Changes in clinical signs after treatment in calves with experimental colisepticemia with Escherichia coli

Background: Colisepticemia is an acute fatal disease in farm animal neonates. Clinical finding of septicemia is non-specific and cannot be differentiated from signs of non-infectious disease or disease with local infection such as diarrhea

Objectives: Evaluation of clinical signs variations in calves with experimental septicemia with Escherichia coli 01 11 :H8

Methods: Colisepticemia was experimentally induced in ten Holstein bull calves after an adaptation period. Vital signs and 7 clinical criteria were recorded from 24 h before septicemia until 48 h after that. Blood culture was performed and treatment was done based on antibiogram from 24 h after challenge

Results: Changes of suckling reflex and shock were not significant. Changes of appetite, dehydration, behavior, standing ability, total score from 24 h before the challenge to 24 h after treatment were significant [p=0.00l]. Fecal consistency altered with treatment [p<0.04]. Heart rate [p=0.04 and p=0.033, respectively], respiratory rate [p=0.009 and p=0.001, respectively] and body temperature [p0.00l and p=0.004, respectively] have significant changes till 24 h after challenge and till 24 h after starting treatment. Blood cultures were positive except for 0 h and 48 h after challenge

Conclusions: The present study indicated clinical signs changed unfavorably following septicemia that were dissolved approximately during 24 h, depending on treatment in appropriate time and drug choice. Thus, a targeted scoring system will be useful in clinical evaluation of septicemia, quantifying the changes procedure and treatment efficacy

[Study of MHC polymorphism and its linkage to IGF1 gene in Khorasan indigenous chicken]

Background: indigenous chickens could serve as precious genetic resources that should be considered in conservation and breeding programs. The Major Histocompatibility Complex [MHC] has a strong association to disease resistance/susceptibility, production and reproduction traits in chicken. Therefore, identifying its polymorphism in populations under selective breeding could be used for selection of disease resistant and higher productive breeds. MHC association with quantitative traits could be a result of its linkage with causative genes controlling these traits. Insulin-like growth factor 1 [IGF1] is a candidate marker for phenotypic traits in chicken which are associated with important production and reproduction features

Objectives: based on this hypothesis, MHC polymorphism and its association to IGF1 gene [as a marker for production traits] were investigated in Khorasan indigenous chicken

Methods: in total, 313 DNA samples that belonged to the Khorasan indigenous chicken were analyzed. LEI0258 microsatellite marker and fragment analysis method was used for MHC genotyping. Single nucleotide polymorphism [SNP] of the IGF1 5'-UTR was detected by restriction fragment length polymorphism [PCR-RFLP] and PstI restriction endonuclease enzyme. Linkage disequilibrium between MHC and IGF1 loci were also determined using SAS/Genetics software and likelihood ratio test

Results: collectively, 25 different alleles [185-493 bp] and 76 genotypes of LEI0258 microsatellite were identified in Khorasan population. Two alleles, A [PstI -] and B [PstI +] and three genotypes [AA, AB and BB] were identified for IGF1 gene. Significant linkage disequilibrium [p=0.0083] was observed between LEI0258 and IGF1 loci in this population

Conclusions: these results indicate a high MHC genetic diversity in Khorasan indigenous chicken as a valuable genetic resource. Results from MHC/IGF1 linkage study confirm the hypothesis that MHC association with production traits could be as a result of MHC linkage with causative genes controlling the traits

Study of cathepsins involved in haemoglobin and vitellin digestion in Rhipicephalus [Boophilus] annulatus larvae by one- and two dimensional zymography

