Production of the phospholipase D and heat-shock protein [HSP]-60 recombinant proteins from Corynebacterium pseudotuberculosis

Background: Caseous lymphadenitis, caused by Corynebacterium pseudotuberculosis, is one of the most important diseases of sheep and goats, causing considerable losses for herd owners. Phospholipase D [PLD] is a potent exotoxin produced by C. pseudotuberculosis and it has been considered as the major virulence factor for this bacterium, possibly contributing to the spread of the bacteria from the initial site of infection to secondary sites within the host. Heat shock proteins [HSPs] are important candidates for the development of vaccines because they are usually able to promote both humoral and cellular immune re-sponses in mammals

Objectives: The aim of this study was the cloning and expression of the PLD and HSP genes of C. pseudotuberculosis

Methods: PLD and HSP[60] genes were cloned into pMAL-c2X vector and recombinant plasmids construct was transformed to DH[5] strain of E. coll. Expression of the proteins was shown by SDS-PAGE and accuracy of the cloned genes was confirmed by nucleo-tide sequence analysis

Results: The transformed E. coll strain DH[5] expressed PLD and HSP60 proteins effectively. The expressed fusion protein was found almost entirely in the soluble form

Conclusions: In the following studies the immunogenicity and protectivity of these recombinant proteins against C. pseudotuberculosis infections can be assessed

[Effect of hydro alcoholic extract of nutmeg on Immune response of rat]

Background: Myristica fragrans is an evergreen aromatic tree cultivated in many tropical countries. Nutmeg, the dried ripe seed of M. fragrans, is a popular spice used in sweet and savory cooking, and a variety of drinks

Objective: Nutmeg has a variety of pharmaceutical effects and in this study its effects on immune responses were evaluated

Methods: Six groups [each group 8] of Wistar rats were treated as follows: Even groups received Nutmeg extract at dose 300 mg per kg of body weight intraperitoneally [IP], daily up to 12 days. Odd groups received PBS instead of nutmeg extract. The rats of groups 1 and 2 were immunized IP with 1.35×109 Sheep RBC [SRBC] in days 1 and 6. The rats of groups 3 and 4, were immunized IP with 1.35×109 SRBC in days 1 and subcutaneously in right foot pad with 2.7×107 SRBC in a volume of 0.1 ml on the day 9. Titer of anti-SRBC antibody in the groups 1 and 2 and lysozyme and alternative complement pathway activity of groups 5 and 6 were evaluated on day 13. The swelling of footpads in groups 3 and 4 were measured at 24, 48, and 72 hours after challenge with SRBC and these rats euthanized on day 13 and theirs foot were examined histopathologically for infiltration of inflammatory cells

Results: The result showed that nutmeg extract significantly increase anti SRBC titer [P=0.005], inhibit inflammatory cells infiltration [P< 0.001] but has not any effect on serum lysozyme or complement activity [P=0.4]

Conclusion: In conclusion nutmeg extract shows a significant suppression on cell mediated immunity and stimulatory effect on humeral immune response to SRBC in Wistar rats

Evaluation of immunopathologic effects of aqueous extract of Echinacea purpurea in mice after experimental challenge with Pasteurella multocida serotype A

In order to assess the immunopathological effects of aqueous Echinacea purpurea extract [EPE] on mice experimentally challenged with Pasteurella multocida serotype A, forty female BALB/c mice were randomly divided into four groups. The groups included a control group [received sterile distilled water 2 times/week for 2 weeks, intraperitoneally and then 100 microl sterile saline intranasally], a PMA group [received sterile distilled water as the control group and after 2 weeks, 5.6 x 10[3] CFU/ml of P. multocida serotype A, intranasally], an EPE+PMA group [received E. purpurea extract intraperitoneally 2 times/week for 2 weeks and then challenged as the PMA group] and an EPE group [received E. purpurea extract as EPE+PMA group and then 100 microl sterile saline intranasally]. After 24 and 48 h post challenge, half of the animals in each group were sacrificed and analyzed for bacterial counts in their lungs and livers, TNF[alpha] serum levels and histapathological changes. The results showed significant differences in lung bacterial counts between PMA and EPE+PMA groups. TNF[alpha] serum level was significantly higher in the PMA group. Histopathological examination revealed infiltration of neutrophils in alveolar septa and hyperemia in the PMA group. In addition, the criteria of bronchopneumonia were partially recovered in the EPE+PMA compared to the PMA group. According to the results, it seems that E. purpurea extract has an immunomodulatory effect and can be used to prevent or control of pneumonia caused by Pasteurella

