Nasal Instillation Research Articles

Excretion of Poliomyelitis Virus by Monkeys Used for Testing Material of Human Origin.

SummaryPoliomyelitis virus was recovered from the colon contents of both rhesus and cynomolgus monkeys used in the routine testing of infectious human stools by means of nasal instillation, in addition to other routes of inoculation, when the interval between the last nasal instillation of infectious human stool and the first appearance of paralysis was 4 days or less. In none of 11 paralyzed rhesus monkeys in which this interval was from 5 to 18 days was virus found in the colon contents. The 4-day period following the last nasal instillation of infective human stools may be taken as the time required for elimination of virus swallowed by monkeys, when no local multiplication occurs. The present data indicate that monkeys, receiving human stool suspensions by the nasal or oral route, are a potential source of infection not only to their caretakers but also to other primates in the same animal quarters unless special precautions are taken to avoid fecal contamination. No evidence was obtained that poliomy...

Strains of Virus of Foot-and-Mouth Disease Recovered from Outbreaks in Mexico. Pathogenicity and Invasiveness

A report on the identification of samples of virus received from Mexico has been made.1Before referring to the general effects of the Mexican strains of virus in cattle exposed to infection by inoculation, by contact with diseased cattle, or by the nasal instillation of virus, something should be said about the animals employed in these experiments.Foot-and-mouth disease is not endemic in Great Britain but is introduced at irregular intervals, outbreaks occurring sporadically. Since the policy of control is by “stamping-out,” that is, elimination of infected and “in-contact” animals by slaughter and burning (or under the exigencies of war, by burial) along with the rigorous application of other disinfection and “stand-still” orders, it is possible only to purchase cattle with a “clean” history as regards foot-and-mouth disease. The cattle used in these experiments were Devon steers between V/2. and 2 years old, in good condition and weighing between 600-800 lbs, and they could be considered as relatively ...

SPECIAL REVIEWS

Francis bacon made the suggestion that in gathering scientific data we ought to be neither like spiders, which spin things out of themselves,nor like ants which merely collect, but like bees which both collect and arrange. With Bacon's advice in mind, I would like to arrange the facts brought out by certain studies which bear on the following questions: (1) How does poliomyelitis virus enter the human body? (2) How is it eliminated? (3) Where is it found outside of man? (4) What are the modes of spread? (5) Why do we have epidemics? Until recently, the portal of entry in man was considered by many to be by way of the nose. This concept was based on the following: (1) Virus was reportedly detected in human nasal mucosa in one out of three attempts;1 (2) Virus had been recovered from human nasopharyngeal washings collected in such fashion that it was impossible to tell which component, nasal or pharyngeal, contributed the virus; (3) the only extra neural site by which rhesus monkeys could be infected had been by nasal instillation. It should be noted that when monkeys are so infected, typical lesions are always found in the olfactory bulbs.2 Moreover,infection by this route in monkeys can be prevented by spraying the nose with certain chemicals such as zinc sulfate.3 What evidence stands against the nose? First, it should be stressed that in the temperate zones most epidemics of poliomyelitis strike during the warm months—not the common season for

STUDIES ON ENDEMIC PNEUMONIA OF THE ALBINO RAT

An attempt was made to reestablish the virus-like agent associated with rat pneumonia by the nasal instillation of the infectious material into supposedly normal young rats from the selected Princeton colony. The incidence of the pneumonia in these animals after the injection of suspensions of pneumonic lungs from either rats or mice was not significantly greater than the incidence under natural conditions. In attempting to account for the refractory state of the immature rats it was found that the agent was widely dispersed through the breeding colony at an early age. Detailed tests of many litters indicated that the agent was acquired shortly after birth by way of the upper air passages as the result of maternal contact. In most instances it was so well tolerated by young rats during the suckling period and for several months thereafter that its presence was recognized only by nasal transfer to mice. There is every indication that the rate of infection of rats in the breeding colony approaches 100 per cent by the time the animals are old enough to be used experimentally.

