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Epidemiology, Molecular Detection, Zoonotic Potential, Heamatology And Chemotherapy Of Cryptosporidiosis In Small Ruminants In Southern Khyber Pakhtunkhwa, Pakistan

By: Naimat Ullah Khan (2011-VA-516) | Dr. Muhammad Hassan Saleem.
Contributor(s): Prof. Dr. Aneela Zameer Durrani | Dr. Nisar Ahmad.
Material type: materialTypeLabelBookPublisher: 2017Description: 196p.Subject(s): Clinical Medicine | Clinical Medicine and Surgery--CMS | Phd. ThesisDDC classification: 2880-T Dissertation note: Cryptosporidiosis is one of the most important parasitic enteric protozoan infection affecting all vertebrates. The current study was designed to determine the percent prevalence of cryptosporidiosis in small ruminants and humans along with associated risk factors. An overall highest percent prevalence of cryptosporidiosis recorded in all four categories of small ruminants were 27.22%, 20.56%, 18.33% and 12.22% in lambs, kids, Sheep and goats respectively. In the current study, 21.55%, 18.33% and15% prevalence of the Cryptosporidium infection was recorded in sheep in District Kohat, Bannu and Lakki Marwat respectively. In the present study, the highest month wise percent prevalence in sheep, was observed in the month of August (36.66%) followed by April (26.66%), June (26.66%), May, July and September (23.33%), February (10.66%), March (10%), November (10%) while the lowest percent prevalence was observed in the month of December and January (6.66%). In sheep, season wise percent prevalence was also studied where highest prevalence was recorded in summer and autumn season (23.33%), followed by spring (20%) while the lowest percent prevalence was found in the winter season (10%). In sheep, age wise percent prevalence was also studied where highest percent prevalence was found at the age of 1 year (22.38%) followed by 1-2 years (18.03%) while the lowest at the age of 2-3 years (13.46%). In sheep, sex wise percent prevalence was also documented where highest percent prevalence was recorded in female (18.80%) followed by male (17.02%) where lowest percent prevalence was recorded. In goats, the percent prevalence of the Cryptosporidium infection was also studied in three selected areas where recorded 6.66%, 11.66% and 18.3% prevalence in District Bannu, Lakki Marwat and Kohat respectively. Similarly, in goats, overall highest month wise percent Summary 154 prevalence was recorded in the month of August (30%), followed by July (23.33%), June (20%), May (16.66%), March and September (13.33%), April and November (10%), January, February and October (3.33%) while the lowest percent prevalence was recorded in December (0%). In the current study, the season wise prevalence was also studied in goats where highest percent prevalence was recorded in the summer season (20.83%), followed by spring (13.33%), autumn (11.66%) while the lowest prevalence was observed in winter season (3.33%). The highest age wise percent prevalence was recorded at the age of 1 year (18.58%) followed by 1-2 years (10.20%) while the lowest at the age of 2-3 years or above (5.95%). According to the sex wise percent prevalence, the highest percent prevalence was recorded in male (12.30%) while the lowest in females (12.17%). The overall highest percent prevalence of the Cryptosporidium infection was also recorded in lambs in three areas where 33.33%, 25% and 23.33% prevalence was recorded in Kohat, Lakki Marwat and Bannu respectively. The highest month wise percent prevalence was recorded in the month of August (46.6%), followed by other months of the year such as July (40%), April, May and June (30%), September and October (26.66%), November and January (20%) while the lowest in the month of February and December (16.66%) in lambs. The Season wise percent prevalence was recorded in lambs where highest percent prevalence was recorded in summer season (36.66 %), followed by spring and autumn (26.66%) while the lowest in winter season (18.33%). According to the age wise percent prevalence in lambs, the highest prevalence was recorded at the age of 1-15 days (38.09%) followed by 16-30 days (29.41%) while the lowest at the age of 31-60 days or above (15.15%). In lambs, the highest sex wise percent prevalence was recorded in females (31.18%) while the lowest percent prevalence was observed in males (22.98%). Summary 155 In kids, overall highest percent prevalence was 20.55% recorded in three selected districts where the highest prevalence was recorded in District Kohat (23.33%), followed by District Bannu (20%) while the lowest in District Lakki Marwat (18.33%). In kids the month wise percent prevalence was also studied where the highest percent prevalence was recorded in May and August (33.33%), followed by June, July and September (26.66%), March, April and October (20%), November and December (13.33%) while the lowest percent prevalence was recorded in the month of the January (6.66%). The Season wise percent prevalence was also recorded in kids, where the highest percent prevalence was observed in the summer season (30%), followed by autumn (23.33%), spring (20%) while the lowest prevalence was recorded in winter season (10%). The highest age wise percent prevalence in kids was also recorded at the age of ≤1-15 days (33.92%), followed by 16-30 days (15.38%) while the lowest at the age of ≥31-60 days or above (13.55%). Sex wise percent prevalence was also determined in goat kids where, the highest percent prevalence was recorded in female (20.98%) followed by male kids (19.19%). To conduct molecular study, 360 fecal samples of sheep were analyzed for presence of the Cryptosporidium oocysts through simple microscopic method first then confirmed by PCR. DNA was extracted with the help of DNA extraction kit (Made in USA, GFC