1. CALF DIARRHOEA
Dr. V.K.Gupta
Senior Scientist,
Medicine Division
I.V.R.I.IZATNAGAR(UP

2. IntroductionIntroduction
• Major cause of economic loss to the cattle
industry
• Leading cause of death in dairy heifer (6.6%)
and beef calves (5.6%) aged < four months
• 45 per cent of the total losses in Dairy industry
in India
• Significant cause of economic loss in cattle
industry with mortality in calves upto 54.58%
• Major cause of economic loss to the cattle
industry
• Leading cause of death in dairy heifer (6.6%)
and beef calves (5.6%) aged < four months
• 45 per cent of the total losses in Dairy industry
in India
• Significant cause of economic loss in cattle
industry with mortality in calves upto 54.58%

3. Cont…Cont…
• Diarrhoea has a variety of causes, infective as
well as non-infective
• Infectious agents play a major role in neonatal
calf diarrhoea
• Diarrhoea has a variety of causes, infective as
well as non-infective
• Infectious agents play a major role in neonatal
calf diarrhoea

4. EtiologyEtiology
 Viral scours
 Rota viruses
 Corona viruses
 Bovine virus diarrhoea
 Bacterial scour
 Enterotoxgenic E.coli (ETEC)
 Clostridium perferingens type B, C and D
 Salmonella spp.
 Proteus spp.
 Pseudomonas spp
 Viral scours
 Rota viruses
 Corona viruses
 Bovine virus diarrhoea
 Bacterial scour
 Enterotoxgenic E.coli (ETEC)
 Clostridium perferingens type B, C and D
 Salmonella spp.
 Proteus spp.
 Pseudomonas spp

5. Cont..Cont..
Protozoa
Eimeria spp.
Cryptosporidium spp
Protozoa
Eimeria spp.
Cryptosporidium spp

6. Predisposing factor
 Inadequate nutrition of the pregnant dam
Inadequate environment for the newborn calf
Inadequate colostrum ingestion
Inadequate immunoglobulin absorption from the
calf’s gut
Poor hygiene and overcrowding
Mixing of different age groups
 Inadequate nutrition of the pregnant dam
Inadequate environment for the newborn calf
Inadequate colostrum ingestion
Inadequate immunoglobulin absorption from the
calf’s gut
Poor hygiene and overcrowding
Mixing of different age groups

7. E. coli
• E.coli has been frequently implicated as the primary
bacterial cause in calf scours
• Isolated enteropathogenic E. coli from 90%
diarrhoeic calves in an organized farm
• Most common cause of diarrhoea in calves aged
upto 4 months in presence of regular deworming
and adequate management conditions
• E.coli has been frequently implicated as the primary
bacterial cause in calf scours
• Isolated enteropathogenic E. coli from 90%
diarrhoeic calves in an organized farm
• Most common cause of diarrhoea in calves aged
upto 4 months in presence of regular deworming
and adequate management conditions

8. Contd..
Five categories of Escherichia coli have been well
associated with diarrhoea in several epidemiological
studies
1)Enteropathogenic E. coli(EPEC)
2)Enteroaggregative E.coli (EAEC)
3)Enterotoxigenic E.coli (ETEC)
4)Enteroinvasive E. coli (EIEC)
5)Shiga Toxin-producing E. coli (STEC)
Five categories of Escherichia coli have been well
associated with diarrhoea in several epidemiological
studies
1)Enteropathogenic E. coli(EPEC)
2)Enteroaggregative E.coli (EAEC)
3)Enterotoxigenic E.coli (ETEC)
4)Enteroinvasive E. coli (EIEC)
5)Shiga Toxin-producing E. coli (STEC)

9. Clinical signs
• Seen in calves < 3-5 days old
• Anorexia, elevated body temperature and depression
with varying degrees of dehydration
• Increased heart rate, watery to semisolid faeces and
dehydration
• Hyperkalaemia, metabolic acidosis and azotaemia
• Hyponatraemia, isokalaemia, hyperchoraemia and
hypoproteinaemia
• Calves from 4 days to 2 month old may manifest with
diarrhea or primarily as dysentery with blood and
mucus in the feces.
• Seen in calves < 3-5 days old
• Anorexia, elevated body temperature and depression
with varying degrees of dehydration
• Increased heart rate, watery to semisolid faeces and
dehydration
• Hyperkalaemia, metabolic acidosis and azotaemia
• Hyponatraemia, isokalaemia, hyperchoraemia and
hypoproteinaemia
• Calves from 4 days to 2 month old may manifest with
diarrhea or primarily as dysentery with blood and
mucus in the feces.

