Prevalence and Chemotherapy of Cryptosporidiosis in Goats using Different Herbal and Allopathic Drugs in Southern Khyber Pakhtunkhwa, Pakistan

Total of 1440 fecal samples (n=40/month/district)) were collected from goats comprising 40 samples per month from each of 3 districts (Bannu, Lakki Marwat and Kohat) of Southern Khyber Pakhtunkhwa for the period of one year. .

Each fecal sample (about 5 g) was directly collected from the rectum of goats using sterile plastic bags in order to ensure the maximum possible sterility. The procedure was approved by institutional ethical committee and care was taken to minimize the pain and number of animals throughout the experiment. All the samples were transported to laboratory and preserved in 10 % formalin at 4 0C till microscopic examination of Cryptosporidium oocysts.

For microscopic examination, about 3 g fecal materials were dissolved in distilled water to make homogenized solution, centrifuged for 1 min at 1500 rpm. Supernatant was removed and sediment was resuspended in the flotation solution of ZnSO4 (44%). The solution was recentrifuged at 1500 rpm for 1 mint. Finally, sediment was examined under microscope and Cryptosporidium oocysts were stained by modified Ziehl- Neelsen (MZN) technique (Casemore and Sands, 1985).

For chemotherapeutic trials a total of 50 goats of same age and weight of either sex naturally infected with Cryptosporidium infection were randomly selected and divided into five groups (A, B, C, D and E) each of 10 heads. The animals in group A, B C and D were treated with Azithromycin, Metronidazole, Allium sativum (garlic) and Paromomycin respectively while Group E was kept as control positive group and no treatment was provided during trial period. All these animals were kept under same management and feeding pattern.

For drug and garlic administration about 1-2 bulbs (50 mg/kg.wt) of Allium sativum were fed orally, while other drugs azithromycin, metronidazole and paromomycin were administered orally as 500 mg, 50 mg, 100 mg per kg body weight per day, respectively, for five consective days.

Overall, 198/1440 (13.75%) prevalence were recorded in study area. Highest prevalence (19.16%) was recorded in district Kohat followed by District Lakki Marwat (12.08%) and lowest in district Bannu (10%) (Table I). The highest month wise prevalence of cryptosporidiosis was recorded during July-August (26%, 27%), followed by June (20%), September (16%), April/May (14%), October (12%), January (11%), March (9%), November (8%), February (7%) and lowest in December (1%).

Our results are in close agreement to a researcher with 17.7% percent prevalence in Nigeria (Danladi et al., 2015). While disagree with some findings of 3.3% in Sokoto and 24.0% in Plateau states (Pam et al., 2013; Faleke et al., 2014). This high prevalence in our study may be due to irregular or limited parasitic infection control program. While, lower prevalence of Cryptosporidium infection in goats may be attributed to the environmental and ecological factors where relative low humidity and short period of rainfall may be recorded (Yu and Seo, 2004).

Means with different superscripts differ at (P < 0.05)

Table II. Cryptosporidium oocysts/g of feces and % efficacy of cryptosporidiosis in goats treated with different selected allopathic drugs and garlic administered over the period of 28 days.

Means with different superscripts differ at (P < 0.05).

Results revealed highest percent prevalence during summer season (22%), followed by autumn (14%), spring (12%) and lowest was (7%) in winter. Overall, age wise percent prevalence of cryptosporidiosis in 1 year old goats was higher than 1-2 (14%) years and 2-3 (8.27%) years or above. Similarly, on sex basis, little higher prevalence (15.66 %) was observed in female than male animals (13.91%) (Table I).

