Conjunctival Scrapings Research Articles

Isolation of Chlamydia psittaci from cats with conjunctivitis.

Chlamydia psittaci was repeatedly demonstrated in stained smears of conjunctival scrapings from a group of cats in a single household and in 5 instances the organism was isolated by yolk sac inoculation of 6-day-old pathogen free, embryonated hen eggs. Thirteen of 15 cats in the cattery developed conjunctivitis at various times over a 9-month period. The outstanding features of the disease were its severity, chronicity and refractoriness to treatment. Prolonged (2 week) treatment with tetracycline was required to effect clinical recovery. Nine of 14 cats in the household developed significant complement-fixing (CF) antibody titres (greater than 128) to the chlamydia group antigen. A single serum from the owner had a titre of 32 although no associated illness was recognized. Of 134 serums collected from random source cats aged 1 month to 16 years, 17 (12.7%) also contained CF antibody to chlamydia group antigen. This is the first report of the isolation of chlamydia from cats with conjunctivitis outside North America and the first report to indicate general incidence figures for chlamydia infection of cats where vaccination is not used.

Chlamydial Infections

Chlamydia trachomatis was first described by Halberstadter and Von Prowazek in 1907, as a characteristic intracytoplasmic inclusion in the conjunctival scrapings from a patient with trachoma. Although the organism was identified as the etiologic agent of trachoma and inclusion conjunctivitis in the neonate, a full understanding of its nature was not feasible until C trachomatis was first successfully propagated in cell cultune in 1965. This technical innovation greatly facilitated investigation of the biology and the epidemiology of chlamydial infections in man. C trachomatis is now recognized to be one of the most prevalent sexually transmitted diseases in the United States today. It is responsible for 40% to 50% of cases of nongonococcal urethritis (NGU) in men and a large reservoir of genital infection, largely asymptomatic, in women. In the United Kingdom, where NGU is a reportable disease, NGU is now more prevalent than gonorrhea and is considered to be the most important male veneral disease. The cervical infection in women is of special importance to pediatricians as the infection may be transmitted to the infant during parturition, leading to both ocular and respiratory disease. A thorough understanding of the microbiology and epidemiology of C trachomatis is an important prerequisite for appreciating the clinical features, diagnostic methods, and management of these infections by pediatricians.

Adult inclusion conjunctivitis. Clinical characteristics and corneal changes.

Twenty-five consecutive cases of adult inclusion conjunctivitis were studied. Diagnosis was based on the finding of typical Halberstaedter-Prowazek inclusion bodies on conjunctival scrapings. Corneal involvement was common (20/25 patients); most frequent was superficial epithelial keratitis (15/25). Other changes seen included subepithelial nummular keratitis (6/25), marginal keratitis (2/25), (4/25). There seemed to be a predilection for involvement of the upper half of the cornea. Conjunctival scarring occurred in one patient only. Associated genitourinary symptoms were spontaneously reported in three patients only; however, on careful questioning, an additional 12 patients were found to have a history of urethritis or cervicitis. A comparison with viral follicular conjunctivitis is made. The location and pattern of keratopathy, associated genitourinary complaints, mucopurulent nature of discharge, and lack of response to standard topical therapy would seem to suggest chlamydial cause. Conjunctival scrapings are very helpful in differential diagnosis between viral and chlamydial conjunctivitis.

Giant Papillary Conjunctivitis after Keratoplasty

Giant papillae of the upper tarsal conjunctiva have been found in vernal conjunctivitis and, more recently, in giant papillary conjunctivitis in patients with contact lenses. A similar syndrome occurred in 11 patients who had undergone penetrating keratoplasty with interrupted nylon sutures. Minimal itching, mild mucous discharge, mild blepharoptosis, and giant papillae of the upper tarsal conjunctiva were associated with exposed suture ends. Removal of exposed sutures led to resolution of the condition. Conjunctival scrapings from three of five patients demonstrated eosinophils. Although the syndrome may have an immunologic basis, the findings are also consistent with a mechanical origin related to exposed nylon sutures.

Vernal and Giant Papillary Conjunctivites. Part 1— Vernal Conjunctivitis

There are many similarities between vernal conjunctivitis and the giant papillary conjunctivites associated with contact lens wear or following cataract surgery. In these articles, the clinical features and pathological morphology of these conditions will be described. Vernal conjunctivitis has a characteristic clinical appearance and the diagnosis can be confirmed by the presence of eosinophils in conjunctival scrapings. The cellular reaction and immunoglobulins associated with the condition indicate that it has an immunological basis.