[Diagnosis of Dirofilaria immitis infection in urban and rural dogs in Ahvaz city by counterimmunoelectrophoresis]

Dirofilaria immitis is a nematode that is highly prevalent in dogs and it can cause dirofilariosis. The disease is transmissible to human, so it is important in terms of public health. The aim of the present study was to diagnose Dirofilaria immitis infection in the dog population of urban and rural areas of Ahvaz by counterimmunoelectrophoresis and compared with knott test. In the present study, serum of 200 dogs [80 urban and 120 rural], were evaluated for detection of Dirofilaria immitis infection, in Ahvaz area, Southwestern Iran. Counterimmunoelectrophoresis and modified knott's test were conducted on all blood samples to trace the antibody and microfilariae. Using counterimmunoelectrophoresis test, 9.5 percent of dogs [19 cases] were infected, but in modified Knott test, positive cases were detected 8 percent [16 cases]. Counterimmunoelectrophoresis test showed three more positive cases [one urban and two rural dogs] compared with the modified Knott test, which probably was due to occult infection. However, the difference was not significant [p>0.05]. Based on the results of counterimmunoelectrophoresis test, 6.25 percent of male and zero percent of female dogs in urban areas and 13 percent of male and 7.6 percent of female dogs in rural areas were infected respectively. The highest prevalence of infection in 6 year-or-more age groups was 23.3 and 14.29 percent in urban and rural dogs respectively. Statistical analysis did not show any significant relationship between infection and factors such as age and sex of the studied dogs [p>0.05]. It is proposed that for more accurate diagnosis of dirofilariosis, counterimmunoelectrophoresis test and the modified knott's test be used simultaneously

[ effect of omeprazole on colostral total IgG absorption in neonatal lambs]

Omeprazole as an inhibitor decreases abomasal acid secretion. On the other hand, acidity would be a determinant for absorption of Immunoglobulins. The aim of this study was to evaluate the effect of increasing abomasal pH due to Omeprazole administration on the rate of intestinal immunoglobulin absorption in newborn lambs. 30 lambs immediately after birth were divided into 6 groups of 5 animals each as follows: Group1; The lambs received colostrum from birth to 84 hours after birth. Group2; The lambs were fed by colostrum + Omeprazole [4mg/Kg] from birth to 84 hours after birth. Group 3; The lambs were given milk for the first 24 hours and after then until hour of 84 by colostrum + Omeprazole. Group 4; The lambs were fed with milk + Omeprazole for the first 24 hours and after then until hour of 84 with colostrum + Omeprazole. Group 5; The lambs received milk for the first 6 hours after birth and after then received colostrums until hour of 84. Group 6; The lambs were fed with milk + Omeprazole in the first 6 hours after birth and after then until hour of 84 by colostrum + Omeprazole. Blood samples were collected at 0,24,48,72 and 96 hours after birth. The total IgG was measured by ELISA method. Serum IgG levels in group 4 showed significant decrease when compared with the control [group 3]. However, no significant difference was shown in the serum IgG levels among groups. The results of this study showed that after birth increase in abomasal pH do not make effect on IgG absorption

serological survey of Leptospiral infection of cats in Ahvaz, southwestern of Iran

Leptospirosis is a zoonosis with numerous reservoir hosts. The disease is caused by infection with various serovars of Leptospira interrogans sensu lato. This study was conducted to evaluate the seroprevalence of leptospiral infection in stray cats in Ahvaz [south-western Iran] from April 2007 to June 2008. Blood samples were collected from 102 stray cats and screened for leptospiral infection using the microscopic agglutination test [MAT]. Five of the 102 cats [4.9%] were serologically positive for at least one serovar of L. interrogans. The greatest number of reactors was for L. interrogans serovar balum [five serum samples]. Antibodies against more than one serovar [namely, serovars balum and australis] were detected in one sample. All positive titers were detected at 1:100 dilution. The prevalence of leptospiral infection was 5.3% and 4.4% in male and female cats, respectively. There was no significant difference in positive titer prevalence between different sexes but prevalence was significantly different between age groups [P = 0.021], as all cats with positive titers were three years of age or more. This is also the first report of infection with L. interrogans serovars balum and australis from cats in Iran

Antigenic detection of cryptosporidium parvum in urban and rural dogs in Ahvaz district, southwestern Iran