10. Salmonella sp.
• S. typhimurium
• S. dublin
• In calves 2-12 weeks old
• Produces enterotoxin causes inflamatory
changes in intestine and bacterimia
• S. typhimurium
• S. dublin
• In calves 2-12 weeks old
• Produces enterotoxin causes inflamatory
changes in intestine and bacterimia

11. Clostridium perfringens
• Type B and C most common cause of calf
enterotoxemia
• Hemorrhagic enteritis with ulceration of the
mucosa
• fetid, blood-tinged diarrhea, dysentery, abdominal
pain, convulsions, and opisthotonos.
• Death may occur in a few hours, but less severe
cases survive for a few days, and recovery is
possible.
• Type B and C most common cause of calf
enterotoxemia
• Hemorrhagic enteritis with ulceration of the
mucosa
• fetid, blood-tinged diarrhea, dysentery, abdominal
pain, convulsions, and opisthotonos.
• Death may occur in a few hours, but less severe
cases survive for a few days, and recovery is
possible.

12. Diagnosis
• Isolation and identification of bacteria• Isolation and identification of bacteria

13. E. coli isolation
Approximately, 1g of faecal sample is inoculated
in 10 ml of Buffered Peptone Water (BPW) and
incubated at 37°C for 18 h
A loopful of enriched inoculum will then be
plated onto Eosin-Methylene Blue (EMB) agar or
MacConkey agar
 Inoculated plates are incubated at 37°C for 24
hours
The presumptive colonies will be subjected to
further confirmation by biochemical
characterization
Approximately, 1g of faecal sample is inoculated
in 10 ml of Buffered Peptone Water (BPW) and
incubated at 37°C for 18 h
A loopful of enriched inoculum will then be
plated onto Eosin-Methylene Blue (EMB) agar or
MacConkey agar
 Inoculated plates are incubated at 37°C for 24
hours
The presumptive colonies will be subjected to
further confirmation by biochemical
characterization

14. Colony characteristics
EMB agar
• Metalic green sheen
MacConkey agar
• Pink colour colony

15. Salmonella isolation
Pre-enrichment in non-selective medium
(buffered peptone water).
 Selective enrichment in Tetrathionate broth
(Müller-Kauffmann) and Rappaport
Vassiliadis soy peptone (RVS) broth.
 Subcultivation on Xylose Lysine
Desoxycholate (XLD) agar and on Brilliant
Green agar (BGA) (or another selective agar
media).

16. Colony characteristics of salmonella
Xylose Lysine Desoxycholate (XLD)
agar
Xylose Lysine Desoxycholate (XLD)
agar
• Red-yellow with black centers• Red-yellow with black centers
Brilliant Green agar (BGA)Brilliant Green agar (BGA)
• red to pink-white colonies
surrounded by brilliant red
zones
• red to pink-white colonies
surrounded by brilliant red
zones

17. Biochemical characterization
Indole test
Each test isolate will be cultured in 3 ml of peptone
water containing tryptophan at 37ºC for 48 h.
One ml of diethyl ether is added, shaken well and
allowed to stand until the ether rises to the top.
0.5 ml of Kovac’s reagent is gently run down the
side of the test tube to form a ring in between the
medium and the ether.
Development of brilliant red colored ring at the
interface will be interpreted as positive test
Indole test
Each test isolate will be cultured in 3 ml of peptone
water containing tryptophan at 37ºC for 48 h.
One ml of diethyl ether is added, shaken well and
allowed to stand until the ether rises to the top.
0.5 ml of Kovac’s reagent is gently run down the
side of the test tube to form a ring in between the
medium and the ether.
Development of brilliant red colored ring at the
interface will be interpreted as positive test