vivantis). The targeted gene of parasite was 18s rRNA which result in amplification of a segment of genomic DNA at 435 bp. The following primers sequence was used for Forward primer: (5-AAGCTCGTAGTTGGATTTCTG- and reverse primers (5-TAAGGTGCTGAAGGAGTAAGG-3. An overall molecular percent prevalence of the Cryptosporidium infection was 24.99% in sheep in three selected zones of southern KPK. The highest molecular percent prevalence was 31.66%, 25% and 18.33% in District Kohat, Bannu and Lakki Marwat respectively. Summary 156 The highest season wise molecular percent prevalence was also recorded where the highest percent prevalence was recorded in the summer (33.33%), followed by autumn (30%), spring (26.66%) while the lowest in the winter season (13.33%). Molecular percent prevalence was higher in females (27.08%) than male (25.53%). On the basis of environmental factors, overall the highest percent prevalence was recorded in the month of August where highest ambient temperature, relative humidity and heavy rain fall was recorded. To find out Zoonotic aspect of the Cryptosporidium infection, the overall highest percent prevalence was recorded in children (16.66%), followed by adults (5.55%). The highest percent prevalence was recorded in diarrhoeic children where direct contact with small ruminants was observed while the lowest prevalence was recorded in those children where no direct or indirect contact was observed. To conduct the therapeutic trials, a total of 50 goats were selected of the same weight and age that were naturally infected by Cryptosporidium under field conditions. All the goats were placed under same feeding and management conditions and randomly divided into five groups such as A, B, C, D and E. All animals in groups A, B, C and D were treated with Azithromycin (10mg/kg b.wt), Metronidazole (50mg/Kg b.wt), Allium sativum (50mg/Kg b.wt) and Paromomycin (100mg/kg b.wt) respectively while Group-E was placed as a positive control group. The highest percent efficacy in reduction of OPG was shown by different drugs such as Paromomycine (91.77%) followed by Metronidazole (78.20%), Allium sativum (77.00%) while the lowest percent efficacy was shown by Azithromycin (59.29%). On the basis of hematological study, lower lymphocytes count was (48.39%) recorded in non-infected animals while higher (54.33%) count was recorded in infected animals. Similarly higher eosinophil count was (6.73%) recorded in infected group while lower (50 %) counts were recorded in non- Summary 157 infected group. Hb level was higher in infected group than healthy animals. PCV level was higher (42.94%) in infected animals while low (34.62%) in healthy animals. Biochemical analysis of the serum showed, higher quantity of total protein, albumin, ALP, Sodium, Potassium, Chloride, Zinc, Copper, Urea and Creatinine was recorded in infected goats while lower quantity was observed in healthy goats.
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Veterinary Science 2880-T (Browse shelf) Available 2880-T
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Cryptosporidiosis is one of the most important parasitic enteric protozoan infection affecting all vertebrates. The current study was designed to determine the percent prevalence of cryptosporidiosis in small ruminants and humans along with associated risk factors.
An overall highest percent prevalence of cryptosporidiosis recorded in all four categories of small ruminants were 27.22%, 20.56%, 18.33% and 12.22% in lambs, kids, Sheep and goats respectively.
In the current study, 21.55%, 18.33% and15% prevalence of the Cryptosporidium infection was recorded in sheep in District Kohat, Bannu and Lakki Marwat respectively. In the present study, the highest month wise percent prevalence in sheep, was observed in the month of August (36.66%) followed by April (26.66%), June (26.66%), May, July and September (23.33%), February (10.66%), March (10%), November (10%) while the lowest percent prevalence was observed in the month of December and January (6.66%). In sheep, season wise percent prevalence was also studied where highest prevalence was recorded in summer and autumn season (23.33%), followed by spring (20%) while the lowest percent prevalence was found in the winter season (10%). In sheep, age wise percent prevalence was also studied where highest percent prevalence was found at the age of 1 year (22.38%) followed by 1-2 years (18.03%) while the lowest at the age of 2-3 years (13.46%). In sheep, sex wise percent prevalence was also documented where highest percent prevalence was recorded in female (18.80%) followed by male (17.02%) where lowest percent prevalence was recorded.
In goats, the percent prevalence of the Cryptosporidium infection was also studied in three selected areas where recorded 6.66%, 11.66% and 18.3% prevalence in District Bannu, Lakki Marwat and Kohat respectively. Similarly, in goats, overall highest month wise percent
Summary
154
prevalence was recorded in the month of August (30%), followed by July (23.33%), June (20%), May (16.66%), March and September (13.33%), April and November (10%), January, February and October (3.33%) while the lowest percent prevalence was recorded in December (0%). In the current study, the season wise prevalence was also studied in goats where highest percent prevalence was recorded in the summer season (20.83%), followed by spring (13.33%), autumn (11.66%) while the lowest prevalence was observed in winter season (3.33%). The highest age wise percent prevalence was recorded at the age of 1 year (18.58%) followed by 1-2 years (10.20%) while the lowest at the age of 2-3 years or above (5.95%). According to the sex wise percent prevalence, the highest percent prevalence was recorded in male (12.30%) while the lowest in females (12.17%).