MULTIPLICATION AND SPREAD OF THE VIRUS OF ST. LOUIS ENCEPHALITIS IN MICE WITH SPECIAL EMPHASIS ON ITS FATE IN THE ALIMENTARY TRACT

1. Beginning at 24 hours after intravenous injection of about 10 million intracerebral LD(50) of virus there was evidence of simultaneous, progressive multiplication in the brain and intestinal tract. 2. When the virus was introduced directly into the brain or the nasal cavities and mouth, none was found in the intestinal tract until there was general centrifugal spread from the central nervous system during the last stages of the infection at 96 or 120 hours after inoculation when the virus in the entire brain had reached a concentration of about 3 billion LD(50). 3. Centrifugal spread began when the virus in the brain reached a concentration of about 400 million LD(50) and virus appeared in the pharynx, tongue, and adrenals before it was demonstrable in the intestinal tract, blood, or viscera such as the spleen, liver, and kidneys. 4. Despite the high concentrations of virus which developed in the intestinal tract following intravenous inoculation, it was not demonstrable in the stools, differing in this respect from Theiler's virus in mice and poliomyelitis virus in human beings and monkeys. 5. No antiviral agent was found in the stools, but the urine of normal mice having a pH of 5.6, inactivated large amounts of St. Louis encephalitis virus. 6. There was no evidence of multiplication in the nasal mucosa of mice which succumbed with encephalitis following nasal instillation of the virus, the course of events being comparable in this respect to the behavior of the M.V. poliomyelitis virus in rhesus monkeys. 7. At the terminal stage of infection the virus content per milligram of tissue was as great in the leg muscles as in the sciatic nerves. Since this was also true for the urinary bladder, heart, lungs, and tongue among other tissues, and since the amount in the blood was too negligible to account for it, it would appear that the virus either accumulated in these tissues by diffusion from the nerve fibers, along which it was spreading from the central nervous system, or that it multiplied in some constituent other than the nerve fibers.

Aspergillus fumigatus Infection in the Eyes of Baby Chicks

THE pulmonary form of Aspergillus fumigatus infection is of frequent occurrence and has been described in many species of birds. Infection of the eyes by this fungus is apparently less well known. Seven such outbreaks occurring during February, March, and April, 1946, will be described.So far as we know, there has been but one report heretofore of infection of the eyes by Aspergillus fumigatus. This is an excellent description by Reis (1940) of two outbreaks occurring in Brazil. Out of 1,000 chicks two to five weeks old, 100 were affected. Sawdust used for litter was determined to be the source of infection.Reis was not able to reproduce the disease by cohabitation. Inoculation of occular catarrh or corneal tissue of diseased animals by occular instillation, nasal instillation, instillation in the eye the cornea of which had been lightly scarified, and instillation beneath the nictitating membrane gave negative results. The .

Aspiration Bronchography

[While it is generally accepted that in skilled hands injection of oil through the cricothyroid membrane or the trachea yields the greatest number of successful bronchograms, many radiologists are reluctant to undertake the injection themselves. They dislike the minor surgical procedure involved and realise that, in addition to its being an ordeal for the patient, there may be unpleasant sequelæ if the injection is faulty. David C. Ritchie, whilst a radiographer at Horton Hospital, observed the method of simple nasal instillation demonstrated by Brock, who uses this route exclusively. Ritchie describes this method below. It has the considerable merit of allowing the attempt to be repeated, if necessary, without subjecting the patient to any great discomfort. A.M.R.] There are, in common use to-day, four main methods of introducing iodised oil into the bronchial tree before bronchography is carried out. These are as follows:— (1) A large bore needle is inserted through the cricothyroid membrance or through the trachea and the oil is injected through it directly into the trachea. (2) The oil is dropped into the trachea over the back of the tongue through a special curved cannula. (3) The injection is made through a rubber catheter which has been passed into the trachea via the nose. (4) The oil is introduced through a gum elastic catheter which has been left in position after bronchoscopy.

