Genital Tract Infection 1

Normal flora in the urethra of female depends on the age: Normal flora in the urethra of Male are, Staphylococcus Coagulase negative, Diphtheroid, Mycobacterium smegmatis. Normal flora in the urethra of female depends on the age: Before puberty Staphylococcus Coagulase negative Diphtheroid During puberty : Lactobacillus Gardnerella vaginalis Streptococcus group B Streptococcus, beta, alpha, and gamma hemolytic Streptococci. Enterobacteriaceae 2

Actually, all of STD “Sores” (ulcers) Syphilis :Treponema pallidum Background Actually, all of STD “Sores” (ulcers) Syphilis :Treponema pallidum Genital herpes (HSV-2, HSV-1) Chancroid - Hemophilus ducreyi

STDs of Concern (continued) Background “Drips” (discharges) Gonorrhea Chlamydia Trichomonas vaginitis / urethritis Candidiasis (vulvovaginal, less problems in men) Bacterial vaginosis

Procedure of collection sample from a patient with Urethritis Urethral specimen without external discharge . Profuse urethral discharge Vaginal swab 5

Cervical swab: is a swab which is taken from the cervix). 6

Culturing: When the samples received in the lab (Urethral swab or vaginal swab or cervical swab), we search for the most common causative agents (Haemophilus ducreyi, Candida albicans, and N. gonorrhea). 7

Therefore we used the following media as primary plates: Blood Agar Chocolate Agar Sabouraud dextrose agar. MacConkey Agar Incubate all media for 24 hours at 35°C. For chocolate agar media used 5-10% CO2 condition.

NON-ULCERATIVE GENITAL INFECTIONS Cervicitis/urethritis most common Neisseria gonorrhoeae Chlamydia trachomatis Patients infected with Neisseria gonorrhoeae or Chlamydia trachomatis in the genital tract may complain of: urethral discharge vaginal discharge dysuria nothing at all

These organisms result in similar clinical presentations although with Neisseria gonorrhoeae,the incubation period tends to be shorter and the symptoms tend to be more severe, but not always.

Neisseria gonorrhoeae Gram negative diplococci ( intra and extra cellular) Fastidious Requires supplements to media (X, V factors) May take 48 – 72 hours to grow in culture Not part of normal flora

Diagnosis For gonorrhea Urethral discharge may be examined by Gram stained 12

Small, gray Translucent, raised Colony morphology on modified Thayer-Martin (MTM) agar Small, gray Translucent, raised

Oxidase test (+) Identification Yellow is a positive result on carbohydrate utilization tests

Chlamydia Trachomatis Obligate intracellular parasite Cell wall components similar to gram negatives, but not seen with gram stain Requires tissue culture in laboratory inclusions seen in infected cells Not part of normal flora

Chlamydia trachomatis: prepare a smear from either urethral swab or cervical swab 16

Trichomonas vaginalis Budding cells Wet mount preparation of vaginal discharge is the most useful for rapid diagnostic test for: Trichomonas vaginalis Budding cells 17

For identification Candida albicans we used germ tube method 18

Procedure For Germ Tube Test A small amount of an isolated colony is suspended in a test tube containing 0.5 ml of rabbit or human plasma or serum. The test tube in incubated at 35°C for no longer than 2 hours. A drop of the yeast-serum or plasma suspension is placed on a clean slide, covered with a cover slip and examined it under microscope (40x)for the presence of germ tubes. The test is not valid if examined after 2 hours. 19

Trichomonas vaginalis Trophozite is the only stage present in the life cycle. They are 7-32µm long by 5-12µm wide.

Symptoms Usually none. Particularly in males. They don’t show symptoms. In females it ranges from: itching, fishy smell frothing/clear/greenish discharge that is profuse from vagina (leukorrhea).

Bacterial Vaginosis Characterized by (polymicrobial) (due to overgrowth of anaerobes and loss of lactobacilli. Gardnerella may be a marker but not the sole cause of disease) Clue cells Homogenous gray discharge. An amine or fishy odor The PH is greater than 4.5 Absence of a predominance of Lactobacillus on Gram stain slide 22

BV Clinical Characteristics (cont.) Clue cells – vaginal epithelial cells so coated with bacteria that the cell borders are obliterated. Were originally described on wet mount

Normal Vaginal Gram Stain

BV 25

ULCERATIVE GENITAL INFECTIONS Herpes Simplex Virus Syphilis Chancroid

In ulcer swab we suspect Haemophilus ducreyi and Treponema pallidum ( wet mount preparation under dark field microscope) 27

Treponema Pallidum Spirochaetes Diagnosis depends on - dark field examination (1o and 2o stages) of material from the skin lesions - serology to look for antibodies

SEROLOGIC TESTS FOR SYPHILIS Non-treponemal antibody tests - uses cardiolipin antigen, not Treponema Pallidum - VDRL (Venereal Diseases Research Lab) - RPR (Rapid Plasma Reagin, also ART) Used as screening tests ,simple, cheaper False positives common, must be confirmed by a specific test

Treponemal tests use whole treponemes or treponemal extracts to detect antibodies TPHA: Treponemal Pallidum Heamagglutination Test.