18. • Indole• Indole

19. Cont..
Methyl Red test
Each test isolate will be inoculated in 5 ml of
sterile MR-VP broth.
After 5 days of incubation at 37ºC, add 5 drops
of methyl red solution.
Methyl Red test
Each test isolate will be inoculated in 5 ml of
sterile MR-VP broth.
After 5 days of incubation at 37ºC, add 5 drops
of methyl red solution.

20. Voges -Proskauer (V-P) test
Grow the presumptive test isolates in 3 ml of
sterile MR-VP broth at 37ºC for 48 h.
Add 0.6 ml of 5% alpha- naphtol and 0.2 ml of
40% potassium hydroxide containing 0.3%
creatine per ml of broth culture.
After proper vortexing, allow the broth to stand
for 5-10 minutes to observe the color formation.
Development of pink-red color indicated positive
result.
Voges -Proskauer (V-P) test
Grow the presumptive test isolates in 3 ml of
sterile MR-VP broth at 37ºC for 48 h.
Add 0.6 ml of 5% alpha- naphtol and 0.2 ml of
40% potassium hydroxide containing 0.3%
creatine per ml of broth culture.
After proper vortexing, allow the broth to stand
for 5-10 minutes to observe the color formation.
Development of pink-red color indicated positive
result.

21. Cont..
Carbohydrate fermentation test:
Perform the test by inoculating 0.2 ml of nutrient
broth culture of test isolates into the tubes
containing sugars such as glucose and lactose.
Incubate for 24 h at 37ºC.
Acid production indicated by the color change
from red to yellow and gas production noted by
the accumulation of gas bubbles in the inverted
Durham’s tube is suggestive of positive result
Carbohydrate fermentation test:
Perform the test by inoculating 0.2 ml of nutrient
broth culture of test isolates into the tubes
containing sugars such as glucose and lactose.
Incubate for 24 h at 37ºC.
Acid production indicated by the color change
from red to yellow and gas production noted by
the accumulation of gas bubbles in the inverted
Durham’s tube is suggestive of positive result

23. Cont..
 Catalase test
Take 3 ml of catalase reagent (3% H2 O2) in a test
tube.
Take single colony of test isolate with a glass rod
and allow to merge in the catalase reagent.
Observe for bubble formation which indicates
positive test.
 Catalase test
Take 3 ml of catalase reagent (3% H2 O2) in a test
tube.
Take single colony of test isolate with a glass rod
and allow to merge in the catalase reagent.
Observe for bubble formation which indicates
positive test.

24. Cont..
Citrate Utilization test
Insert each test isolates into the butt of the
Simmon’s Citrate slant using an inoculation needle
and pull out of the butt by streaking against the slant
in a zig-zag pattern.
Incubate the tubes for 24 h at 37ºC
The colour change of the medium from green to
blue is indicative of positive reaction.
Citrate Utilization test
Insert each test isolates into the butt of the
Simmon’s Citrate slant using an inoculation needle
and pull out of the butt by streaking against the slant
in a zig-zag pattern.
Incubate the tubes for 24 h at 37ºC
The colour change of the medium from green to
blue is indicative of positive reaction.

25. Biochemical characteristic of E. coli
Ferments glucose and lactose
produces gas
Positive for Indole and Methyl red test
Negative for Voges -Proskauer (V-P) and citrate test
• IMViC test E. coli → + + - -
Ferments glucose and lactose
produces gas
Positive for Indole and Methyl red test
Negative for Voges -Proskauer (V-P) and citrate test
• IMViC test E. coli → + + - -

26. Biochemical characteristic of Salmonella
Does not ferment lactose
IMViC test → - + - +
Does not ferment lactose
IMViC test → - + - +

27. Treatment
 Fluid therapy for water and electrolyte
replacement and correction of acid-base
disturbances, alteration of the diet, and
antimicrobial and anti-inflammatory therapy.
 Fluid therapy for water and electrolyte
replacement and correction of acid-base
disturbances, alteration of the diet, and
antimicrobial and anti-inflammatory therapy.