STUDIES ON ENDEMIC PNEUMONIA OF THE ALBINO RAT

A specific disease entity was established in white mice by the nasal instillation of lung and exudate suspensions from each of 14 adult albino rats affected with endemic pneumonia. The induced disease was characterized by pneumonia (96 per cent), otitis media (94 per cent), and rhinitis (30 per cent). It progressed slowly after an incubation period of 7 to 14 days and was attended by a variable mortality which reached 33 per cent in 22 weeks. Maintenance of the disease was regularly accomplished by direct transmission from mouse to mouse but only by the nasal route and only from the three foci of involvement. It was also transmissible by direct contact but required a prolonged period of exposure. The relation of the disease to infectious catarrh, which it closely resembles, is discussed.

STUDIES ON THE NATURE OF THE VIRUS OF INFLUENZA

The pathogenic agent of influenza A has been sedimented from infected extra-embryonic fluids of the developing chick, embryo by ultracentrifugation. Material so obtained contains two fractions resolvable in the analytical centrifuge cell. The first, a homogeneous fraction, showed a sedimentation constant S(20) = 20 to 31 x 10(-13). The second showed a sedimentation constant S(20) = about 800 +/- 100 x 10(-13), was much less homogeneous than the first, and was shown to consist principally of aggregated particles of the more disperse fraction. Both fractions contained the virus in essentially equal amounts per unit of protein weight as calculated from nitrogen determinations. Electron micrographs of the isolated virus protein indicated that the predominating unit is roughly spherical in shape and has a modal particle diameter of about 11 mmicro, in good agreement with the sedimentation data in indicating a molecular weight of about 650,000. Approximately 300 of the particles having molecular weight of 650,000 were present in the minimal dose producing infection in mice after nasal instillation. The influenza A virus may now be regarded as one of the smallest pathogenic agents thus far isolated. Preliminary analyses indicate that it is also one of the least complex, being composed principally of nucleoprotein.

RECIPROCAL TRANSMISSION TESTS WITH INFECTIOUS CATARRH OF CHICKENS, MICE, AND RATS

Infectious catarrh of chickens (fowl coryza of slow onset) was not transmissible to mice or rats by nasal instillation of the specific coccobacilliform bodies. Exudates were also inactive in both rodents on foot pad injection. The infectious catarrhs of the mouse and the rat were reciprocally transmissible by the nasal injection of exudates or tissue cultures of the respective coccobacilliform bodies and by direct contact. Exudates and cultures also produced an arthritic reaction in both hosts on foot pad injection. The coccobacilliform bodies of mouse catarrh were innocuous in chickens on nasal instillation, whereas those of rat catarrh were established locally but were maintained for only two passages. In the opposite host each of the two rodent forms of infectious catarrh reproduced the typical features of the naturally acquired disease, a highly fatal pneumonia being characteristic of the mouse but not of the rat.

