Calf diarrhoea
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Transcript of Calf diarrhoea
CALF DIARRHOEA
Dr. V.K.GuptaSenior Scientist,Medicine DivisionI.V.R.I.IZATNAGAR(UP
Introduction Introduction
• 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%
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
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
Cont..Cont..
ProtozoaEimeria spp. Cryptosporidium spp
ProtozoaEimeria spp. Cryptosporidium spp
Predisposing factor
Inadequate nutrition of the pregnant damInadequate environment for the newborn calfInadequate colostrum ingestion Inadequate immunoglobulin absorption from the
calf’s gutPoor hygiene and overcrowdingMixing of different age groups
Inadequate nutrition of the pregnant damInadequate environment for the newborn calfInadequate colostrum ingestion Inadequate immunoglobulin absorption from the
calf’s gutPoor hygiene and overcrowdingMixing of different age groups
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
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)
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.
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
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.
Diagnosis
• Isolation and identification of bacteria • Isolation and identification of bacteria
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
Colony characteristics
EMB agar
• Metalic green sheen
MacConkey agar
• Pink colour colony
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).
Colony characteristics of salmonella
Xylose Lysine Desoxycholate (XLD) agarXylose 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
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
• Indole • Indole
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.
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.
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
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.
Cont..Citrate Utilization testInsert 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 37ºCThe colour change of the medium from green to
blue is indicative of positive reaction.
Citrate Utilization testInsert 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 37ºCThe colour change of the medium from green to
blue is indicative of positive reaction.
Biochemical characteristic of E. coli
Ferments glucose and lactose
produces gas
Positive for Indole and Methyl red testNegative for Voges -Proskauer (V-P) and citrate test
• IMViC test E. coli → + + - -
Ferments glucose and lactose
produces gas
Positive for Indole and Methyl red testNegative for Voges -Proskauer (V-P) and citrate test
• IMViC test E. coli → + + - -
Biochemical characteristic of Salmonella
Does not ferment lactose
IMViC test → - + - +
Does not ferment lactose
IMViC test → - + - +
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.
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.
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.
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
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
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).
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
Crypto. oocyst
• n
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.
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.
EIMERIA
Eimeria bovis E. zuerniiBloody, 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. zuerniiBloody, 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.
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
Diagnosis
• Faecal smear
• Faecal smear
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
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
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