28. Protozoa
CRYPTOSPORIDIOSIS
•19 species and 40 genotypes of Cryptosporidium.
•C parvum is a common cause of calf diarrhea
•Cryptosporidial oocysts have been detected in the
feces of 70% of 1- to 3-wk-old dairy calves.
•Infection can be detected as early as 5 days of age,
with the greatest proportion of calves excreting
organisms between days 9 and 14.
CRYPTOSPORIDIOSIS
•19 species and 40 genotypes of Cryptosporidium.
•C parvum is a common cause of calf diarrhea
•Cryptosporidial oocysts have been detected in the
feces of 70% of 1- to 3-wk-old dairy calves.
•Infection can be detected as early as 5 days of age,
with the greatest proportion of calves excreting
organisms between days 9 and 14.

29. Cont..
• Many reports associate infection in calves with
diarrhea occurring at 5–15 days of age.
• Immunocompromised animals are more
susceptible to clinical disease than
immunocompetent animals
• source of cryptosporidial infection is oocysts
that are fully sporulated and infective when
excreted in the feces.
• Many reports associate infection in calves with
diarrhea occurring at 5–15 days of age.
• Immunocompromised animals are more
susceptible to clinical disease than
immunocompetent animals
• source of cryptosporidial infection is oocysts
that are fully sporulated and infective when
excreted in the feces.

30. Clinical findings
• Mild to moderate diarrhea that persists for several
days regardless of treatment
• Feces are yellow or pale, watery, and contain
mucus
• In most cases, the diarrhea is self-limiting after
several days
• The persistent nature of the diarrhea leads to a
marked energy deficit in these circumstances, and
the calves die of inanition at 3–4 wk old
• Mild to moderate diarrhea that persists for several
days regardless of treatment
• Feces are yellow or pale, watery, and contain
mucus
• In most cases, the diarrhea is self-limiting after
several days
• The persistent nature of the diarrhea leads to a
marked energy deficit in these circumstances, and
the calves die of inanition at 3–4 wk old

31. Diagnosis
• Detection of oocysts by examination of fecal
smears with Ziehl-Neelsen stains, fecal
flotation techniques, ELISA, fluorescent-
labeled antibodies, a rapid
immunochromatographic test, and PCR
• Detection of oocysts by examination of fecal
smears with Ziehl-Neelsen stains, fecal
flotation techniques, ELISA, fluorescent-
labeled antibodies, a rapid
immunochromatographic test, and PCR

32. Wet smear technique
• Take a clean glass slide , add one drop of
water and mix with a loop full of faeces.
• Add one drop of Malachite green on it.
• Cover with cover slip and observe under
microscope (40X).
• Take a clean glass slide , add one drop of
water and mix with a loop full of faeces.
• Add one drop of Malachite green on it.
• Cover with cover slip and observe under
microscope (40X).

33. Modified Ziehl-Neelsen staining
Make a feacal smear on glass slide
↓
Heat fix it then fix with alcohol
↓
Cover the smear with carbol fuchsin for 30 mins
↓
Wash with distilled water
↓
Destain with acid alcohol
↓
Wash with dist. Water
↓
Counter stain with methylene blue for 5 mins
↓
Wash , dry and observe under oil immersion objective
Make a feacal smear on glass slide
↓
Heat fix it then fix with alcohol
↓
Cover the smear with carbol fuchsin for 30 mins
↓
Wash with distilled water
↓
Destain with acid alcohol
↓
Wash with dist. Water
↓
Counter stain with methylene blue for 5 mins
↓
Wash , dry and observe under oil immersion objective

34. Crypto. oocyst
• n

35. Control
Calves should be born in a clean environment,
and adequate amounts of colostrum should be
fed at an early age.
Calves should be kept separate without calf-to-
calf contact for at least the first 2 wk of life,
with strict hygiene at feeding
Diarrheic calves should be isolated from
healthy calves during the course of the
diarrhea and for several days after recovery.
Calves should be born in a clean environment,
and adequate amounts of colostrum should be
fed at an early age.
Calves should be kept separate without calf-to-
calf contact for at least the first 2 wk of life,
with strict hygiene at feeding
Diarrheic calves should be isolated from
healthy calves during the course of the
diarrhea and for several days after recovery.