Guinea-pig inclusion conjunctivitis as a model for the study of trachoma: clinical, microbiological, serological, and cytological studies of primary infection.

The course of primary ocular infection with guinea-pig inclusion conjunctivitis agent was followed in 2 groups of animals. One group of fully grown animals was repeatedly scraped; the other of small animals was used on 1 occasion only and scraped after clinical examination. The intensity of conjunctival inflammation was measured, conjunctival scrapings were taken, and the numbers of polymorphonuclear cells, mononuclear cells, and epithelial cells containing chlamydial inclusions were counted, and the level of antibodies in serum was measured. It was found that inflammation of the conjunctiva lasted for about 30 to 40 days, and the clinical features (oedema, hyperaemia, papillary reaction) were very similar in the 2 groups. Inclusions and polymorphonuclear cells were found for up to 21 days, and mononuclear cells were found on days 7 to 25. Serum antibodies were first detected on day 10 and reached a peak on day 21. The intensity of inflammation was significantly higher on day 2 in the animals which had been scraped. After this the severity of the inflammation and the course of disease were similar in the 2 groups.

Comparative sensitivity of fluorescent antibody staining of conjunctival scrapings and irradiated McCoy cell culture for the diagnosis of hyperendemic trachoma.

The sensitivity of an indirect fluorescent antibody (FA) staining technique for detecting chlamydial inclusions in scrapings from the whole conjunctiva (upper tarsus, upper fornix, and lower lid) was compared with the sensitivity of culture in irradiated McCoy cells for the diagnosis of hyperendemic trachoma. In a group of 211 patients with various grades of active trachoma from the Bandar Abbas area of Southern Iran 42 patients were positive for chlamydiae by either method. There was little difference between the rates of positivity of FA staining of the scrapings from the whole conjunctiva (28 positives) and culture in irradiated McCoy cells (32 positives). In the patients included in this study chlamydial inclusions were detected in 15 eyes by examination of FA stained scrapings taken from the upper tarsal conjunctiva, whereas inclusions were detected in 40 eyes by the additional examination of scrapings taken from the upper fornix and lower lid (P less than 0.001). The examination of FA stained scrapings taken from the whole conjunctiva and spread as a single but larger smear may provide a satisfactory alternative to cell culture methods for the diagnosis of trachoma, particularly for field studies when cell culture facilities are not available.

1. Masquerade Syndrome

A 52-year-old man experienced chronic blepharoconjunctivitis of the right eye of the three years' duration that was unresponsive to medications. Conjunctival cytologic examinations suggested carcinoma and this was confirmed by biopsy specimens of the conjunctiva. However, only a full-thickness eyelid biopsy established that the tumor was a sebaceous carcinoma. We discuss the importance of considering this condition in all cases of unilateral, persistent, or recurrent eyelid inflammation resistant to the usual therapy; the diagnostic value of conjunctival scrapings and full-thickness lid biopsy; and the necessity of interpretation of the biopsy material by a pathologist experienced with, and alerted to look for, this neoplasm. Early recognition of sebaceous carcinoma followed by prompt adequate therapy should improve the survival of patients with this rare but malignant tumor.

Sensitivity of immunoperoxidase and immunofluorescence staining for detecting chlamydia in conjunctival scrapings and in cell culture.

The sensitivities of Giemsa, immunofluorescence, and immunoperoxidase staining for the detection of Chlamydia psittaci inclusions in conjunctival scrapings and in irradiated McCoy cell monolayers were compared. Conjunctival specimens were obtained from a cat colony in which a trachoma-like disease, feline chlamydial keratoconjunctivitis, was endemic. The two immunochemical techniques were found to be of equal sensitivity and 50% to 100% more sensitive than Giemsa stain. Permanent preparations of immunoperoxidase stained material can be made and can be read using a simple light microscope. These features make the technique more useful than immunofluorescence staining, which gives temporary preparations that must be examined with a specialised fluorescence microscope.

