Kentucky’s Female-Led Bacteriology Laboratory

State Bacteriologist, Assistant, Microscopists, and Mailing Clerk. Image from the Bulletin of the State Board of Health of Kentucky.

Dr. Lillian South (left) and Miss Cecilia Barthelme in 1938. Image from the Courier-Journal.

In 1910, Kentucky’s General Assembly established the Commonwealth’s first State Laboratory of Bacteriology in a tiny room in Bowling Green at St. Joseph’s Hospital. The room cost the state nothing and was also furnished by the hospital. Dr. Lillian H. South was voted, unanimously, as the State Bacteriologist making her among the first women in the country to head such a laboratory at the state level. This was years before the 19th Amendment was ratified affording the women the right to vote. As the days and weeks progressed, word of mouth informed many Kentucky physicians that the laboratory existed for the purpose of assisting medical professionals in developing patient diagnoses. The laboratory also held another important role and that was in identifying contagious and infectious diseases with a goal to prevent them and to furnish sera and other scientific aids in their treatment. Interestingly, any physician, public health official, or citizen of Kentucky could utilize the lab’s services “without a cent of cost.” Specimens were being sent to the lab at a rapidly growing pace and the increase in the need for microscopists meant that the tiny one-room laboratory would need to expand soon.

State Bacteriological Laboratory with Dr. Lillian South and Stenographer. Image from the Bulletin of the State Board of Health of Kentucky.

Prof. Wickliffe Rose in 1910. Image from the Houston Post.

It was around this time when Professor Wickliffe Rose, the Executive Secretary of the Rockefeller Sanitary Commission, made several visits to Kentucky’s State Bacteriological Laboratory. As a separate entity to the Rockefeller Foundation, the Rockefeller Sanitary Commission was established in 1909 to become a think-tank of sorts to help prevent the rapid spread of hookworm diseases throughout the American South. Though hookworm disease was endemic in the South it was also very curable. As such, the Rockefeller Sanitary Commission has had dual purposes and that was to promulgate the establishment of public health networks in the South. For several reasons, public health in the American South had been long neglected which made curable diseases like hookworm endemic. When a contagious or infectious disease became endemic in an area or region, particularly one that was ill-equipped to handle it, it would often become overwhelming for local physicians to treat the disease(s). As such, a curable disease would sometimes become fatal or cause irreversible damage to the body.

After several visits to Kentucky’s State Laboratory of Bacteriology, the Rockefeller Sanitary Commission approved funding to build a new, free-standing laboratory that would also have living quarters for Dr. Lillian South. With Dr. South living adjacent to the laboratory she would be able to receive specimens, start cultures, and distribute emergency supplies of serums and vaccines day or night. Additionally, the Commission provided salaries for an additional twelve microscopists, all of whom were women, four full-time Sanitary Inspectors with microscopes, stereopticons, and other publicity outfits, for special investigations as to the prevalence of, and to furnish free treatment for, hookworm disease and other sickness caused by intestinal parasites, and to arouse popular interest in restricting the occurrence and spread of these and all other preventable sicknesses, expending for three years in this work, until it was well underway.

The First Four Years

In its first four years of operation, the State Laboratory of Bacteriology examined 237,328 specimens, the majority of those being for hookworm and other intestinal parasites. Additional specimens received were for tuberculosis, typhoid fever, influenza, meningitis, dysentery, malaria, gonorrhoea, rabies, diphtheria, pneumonia, and syphilis.

  • 68,085 persons with intestinal parasites were treated
  • 315 persons bitten by rabid animals were immunized against hydrophobia
  • 11,221 packages of diphtheria, typhoid, meningitis, tetanus and other serums, bacterins, and vaccines were provided to health and fiscal officials and physicians of every county in the Commonwealth at wholesale cost

Benefactors of the State Laboratory of Bacteriology came from every walk of life. Many came from “intelligence and appreciative classes” but the importance of no fees to any citizen can never be played down; no expense to any family to have this aid for his or her physician in times of emergencies, including threats to health and life, meant that lives could be saved and irreversible harm to the body avoided at a level never before seen. Still, it was important to campaign awareness for the laboratory; those in the medical profession, club women, teachers, and the press worked to make possible the life-saving work offered by the laboratory. The life-saving work wouldn’t have been possible without the hard work and dedication of the skilled and proficient laboratory staff. A group of educated, ambitious women, “anxious to do things for the work’s sake,” to earn a promotion, the spirit of emulation was so pronounced, and produced such results, that requests for their services began coming in from other State Laboratories. Two of the employees were selected by the International Health Commission to conduct its Health Exhibit and Demonstrations at the Panama Expositions held in San Francisco.