Table II shows efficacies of different chosen drugs on the basis of reduction in the OPG (oocyst per gram) of feces after treatment. The efficacy of Azithromycin was 13.90 %, 29.07 %, 44.87 % and 59.29 % on 7th, 14th, 21th and 28th day post treatment. Statistically, non-significant difference in OPG reduction was recorded in relation to time (P>0.05). Similarly, a single dose of Metronidazole caused a significant decrease in oocyst per gram of feces on 7th day post treatment (P<0.05). On day 7 post treatment 25.74% decrease was observed. Percent efficacy of Metronidazole was 46.70%, 68.14%, and 78.20 % recorded on 14 th, 21 th and 28 th day respectively. Similarly, a single dose of Allium sativum caused a significant decrease in OPG on 7th day post treatment and onward. On day 7th post treatment (27.06%) reduction was recorded in OPG. Similarly, percent efficacy of Allium sativum was 49.70 %, 62.24 % and 77.00 % on 14th, 21th and 28th day respectively. Similarly, significant (P<0.05) decrease was observed in OPG count on 7th day post treatment. Percent (%) efficacy of paromomycin calculated on 7th, 14th, 21th and 28 th day was 51.77%, 67.08%, 87.54 % and 91.77 %, post treatment, respectively.

Similar findings were reported Masood et al. (2013) in calves during summer season followed by autumn, spring and the lowest in winter season. The Cryptosporidiosis is worldwide in distribution and commonly found in all wet and warm seasons (Jafari et al., 2012). Our results are consistent with other researchers that strong correlation between the warm and wet seasons of the year and the prevalence of infection rate may be present.

Akinkuotu et al. (2015) correlated age of the animals with Cryptosporidiosis infection where highest prevalence was recorded in pre-weaned kids (62.7%) followed by Post weaned kids (30.4%) and lowest in adults (22.5%). Highest percent prevalence was found in young animals of 1 year or below. Our findings contrast to these findings that may be attributed to the differences in hygienic conditions, environmental factors such as heavy rain fall, high relative humidity, high temperature, study design, immune status of animal, feeding and watering management conditions. Although overall infection was observed in both sexes and little higher prevalence (15.66 %) were observed in female than male (13.91%) in three selected districts of Southern KPK. These findings are in line with Pam et al. (2013) who recorded 25.3 % prevalence of Cryptosporidium in female and 22.7% in male and not in line with (Akinkuotu et al., 2015) where 47.5% and 29 % prevalence was recorded in female and male animals. The constancy of higher percent prevalence rates in females than male in goats may be due to hormonal disturbance during pregnancy and lactation stress.

A single dose of Azithromycin (10 mg/ kg b.wt) was used and OPG count was recorded as 13.90 %, 29.07 %, 44.87 % and 59.29 % on 7th, 14th, 21th and 28th day post treatment as supported in previous similar nature of work (Elitok et al., 2015).

In current study, a single dose of Metronidazole used against group B naturally infected with Cryptosporidiosis caused a significant decrease in oocyst per gram (OPG) of feces from 7th day post treatment and onward (P<0.05). At day 7 post treatment, 25.74% decreases was observed in oocyst count. Our findings are very close to Masood et al. (2013) that Metronidazole is highly effective in reducing oocysts burden.

Allium sativum caused significant decrease in OPG of feces on 7th day post treatment and onward. At day 7th post treatment (27.06%) reduction was recorded in OPG. Similarly, percent efficacy of Allium sativum was 49.70%, 62.24% and 77 % on days 14, 21 and 28, respectively. Our results corroborate with Masamha et al. (2010) that garlic disrupts the normal physiology of the parasite like movement, absorption of food and reproductive activities, resulting in the reduction of burden. Similarly, the phagocytic activities and natural killer cell activity may be increased during the garlic treatment period as reported by (Sutton and Haik, 1999).

The current study confirms the Cryptosporidium infection in goats in southern parts of KPK, Pakistan and validates the traditional use of Allium sativum as therapeutic agent. Furthermore, it might be a good choice against different protozoal/bacterial diseases as local remedy. Further experimentation and investigation may be carried out in combination with different drugs/ plants in order to explore its antimicrobial potentional. Awareness campaign among public about the zoonotic nature of the disease, good man­agement practices including proper hygiene maintenance are more appropriate tools for the control of Cryptosporidium infection.

The authors declared that this study has received no financial support.

The authors declare there is no conflict of interest.