The overall highest percent prevalence of the Cryptosporidium infection was also recorded in lambs in three areas where 33.33%, 25% and 23.33% prevalence was recorded in Kohat, Lakki Marwat and Bannu respectively. The highest month wise percent prevalence was recorded in the month of August (46.6%), followed by other months of the year such as July (40%), April, May and June (30%), September and October (26.66%), November and January (20%) while the lowest in the month of February and December (16.66%) in lambs. The Season wise percent prevalence was recorded in lambs where highest percent prevalence was recorded in summer season (36.66 %), followed by spring and autumn (26.66%) while the lowest in winter season (18.33%). According to the age wise percent prevalence in lambs, the highest prevalence was recorded at the age of 1-15 days (38.09%) followed by 16-30 days (29.41%) while the lowest at the age of 31-60 days or above (15.15%). In lambs, the highest sex wise percent prevalence was recorded in females (31.18%) while the lowest percent prevalence was observed in males (22.98%).
Summary
155
In kids, overall highest percent prevalence was 20.55% recorded in three selected districts where the highest prevalence was recorded in District Kohat (23.33%), followed by District Bannu (20%) while the lowest in District Lakki Marwat (18.33%). In kids the month wise percent prevalence was also studied where the highest percent prevalence was recorded in May and August (33.33%), followed by June, July and September (26.66%), March, April and October (20%), November and December (13.33%) while the lowest percent prevalence was recorded in the month of the January (6.66%). The Season wise percent prevalence was also recorded in kids, where the highest percent prevalence was observed in the summer season (30%), followed by autumn (23.33%), spring (20%) while the lowest prevalence was recorded in winter season (10%). The highest age wise percent prevalence in kids was also recorded at the age of ≤1-15 days (33.92%), followed by 16-30 days (15.38%) while the lowest at the age of ≥31-60 days or above (13.55%). Sex wise percent prevalence was also determined in goat kids where, the highest percent prevalence was recorded in female (20.98%) followed by male kids (19.19%).
To conduct molecular study, 360 fecal samples of sheep were analyzed for presence of the Cryptosporidium oocysts through simple microscopic method first then confirmed by PCR. DNA was extracted with the help of DNA extraction kit (Made in USA, GFC vivantis). The targeted gene of parasite was 18s rRNA which result in amplification of a segment of genomic DNA at 435 bp. The following primers sequence was used for Forward primer: (5-AAGCTCGTAGTTGGATTTCTG- and reverse primers (5-TAAGGTGCTGAAGGAGTAAGG-3. An overall molecular percent prevalence of the Cryptosporidium infection was 24.99% in sheep in three selected zones of southern KPK. The highest molecular percent prevalence was 31.66%, 25% and 18.33% in District Kohat, Bannu and Lakki Marwat respectively.
Summary
156
The highest season wise molecular percent prevalence was also recorded where the highest percent prevalence was recorded in the summer (33.33%), followed by autumn (30%), spring (26.66%) while the lowest in the winter season (13.33%). Molecular percent prevalence was higher in females (27.08%) than male (25.53%). On the basis of environmental factors, overall the highest percent prevalence was recorded in the month of August where highest ambient temperature, relative humidity and heavy rain fall was recorded.
To find out Zoonotic aspect of the Cryptosporidium infection, the overall highest percent prevalence was recorded in children (16.66%), followed by adults (5.55%).
The highest percent prevalence was recorded in diarrhoeic children where direct contact with small ruminants was observed while the lowest prevalence was recorded in those children where no direct or indirect contact was observed.
To conduct the therapeutic trials, a total of 50 goats were selected of the same weight and age that were naturally infected by Cryptosporidium under field conditions. All the goats were placed under same feeding and management conditions and randomly divided into five groups such as A, B, C, D and E. All animals in groups A, B, C and D were treated with Azithromycin (10mg/kg b.wt), Metronidazole (50mg/Kg b.wt), Allium sativum (50mg/Kg b.wt) and Paromomycin (100mg/kg b.wt) respectively while Group-E was placed as a positive control group. The highest percent efficacy in reduction of OPG was shown by different drugs such as Paromomycine (91.77%) followed by Metronidazole (78.20%), Allium sativum (77.00%) while the lowest percent efficacy was shown by Azithromycin (59.29%). On the basis of hematological study, lower lymphocytes count was (48.39%) recorded in non-infected animals while higher (54.33%) count was recorded in infected animals. Similarly higher eosinophil count was (6.73%) recorded in infected group while lower (50 %) counts were recorded in non-
Summary
157
infected group. Hb level was higher in infected group than healthy animals. PCV level was higher (42.94%) in infected animals while low (34.62%) in healthy animals. Biochemical analysis of the serum showed, higher quantity of total protein, albumin, ALP, Sodium, Potassium, Chloride, Zinc, Copper, Urea and Creatinine was recorded in infected goats while lower quantity was observed in healthy goats.

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