VITAL STAINING PROPERTIES OF NEUTRAL RED

Vital staining by instillation in the conjunctival sac of 1% neutral red in 54 normal and 153 differently affected eyes was studied by slit lamp examination. Of these, 134 eyes were after-stained by a mixture of 1% tetrazolium and 1/4% alcian blue and 73 eyes were after-stained by a mixture of 1% rose bengal and 1% fluorescein. Microscopy of a further 10 conjunctival scrapings and 40 mucous conjunctival threads disclosed neutral-red-stained inclusion bodies in the cytoplasm of the epithelial cells and the granulocytes. In the slit lamp the cornea and the conjunctiva were seen to be stained only rarely and, if so, poorly by neutral red, both in normal and affected eyes. Better staining was obtained with tetrazolium, and the best with rose bengal. The plica semilunaris, Marx' line, and the mucous thread were stained by neutral red, more intensely by rose bengal, and the least by tetrazolium. The inferior fornix and the tarsus were rarely stained by any of the three dyes. Neutral red seems to offer no diagnostic advantages over the vital stains with which the dye has been compared in the present study. More particularly we found no pathological processes that were stained more intensely by neutral red than by the other vital stains.

Immunity to vaginal reinfection in female guinea pigs infected sexually with Chlamydia of guinea pig inclusion conjunctivitis.

Guinea pig boars were inoculated intraurethrally with the chlamydial agent of guinea pig inclusion conjunctivitis (GPIC). At the heights of their urethral infections, they were caged with sows in estrus. Whereas some of the sows had not been previously exposed to GPIC agent, others had received an intravaginal inoculation 5 to 8 weeks earlier. Those sows for which infected boars provided the first exposure were challenged by intravaginal inoculation 5 to 8 weeks later. Vaginal and conjunctival scrapings were taken regularly and stained for chlamydial inclusions. Titers of serum anti-GPIC antibodies and of vaginal secretory IgA anti-GPIC antibodies were determined by immunofluorescence. Our results show for the first time that a sexually acquired vaginal GPIC infection induces immunity to manual reinfection of the vagina. Because of the high incidence of secondary conjunctival infections among the vaginally infected sows, we could not provide a sound statistical basis for our tentative conclusion that manual infection of the vagina induces immunity to sexual reinfection. The results of our antibody titrations confirm previous work showing that vaginal GPIC infection induces formation of both serum antibody and vaginal secretory immunoglobulin A antibody.

Echovirus type 7 isolated from conjunctival scrapings.

Echovirus type 7 was isolated from 7 conjunctival scrapings obtained during an epidemic of meningoencephalitis caused by the same virus. The patients suffered from conjunctivitis or keratoconjunctivitis, and also had gastrointestinal symptoms, fever, headache and lymphadenopathy early in their illness. Two characteristics of the isolated strains were not in agreement with those of the standard echovirus type 7: they adapted only slowly to cell cultures, and no viral hemagglutinin for human red blood cells could be demonstrated. Our data suggest an etiologic association of echovirus 7 with eye disease.

Diagnosis of herpes simplex virus infection by immunofluorescence

The utility of the indirect immunofluorescent antibody (IFA) technique for diagnosis of herpes simplex virus (HSV) infection was examined by testing specimens for this agent from 31 patients with encephalitis or meningitis, 17 with conjunctivitis, 19 with genital disease, and 1 with genital disease and meningitis. Brain biopsy tissue from four patients with encephalitis was positive by IFA and virus culture for HSV. Leukocytes in cerebrospinal fluid from these four patients and one with HSV meningitis were also positive by IFA, but virus isolation attempts on the fluid were all negative. Conjunctival scrapings from two patients with conjunctivitis were positive for HSV by both IFA and virus culture. Eleven of 12 culture-positive lesions of herpes progenitalis were positive by IFA, and 1 dark field-positive syphilitic chancre was also positive for HSV by both IFA and culture. Evidence for specificity of the results was provided by internal controls in each test and negative results from patients with other diagnoses. Thus, the IFA technique constituted a rapid, sensitive, and specific diagnostic method for the diagnosis of HSV infections.

急性出血性結膜炎ウイルスに関する研究

Over a period of four months from September to December in 1972, acute hemorrhagic conjunctivitis (AHC) was prevalent in the Kuwana area of Mie Prefecture. An attempt was made to isolate AHC virus from 111 patients in the monkey kidney cell culture. The isolation rates from conjunctival scrapings, throat swabs, and feces were 25.2%, 5.1%, and 3.8%, respectively. An optimal temperature was 33°C for the propagation of AHC virus. Some strains were grown well at 37°C, as well as at 33°C. On the basis of these results, it is quite probable that the infection of AHC virus is generally transmitted from feces to mouth in nature, as well as that of common enterovirus.

