Toxoplasma gondii is one of the most prevalent intracellular parasites and is threatening the health of both humans and animals, therefore causing incalculable economic losses worldwide. Vaccination is thought to be an efficient way of controlling toxoplasmosis. T. gondii microneme protein 11 (MIC11) is a soluble microneme protein which is presumably considered facilitating the early stage of cell invasion. To evaluate the protective efficacy of T. gondii MIC11, in the present study, a new DNA vaccine-encoding the α-chain of T. gondii MIC11 was constructed using the pcDNA3.1 vector. Expression of MIC11 from this vector was confirmed by indirect immunofluorescence assay following transfection into baby hamster kidney (BHK) cells. Intramuscular immunization of BALB/c mice with pcDNA/MIC11 was carried out to evaluate the immune responses by serum antibodies titers, lymphoproliferation assay, and cytokines assay. The protective efficacy was evaluated by survival rate in mice after challenging with highly virulent strain of T. gondii. The results demonstrated that this vaccination elicited significant humoral responses and T. gondii lysate antigen (TLA)-stimulated lymphoproliferation (p < 0.05). Compared to controls, the pcDNA/MIC11 immunized mice had high production of IFN-γ, IL-12, and IL-2 (p < 0.05), but not IL-4 (p > 0.05), indicating that a predominant Th1 type response was developed. The vaccination also increased the survival rate of immunized mice when they were challenged with a lethal dose of tachyzoites of T. gondii RH strain. These data suggest that T. gondii MIC11 is a reasonable vaccine candidate deserving further studies, and pcDNA/MIC11 is a potential strategy for the control of toxoplasmosis.
BackgroundApicomplexan parasites of the genus Babesia, Theileria and Plasmodium are very closely related organisms. Interestingly, their mitochondrial (mt) genomes are highly divergent. Among Babesia, Babesia orientalis is a new species recently identified and specifically epidemic to the southern part of China, causing severe disease to water buffalo. However, no information on the mt genome of B. orientalis was available.MethodsFour pairs of primers were designed based on the full genome sequence of B. orientalis (unpublished data) and by aligning reported mt genomes of B. bovis, B. bigemina, and T. parva. The entire mt genome was amplified by four sets of PCR. The obtained mt genome was annotated by aligning with published apicomplexan mt genomes and Artemis software v11. Phylogenetic analysis was performed by using cox1 and cob amino acid sequences.ResultsThe complete mt genome of B. orientalis (Wuhan strain) was sequenced and characterized. The entire mt genome is 5996 bp in length with a linear form, containing three protein-coding genes including cytochrome c oxidase I (cox1), cytochrome b (cob) and cytochrome c oxidase III (cox3) and six rRNA large subunit gene fragments. The gene arrangement in B. orientalis mt genome is similar to those of B. bovis, B. gibsoni and Theileria parva, but different from those of T. orientalis, T. equi and Plasmodium falciparum. Comparative analysis indicated that cox1 and cob genes were more conserved than cox3. Phylogenetic analysis based on amino acid sequences of cox1, cob and cox1 + cob, respectively, revealed that B. orientalis fell into Babesia clade with the closest relationship to B. bovis.ConclusionsThe availability of the entire mt genome sequences of B. orientalis provides valuable information for future phylogenetic, population genetics and molecular epidemiological studies of apicomplexan parasites.
A cross-sectional study was conducted in desert environment of Jhang (Pakistan) from November 2008 to October 2009 on she-camels kept under pastoralist conditions to determine the prevalence of mastitis, impact of risk factors, and isolate the dominant mastitis-causing bacteria on total of 150 lactating she-camels by using clinical examination and surf field mastitis test. From the 150 she-camels examined, 69 (46%) were positive for mastitis at animal level, 12 (8%) clinical, and 57 (38%) subclinical. Age, parity number, stage of lactation, breed, production system, hygiene of milking process, and presence of lesion on udder/teat were found significantly associated (p<0.05) with the prevalence of mastitis in she-camels. There was the lowest prevalence (33.33%; 15 of 45) of mastitis in she-camels of 5-7 years of age, while the highest (80%; 12 of 15) in the animals aged between 14 to 16 years. Stage of lactation significantly affected (p<0.05) and was found to be associated with the prevalence of mastitis being the highest (54.55%; 18 of 33) during the initial stage of lactation (0 to 1 month) followed by last 2 months (10-12 months) as 54.17% and mid-stages (1-3 and 3-10 months) of lactation as 28.57% (6 of 21) and 37.50% (9 of 24), respectively. According to breed of camels, it was noted that the prevalence of mastitis affected significantly (p<0.05) being the highest in crossbred (Desi × Mareecha) as 51.39% (37 of 72) followed in order by Mareecha and Desi as 43.14% and 37.04%, respectively. Staphylococcus (42.19%) and Streptococcus (15.63%) genera were the dominant isolates identified. Good hygiene in milking process, milking clinically infected she-camels at last, culling chronic mastitis carriers, treating clinically infected she-camels, and dry period therapy could reduce the prevalence of contagious mastitis in the study area.
This paper provides an estimation of the point prevalence of fascioliasis and its economic impact in terms of increased milk yield after chemotherapy of a bovine population from the district of Toba Tek Singh, Punjab, Pakistan. A total of 2400 cattle and buffaloes were examined quantitatively using the McMaster egg-counting technique. Infected cattle and buffaloes (50 of each) were randomly selected and each divided into two groups of 25 animals. Groups A (buffaloes) and C (cattle) were treated with oxyclozanide (orally, 16.6 mg kg- 1 body weight). Groups B and D served as negative controls for buffaloes and cattle, respectively. Pre- and post-treatment milk yield was recorded to determine if there were any changes in milk yield after treatment. Of 2400 faecal samples analysed, 654 (27.25%) were positive for Fasciola spp. with a mean number of eggs per gram (EPG) of 503.2. The point prevalence and worm burden of fascioliasis was significantly higher (OR = 2.13; P < 0.05) in buffaloes (34.58%; 415/1200; mean EPG maximum likelihood = 521.4) as compared to that of cattle (19.92%; 239/1200; mean EPG maximum likelihood = 415.8). Among the parasite species, F. gigantica (19.88%; 477/2400) was predominant (OR = 3.12; P < 0.05) as compared to F. hepatica (7.38%; 177/2400). An average daily increase of 0.67 and 0.87 litres of milk, with 0.41% and 0.37% more fat per animal, was observed in oxyclozanide-treated buffaloes and cattle, respectively. The economic value of reduced production of infected animals was estimated as US$0.33 and 0.32 per animal per day for cattle and buffaloes, respectively.
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