50 of IM is 2 times that of CM for any given study. There is a definite therapeutic difference between IM and CM, but that difference is not well defined at this time. TB incidence is very low in CM, an interesting observation since TB in the human population in china is high. Do have a significant measles problem in purchased CM's. DENTISTRY (Cheek of monkey) þ þ Disto-facial À¿ Facial ÚÙ Mesio-facial À¿ ÚÙ À¿ ÚÙ þÍÍÍÍÍÍÍÍÍÍÍÍÍÍÍþ º º (Tail of monk)þÄÄÄÄĶ Tooth ÇÄÄÄÄÄÄÄÄþ(Front of monk) º º þÍÍÍÍÍÍÍÍÍÍÍÍÍÍÍþ ÚÙ À¿ Disto-lingual ÚÙ Lingual À¿ Mesio-lingual ÚÙ À¿ þÄÙ (Tongue) þ Initial exam: Facial view is first, laterals are next (Extraoral, then intraoral). Often will find abscesses in mandible, palpate tissues of mouth extensively. Young Primates: - No pre-molars (only 1 molar) - Lots of space between teeth - Much softer tan permanent teeth - Grind down faster - Develop caries, etc easier Thumb sucking causes malocclusion and finger deformity. Associated behavior includes hair playing, ear or fanny plucking with non-sucking fingers. Very characteristic. Instruments needed for dental exam: - Mirror - Explorer - Williams periodontal probe Periapical abscess may or may not be related to affected tooth. Be sure and check other teeth well. With abscessed tooth often have ancillary cyst. You need to pull tooth and curette to bone. Leave no soft tissue un-curetted. Dry sockets occur fairly frequently. A dry socket is an osteitis in alveolar bone. Curette the socket until it bleeds freely. Be sure and place a clot in the tooth socket, even if it is necessary to draw IV blood and place in socket. Wait to observe clot. Suture over socket to prevent dry socket. Recheck teeth at 48 hours post-operative. Dissolution of clot indicates deep seated bacteria and you need to replace the clot. Suturing the gum line helps hold the clot in place. Try not to extract, if at all possible. Extraction results in migration which can result in malocclusion. Cut teeth off at gum line or (as will be described in an up coming paper) cut off below gum line, but leave the root in place. Permanent maxillary teeth come in in front of deciduous teeth. Permanent mandibular teeth come in behind deciduous teeth. If decay of the deciduous tooth affects the pulp, then exfoliation and root resorption WILL NOT OCCUR. Eventual outcome will be fractured deciduous tooth, retained deciduous, periodontal disease, osteomyelitis, maxillary resorption. Periodontal disease is an IMMUNOLOGIC disease. There are no bacteria found in the affected tissues, only in the spaces. The problem is the exotoxin, that causes periodontal disease. There is no cure of periodontal disease, only control. The gingival fluid flushes the crypt spaces out. If the flow decreases, then the problem gets worse. PERIOTRON: Paper to measure fluid produced by tooth sulcus. You can run complete chemistries off of the fluid. This fluid has the same constituent as sera, only it is more concentrated and has lots of PMN's. Periodontal probing ... Tip of the probe always touches the tooth, never into the tissue. Bleeding while probing indicated gingivitis, which may or may not lead to periodontal disease. There are such things as genetic resistance to periodontal disease or tooth decay or both. Gingivitis ... inflammation of the collar around the tooth. PELLICLE: Saliva film over teeth. Bacteria floats in the pellicle. Plaque forms in the pellicle and attaches to the tooth surface. This occurs within 5 minutes after brushing. All personnel should use maximum protection when cleaning teeth. Remember ... you are actually aerosolizing blood. Use 2 masks, sealed with eye protection, hood, clothes, etc. Plaque and calculus accumulation indicate animal not using teeth to eat. Usually the result of disease. Fix the disease process, the animal will start eating again, and the teeth will clean themselves. Normal eating generally keeps teeth clean. Use lidocaine with 1:100,000 epinephrine to control bleeding. Decrease risk of abscess or dry socket if first clean calculus off teeth, then extract. Extraction without prior cleaning will drive calculus into alveolar tissues. Periodontal disease is a more severe than decay. You cannot tell decay only by seeing a dark spot. Press