ABSTRACT
VP2 gene coding region of a vaccinal strain [D78] of infectious bursal disease virus [IBDV] was cloned in a eukaryotic expression vector, pSec Tag2A. The gene was placed downstream of Ig kappa chain leader sequence, under the control of human cytomegalovirus [hCMV] immediate early enhancer and promoter. The construct pSec Tag2A-VP2 was transferred in COS-7 cell line and the expression and secretion of VP2 was assessed by dot blotting and antigen capture ELISA. The antibody used in the immunological assays was a neutralizing monoclonal antibody [1A6] against VP2. Positive reaction with the antibody indicated the construct was functional with respect to expression and secretion of a native VP2
ABSTRACT
In order to investigate the prevalence of Mannheimia haemolytica infection in cattle, nasal and nasopharyngeal swabs and blood samples were obtained from 250 cattle after slaughter at Ahvaz [southwestern Iran] abattoir. Nasal and nasopharyngeal swabs were cultured on blood agar and incubated at 37 [degree sign] C for 24-48 h. The suspected bacterial cultures were processed for isolation of M. haemolytica following routine bacteriological techniques. Sera were tested by indirect hemagglutination test [IHA] to reveal antibodies against the organism. M. haemolytica was isolated from 1.6% of the samples cattle. Statistical analysis showed that there was no relationship between age and sex with bacterial infection. Serological studies showed that 71.6% of tested sera contained antibody [titer >/= 1/16] against M. haemolytica. There was no association between age and sex with serological results
ABSTRACT
Tetanus is a disease caused by tetanus toxin, a potent inhibitor for the release of inhibitory neurotransmitter in the central nervous system that causes spastic paralysis. Fragment C [52 kD] of this toxin is responsible for binding to the neuronal membrane. For this reason, and also its non toxigenic and immunogenic nature, this fragment might be ideal for new vaccine development. Presently, with respect to the incidence of disease in neonates and animals and the side effects of toxoid vaccine, designing a more effective and efficient vaccine for prevention of this disease is crucial. A segment of Clostridium tetani DNA corresponding to C fragment of tetanus toxin was amplified using polymerase chain reaction. This fragment was cloned into expression vector pMalc2x, under the control of the lac promoter. Expression of this plasmid in Escherichia coli was confirmed by western blotting. In this study, the vector had a strong promoter to allow high level expression of C fragment. Based on our results it appears that this recombinant plasmid may be suitable for the production and development of recombinant vaccine and also has many other applications, such as construction diagnostic kits, production hyperimmune antiserum for serotherapy and as a vehicle for drug delivery to CNS
ABSTRACT
Kruppel-like factors [Klfs] are highly related zinc-finder family of transcription factors implicated in the regulation of the eukaryotic cellular growth and differentiation of a diverse set of cells in mammal. Using RT-PCR technique, a 456 bp cDNA fragment encoding N-terminus part of a Klf2b was isolated from the skin mucosa of common carp [Cypritnus carpio] using two degenerative oligonucleotide primers. Use of this fragment as a probe allowed the isolation of a larger cDNA clone through the searching of the GenBank expressed sequence tag database. The size of the amplified product is 1157 bp, which encodes a polypeptide of 274 amino acid residues with a predicted molecular mass of 30.359 kDa and theoretical pI of 4.88. The deduced amino acid sequence exhibited 79, 54, and 53% identity to the homologous Klf2b identified from zebrafish Anio rerio, Spotted Green Pufferfish Tetraodon nigroviridis and Atlantic salmon Salmo salar, respectively. The common carp protein is 50% similar to Klf2 orthologues in African clawed frog Xenopus laevis, 44% in chicken Gallus gallus and is 30% similar to the mammalian Klf in house mouse Mus musculus
ABSTRACT
Ten female lambs of 7-month-old were divided into two equal groups and raised under a helminth-free conditions. Animals in group 1 were immunized two times by whole gut homogenate [WGH] of Haemonchus contortus emulsified in Freund's adjuvant. In group 2 [control], animals were injected by phosphate buffered saline emulsified in the same adjuvant. Animals were challenged by 10000 third-stage larvae [L3] of Haemonchus contortus on day 33 after the first immunization and then humanely killed on day 42. Animals were tested for serum antibody and eggs per gram of faeces [EPG] throughout the study and nematodes in their abomasom were counted after necropsy. The results indicated that animals immunized with WGH showed a higher level of serum antibodies. A significant difference was observed in mean optical density of sera in ELISA between the two groups [P<0.05] and a 77 and 78% reduction in EPG and nematode counts at necropsy, respectively [P<0.05]
Subject(s)
Female , Animals , Sheep , Immunization , Enzyme-Linked Immunosorbent Assay , VaccinationABSTRACT
Exsheathing fluid [EF] and excretory-secretory products [ES] of infective third-stage cultured larvae of Ostertagia circumcincta were analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis [SDS-PAGE]. Five and seven predominant proteins were found in the EF and ES products, respectively. Immunoblotting by sheep pre-infection serum did not react with any of the EF and ES proteins, but the post-infection serum recognized four proteins of 44.5, 41.5, 38 and 24 kDa of the ES products. None of the EF products was recognized by the post-infection serum. Protectivity of the four proteins remains to be determined
ABSTRACT
Definitive diagnosis of brucellosis is made by isolation of the causative agents, which is a time-consuming procedure. To evaluate the efficacy of Dot-ELISA for detecting brucellae in clinical samples, 94 different specimens taken from animal origin were cultured on brucella selective culture media and colonies were identified biochemically. The specimens were also examined after centrifugation by Dot-ELISA using a specific anti-brucella antibody, a suitable peroxidase conjugate and substrate. Of the 94 samples, 5 [5.31%] were positive in Dot-ELISA and 4 [4.25%] had positive cultures. In comparison with culture, the sensitivity and specificity of Dot-ELISA for detection of brucellae in the samples was 80 and 100%, respectively. There was 98.9% agreement between the two tests. The results indicated that Dot-ELISA is a good and rapid test with acceptable sensitivity and specificity for detection ofBrucella spp. in aborted fetal stomach contents
ABSTRACT
To develop an ELISA technique for the detection of antibodies against Babesia ovis, the infected erythrocytes were lysed and the supernatant soluble antigen, after sonication and ultracentrifugation of the lysate was used as antigen. Optimal dilution of the antigen was determined by checkerboard titrations, using positive and negative control sera. A correlation of 85% was observed between the results of the developed ELISA and IFA techniques. To study the seroprevalence of Babesia ovis in Khouzestan province, south of Iran, a total of 1000 sheep sera were collected from different areas of the province and tested against Bahesia ovis using the ELISA technique developed. The results showed an average seroprevalence of 47.5% in the province. Our results indicated a significant increase of the seroprevalence by advancement of age of the animals. There was no significant difference between the seroprevalence of female and male sheep