Cryptosporidium parvum is a zoonotic protozoan parasite with a wide range of vertebrate hosts. The present study was conducted to determine the prevalence of Cryptosporidium parvum in urban and rural dogs of the Ahvaz area. Faecal samples were collected randomly from 93 dogs between May 2005 and September 2007. The studied dogs were divided into two groups [urban and rural] and based on age into three groups [<6 months, 6 months -3 years and >3 years]. The results were analyzed by using Chi-square analysis and Fischer's exact test. Prevalence to Cryptosporidium parvum antigens was 4.3% [4 of 93] by means of ELISA, indicating that this antigen is present in the ecosystem. The infection was more prevalent in rural dogs [6.4%; 3 of 47] in comparison with urban dogs [2.17%; 1 of 46], nevertheless, there were no significant differences between the different groups [P?0.05], but the infection was more prevalent in diarrheic dogs [17.65%; 3 of 17] compared with non-diarrheic dogs [1.3%; 1 of 76], and the difference was significant [P=0.019]. Infection was not significant in the different age groups [P>0.05]. Concurrent detection of cryptosporidium parvum with canine distemper [one sample] and parvovirus [one sample] were shown in the studied dogs. Modified Ziehl-Neelsen staining was also carried out and the prevalence of infection was 2.15% [2 of 93]. The use of ELISA allowed the detection of more positive cases than light microscopy. This study showed that Cryptosporidium parvum can be a risk factor, particularly for those dogs in contact together in the population of urban and rural dogs

Antigenic detection of Feline Panleukopenia virus [FPV] in diarrhoeic companion cats in Ahvaz area

The present study was carried out for the antigen detection of Feline panleukopenia virus [FPV] in diarrhoeic cats referred to the Veterinary Clinic of the School of Ahvaz University, in southwest Iran. Faecal samples were collected from 67 diarrhoeic household cats during 2005 to 2007. According to the age and clinical signs, the cats were divided into two groups; <6 months and >6 months, hemorrhagic and non hemorrhagic diarrhoea, respectively. Faecal samples were tested by immunochromatography assay test and 34% of cats were found positive to FPV antigen. The infection was more prevalent in cats less than 6 months [37%] compared with animals older than 6 months of age [31%]. No significant differences were observed between different clinical signs, age and sex of the animals [P>0.05]. The affected cats had no history of vaccination against Tri-cat, but in the healthy cat population, 18% were vaccinated. The difference between the two groups was significant [P<0.05]

serological survey on Brucella canis in companion dogs in Ahvaz

Canine brucellosis is a zoonotic infectious disease caused by Brucella canis. This bacterium can be transmitted to humans as well as other dogs. It is a significant cause of reproductive failure, predominantly in kennels. The aim of this study was to determine the seroprevalence of Brucella canis infection in companion dogs referred to the Veterinary Hospital of Shahid Chamran University of Ahvaz in the southwest of Iran. A total of 102 blood samples were obtained from dogs between 2006 and 2008. Sera were examined by Immunochromatography assay. The studied dogs were divided into two age groups [1-5 years and >5 years] and based on clinical signs [related signs to brucellosis such as scrotum dermatitis, diskospondylitis, lymphadenitis, abortion and infertility] into two groups also. Prevalence to Brucella canis antibodies in these dogs was 4.90% [5 of 102]. The infection had more prevalence in dogs above 5 years [9.3%; 4 of 43] in comparison with dogs less than 5 years [1.69%; 1 of 59], but the difference between the two age groups was not statistically significant [P>0.05]. There was no significant difference between the different sexes either [P>0.05]. Nevertheless, the difference for related signs to brucellosis was significant between the groups [P=0.018]. Three out of 14 cases [21.4%] which had clinical signs and two out of 88 cases [2.3%] which had no clinical signs were seropositive. This study showed that antibody against Brucella canis is present among the companion dog population of the Ahvaz area and preventive measures should be taken to control pathogenic bacteria

first report of concurrent detection of canine parvovirus and coronavirus in diarrhoeic dogs of Iran

Two male dogs at the age of 3 and 5 months of Doberman pinscher and German shepherd breeds were referred to the Veterinary Hospital of Ahvaz University with clinical signs of depression, vomiting, haemorrhagic diarrhoea, profound dehydration, fever and anorexia. The affected dogs had not the history of vaccination. Concurrent infection of canine parvovirus and coronavirus was detected in dogs by means of immunochromatography assay. The haemogram showed lowered white blood cell counts as leukopenia, neutropenia and lymphopenia. The dogs received supportive treatment to correct the life-threatening dehydration and prevention of secondary bacterial infections. Despite treatment, one dog [Doberman pinscher] died within 3 days after the onset of clinical signs, and the second dog recovered after 4 days. To the best of our knowledge, this is the first report of concurrent infection of canine parvovirus and coronavirus in diarrhoeic dogs in Iran

