Introduction: Toxoplasmosis is a widespread zoonosis that causes significant economic losses due to abortions and other developmental disabilities in livestock animals. The objectives of the current study were to determine the prevalence and associated risk factors of Toxoplasma gondii infection in cattle and buffaloes in northern parts of Punjab, Pakistan, where no such work previously existed. Methodology:Serum samples obtained from 400 cattle and 422 buffaloes present on different private and government-owned farms were tested for anti-Toxoplasma IgG and anti-Toxoplasma IgM antibodies by using enzyme linked immunosorbent assay (ELISA). Additional data, including water sources, hygienic status at the farm, management practices, size of the herd, and presence of cats in the vicinity of the farm were obtained using a questionnaire in surveys and interviews. Results: The overall prevalence of infection was 19.75% (79/400) in cattle and 15.16% (64/422) in buffaloes. IgG antibodies were found in 75 (18.75%) cattle and 58 (13.74%) buffaloes, while IgM antibodies were found in 9 (2.25%) cattle and 10 (2.37%) buffaloes. Seroprevalence was significantly higher in females and older animals of both species. Seroprevalence was found to be associated with poor hygienic conditions, extensive management practices, and presence of cats in the surrounding areas. No difference of seroprevalence was observed with respect to different breeds, location, water source, and herd size. Conclusion: The present study found that T. gondii is prevalent in large ruminants in northern Punjab, and may have important implications for the livestock industry and public health.

OBJECTIVESThe health transformation program was a recent reform in the health system of Iran that was implemented in early 2014. Some of the program’s important goals were to improve the equity of payments and to reduce out-of-pocket (OOP) payments and catastrophic health expenditures (CHE). In this study, these goals were evaluated using a before-and-after analysis.METHODSData on household income and expenditures in Guilan Province were gathered for the years 2013 and 2015. OOP payments for outpatient, inpatient, and drug services were calculated, and the results were compared using the propensity score matching technique after adjusting for confounding variables. Concentration indices and curves were added to quantify changes in inequity before and after the reform. The incidence of catastrophic expenditures was then calculated.RESULTSOverall and outpatient service OOP payments increased by approximately 10 dollars, while for other types of services, no significant changes were found. Inequity and utilization of services did not change after the reform. However, a significant reduction was observed in CHE incidence (5.75 to 3.82%).CONCLUSIONSThe reform was successful in decreasing the incidence of CHE, but not in reducing the monetary amount of OOP payments or affecting the frequency of health service utilization.

BackgroundNewcastle disease (ND) is one of the most deadly diseases of poultry around the globe. The disease is endemic in Pakistan and recurrent outbreaks are being reported regularly in wild captive, rural and commercial poultry flocks. Though, efforts have been made to characterize the causative agent in some of parts of the country, the genetic nature of strains circulating throughout Pakistan is currently lacking.Material and methodsTo ascertain the genetics of NDV, 452 blood samples were collected from 113 flocks, originating from all the provinces of Pakistan, showing high mortality (30–80%). The samples represented domesticated poultry (broiler, layer and rural) as well as wild captive birds (pigeons, turkeys, pheasants and peacock). Samples were screened with real-time PCR for both matrix and fusion genes (1792 bp), positive samples were subjected to amplification of full fusion gene and subsequent sequencing and phylogenetic analysis.ResultsThe deduced amino acid sequence of the fusion protein cleavage site indicated the presence of motif (112RK/RQRR↓F117) typical for velogenic strains of NDV. Phylogenetic analysis of hypervariable region of the fusion gene indicated that all the isolates belong to lineage 5 of NDV except isolates collected from Khyber Pakhtunkhwa (KPK) province. A higher resolution of the phylogenetic analysis of lineage 5 showed the distribution of Pakistani NDV strains to 5b. However, the isolates from KPK belonged to lineage 4c; the first report of such lineage from this province.ConclusionsTaken together, data indicated the prevalence of multiple lineages of NDV in different poultry population including wild captive birds. Such understanding is crucial to underpin the nature of circulating strains of NDV, their potential for interspecies transmission and disease diagnosis and control strategies.