the probe into the tooth, if it is soft and gets stuck; it is decay. Periodontal disease is generalized; decay is a point disease that spreads next door. Teeth loss is the end result of periodontal disease and or dental disease. "NOT AGING" Periodontal disease is progressive. So see disease in old age but that is only result of life long problem. Gingival margin develops hemangioma like appearance in diabetes. In AIDS in humans, Ulcerative gingivitis (ANUG) is a big problem. This also can occur in SIV infected monkeys. Secondarily, septicemia, lung and kidney abscesses, caused by fusobacterium and a spirochete. Gingival hyperplasia ... The cause is unknown. Possibly it is a traumatic problem. Often it will result in gingivitis. The hyperplasia should be cut off. Good results will occur. EPULIS: Limited hyperplasia. A generalized term referring to many syndromes. The result of an irritant (Calculus, decay, root tip). Often the result of canine extractions in baboons. Cut the epulis at the surface, remove the irritant, leave it open. (This is the only time you leave a dental procedure open). This and ANUG are the only time find organisms in tissue. Upper canine and the lower premolar have a scissor - like action. Rounding the canine tip is of little or no value; you should also shave the reverse side of the canine. A good method for canine removal is to: 1) Cut the canine off low. 2) Apply Calcium hydroxide 3) Dental varnish 4) Place amalgam 5) Don't put amalgam next to pulp! Calcium hydroxide stimulates pulp tissue to produce dentin to seal pulp chamber. Varnish keeps a space between amalgam and pulp cavity. Varnish will be dissolved as dentin grows. Dentin oxidizes amalgam and results in sealing the tooth. Use TOOTH SEALANT (commonly done for human kids these days) to prevent decay or severe calculus build - up. Color the plastic so that you can visually identify when the sealant is gone and when it needs to be replaced. Wonderful stuff. Simple to apply. Doesn't hurt the animal at all. SIMIAN HEMORRHAGIC FEVER: SHF and measles are probably the 2 greatest killers of primates, i.e. do not survive. July 64 ... Russia, 62 monkeys become ill. All die. Baboons, patas, rhesus, cynos. Transmission by needles, scalpel. October 64 ... 222 of 223 monks at NIH die. Rhesus, cyno, stumptail, baboons, and patas. Spread via tatoos needles and catch gloves. October 67 ... 500 die in Davis, Calif. Rhesus, cynos, patas. Never isolated a virus. November 72 ... NIH, Rockville, 212 Rhesus and Patas die. In a study done later it was found that Patas sera given to rhesus caused the disease. However, it was not thought that direct contact spread the disease. 1975 ... 110 dead cynos and patas. 1976 ... 46 die when newly arrived from the philippines. Patas affected. It is an RNA virus. Flavivirus. It is not aerosol transmitted. Chlorox easily kills, but it is heat resistent. The virus must enter parentally. No oral transmission. It does not replicate in any lab animals other than monks. It is not passed via sexual contact (no vertical transmission). Interferon is given 8 hours pre - infection will protect. Probably it is not a problem in humans. It doesn't grow in chimpanzees. Macrophages are the target organ. Chronic carriers include African Green, Baboon, Patas. Conclusion: Originates in African species, mechanically transmitted, animal contact. It has been experimentally transmitted via biting insects (bugs and flies). It has been (antibodies) in wild caught Mus musculus at New Mexico Primate Res. Center following thier recent epidemic. Are rodent reservoirs? I hope not. Immunosuppression is important. Best method for screening for SHF is Rhesus macrophage test. Many chronic carriers do not have antibodies. March - May 1989. NMRI. 50 monkeys imported from the philippines. All die. The disease spread throughout the facility. Bad quarantine decisions contributed to disease outbreaks. Factors leading to the problem: 1) Unloaded at the dock in protocol research area. 2) The monkey sat on the dock in the hallway leading to the research rooms for one hour before moving to quarantine. 3) Nets were moved from room to room (they were disinfected, but apparently not good enough) 4) There was common movement in all animal study rooms. 