STUDIES IN RODENT POLIOMYELITIS

1. Murine SK poliomyelitis virus has been transferred from mouse to guinea pig with the establishment of a fixed strain of cavian passage virus. 2. The disease thus produced in guinea pigs is characterized by the occurrence of flaccid paralysis. Typical poliomyelitic lesions are found in the anterior horn of the spinal cord. 3. Guinea pigs are susceptible to infection with murine virus by the intracerebral, intravenous, intraperitoneal, and subcutaneous route; cavian passage virus produces paralysis only upon intracerebral or intravenous injection. Neither virus paralyzes guinea pigs by feeding or nasal instillation. 4. The potency of the virus (murine or cavian) in guinea pigs is considerably lower than in mice and compares with the titer of the original SK strain in monkeys. In paralyzed guinea pigs the virus is found only in the central nervous system and not in extraneural sites, such as blood or abdominal viscera. 5. Attempts to cultivate cavian passage virus in tissue culture have yielded evidence of some in vitro propagation but no passage virus has as yet been obtained by this method. 6. Cross neutralization tests with cavian passage virus in guinea pigs and with murine virus in mice have established the serological identity of the two viruses. Inactivation of cavian passage virus in guinea pigs by poliomyelitis-convalescent monkey sera is irregular. Complete neutralization has been obtained with a concentrated poliomyelitis horse serum. 7. Resistance to reinfection with potent virus can be demonstrated in convalescent guinea pigs as well as in guinea pigs which have survived a symptomless infection with either murine or cavian virus. This immunity is demonstrable by the power of the serum of such animals to neutralize the virus in vitro and by the ability of nerve tissue to dispose in vivo of the infectious agent. 8. Cavian passage virus has a limited pathogenicity for rhesus monkeys. Of a total of 35 monkeys injected intracerebrally with guinea pig passage virus 26 failed to respond with any manifest symptoms of disease; 8 monkeys showed various signs of definite involvement of the central nervous system consisting of tremor, convulsions, facial palsy, and localized pareses; 1 monkey developed typical flaccid paralysis. 9. Following injection with cavian virus the virus may be recovered from the tissues of normal monkeys but not from the tissues of convalescent monkeys shortly after a paralyzing attack of poliomyelitis due to SK or Aycock virus. 10. Immunization of monkeys with early cavian passage virus by the subcutaneous route has given no clear-cut evidence of protection against intracerebral reinfection with SK poliomyelitis virus. Neither has there been any evidence of effective interference in monkeys injected intravenously with early cavian passage virus and intracerebrally with RMV poliomyelitis virus. 11. The bearing of the experimental data upon the epidemiology of the human disease is discussed.

THE BEHAVIOR OF POX VIRUSES IN THE RESPIRATORY TRACT

Fowl pox virus from active skin lesions was established in the upper respiratory tract of normal chickens by nasal instillation and maintained for 12 successive passages. The nasal infection was not communicable by direct contact but did afford protection, for at least 6 weeks, against subsequent development of the virus in the skin. Multiplication of the virus in the nasal passages was only irregularly attended by specific mucosal changes and was not accompanied by the vigorous counter-reaction engendered by the causal agents of roup. The same strain of virus on propagation in embryonated eggs also survived and multiplied in the nasal tract but with somewhat reduced activity, the 34th egg transfer failing to afford complete protection. Nasal instillation in mice was followed only by a reaction in the lung from which the virus was recoverable through the 7th day.

INFECTIOUS CATARRH OF THE ALBINO RAT

A disease syndrome referred to as infectious catarrh, encountered under natural conditions of exposure in a rat colony, was transmitted to selected animals and maintained for 5 years by nasal instillation or contact. During this period 37 passages were made in 156 rats, the rates of pneumonia, otitis media, and rhinitis being 28, 63, and 87 per cent, respectively. After the 12th passage, Brucella bronchiseptica and Actinobacillus muris (B. actinoides var. muris), which were originally present, were no longer cultivable from infected rats. By reason of the maintenance of infectious catarrh in the absence of the latter and also because of its non-invasiveness on nasal instillation, it is now believed that Actinobacillus muris is of no direct etiological significance.

INFECTIOUS CATARRH OF THE ALBINO RAT

Evidence is presented that infectious catarrh of the rat is caused by Gram-negative coccobacilliform bodies which resemble the agents of fowl coryza and mouse catarrh. These bodies were demonstrable microscopically in at least 90 per cent of the exudates from the nasal passages and middle ears. The characteristic syndrome was reproduced in selected rats by the nasal instillation of pure tissue culture growths and was maintained for 10 successive passages by the injection of exudate. The experimentally produced disease was also communicable by direct contact. The biological characteristics of the specific bodies are discussed in relation to the organisms of the pleuropneumonia group.

