1.
Pcr-Based Diagnosis Of Canine Parvovirus In Dogs
by Farhan Towakal | Prof.Dr.Masoos Rabbani | Prof.Dr.Khushi Muhammad | Prof.Dr.Zafar | Faculty of Veterinary Sciences.
Material type: ; Format:
print
Publisher: 2007Dissertation note: Canine parvovirosis, caused by a haemagglutinating canine parvovirus (CPV), one of the most important acute viral infectious diseases of pups, had been prevalent in the country. In the present study, 40 faecal samples, from clinically suspected cases of parvovirus diseased dogs and stray dogs were collected. The samples were diluted and centrifuged to collect the supernatant. Being a haemagglutinating (HA) virus; the pre-filtered supernatant from all suspected samples was checked for any HA activity using 1% washed chicken erythrocytes. Out of total of 40 samples, 30 samples were found HA positive. All the HA positive samples and samples found negative were processed for extraction of viral DNA with Genomic DNA purification kit. The net isolated DNA samples were subjected to Polymerase Chain Reaction (PCR) by using specific primers (developed against the variable genomic region of the capsid protein of the DNA) of the CPV. One of the selected primer pair designated as (37, 38) amplified the genornic region present in the CPV-2, the other primer pair designated as (39, 40) that amplified the region present in the CPV-2b.Optimization of PCR was done for the molecular level diagnosis of canine parvovirus and this technique was previously not available or standardized in local conditions of Pakistan. With the use of primer pair (37, 38) best results were found on following optimizing conditions like annealing temperature 55°C, primers concentration (reverse and forward) 25pmoles, Magnesium Chloride concentration 2mM, DNA(Template) volume Spl, Taq DNA polymerase(12.5U) and 2001iM of each dNTPs. For the primer pair (39,40) best results were found on following optimizing conditions like annealing temperature 5 5°C, Magnesium Chloride concentration 1.5mM, primers concentration (reverse and forwid) 20pmoles, DNA(Template) volume 6il, Taq DNA polymerase(12.5U), 2001.iM of each dNTPs. The PCR products were analyzed and compared for their banding pattern of 681bp and 427bp amplified by the primer pairs (37, 38) and (39, 40), respectively, against the standard DNA ladder (1 OObp) by using horizontal agarose gel electrophoresis. One of the HJA positive samples was not amplified by the PCR. The results of this study showed that the specificity of PCR reaction as compared to the HA test and the presence of the CPV-2 and 2b in Pakistan as the prevalent pathogenic strains. This study has been a first step for the molecular characterizationldiagnosis of canine parvovirus in local conditions of Pakistan. It is hoped that this study will pave the way for further advanced studies on this topic.
Availability: Items available for loan: UVAS Library [Call number: 0972,T] (1).
3.
Prevalance And Anthelmentic Activity Of Indigenous Plants Against Trichostrongylus Of Sheep In District Zhob
by Nasib Ullah | Dr.Muhammad Lateef | Prof.Dr. Azhar Maqbool | Prof.Dr.Zafar | Faculty of Veterinary Sciences.
Material type: ; Format:
print
Publisher: 2010Dissertation note: A Trichostrongylid is gastrointestinal nematode which causes the heavy economic losses to the livestock particularly sheep. A total 240 gastrointestinal tracts of sheep from district Zhob abattoir were collected. These samples were isolated and identified for trichostrongylid nematodes. The overall prevalence was 50%(120/240), of which 39.1% in male and 60.8% in female sheep were recorded at slaughter-house. Plants were collected from area of district Zhob. These plants were Identified and authenticated by botanist .The crude aqueous methanolic extract of the plants were used for in-vivo studies. Eighty sheep of either sex, aged between three to six months and naturally infested with Trichostrongylid nematodes (including trichostrongylius spp. I-Iaemonchus contortus, cooperia etc) were selected and managed separately for the experiment. These sheep were divided into 4 groups A ,B,C and D. Group A was contained 10 sheep and was untreated and considered as control. Thirty (30) were kept in Group B, were further sub divided into three equal groups i.e. BI, B2 and B3 and treated with different levels of Chenopodium album @ 1, 2 and 3 g/kg body weight respectively. Group C having thirty (30) was also sub divided into three equal groups i.e. Cl, C2 and C3 and treated with different levels of Artemisia brevifolia @ 1, 2 and 3g/kg body weight respectively. Group D was treated with Levamisole @ 7.5 mg/kg body weight. Faecal egg count reduction was criterion for evaluation. Faecal samples were collected before treatment on day 0 and on day 3, 5, 7, 10 and 14 post treatments for EPG count.