5) Animal to animal contact in the quarantine area. 6) Use of plastic gloves between cages were not changed. 7) Positive pressure of rooms 8) Sewage back - up into quarantine rooms and study rooms 9) Common hallway between quarantine rooms and study rooms 10) In corn cribs, insects travel from crib to crib. In 60 days, an additional 120 monkey died in the corn cribs. The corn cribs were infected by animal movement out of the study rooms and into the breeding cribs prior to the observance of clinical signs (incubation = 5 days). CLINICAL SIGNS OF SHF Lethargy Anorexia Epistaxis Hemorrhage Photophobia Rough Hair Ataxia Fragmentation of RBC's Proximal duodenal hemorrhage (PATHOGNOMONIC) All chemistries are high. ALT, AT, LDH, CPK, GGT. A test called FDP (Fibrin Degradation Product) is used to indicate presence of SHF. If FDP turns positive, you've got problems. VERO cells = African Green Kidney Cells HISTO: Aggregation of fibrin in glomeruli and tubules. Minimal to no liver changes. LAB TEST: Cell culture (kills MA-104 cells but not VERO cells). IFA not good because no survivors (Macaques) i.e. no antibodies. IFA does work in patas (a few survive). Serious cause of disease in Aotus: 1) Hypertrophic form cardiomyopathy 2) Dilative form cardiomyopathy Clinical signs: Sudden death, respiratory distress, syncope, ascites Columbian karyotypes (2, 3, and 4) most often affected. CASE REPORTS LEPROSY 32 year old chimp with clean nasal discharge, ulcerative lesions on hands, feet, and scrotum. Nodular lesions were seen on the face also. CBC : mild leukocytosis; SMAC normal. TB : negative Skin H & E: Chronic active granuloma AFB stain: Mycobacterium leprae lepromin skin test was positive DISCUSSION NOTES ON LEPROSY: This disease is probably out there in much higher percentages than we know. It can imitate many conditions, so it is worth checking on in chronic confused diagnostic dilemmas. 1 - 30 year incubation. Less than 5% of the contacts acquire leprosy. Transmitted via nasal discharge. Does infect Rhesus, African Green, Sooty Mag. VITAMIN C DEFICIENCY 3 and 4 year old rhesus in breeding facility corrals. Signs were very slight at first, consisting of painful movement, hair loss, radiograph swelling, epiphyseal separation, small fractures, gum hemorrhages, joint hemorrhages, inhibition of macrophage function. Recovery a week after supplement started. Treatment: Oranges, tang, dust feed with powdered vitamin C. DISCUSSION NOTES ON VITAMIN C: Early epidemiology of this disease resembles an infectious agent. The rear limbs are most often affected first. It is age related with <1 year and > 5 years almost immune (until it really gets bad). Why? The affected ages are in a growth phase requiring fairly high levels of vitamin C. Age of final epiphyseal closure is 6 years. LISTERIOSIS 9 stillbirths and 2 post natal deaths due to listeriosis in a breeding colony of rhesus. Only sign was gingival lesions of hemorrhage at tooth eruption sites of immature animals. Placenta and fetal were cultured. SHIGELLA FLEXNERI Less than 100 microbes can produce clinical disease. Transmitted by fecal - oral route. Also flies, or fomites. ?Humans and primates without signs are sources of the disease. Resistance to most all antibiotics has been observed. A new medication called ENROFLOXACIN (Baytril) from Haver Pharmaceutics was used with success. This medication is approved for the dog but has restrictions on the use in puppies. There has been no evidence of a contraindication in pregnant females but conservative therapy should be used in the neonate. Dose at 5 mg/kg q 24 hours by nasogastric tube or IM injection for 10 days. An alternative medication which may be hard to get but is effective, is NORFLOXXICILlIN. HERPES B - AN UPDATE OF THE LAST YEAR. Case 1. Animal caretaker scratched approximately on 7 May 1989. On 7 June 1989 presented with lower limb weakness, lower back spasm. Prescribed muscle relaxers and bed rest. On 13 June 1989, re-presented with high fever plus continuing signs of 7 June. Prescribed antibiotics, anti-inflammatory agents and rest. That afternoon he went into respiratory arrest and had copious amount of oral foam. The father gave mouth to mouth resuscitation until