THE OLFACTORY BULBS IN HUMAN POLIOMYELITIS

It was shown in a previous report¹that in experimental poliomyelitis one or both of the olfactory bulbs exhibited characteristic pathologic changes, that is, neuronal necrosis, neuronophagia and perivascular and meningeal cellular infiltration, when the virus invaded the nervous system by the olfactory pathway but not after invasion by other pathways (subcutaneous, intrasciatic, intraocular, tonsillopharyngeal and intracerebral). Despite the fact that in recent years there has been a strong tendency to regard the olfactory pathway as the usual portal of entry for the virus in man, there is actually no direct evidence for such an assumption. It was, therefore, desirable to determine whether or not the olfactory bulbs in human poliomyelitis showed changes similar to those found in experimental poliomyelitis which was induced by nasal instillation of the virus. Smith's²report, in 1934, on 56 olfactory bulbs from about 40 cases was inconclusive, partly because the statement that

Effect of Foster-Nursing upon Inborn Resistance of Mice to St. Louis Encephalitis

SummaryIn strains of mice selectively bred for resistance or susceptibility to St. Louis encephalitis virus, foster-nursing exerted no effect upon the course of disease or mortality-rate following nasal instillation of virus.

THE BEHAVIOR OF POX VIRUSES IN THE RESPIRATORY TRACT

Recovery from the transient pulmonary reaction which accompanies the nasal instillation of variola virus in mice was followed by a measurable protection against the homologous virus and also against vaccinia. Variola virus which regularly survived in the lung of normally susceptible mice through the 5th day was noticeably reduced in titer on the 3rd day in the lung of recovered animals, and usually eliminated by the 5th day. Vaccinia virus produced a less severe systemic reaction in recovered mice and its titer in the lung was significantly reduced on the 5th day. The residence of both viruses in the lung of recovered mice was attended by pathological changes, visible macroscopically.

An unusual case of ephedrine poisoning

Summary Report is made of a case of a 5-month-old child showing symptoms of ephedrine poisoning within thirty-six hours following the nasal instillation of 2 gr. of the drug. The clinical picture showed a combination of features common to atropine poisoning with those commonly observed in strychnine poisoning.

Experimental Infection of Dermacentor Andersoni Stiles with the Virus of Lymphocytic Choriomeningitis

ALTHOUGH only 4 years have elapsed since Armstrong and Lillie 1 showed that lymphocytic choriomeningitis is an infectious disease caused by a virus, the evidence indicates that the etiologic agent can be transmitted in several ways. Traub 2 has shown that the virus is discharged in the urine and nasal secretions of infected mice: Findlay and Stern 3 SUCceeded in infecting 3 of 20 mice by nasal instillation of the virus. Finally Wooley, Armstrong, and Onstott 4 have been able to transmit infection occasionally by instilling the virus into the urethra or vagina of monkeys. They have also demonstrated the virus in the seminal fluid and testicular tissue of infected animals. On the basis of these findings, together with the fact that antibodies against the virus were found by them more commonly in adults and especially in criminal and merchant seamen classes, these investi-

A FURTHER CONTRIBUTION TO VITAMIN C THERAPY IN EXPERIMENTAL POLIOMYELITIS

1. Multiple paralytic doses of poliomyelitis virus (RMV strain), when brought together with small amounts of synthetic ascorbic acid in vitro, are rendered non-infectious as determined by intracerebral injection of such mixtures into rhesus monkeys. 2. Vitamin C administration to monkeys infected intranasally with the RMV strain produces results which differ in accordance with the technique employed for nasal instillation. With an infection of maximum severity, induced by flooding the nasal portal of entry with large amounts of virus, vitamin C administration fails to exert any demonstrable influence on the course of the disease. With a less forceful method of droplet instillation, the picture of the disease in control animals becomes so variable that the results cannot be easily interpreted; but the available data suggest that vitamin C treatment may be a factor in converting abortive attacks into an altogether non-paralytic infection. 3. The administration of synthetic vitamin C to monkeys infected intracerebrally with small doses of the RMV strain gives results comparable to those previously obtained with this substance in monkeys infected intracerebrally with the Aycock strain of poliomyelitis virus. 4. The implications of the findings are discussed.