Enzymatic digestion of proteins in ticks is a complex process and the study of functional proteomics of these enzymes can help to select them as possible vaccine candidates. Blood protein changes [e.g. haemoglobin to vitellin] occur in female mature ticks. Vitellin digestion, as an amino acid and energy source, is one of the vital and important processes in development and evolution of tick eggs and larval stage of unengorged ticks. Several studies reveal a network of proteolytic enzymes involved in haemoglobin and vitellin digestion. These enzymes are mostly cysteine and aspartic peptidases. The aim of this study was the detection of the cathepsins in Rhipicephalus [Boophilus] annulatus larvae extract. In the current research, cysteine proteases extracted from Rhipicephalus [Boophilus] annulatus larvae were studied by one- and two-dimensional zymography. Findings from one dimensional zymography showed a transparent band with 28 KDa. In two-dimensional zymography transparent area are seen in the dark gel background distributed in 21 to 65 KDa zones related to cathepsins. When DTTwas added to incubation buffer [10 mM acetate buffer, pH= 4], the proteolytic activity of some enzymes was increased and appeared as more clear transparent bands in one-dimentional zymography compared with samples incubated in buffer without DTT. As the pH of incubation buffer was acidic and adding DTT resulted in increased activity of the enzymes, therefore, some of these proteolytic enzymes are assumed to be cysteine proteases

Comparison of immunochromatographic rapid test with molecular method in diagnosis of canine parvovirus

Canine parvovirus [CPV] infection is one of the most common causes of infectious gastroenteritis in dogs and is a highly contagious, often fatal disease. The original virus[CPV type 2] has had some mutations since its emergence and new variants [CP V-2a, 2b and 2c] have been reported from many countries all around the world. Early diagnosis and treatment can profoundly affect the disease outcome. To compare the ability of Immunochromatographic [1C] test to detect CPV infection in 50 PCR positive samples [n=50] with regard to virus strains. 50 rectal swabs [n=50] were prepared from suspicious dogs and subjected to PCR and 1C test respectively. The sensitivity of 1C test in PCR positive samples was 84% [42 out of 50 samples] and the positive predictive value of the test was 100%. Using PCR, CPV strains in our study were 2a [18/50,36%] and 2b [32/50,64%] with the predominance of 2b strain. 1C test was also able to diagnose 15/18 [83.3%] of CPV-2a and 27/32 [84.3%] CPV-2b strain positive samples, which means 1C test can detect CPV infections caused by both virus strains [2a and 2b], without significant difference. This study shows that 1C test results are relatively reliable for diagnosing CPV infection in daily veterinary practice and the test is able to diagnose both CP V-2a and CP V-2b which are prevalent strains in Iran

Use of N-trimethyl chitosan for intranasal delivery of DNA encoding M2e-HSP70c in mice

Influenza outbreak has become a great lifethreatening disease in the world. Nasal vaccines can induce systemic IgG and mucosal IgA antibody responses, which establish two layers of immune defense against the infectious pathogens like influenza. Mucosal vaccines must overcome several limitations, including the mucociliary clearance and inefficient uptake of soluble antigens. Therefore, nasal vaccines require potent adjuvants and delivery systems. In this study we evaluated the effect of N-trimethyl chitosan [TMC] as a potent vehicle for DNA encoding M2e/HSP70c in order for intranasal administration in mice. Ectodomain of the conserved influenza matrix protein 2 [M2e], which has been found to induce heterosubtypic immunity, was fused to HSP70359-610 or C-terminus of Mycobacterium tuberculosis HSP70 [HSP70c] in pcDNA3.1 vector [pcDNA/M2e-HSP70c] and then encapsulated into a derivative of chitosan, N-trimethyl chitosan [TMC]. After encapsulation of the plasmid, physical properties of the particles were investigated using Zetasizer[R] 3000 the particles were then administered through the intranasal delivery in BALB/c mice. It was found that the particles had a size ranging between 90-120nm and positive surface charge. The intranasal immunization with M2e-HSP70c+TMC in BALB/c mice significantly induced higher M2e specific IgG than those induced in control groups [pcDNA/M2e-HSP70c without TMC, pcDNA/M2e, bearing M2e alone, and PBS]. The present study showed that the encapsulation of M2e/ HSP70c into N-trimethyl chitosan [TMC] could strongly induce the humoral immune response against the M2e-HSP70c plasmid without lowering the adjuvant efficacy of HSP70c