Economic Impact

Though the Laboratory, in its first few years, made significant public health impacts on Kentuckians, it was often underappreciated. Without the State Laboratory of Bacteriology, the cost to physicians and public health officials of the aforementioned procedures ranged from $2.00 to $10.00; if an allowance of only $1.00 for each examination was made in or by the laboratory the cost would equal $237,328. The fee for each treatment provided by local physicians in private practice would range from $3.00 to $10.00; allowing $1.00 for each given here would equal to $68,085.00. The fee for immunization against rabies in Pasteur Institutes ranged from $300.00 to $500.00; allowing for $200 each for those done in Kentucky would equal $63,000. Customary retail prices of serums, bacterins, and vaccines sent out would have been $33,493.00; prices paid by physicians and public health officials paid only $8,249.60 a total savings of $24,993. During its first four years of inception the total savings to physicians, public health officials, counties, and citizens was the equivalent of $393,406.00 or $98,366.50 annually.

Work of the Laboratory

The early work of the laboratory was broken down into ten areas/categories:

  1. Examination of sputum and various discharges for Tubercle bacilli, influenza, pneumococci, and mixed infections
  2. Examinations of throat cultures for diphtheria, membraneous croup, and septic sore throat
  3. Widal Reaction of the blood and the Diazo Reaction of urine for typhoid fever
  4. Examination of the feces for intestinal parasites
  5. Gonorrhoea
  6. Spirochetes
  7. Rabies or Hydrophobia, and free Pasteur treatment for the prevention of Rabies or Hydrophobia
  8. Examination of urine and feces for typhoid carriers
  9. Distributing stations for diphtheria antitoxin, vaccines, bacterins, and sera, at reduces cost.
  10. Urinalysis


To test for tuberculosis an examination of sputum, pus, exudates, feces, and urine were often utilized to look for the presence of tubercle bacilli. The laboratory used Dr. Kinyoun’s modification of the Uhlenhuth Method of examining sputum. This method involved dissolving the sputum in antiformin which was an alkaline solution of the hypochlorites; this would dissolve all other bacteria except for acid fast bacilli. This also required the use of an electric centrifuge and the Rickards sputum bottle shaker. The latter was designed by Dr. B. R. Rickards and worked to provide an effective tool for routine laboratory work in the examination of sputum. The sputum was vigorously shaken until being thoroughly dissolved.


Diphtheria is due to the Klebs-Loeffler Bacilli, which are small rods of variable shapes and sizes that also grew readily on specially prepared blood serum, and, when stained, showing characteristic granular or barred appearance. As a very contagious disease, there were frequent break-outs in epidemics of considerable importance, especially in schools and other institutions in which a large number of children congregated. Throat cultures were the only reliable method of diagnosis at the time. To obtain a good specimen, a sterile cotton swab on the end of a small stick was sent out by the laboratory to the physician. The physician would rub the sterile cotton swab on the white patch in the throat and red and swollen mucous membranes of the fauces, pharynx, or posterior nares. One the specimen was received by the laboratory it was firmly rubbed over the surface of the blood serum and incubated at body temperature over night; the next morning the specimen was examined and the report sent immediately to the physician that ordered it. It was noted that, “in suspicious cases valuable time should not be lost in waiting for a report from the laboratory, which takes twelve to twenty-four hours to complete, but immediate administration of antitoxin should be instituted.” Confirmed positive cases were to be re-tested every five or six days after the disappearance of the white membrane to ensure that the patient is no long contagious. In order to return to school, the physician must have at least two negative reports received from the laboratory.

Diphtheria collection/mailing kit. Image from the Bulletin of the State Board of Health of Kentucky.