[Serological study of Neospora caninum infection in cattle from Ahvaz area, Iran]

In order to investigate the seroprevalence of Neospora caninum infection in cattle in Southwestern Iran, blood samples were collected from Holstein [121 animals] and cross-breed [436 animals] cattle from three farms and seven areas of Ahvaz, respectively. All of the Holstein cattle were >/= 4 years old but cross-breed cattle were from different age groups [< 2.2-4.5-6 and > 6 years old]. Sera were examined by commercial ELISA kit. Anti - N. caninum antibodies were detected in 117 [21%] sera out of 557 tested. A Significant difference was found between Holstein [53.71%] and cross-breed [11.93%] cattle although there were not any significant differences between age groups

[Seroepidemiology of canine leptospirosis in Ahvaz, Iran]

The presence of dogs has been identified as a risk factor for human Leptospirosis. The aim of this study was the survey of the current state of leptospiral infection in dogs in Ahvaz. Sera from 100 rural dogs of villages around Ahvaz and 49 urban dogs referred to veterinary hospital of shahid Chamran University were detected for antibodies against serovars canicola, icterohaemorrhagiae, grippotyphosa,ballum,hardjo andpomona by microscopic agglutination test. Out of the 149 dogs investigated, 8[5.4%] were serologically positive against at least one serovar of leptospira. Titers >/= 1:100 were detected in 7 rural [7%] and one urban [2.04%] dogs. The predominant titers were directed to serovars hardjo [44.5%] followed by ballum [22.2%], icterohaemorrhagiae [22.2%] andgrippotyphosa [11.1%]. Mixed infection was detected only in one dog. Serovars canicola and pomona were uncommon in dogs from Ahvaz. As expected, a very low prevalence of infection was found in urban pet dogs but in contrast low prevalence of infection in rural shepherd and/or stray dogs with poor level hygiene was unexpectable. However, the sources of infection in these dogs were not clear and this is the first report of infection with serovar hardjo from dogs in Iran

[Antigenic detection of Canine Coronavirus in diarrheic dogs in Ahvaz]

The Purpose of this study was to evaluate the prevalence of Canine Coronavirus [CCV] in diarrheic dogs of Ahvaz district. Fecal samples were collected from 58 diarrheic dogs between 2005 and 2006. Dogs were grouped by age [less than and above 6 months], sex, breed, geographical location [urban and rural] and quality of diarrhea [hemorrhagic and non-hemorrhagic] to determine whether these factors were associated with coronavirus infection, using Fischer's exact test. Prevalence to CCV antigens in these dogs was 3.45% [2 of 58] by means of immunochromatography assay [IC] indicating that this virus is present in the environment. Though the infection had more prevalence in dogs less than 6 months, rural dogs and German shepherds, but there were no significant differences between different sexes, age groups, breeds, location and quality of diarrhea [p>0.05]. CBC in 2 dogs that were affected with coronavirus infection was normal. This study that is the first report on prevalence of CCV in Iran showed that CCV infection in the Ahvaz dogs may be attributed as a cause of viral diarrhea in dogs. A vaccine is recommended, especially for open population and kennel dogs that are at a high risk for contacting the virus

Seropervalence of turkeys chlamydiosis in Khuzestan province

In order to investigate seroprevalence of turkey's chlamydiosis in Khuzestan province, 270 turkeys, of different sex and ages from different parts of Khuzestan province were bleed and their sera were tested for antibodies to Chlamydophila psittaci using an enzyme-linked immunosorbent assay [Immunocomb-ILS]. According to results the seroprevalences of chlamydiosis were 58.9% in tested turkeys. Chi square test showed that the prevalence were significantly lower in young, under 6 months [25%] than adult's turkeys [66%] and in cold [68.9%] than warm [49.3%] months [p < 0.005]. In Khuzestan, highest [85.7%] and lowest [31.42%] prevalences were in Shoshtar and Haftgel respectively. It is inferred that C. psittaci is widespread in turkey's populations in Khuzestan and further attempts for definitive diagnosis by isolation of its etiologic agents are recommended

Comparison of PCR and bacterial culture methods for diagnosis of dairy cattle's subclinical mastitis caused by staphylococcus aureus