The results of EPG for animals in group B1 at day 0 was 1325. This rate was reduced to 1250, 1125, 995 and 702 at day 3, 7, 10 and 14, respectively. Similarly EPG of sheep in B2 were 1280 at day 0 and reduced to 1205, 1202, 1001 and finally to 690 at day 3, 7, 10 and 14 respectively. Best results in B group against the nematodes were in B3 in which Chenopodium album was used 3g/kg bw. The results were 1250, 1231, 1145, 590 EPG at day 0, 3, 7, 10 and 14. The sheep in group Cl showed 1203 EPG at day 0, when treated with Ig/kg bwArtemisia brevifolia, the EPG was reduced to 1173, 1115, 700 and 528 on day 3, 7, 10 and 14 respectively. Second level of treatment C2 of
Artemisia brevifolia which was given @ 2g/kg bw initially contained reduced to 1202 EPG, on day 3, 7, 10 and 14 the EPG counts were 1020, 631, 546 and 459, respectively.
Highest dose of Artemisia brevfolia was 3g/kg body weight to sheep in group C3. On day zero the EPG count was 1196. On day 3, 7, 10 and 14th day the EPG count decreased to 1079, 905, 528 and 396 respectively. The sheep in group D, treated with recommended dose of Levamisole showed 1138 EPG prior to medication, which reduced remarkably to 681, 536, 357 and 147 on day 3, 7, 10 and 14 respectively. Although no untoward effects of plants were observed but best EPG reduction results (87.08 %) were observed in Levamisole as compared to treatment of 3gm/kg b.w Chenopodium album (51 .03 %) and Artemisia hrevfolia 3g/kgbw (66.88 %).
CONCLUSIONS
A wide variety of plants are naturally available in the Indo-Pakistan subcontinent which possess narrow or broad spectrum anthelmintic activities. No doubt this is true in other regions of the world as well where gastrointestinal parasitism is an important problem in livestock keeping, and the availability of commercial drugs may be limited. Conventionally, trichostrongylids has been tackled with use of synthetic anthelmintic but owning to development of anthelmintic resistance against major groups of anthelmintics viz., benzimidazole, Levamisole and avermectins, people are looking for alternatives to synthetic chemicals.
The phytochemical analysis of these plants and controlled anthelmintic trials along with contemporary knowledge of parasite control strategies may offer new opportunities for effective and economical control of parasitic diseases. So these plants can be better alternative for synthetic chemicals.
Quality control extracts of Artemisia brevifolia and Chenopodium album or possibly isolated bioactive compounds could be a promising alternative to conventional anthelmintics fbr the treatment of gastrointestinal trichostrongylids of small ruminants in the future. Such a treatment could be used in control strategies against gastro intestinal nernatodes in organic and conventional production systems. Further research is needed for studies on the bio active constituents, as well as on the reproducibility, dosage, application regime. toxicity and effectiveness of Artemisia brevifolia and C'henopodium a/bums in other host species and against other economically important gastro intestinal nematodes species.
RECOMMENDATIONS
It is recommended that further research could be carried out on large sample size in different seasons of the year and large number of plants, identification of active principles of plants with proven anthelmintic activity, standardization of dose and toxicity studies for drug development. In addition to this, large number of samples of the same plant from different geographic areas should be subjected to experimentation keeping in view the possibility of differences in chemical composition of soils.
Availability: Items available for loan: UVAS Library [Call number: 1075,T] (1).