[Effect of climatic factors on canine Lime borreliosis in Iranian Caspian sea littoral provinces]

Lyme borreliosis is a worldwide zoonotic disease caused by spirochetes of the Borrelia burgdorferi sensu lato complex. There are no reports on this subject in dogs from Iran. Determining the serologic prevalence level of produced antibodies against Borrelia burgdorferi sensu lato complex in three Caspian littoral provinces of Iran and studying the effect of climatic risk factors on it are the first aims of this study. During the period from July to September 2009 a seroepidemiological study was conducted on 273 dogs in three Caspian provinces of Guilan, Mazandaran and Golestan, Iran's known habitats of tick [Ixodes ricinus]. In order to study the correlation between infection distribution and climatic factors by geographic information system [GIS], geographic position of seronegative and seropositive dogs was overlaid on climatic maps of Guilan, Mazandaran and Golestan provinces. Multivariate regression model and correlation matrix analyses were used for statistical analysis. From 273 serum samples in the whole studied area, 22 [8.1%] showed antibodies against B. burgdorferi sensu lato complex. The seroprevalence of B. burgdorferi sensu lato in provinces of Guilan, Mazandaran and Golestan were 0.0% [0.91], 2.2% [2.91] and 22% [20.91], respectively. Mean annual temperature had positive and significant correlation with B. burgdorferi sensu lato complex seroprevalence in sampled dogs of the three north provinces [p<0.05]. Regarding the seroprevalence of Lyme borreliosis in dogs of three Caspian provinces of Iran, more attention must be paid to this disease, especially in Golestan province. This is the first study on the role of climatic factors in canine Lyme borreliosis in Iran

[ effect of parenteral administration of erythromycin on Immunoglobulin G absorption in neonatal calves]

Motilides mainly erythromycin have a great ability to increase abomasal emptying rate. The aim of this study was to evaluate the effect of erythromycin as a prokinetic agent on abomasal emptying rate and Immunoglobulin G absorption in neonatal calves. Twelve holstein neonate calves were divided into two groups [treatment and control] of 6 Calves each. One hour after birth, treatment and control groups were injected by erythromycine [8.8 mg/kg; IM] and normal saline [IM]. After 30 minutes, calves were fed by 3 liters of colostrum using esophageal tube. Venous blood samples for determination of plasma IgG were obtained at 1, 3, 6, 9, 12, 18, 24, 36, 48 hours and 5 and 7 days after birth. The results showed that administration of erythromycin caused a significant increase in the serum IgG level [20.394 mg/mL], compared to the control group [15.021 mg/mL]. This study revealed that erythromycin increases the serum IgG level probably through abomasal emptying acceleration

[Study on P53 gene alterations [5 and 6 exons] in bovine urinary bladder tumors]

P53 is a tumor suppressive gene which frequently mutates in tumors of animals and human. This gene commonly mutates in urinary bladder tumors of human beings. Urinary bladder tumors have occurred in cattle with bovine enzootic hematuria [BEH]. The aim of present study was to evaluate P53 mutations in 15 samples of different bovine urinary bladder tumors by the PCR-SSCP technique. Fifteen paraffin embedded blocks were selected from different kinds of bovine urinary bladder tumors. DNA was extracted from the samples and PCR was done by using specified primers for 5 and 6 exons. After electrophoresis, the PCR products were assessed by the SSCP method, and samples with changes in electrophoresis patterns were selected and sequenced. Results showed that there are intronic alterations of the P53 gene in cattle with urinary bladder tumors. There were no changes in the electrophoretic pattern of exons 5 and 6, but on each side of the designed primers for exon 6, there was a part of introns 5 and 6. The samples, including Hemangioma, Papilloma and Carcinoma in situ with electrophoretic changes, showed nucleotide T deletion with number 9332 in intron 6 after direct sequencing. Intronic mutations can be a predisposition for developing cancers. It is possible that some of urinary tumors are inducted by P53 mutations in intronic zone

Development and ELISA-based detection of anti-M2e IgY antibodies using an encoding plasmid for M2e-Hsp70 c-terminal gene