To compare PCR and bacterial culture methods for diagnosis of subclinical mastitis caused by Staphylococcus aureus, 100 milk samples from cattle with subclinical mastitis and 20 samples from healthy cattle were collected and tested. The samples were cultured on selective blood agar and bacteria were identified by standard methods. DNA extracted from samples was subjected to PCR reaction with species specific primers and PCR products were analyzed by agarose gel electrophoresis. Based on the PCR results the prevalence of subclinical mastitis due to S. aureus was 25%. In the bacteriological culture of single milk sampling, S. aureus was isolated from the same samples being positive in PCR. A correlation of 100% was found between PCR and single milk sampling culture method by Mc Nemar test. All of the CMT negative samples were also negative in culture and PCR methods. The results of this study indicate that the PCR reaction is sensitive and specific for diagnosis of S. aureus in subclinical mastitis and can detect this pathogen in milk samples at species level in few hours

prevalence of mycobacterium paratuberlosis infection in ileocecal valve of cattle slaughtered in ahvaz abattoir, Southern Iran

To investigate the prevalence of Mycobacterium paratuberculosis [M avium subsp paratuberculosis infection among cattle slaughtered in Ahvaz abattoir, samples were taken from 250 cattle. Before slaughter. sex and age were assessed. Thereafter, samples were taken from ileocecal valve, rectum mucosa, ileocecal lymph node and stool. All smears which taken from these samples were stained by Ziehl-Neelsen method and examined by light microscopy. At first, ileocecal samples were examined; other samples were examined when ileocecal sample was positive. Out of the 250 cattle, 5 [2%] were positive. With the exception of one sample in which the organism was observed in both ileocecal valve and ileocecal lymph nodes in other positive samples the bacteria was observed in only ileocecal valve smear

Comparison of Dot-ELISA with microbial culture for detection of Brucella spp. in clinical specimens

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

Serodiagnosis of sarcocystosis in water buffalo [Bubalus bubalis] by ELISA test and its comparison with digestion method

To compare the ELISA test with digestion method for dignosis sarcocystosis in water buffalo and estimate its sensitivity and specificity. Cross sectional study.Three hundred slaughtered water buffaloes in Ahwaz abattoir.Blood and oesophageal muscles were examined. Oesophageal muscles were examined for sarcocystis by both macroscopic and microscopic [digestion method] examination.Then the ELISA test were designed and compared with digestion method.Finally its sensitivity and specificity were determined. Sensitivity, specificity and 95% confidence intervals were determined by comparing the results obtained by the ELISA assay and digestion method. Me nemar test were used for comparing the percentage of positive cases and their correlation. While specific anti-sarcocytis antibodies were detected in 54.3% of cases macroscpic and microscopic infection were 20% and 57%, respectively. Furthermore, while the positive results of the macroscopic examination significantly differed from those values of the digestion and ELISA methods [P<0.01], no differences were observed between the positive results of the microscopic infection [digestion method] and the ELISA test [P>0.05]. Conclusion: ELISA test providing an effective and reliable means for detecting sarcocystosis in naturally infected water buffaloes. Its simplicity and ease of performance makes it particulary suitable for using in large-scale epidemiological surveys of livestock

Diagnosis of experimentally infected sheep hydatidosis through antigen detection in serum and urine, with coagglutination [Co.A] test

To evaluate coagglutination test in the serum and urine of sheep for diagnosing of hydatidosis. Experimental study. Two rabbits, three dogs and nineteen sheep. Ovine hydatid cysts from affected livers and lungs, were collected from Ahwaz abattoir [Khozestan province, Iran]. The hydatid fluid [HF] and protoscolces were aseptically obtained in lab. Hydatid fluid was centrifuged and injected to rabbits in two steps. After then, rabbit hyperimmune sera were collected. Furthermore, each dog was given 15,000 viable protoscoleces. Less than two months later, dogs were autopsied after euthanasia and all Echinococcus granulosus worms were collected and their eggs were released. Almost, 2000 eggs were orally administred to each [N= 13]. The six other sheep were kept as control. All sheep were bled each week and their urine samples were collected fortnight. All sera and urine samples were examined with coagglutination [Co.A] test. While sensitivity of coagglutination test, was nil during five weeks of post-infection [p.i.], its values showed a biphasic pattern. While, it increased up to 23% in the sixth week and after then up to 100% in the 12th and 13th week of p.i. it decreased in the following weeks. Specificity of test was 100% throughout the experiment. While examination of urine in the affected sheep resulted in positive reaction from 6th week of p.i, its sensitivity and the sensitivity gradually increased up to 100% at 12th week of p.i. Furthermore, specificity of the test for urine of non-infected sheep remained 100%. These results suggest that the time of appearance of hydatid antigens in serum and urine is approximately alike. While positive results are very valuable, negative ones do not rule out hydatidosis