Micro Cell Culture Method for Isolation of Chlamydia trachomatis

Presterilized mictotiter plates (96 wells) with BHK-21 cells on 5-mm cover slips were successfully used for cell culture isolation of trachoma from 15 infected conjunctival scrapings.

Micro Cell Culture Method for Isolation of Chlamydia trachomatis

Presterilized mictotiter plates (96 wells) with BHK-21 cells on 5-mm cover slips were successfully used for cell culture isolation of trachoma from 15 infected conjunctival scrapings.

Follow-up studies in neonatal inclusion conjunctivitis.

Seven infants with documented neonatal inclusion conjunctivitis were studied at 7 to 11 months of age. All had received topical tetracycline or sulfacetamide therapy for 10 to 14 days at the time of initial diagnosis. None had recurrence of disease or persistent clinical symptoms. At follow-up, inflammatory cells were present in conjunctival scrapings but Giemsa and indirect fluorescent antibody stains revealed no inclusions. No child had conjunctival scars but two had minimal scarring of the corneal limbus as described in previous investigations on inclusion conjunctivitis.

PANDEMIC OF NEW TYPE OF CONJUNCTIVITIS

A new type of conjunctivitis was prevailing in a pandemic fashion all over South-East Asia and Japan in 1970 and 1971. The disease is characterised by severe subconjunctival hæmorrhage, but its clinical course is usually benign and complete recovery comes within a fortnight. A cytopathic agent has been isolated by primary human embryonic kidney cell culture of conjunctival scrapings or swabs taken from patients with acute hæmorrhagic conjunctivitis (A.H.C.) in three different outbreaks in Japan. One of the earliest isolates which was chosen for detailed study showed general characteristics of enterovirus, but it was not neutralised by intersecting pools of antisera against known enteroviruses, and it was not pathogenic to suckling mice. It is suggested that the agent is an enterovirus of a new serotype. 10 pairs of acute and convalescent sera were tested, and there were definite neutralising antibody rises against this agent in all of them. No specific bacteria were found from cultures of conjunctival swabs. It is concluded that A.H.C. in Japan is caused by a hitherto unknown type of enterovirus and that outbreaks in other countries may be due to a similar agent.

Sensitivity of trachoma agent to streptomycin and related antibiotics.

For isolating trachoma (TRIC) agent from conjunctival scrapings, both streptomycin and neomycin were effective in preventing bacterial contamination, but at high concentrations neomycin was rather more inhibitory to TRIC agent. Prolonging storage of scrapings with neomycin at 4 degrees from 30 min. to 24 hr. reduced the bacterial contamination rate, but also diminished the chance of isolating TRIC agent at the 1st passage. Two freshly isolated TRIC agents differed in their susceptibility to neomycin and streptomycin. Kanamycin and framycetin appeared to be less suitable than streptomycin for use in isolation of TRIC agent. In isolation studies, the possibility of inhibiting TRIC agent by high concentrations of antibotics, including streptomycin, should be borne in mind.

Criteria for measuring the efficacy of trachoma vaccines in baboons.

Trachoma vaccines are usually assayed by testing their ability to protect monkeys or baboons against subsequent challenge of the conjunctiva with a pathogenic strain of trachoma/inclusion conjunctivitis (TRIC) agent. In such experiments the course of infection in vaccinated baboons was compared in terms of arbitrary scores assigned to a range of clinical signs, and of counts of TRIC inclusions in conjunctival scrapings. Analysis of many such scores indicated that after a large challenge dose of strain MRC-4s, the scores for signs of inflammation reached their maximum earlier than the follicle score; the inflammation score was closely related to the number of inclusions, whereas the follicle score was not. With this system, the optimum periods for eliciting differences between vaccinated and control measures varied according to the sign used; it was later for follicles than for inflammation or inclusions. For assessing the influence of vaccination, the mean of the inflammation scores read weekly for the first 3 weeks after challenge and the mean inclusion score over the same period were equally satisfactory, and either was rather better than the mean of three follicle scores taken over the period 3-6 weeks.For assessing the influence of vaccines or therapeutic agents on experimental trachoma it is important to determine which signs discriminate best between treated and control animals, and the optimum times for measuring them.