paramedics arrived. Prior to going unconscious he said he had had a bite on the shoulder. He was then quadriplegic and quickly became comatose. Life support was removed based on NMRI findings of 20 June. He died 30 minutes later. The initial bite site continued to grow virus up to death. This has been characteristic of exposures thus far. This isolate in Michigan was the most neurovirulent of all isolates thus far. This patient, as with other before him, had a hemorrhagic conjunctivitis (Almost a pathognomonic lesion?). Case 2. On 20 June this patient entered the hospital experiencing lower limb weakness. This patient was from the same facility as patient 1. He was placed on Acyclovir and eventually released from the hospital. Case 3. The bite of this patient culture positive. He was placed on Acyclovir and released. He has remained consistently negative for subsequent cultures. Case 4. Symptomatology was correct, but the virus was never cultured. CSF profile did indicate B virus was present. Patient was treated and has been released on medication. Case 5. The father of the patient in case 1. Has remained negative by all test for exposure to B virus. Continues to be monitored. DISCUSSION NOTES OF B VIRUS: 1) Prior exposure to Herpes simplex does not provide protection to B virus as was earlier considered. 2) New medications are now becoming available: a) FEAU and FIAU (Flurouricils) are 100 fold more effective than acyclovir in laboratory test. Toxicity is a problem, but probably not ass severe as originally thought. These are experimental drugs at NIH presently. b) DAPG ... Cytovine or Cyclovir is a second generation product. It is a better drug that Acyclovir but has more clinical side effects. It is administered IV as inpatient. c) Acyclovir is now a licensed drug under the name Syntex. 3) In a study underway at NIH, of 160 human sera samples collected from caretakers and others associated with primates, 3 have been found to have definite antibodies to B virus and 5 have been termed suggestive positives. The work is not complete, and repeat testing will be performed before the results are released to the common man. 4) FDA is the enforcing agency for B virus related items (they also do salmonellosis in turtles and psittacosis in chickens; all else belongs to USDA). This is under the Public Health Service Act. The word from the FDA guys at the meeting was ... it is time to pay the piper! They have recent sent a letter to the folks in Michigan fining the individual supervisors and the institution for discrepancies. See attachment. 5) The Criminal Enforcement Act of 1984 (PL 98 - 596) list approved fine for wrong doer. For those interested fines are as follows: If you are negligent and a human is harmed: $100,000.00 and a year in jail. If you are negligent and a human is killed: $200,000.00 plus jail time. If your institution is guilty of harm: $200,000.00 If your institution is guilty of death: $500,000.00 6) Informed consent will help, but it may still get tough. Informed consent DOES NOT pertain to the wife of the injured caretaker. Be sure personnel being brief understand what it is we are talking about. Some have recently complained, that they only had a 4th grade education and didn't understand the big words. Make it simple. "It will kill you." Get legal people to sit in on staff meeting when discussing the hazards. Make it an annual requirement for review. In other words, do all you can to assure everyone is aware, at their level of understanding, what B virus is and how is can affect them. Then sit back and worry. Anyone want to bump up your AVMA liability insurance? 7) The same business applies to transport of infected primates. FDA says that you had better ship only those you are reasonably sure are free of disease / not shedding. Although it is not yet required, a statement by the veterinarian saying that these monkey are from B-virus free colonies, but they appear clinical healthy may be in our future. As soon as you do that, expect the airlines to refuse shipment (some folks are doing so now ... Burlington) 8) As a therapeutic approach, one may consider `cutting out' the virus from the infected finger; B virus moves at 3 mm a day based on lab studies of Dr. Hilliard