36. Cont..
• Hyperimmune bovine colostrum can reduce
the severity of diarrhea and the period of
oocyst excretion in experimentally infected
calves.
• Many research groups have attempted to
develop effective vaccines against
cryptosporidia. Unfortunately, to date,
vaccinations have not been effective.
• Hyperimmune bovine colostrum can reduce
the severity of diarrhea and the period of
oocyst excretion in experimentally infected
calves.
• Many research groups have attempted to
develop effective vaccines against
cryptosporidia. Unfortunately, to date,
vaccinations have not been effective.

37. EIMERIA
Eimeria bovis
E. zuernii
Bloody, mucoid diarrhea is often seen 1-3 days
before 1st
oocysts are shed. Prepatent period
16-17 days, peak oocyst production at 3
weeks. Patent period is 2-3 weeks, then self
limits.
Eimeria bovis
E. zuernii
Bloody, mucoid diarrhea is often seen 1-3 days
before 1st
oocysts are shed. Prepatent period
16-17 days, peak oocyst production at 3
weeks. Patent period is 2-3 weeks, then self
limits.

38. Clinical Signs
• Bloody, mucoid diarrhea: epithelial mucosal lesion,
dehydration, depression, tenesmus, occasional rectal
prolapse
• Seasonality: Disease most common in fall, least
summer; a highly pathogenic form of the disease
‘winter coccidiosis’ of unclear epidemiology occurs
• Acute death by 5-7 days: Others develop secondary
enteritis, pneumonia, a few linger in poor condition
and are culled
• Bloody, mucoid diarrhea: epithelial mucosal lesion,
dehydration, depression, tenesmus, occasional rectal
prolapse
• Seasonality: Disease most common in fall, least
summer; a highly pathogenic form of the disease
‘winter coccidiosis’ of unclear epidemiology occurs
• Acute death by 5-7 days: Others develop secondary
enteritis, pneumonia, a few linger in poor condition
and are culled

39. Diagnosis
• Faecal smear• Faecal smear

40. Diagnosis
Gross pathology findings in colon and rectum.
Evidence of hemorrhagic enteritis with frank blood, mucous
Gross pathology findings in colon and rectum.
Evidence of hemorrhagic enteritis with frank blood, mucous

41. Treatment
• nChemotherapeutic agents Treatment Prevention
Sulfadimidine
(sulfamethazine)
140 mg/kg BW orally daily
for 3 days
In feed 35 mg/kg BW for 15
days
Amprolium 10 mg/kg BW daily for 5
days
In feed 21 mg/kg BW for 21
days
Monensin 3 mg/kg BW for 20 days In feed 33 g/tonne for 31
days
Lasalocid 3 mg/kg BW/ day In feed 40 mg/kg of starter
from 3 days to 12 weeks

42. Control
• Treat to prevent incubating new cases and reduce
oocyst shedding ‘multiplier effect’; clinical signs are
seen after damage is done
• Treat until infections self-limit and/or immunity builds
• Oocysts live for 1 year at 4o
C, resist mild freezes;
sunlight and dry heat kills oocysts in 4 hours
• Provide clean dry conditions, reduce stress, crowding
• Do not feed off ground or follow probable heavy
contamination
• Treat to prevent incubating new cases and reduce
oocyst shedding ‘multiplier effect’; clinical signs are
seen after damage is done
• Treat until infections self-limit and/or immunity builds
• Oocysts live for 1 year at 4o
C, resist mild freezes;
sunlight and dry heat kills oocysts in 4 hours
• Provide clean dry conditions, reduce stress, crowding
• Do not feed off ground or follow probable heavy
contamination