Typhoid Fever

Two types of tests were often utilized to attempt to best find the typhoid bacilli. The first is Ehrlich’s Diazo reaction and the Widal reaction, secured by mixing a small blood sample from the patient with some known typhoid fever germs where there would be a definite clumping of the germs typical of this disease. Ehrlich’s Diazo reaction of the urine was a particularly satisfactory method as the reaction could appear early in the disease process, usually by the fifth day. It was recommended that physicians, in suspect typhoid fever cases, send in a sample of urine during the first week and, at the beginning of the second week, send in a sample of blood. At the time, the Widal agglutination test was confirmatory of typhoid in 95 percent of cases; fifty percent of the cases showed the positive reaction on or about the eighth day, 25 percent of cases during the second week, and in 20 percent during the third week; for about 5 percent of cases, it did not show until as late as the fourth week. As such, the laboratory reiterated that these results were to be used as an adjunct in aiding diagnosis and should never be used in place of bedside care with an experienced physician. Negative results in the early stages was not solid proof that the patient was free of the typhoid bacilli.

Typhoid bacilli collection/mailing kit. Aluminum plate with a depression for drop of blood. Needle for making puncture. Look of wire for transferring drop of blood from lobe of ear or finger to aluminum plate. Image from the Bulletin of the State Board of Health of Kentucky.

Two primary typhoid vaccines distributed by the laboratory. Images from the Bulletin of the State Board of Health of Kentucky.


Around 1915 there were an estimated 300,000 cases of gonorrhea in Kentucky. The State Board of Public Health and the State Laboratory of Bacteriology considered it important to add the examination of this disease to public health work. Gonorrhea is transmissible in all its stages and is due to the gonococcus or bacillis of Neisser. It is coffee-shaped, occurs in pairs, with the inner surfaces being flattened and separated from one another by narrow intervals. The “vulgar” term to reference gonorrhea during this time and later was “clap.” Additionally, it was the cause of a large part of blindness as the eyes of newborns were especially susceptible. The gonococcus was demonstrated in the pus of acute gonorrhea and ophthalmia neonatorum, it was found in the urethral discharges of gonorrhea from the beginning to the end of the disease, and often for many months or years after the supposed recovery. The organisms are largely found within the pus cells or attached to the surface of the epithelial cells. In the acute stages in women, the bacteria were more readily demonstrated in a specimen obtained by milking the urethra.

Gonococcus collection/mailing kit. Image from the Bulletin of the State Board of Health of Kentucky.


Syphilis is due to the spirochaete pallida, found in all syphilitic lesions, but most abundantly in the chancre, mucous patch, and in condylomata (genital warts). According to several Kentucky physicians at the time, the examination for spirochetes was usually unsuccessful; largely because few specimens were properly obtained. To compound the issues, the organisms were found to not grow in the presence of air and thus they were never found in surface secretions. The State Laboratory of Bacteriology recommended that the surface of a superficial lesion (e.g. a chancre) should first be washed with soap and water and then gently scraped with a scalpel until lymph, as free from blood as possible, began to exude. Spirochetes were found most abundantly by scraping the indurated edges of the sore; the Wasserman reaction would not appear before the third or fourth week. When the laboratory received the specimen, the slide was passed through a flame to fix the smear. It was then kept in a jar containing a fresh solution of 10 percent silver nitrate for 24 hours. If the specimen was positive it should the spirochetes as delicate corkscrews in shape, stained black. Spirochetes in a specimen was considered conclusive evidence that the patient had syphilis and thus treatment could begin immediately.

Spirochete collection/mailing kit. Image from the Bulletin of the State Board of Health of Kentucky.


Rabies is one of the oldest and more recognizable infectious diseases. Written about by Aristotle, Celsus, and Galen, little had been accomplished in terms of curative interventions. It wasn’t until 1880 when Louis Pasteur discovered that rabies may be prevented by injecting via hypodermic needle trace doses of the poison very early on in the disease and gradually increasing the dose day by day until the person became so inoculated that he or she would not contract the disease. In 1903, Dr. Adelchi Negri described certain bodies (later known as Negri bodies) in large nerve cells as a definite means to establish the diagnosis of rabies or hydrophobia. Negri bodies were found in certain sections of the brain and thus the State Laboratory of Bacteriology requested the heads of animals suspect of having rabies. At the time, Dr. Negri believed that, in addition to being a positive sign for rabies but that they were “probably animal parasites and cause it.” Rabies was essentially found in all parts of the world and, in 1915, occurred most frequently in dogs, cats, wolves, horses, cows, pigs, skunks, deer, and man.

