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Role of Cryptosporidial Infection F. Osman and Ali Sadiek

Role of Cryptosporidial Infection F. Osman and Ali Sadiek



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Article published in 13th Cong. Fac. vet. med. Assiut Univ. 23-25 Nov. 2008
Article published in 13th Cong. Fac. vet. med. Assiut Univ. 23-25 Nov. 2008

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Animal Health Research Institute-New Valley
Role of Cryptosporidial infection as a pathogen of neonatal calf diarrhea in Assiut GovernorateByF. A. Osman and A. H. Sadiek*
 Dept. of Animal Medicine, Faculty of Vet. Medicine- Assiut University
The present study was conducted on 220 neonate diarrheic calves aged 1-60 days, during the period from November 2007 to April 2008, among of which 200 calves, were suffering profuse diarrhea. Complete clinicalexamination of diarrheic calves was carried out. Fecal and blood sampleswere collected for detection of 
by both direct standardfecal examination and by modified Ziel- Neelsen staining. Variabledegrees of mucoid, blood stained profuse diarrhea, apathy, loss of suckling affinity, straining, dehydration and recumbency were the mainsigns observed. The prevalence of cryptosporidaial infection amongdiarrheic calves in this study was 21 %. The highest rate of infection wasin calves aged 10 - 30 days (73.8 %), while the lowest rate of infectionwas in calves aged 1 - 10 days (7.14 %). Mixed infection with coccidial
was noticed in 7 cases (16.8 %) and with
in 2 cases (4.8%). The co-existent diarrhea and dehydration were associated withhemoconcentration which is represented by significant increase in PCV, blood urea nitrogen (BUN), hyponatremia, hypochloremia, hyperkalemiaand hypoglycemia. Significant increase ( p<0.01) in TWBCs may suggestan associating bacterial infection. Some infected calves was complicatedwith bronchopneumonia, among of which 2 cases has been died. It could be concluded that
is recognized as an important primaryor secondary pathogen in neonates diarrheic calves, resulting in severeeconomic losses in neonates. Other causes such as bacteria and viruscould not be excluded.
Keyword : Neonate calves, Cryptosporidium, Diarrhea
 Neonatal calf diarrhea is considered one of the most common andeconomically devastating conditions encountered in raising calf industry.Diarrhea in neonate calves is facilitated by many hygienic, nutritional,environmental and managemental stress factors in addition to bacteriasuch as E
. coli, rota and corona
viruses (Mackenzie et al., 1994).
is considered the most important protozoan causing
neonate calf diarrhea and case fatalities either alone or in association withother bacteria, viruses and coccidia especially in the first 20 days of life(O'Donoghue, 1995 and Ambroise, 2000).
C. parvum
is an intracellular protozoan causing enteritis anddiarrhea in cattle, sheep, goat and man, the first report on bovine
was published in 1971. Now
has been identified asone of the primary etiological agents of neonatal calf diarrhea (Tzipori,1985 a & b; Janoff and Reller 1987; and Naciri et al., 1999). The parasitecan be transmitted by the direct fecal oral rout or through ingestion of contaminated, milk or milk replacer, water or grain (Kirkpatrick 1985).Herd level prevalence of 
C. Parvum
has been reported to range from 13to 100% (Wade et al, 2000 and Santin et al, 2004).The most prominent signs of Cryptosporidiosis in calves aredepression, anorexia, apathy, abdominal pain, fever and mainly diarrheaaccompanied by the shedding of a large number of oocysts, dehydrationand /or poor condition. Calves most often recovers spontaneously within1-2 weeks even though there is a large variation between individuals inhow they respond to and recover from infection (Tzipori,1985 a andMackenzie et al., 1994). Concomitant infection with other enteric pathogens can aggravate the clinical signs and prolong the duration of disease. Morbidity and mortality can be very high in calves less than 2weeks old (Mackenzie et al., 1994). The objectives of the present studywere to investigate the prevalence and significance of 
C. Parvum
inneonatal diarrheic calves and the extent of its concurrent clinical andlaboratory consequences.
Materials and methods
1- Animals and samples.
A total no. of 220 neonate Friesian calves aged 1-60 days wasselected from different localities in Assiut Governorate during the periodfrom Nov. 2007 – April, 2008 and used for this study, among of which200 calves were suffering variable degrees of enteritis, diarrhea anddehydration. Twenty calves were proved to be clinically and laboratoryhealthy and kept as control group (tab. 1). Diarrheic calves wereclassified according to their ages into three groups group I: 1- 10 days,group II: 10 -30 days, and group III: 30 -60 days.Sampling was collected from all calves during the above mentioned period to reduce any potential confusing effects of season on the prevalence of cryptosporidium as follow:a- Fecal samples were collected directly from calves and immediately placed in a screw caped bottles. The samples were labeled with the age of calves, duration and intensity of diarrhea and the associating clinicalsigns and sent to the laboratory.
 b- Two blood samples were collected from jugular vein, in clean sterilecentrifuge tubes. The first blood sample was collected with anticoagulant(sodium salt of EDTA) for estimation of TRBCs, WBCs and PCV %. Thesecond blood sample was collected without anticoagulant and stored for  biochemical study.
 2- Clinical examination.
Clinical examination of diarrheic calves was carried out according toRosenberger, (1979) with special reference to description of diarrhea, body temperature, suckling affinity, posture and vitality of calves as wellas signs and degree of dehydration.
 3- Parasitological examination.
Samples were stored at 4
c in the laboratory and processedimmediately at the same day of collection. The standard concentrationfloatation technique was initially applied on all samples. For each sample,1g of feces was processed using sugar solution (SG 1.33) as the flotationmedium to recover 
C. parvum
s and other parasites. Calveswere considered positive, if more than
10 oocyst 
s /HPF with the correctmorphology (i.e. optical properties, internal structure, size, shape) werefound. The modified acid- fast
 Ziehl –Neelsen
stain is most reliably andtraditionally used to detect the presence of 
 (Webster et al., 1998) as follow:1- Prepare a thin smear by spreading a small amount of feces on slidewith a wooden dowel.2-Fix with gentle heat until the slide is uncomfortable to the touch andcool the slide before proceeding.3- Stain with carbol- fuchsin solution for 3 minutes.4- Wash in running water until no more stain appears in the wash water .5- Rinse with decolorizer until no more stain appears in (decolorizer caneasily made by adding 3 ml HCl to 97 ml 95% ethanol).6- Rinse gently in running tape water.7- Counter stain with the Methylene blue (0.03%) stain for 30 sec.8- Wash gently under running water, stand the slides on end and air dry.9- Oil immersion lenses are corrected for use with cover slide. Permanentreference slide can be made by dipping the dry stained slide in xyline.
Hematological and biochemical analysis.
The total RBCs, total WBCs, Hb concentration and PCV wereestimated according to Coles (1986). Blood glucose, blood urea andcreatinine levels in blood sera were estimated using test kits supplied by bio–Meraux. Serum sodium and potassium levels were estimated photometerically by a flam photometer (Corning Model 400 EnglandESSEX). Blood serum chloride level was estimated using chlorideanalyzer (corning Model 925, England Essex).

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