The use of IgYs in a variety of methods in different areas of research, diagnostics, medical application and biotechnology should be considered widely. Development of antibodies against extra cellular domain of influenza M2 [M2e] protein in egg yolk of laying hens. A Fusion construct harboring C-terminal of bovine heat shock protein 70 [Hsp70] and influenza M2e coding genes was injected to laying hens. Serum and egg yolk antibodies were screened for the presence of anti-M2e antibodies by indirect enzyme-linked immunosorbent assay [ELISA]. Anti-M2e antibodies were detected in egg yolks and sera of injected hens from 13 and 7 days post injection [PI], with the peak titer detected on 41 and 35 days PI, respectively. Anti-M2e IgY titers could be an index for expression potential of pcDNA3.1-M2e-HspCterminal construct in laying hens. This construct could be considered as a promising tool in production of anti-M2e polyclonal, monospecific IgY antibodies. Such anti-M2e antibodies could be exploited for influenza diagnostic and therapeutic measures

[ effect of different levels of cadmium on the histopathological changes and the rate of lymphoid cells apoptosis of bursa of fabricius in broiler chickens]

Cadmium toxicity can cause kidney failure, liver damage and a weakened immune system in experimental and naturally occurring toxicities. This study was designed to investigate the effects of cadmium [Cd] on the histology and the rate of lymphoid apoptosis in the bursa of fabricius of chicken. One-hundred 20-day-old male Ross broilers were purchased and randomly divided into four groups. The control group [C] received no Cd, whereas groups 1, 2, and 3 had rations administered containing 25, 50 and 100 ppm cadmium as CdCl, respectively. At days 14, 28 and 42, seven chicks from each group were randomly selected and sacrificed. The bursa of Fabricius of each chick was removed, weighed, fixed in 10% buffered formalin and processed for histopathology and assessment of the rate of lymphoid cells apoptosis. The apoptotic cells were demonstrated in paraffin embedded tissue sections using the TUNEL[terminal oxynucleotidyl transferase-mediated dUTP nick end labeling] method. The concentration of Cd in the liver samples was measured by atomic absorption. Areverse correlation between the levels of Cd in the rations and the body weight of the chickens [p < 0.01] was found. The concentration of Cd in the liver showed a positive correlation with the levels of Cd in the rations [p < 0.01]. The number of apoptotic lymphoid cells was significantly increased in those groups receiving higher levels of Cd [especially groups 2 and 3] [p<0.01]. Morphologically, plicas and lymphoid follicles of groups 2 and 3 were smaller than of the control group. In the histological analysis they were found to be hypocellular and some of them were edematous. Compared to the control group, there was an increase in the number of intraepithelial cysts in groups 2 and 3 at days 28 and 42. In addition, atrophic changes of bursal paranchyma were observed in group 3 after 42 days. It can be concluded that under experimental conditions the higher concentrations of Cd in the rations [50 and 100 ppm]has detrimental effects on the bursa of Fabricius of chickens

Detection of tumor antigens in bovine viral lymphosarcoma by immunoprecipitation method

In this study one lymph node from cow with viral lymphosarcoma and one lymph node from healthy cow were analyzed by three different methods for protein extraction, include simple homogenizing in PBS, sonication and phenol extraction. Tumor antigen detection relies on immunoprecipitation followed by SDS-PAGE. In this experiment Streptococcus pyogenes was used for purification of immune complexes. Eletrophoretic patterns were obtained using reduced and non reduced buffers. In PBS homogenization 3 distinct antigens [39, 32 and 30 kDa] were observed. When sonication or phenol extraction were used, 5 [72, 48, 42, 32 and 30 kDa] and 6 [104, 77, 54, 32, 30 and 26, 5 kDa] distinct antigens were observed, respectively. This study showed that the ability of antigen detection mainly depends on protein extraction procedure and antigen-antibody equilibrium achieved by quantification of precipitins. Immunoprecipitation could be used successfully for study of tumor antigens in bovine leukosis