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Toxoplasmosis is one of the most common zoonotic protozoal diseases. Recent advances in biotechnology have produced recombinant protein, which are immunogenic, and progress in nano-pharmaceutics has generated encapsulated protein in nanospheres, which are suitable for vaccine delivery. DNA was extracted from Toxoplasma gondii oocysts and was confirmed through nested PCR and sequencing. The 1665 bp of ROP18 was cloned into the easy vector system: pGEM-T by the T-A cloning method. DH5α bacteria were transfected with pGEM-ROP18. ROP18 was subcloned from pGEM-ROP18 into pET28-ROP18. BL21 bacteria were transfected with pET28-ROP18. Thus, rROP18 protein was expressed in BL21 bacteria by induction at different concentrations of isopropyl β-D-1-thiogalactopyranoside. Protein expression was confirmed through SDS-PAGE and Western blotting. The immunoblot of rROP18 was recognized by anti-HIS antibodies and sera from infected mice at 67 kDa. Recombinant ROP18 protein was encapsulated in nanoparticles with PLGA and was characterized through scanning electron microscopy. Intraperitoneal immunizations with rROP18 protein and intranasal immunization of nanospheres were carried out in mice, and the immune response was detected by ELISA. Results showed that rROP18 in nanospheres administered intra-nasally elicited elevated responses of specific IgA and IgG2a as compared to groups inoculated intra-nasally with rROP18 alone, or injected subcutaneously with rROP18 in montanide adjuvant. It was concluded that nanospheres of ROP18 would be a non-invasive approach to develop vaccination against T. gondii. Further experiments are needed to determine the cellular response to these nanospheres in a mouse model for chronic toxoplasmosis.

In the present study, sterilized rice bran was fermented for 48, 72 and 96 hours by Aspergillus flavus and proximate analysis indicated that fermentation changed nutritive value of the rice bran. Four experimental feeds were fed to 4 groups of day old broiler chicks for 6 weeks to evaluate their effects on broiler performance (weekly feed intake, weight gain, FCR and mortality). During first 2 weeks, birds of group D showed significant difference (P less than 0.05) in feed intake and weight gain as compared to groups A (control), B and C. All groups did not show significant differences in feed conversion ratios (FCR). During 3rd week of trial, chickens of Group D showed significant difference (P less than 0.05) in feed intake, weight gain and FCR as compared to group A. During last 3 weeks, birds in group D showed significant difference (P less than 0.05) in feed intake, weight gain and FCR as compared to birds of groups A, B and C. Group D showed significantly higher weight gain and better FCR (P less than 0.05) as compared to groups A, B and C. On the whole, broiler chickens fed on 96 hours fermented rice bran showed best growth performance. In-vivo phytase activity was determined in ash of tibia bones collected from selected birds in each group. Group D showed significantly high percentage (P less than 0.05) of ash and phosphorus in tibia bones as compared to groups A (control), B and C. In conclusion, fermented rice bran showed beneficial effect on broiler performance and could be used in poultry feed as phytase source.

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The linkage disequilibrium (LD) is an important tool to study quantitative trait locus (QTL) mapping and genetic selection. In this study, we identified the extent of linkage disequilibrium (LD) in Sahiwal (n = 14) cattle using the bovine high density single-nucleotide polymorphisms (SNPs) BeadChip. After data filtering, 500,968 SNPs comprising 2518.1 Mb of the genome, were used for the LD estimation. The minior allele frequency (MAF) was 0.21 in a substantial proportion of SNPs and mean distance between adjacent markers was 4.77 ± 2.83 kb. The overall mean LD between adjacent markers was 0.18 (r 2 ) and 0.55 (|D'|), respectively. The LD (r 2 ) values reduced with the increase in distance between adjacent markers from 1 kb (0.35) to 100 kb (0.12) and |D'| specified distinct decay of the LD. Chromosomes 1, 27, 28 and 29 presented the LD at some distance between markers. The extent of LD was higher, except these four chromosomes, for markers separated by 20 kb. At < 3 kb distance, the upper value of the linkage disequilibrium (LD) was observed at 0.30. High level of the linkage disequilibrium (LD) between markers was observed at high minor allele frequency (MAF) threshold (0.15), at the short distance between markers. The results of this study revealed that the Bovine high density SNPs BeadChip will be informative for the estimation of breeding value in Sahiwal cattle.