A fascinating component of rabies is its incubation period and variability. In animals, it was seldom seen under ten days but could also extend over several months. The majority of cases in humans occurred at the end of the third month after being bitten. Up to 1915, the earliest case in which the shortest time in which human hydrophobia had developed was fourteen days after being bitten by an infected dog. The longest on record at the time was seven months. Incubation of rabies in humans is also affected by various factors; first is the location of the bite. Bites on the hands and face or other exposed parts of the body with rich nerve supplies were more susceptible to result fatally if not treated immediately. This was because the ‘poison’ was supposed to travel along the nerve trunks. Another factor is the quantity of poison or virus introduced; bites through clothing tended to be less dangerous as fewer quantities of the virus were introduced. In 1886, a study showed that the average case developed seventy-two days after being bitten.

Group of patients that had contracted rabies who were being given free treatment at the State Board of Health at Bowling Green. The horse, who also contracted rabies, had been treated as well. Image from the Bulletin of the State Board of Health of Kentucky.

Symptomatology in animals fell into one of two categories; furious or excited and quiet, silent, paralytic. Changes in animal behavior were frequently noted (e.g. loving, playful animals becoming irritable and biting unprovoked). The paralytic form of rabies, also known as hydrophobia, was frequently found in Kentucky. That is, animals may act as if they have a bone or object stuck in their throat; the pet owner would proceed to help the animal out and would end up getting bit; in addition to dogs this was also noted in cows or mules that also had rabies. Since the time of Celsus, the most commonly used treatment was cauterization of the wound by using caustic fuming nitric acid. Carbolic acid was less effective and silver nitrate proved to be useless. Pasteur treatment was often used and utilized by the State Board of Health of Kentucky as a preventative treatment. They were furnished by the United States Public Health Service and, because they could not be stored, had to be ordered fresh for each patient. There was a major stipulation to this as patients had to receive the treatment under the supervision of the State Bacteriologist, Dr. Lillian South. In order to get the treatment for free, the patient had to arrange to stay in Bowling Green for twenty-one days; room and board started at $3.50 per week. Special arrangements were made to care for children who had to travel without their parents. For those unable to stay away from their home for twenty-one days, treatment could be sent to local physicians, if ordered through the local health officer, for the cost of $35.00.

Rabies vaccine and treatments used in Kentucky. Image from the Bulletin of the State Board of Health of Kentucky.


Malarial organisms in a red blood corpuscle. Image from the Bulletin of the State Board of Health of Kentucky.

Malaria is due to a parasite called plasmodium malaria and transmitted from one human to another typically by the mosquito (anophiles culex). A mosquito biting a person who is infected with the parasite, after right to fourteen days, would then become viable to infecting other persons it bit. In 1915 there were three types or classifications of malaria; tertian, quartan, and aestivoautomnual. Each type had its own corresponding variety of organisms as well as symptoms. The parasites would appear in the blood within five to twenty-one days of being bitten and could retain infectious status over a long period of time. It was noted that the parasites could possess a new vigor even after an extended dormant period following active treatment. As such, the best way to identify the pathogen was through microscopical examination of the blood in patients with suspected cases. Treatment required the administration of at minimum twenty grains of quinine every day for five days followed by ten grains every fifth day, as from thirty to sixty days relieves most cases and causes the parasites to disappear from capillary circulation. As such, it was uncommon for the organism to be found after administration of quinine.

Malaria collection/mailing kit. Image from the Bulletin of the State Board of Health of Kentucky.

Hookworm, Roundworm, Pinworm, and Other Parasites

Biological Products Stocked by the Kentucky State Bacteriological Laboratory

Urinalysis Form

Contributed by Shawn Logan |

⁘ Works Cited ⁘

  1. The Lexington Leader, 9 December 1949
  2. The Lexington Leader, 12 December 1952

  1. Chapin, Charles V. “Variation in Type of Infectious Disease as Shown by the History of Smallpox in the United States 1895-1912.” The Journal of Infectious Diseases 13, no. 2 (1913): 171-96.
  2. Curtis, Henry S. “The Rockefeller Sanitary Commission.” The Journal of Education 74, no. 7 (1911): 176.
  3. Tournès, Ludovic, and Giles Scott-Smith, eds. Global Exchanges: Scholarships and Transnational Circulations in the Modern World. New York: Berghahn Books, 2018.
  4. Kentucky. State Board of Health. Bulletin of the State Board of Health of Kentucky. Louisville: State Board of Health of Kentucky.
  5. The Courier-Journal (Louisville, Kentucky), 14 April 1938, p. 2.
  6. The Houston Post (Houston, Texas), 7 April 1910, p. 5.

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