Thursday, July 2, 2020

Leptospira - epidemiology, laboratory diagnosis, prophylaxis, treatment

Epidemiology

      Leptospirosis- most widespread of zoonoses

      Present in all continents, except Antartica

      Pathogenic leptospires survive for long periods in (the convoluted tubules of) the kidneys in natural hosts, multiply and are shed in urine

      Animal carriers excrete upto 100 million leptospires per ml of urine

      Infected urine contaminates mud/water that is neutral or slightly alkaline-leptospires survive for weeks

      People come into contact with such water-leptospires enter the body through abraded skin or mucosa and initiate infection

      Certain occupational groups-agricultural workers in rice/cane fields, miners, sewage cleaners exposed to infection-more common in them

      Leptospires can be shed through milk of lactating animals-die rapidly in milk

      Human infection through milk unknown

      Not shed in saliva-animal bites not infectious;   Arthropods do not transmit the infection

      Animals as carriers- rats-ubiquitous, carry the most pathogenic serotype icterohemorrhagiae

      Field mice carry grippotyphosapigs carry pomonadogs carry canicola serotypes

      Generally nonpathogenic in the reservoir animal

      Of veterinary importance since infection in cattle/pigs cause economic loss

      Infection among animals- transmitted by contaminated water/fodder

      Human beings are an aberrant/end host- no transmission to other humans

      Was a rural disease of agricultural workers-  Now, an urban problem in developing countries- overcrowding, insanitation, increasing rat population and habit of walking barefoot

Laboratory diagnosis 

Leptospires are seen in the blood during the acute phase of the disease but can seldom be demonstrated after 8-10 days. They persist in the internal organs, and most abundantly in the kidneys, so may be demonstrated in the urine in the later stages of the disease.

v  Microscopic Demonstration in blood/urine

v  Isolation in culture

v  Inoculation in Guinea pigs

v  Serological tests

Examination of Blood

      Helpful in early stages of the disease (before antibiotics are given)

      Leptospires disappear from blood after the first week

1) Examination of blood under dark field microscope/by immunofluorescence; but of little practical value

2)Three or four drops of blood are inoculated into EMJK or similar medium- incubated at 370C for two days and left at room temperature in the dark for two weeks

      Samples from the cultures are examined every third day for the presence of Leptospires under dark ground illumination

      Primary isolation may be delayed and may take many weeks to months

      Chances of isolation are increased by culturing blood daily at the early stage of the disease

      Leptospires may be isolated from CSF also

3)      The blood from the patient is inoculated intraperitoneally into young guinea pigs

With virulent serotypes- icterohemorrhagiae, the animals develop fever, die within 8-12 days with jaundice and hemorrhage into the lungs and serous cavities

With other serotypes- canicola or pomona, animal may not become ill-Leptospires should be demonstrated in the peritoneal fluid, by blood culture or by serology

From the third day after inoculation, the peritoneal fluid is examined daily under dark ground illumination

When Leptospires are detected the blood withdrawn by cardiac puncture is inoculated into culture media

 Urine Examination

  • Leptospires appear in the urine in the second week of the disease and intermittently thereafter for 4-6 weeks
  • Urine should be examined immediately - Leptospires readily undergo lysis in acid urine

1)      Centrifuged deposit of urine is examined under dark field microscope

2)      Direct culture of urine seldom successful due to contamination but isolation is possible by inoculation into guinea pigs

Serological tests

  • Antibodies appear in serum towards the end of the first week of the disease, increase till the fourth week, then reduces.
  • Antibodies/Agglutinins are demonstrable years after the infection
  • Serological tests can be- broadly reactive genus specific screening tests and the serotype specific tests

v  Genus specific/ Broadly reactive tests -identify Leptospiral infection without identifying the exact serotype

      The antigens for these tests are prepared from the nonpathogenic L. biflexa Patoc 1 strain

     Sensitized erythrocyte lysis (SEL), complement fixation, agglutination and indirect immune florescence, ELISA

      ELISA to detect IgM and IgG separately, to determine the stage of infection can be done

      Simple and rapid dip stick assays for detecting Leptospira specific IgM antibody in human sera available

v  Serotype specific tests

      Identify the infecting serotype by macroscopic and microscopic agglutination tests

      Macroscopic agglutination tests done with inactive formalinized suspensions of Leptospiral serotypes with serial dilutions of the test serum

   Microscopic agglutination tests (MAT) use live cultures of different serotypes and agglutination is observed under the low power dark field microscope- more specific- done in reference laboratories

Prophylaxis

      Leptospirosis results from contact of skin or mucosa with contaminated water

Ø  Rodent control

Ø  Disinfection of water

Ø  Wearing of protective clothing

      Vaccination of high risk groups such as agricultural workers encouraged.  Immunity is serotype specific

Ø  Vaccination of dogs, cattles, pigs

Therapy

      Sensitive to penicillin and tetracyclines

      Treatment should be started early in the course of the disease

      Penicillin given as IV : 1-2 million units 6 hourly for 7 days, for serious cases

       A mild Jarish Herxheimar reaction seen in some

      Doxycycline 200 mg orally given once in a week effective for prophylaxis


      Jarish Herxheimar reaction - common detoxification response of the body to the increased toxins released during a treatment for pathogens

      the amount of toxins released into the circulation is more than what our body can safely handle quickly

      lot of stress on internal organs, especially the colon, liver and kidneys that are involved in filtering toxins in the body


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