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Calf diarrhoea

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Calf diarrhoea

  1. 1. CALF DIARRHOEA Dr. V.K.Gupta Senior Scientist, Medicine Division I.V.R.I.IZATNAGAR(UP
  2. 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. 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. 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. 5. Cont..Cont.. Protozoa Eimeria spp. Cryptosporidium spp Protozoa Eimeria spp. Cryptosporidium spp
  6. 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. 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. 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. 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. 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. 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. 12. Diagnosis • Isolation and identification of bacteria• Isolation and identification of bacteria
  13. 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. 14. Colony characteristics EMB agar • Metalic green sheen MacConkey agar • Pink colour colony
  15. 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. 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. 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. 18. • Indole• Indole
  19. 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. 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. 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
  22. 22. 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.
  23. 23. 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.
  24. 24. 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 → + + - -
  25. 25. Biochemical characteristic of Salmonella Does not ferment lactose IMViC test → - + - + Does not ferment lactose IMViC test → - + - +
  26. 26. 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.
  27. 27. 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.
  28. 28. 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.
  29. 29. 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
  30. 30. 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
  31. 31. 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).
  32. 32. 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
  33. 33. Crypto. oocyst • n
  34. 34. 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.
  35. 35. 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.
  36. 36. 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.
  37. 37. 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
  38. 38. Diagnosis • Faecal smear• Faecal smear
  39. 39. 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
  40. 40. 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
  41. 41. 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

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