Comparison of three immunodiagnostic tests for experimental Heterophyes heterophyes infection in dogs

The aim of this study was to compare the performance of three in-house diagnostic tests, i.e. counter current immunoelectrophoresis (CCIE), intradermal (ID) and indirect fluorescent immunoassay (IFI), for the diagnosis of Heterophyes infection. One hundred and twenty puppies were randomly divided into eight groups (n=15/group). Heterophyes heterophyes infections were established in these puppies by administering a dose of 50 H. heterophyes encysted metacercariae/puppy by gavage. Forty puppies of similar age and sex were divided into eight groups, of five puppies each and were used as negative controls. Sera pooled from separate infected and uninfected groups were tested against H. heterophyes antigens, weekly for 8 weeks post-infection (PI). The ID assay detected infected puppies sooner than any of the serological tests. Sero-conversion was first detected 2 weeks PI by ID assay and 1 week later by CCIE and IFI assays. ID test performed well till the end of the experiment (sensitivity and specificity: 100% and 90%, respectively). Both IFI and CCIE assays had a sensitivity of 40% and 20%, respectively for early detection of antibody, which was less sensitive than ID but both assays were more specific (100%) than the ID assay. The lowest agreement was between ID and IFI tests (40.3%), whilst the highest was observed between CCIE and IFI tests (67.2%). Of the three evaluated methods, the ID test could be recommended for scientific and laboratory diagnosis of heterophyosis in naturally infected animals. However, since none of the investigated method are optimal (i.e, offers 100% specificity and sensitivity), the choice of test employed must depend on the aim of the study.     

Comparison of intramuscular and footpad subcutaneous immunization with DNA vaccine encoding HSV-gD2 in mice

Herpes simplex virus type 2 is the most common infectious agent in humans that causes genital herpes disease and vaccination is a desirable method to prevent herpes infections. An effective therapeutic vaccine will need to elicit virus-specific immune responses. The route of immunization has important role in immune responses. In this study, DNA vaccine encoding glycoprotein D of herpes simplex virus type 2 (HSV-gD2) was prepared and injected via intramuscular and footpad routes to determine the optimal method of delivery for immune stimulation. The control manipulation of immune response by concerning route of administration is highly appreciated issue by researches. Although DNA vaccine containing HSV-gD2 is effective in both intramuscular and footpad injection routes, the latter could induce significantly higher cellular responses against HSV-2.     

A comparative study on behavioral,physiological, and adrenal changes in buffaloes during the first stage of labor with normal and difficult parturition

The present study was primarily carried out to trace the behavioral, physiological, and adrenal changes during the first stage of labor in cases of dystocia, either in parturient buffalo heifers or cows, and their clinical significance. Of the 118 investigated buffalo herds, 40 cows (dystocia = 20 and eutocia = 20) and 40 heifers (dystocia = 20 and eutocia = 20) were used in this study. Cows approaching actual birth were transferred to a calving box with straw bedding. Behavior of the studied animals was recorded from the time that the animal was moved to the calving box until the emergence of the fetal limbs in the vulval lips. If no progress in parturition was observed within two hours after the rupture of fetal sacs (end of the first stage), these cases were considered as suffering dystocia. As the fetal limbs emerged from the birth canal, animals were examined clinically to determine their average pulse and respiratory rates as well as body temperature. Blood samples were collected to determine the cow/heifers cortisol level. Results of the present study indicated that dystocia was accompanied by disturbed physiological status and cortisol levels. Practical monitoring system of parturient animals, through observing behavioral, physiological, and adrenal aspects during calving, should be regarded to expect dystocia and provide help for the animal in the appropriate time.     

Bacillary hemoglobinuria in Japanese black cattle in Hiroshima, Japan: a case study

Three Japanese Black cows housed with 6 other cows exhibited main clinical symptoms of severe hemoglobinuria. Hematological analyses conducted after antibiotic therapy demonstrated severe anemia, and biochemical analyses indicated both severe hemolysis and disruption of hepatic function. One of the three cows died. Based on the above analyses and observation of typical clinical symptoms, a speculative diagnosis of bacillary hemoglobinuria was made, and immediate high-dose antibiotic treatment improved the general conditions of the surviving animals. Blood samples from the other 2 cows were collected sequentially after antibacterial therapy. Clostridium haemolyticum was detected by a nested polymerase chain reaction analysis of the blood samples. The cows were diagnosed with the second recorded occurrence of bacillary hemoglobinuria in Japan.     

Isolation and partial characterization of excretory/secretory antigens of Gastrothylax crumenifer

The present study was carried out to identify the excretory/secretory (E/S) antigens of the rumen infecting digenetic trematode Gastrothylax crumenifer that may be useful for the immunodiagnosis of rumen amphistomosis particularly during the pre-monsoon season during which this rumen parasite stops shedding eggs. The in vitro released E/S proteins were purified on a Sephadex G-200 column. The gel filtration profile revealed three distinct fractions F1-F3 where F1 and F3 appeared as sharp peaks while the F2 fraction was dispersed. The antibody titre against each of the purified E/S fractions was determined by ELISA using anti-whole E/S polyclonal antibodies raised in rabbit. Among the three fractions, the antibody titre against F1 was highest (1:12,800) whereas IgG titre was very low (1:50) for fraction F2 and F3 (1:100). Of the total polypeptides resolved on gradient SDS-PAGE, only a few antigenic polypeptides were detected in each fraction with hyperimmune anti-serum as revealed by Western Blot analysis. However, a 33 kDa antigen detected in each fraction appeared to be immunodominant which could be exploited for the diagnosis of the pouched amphistome.     

Evaluating the correlation between adrenal gland dimensions and aortic diameter in healthy dogs

In the present study, the relationship of normal adrenal dimensions with weight, aortic diameter, age and sex was evaluated. Moreover, a formula for estimating the normal dimensions of adrenal gland was provided. Thirty‐two intact adult dogs that were considered healthy based on history, physical examination, routine blood works and specific adrenal tests as well as absence of any abnormal ultrasonographic findings were included. Adrenal glands and abdominal aorta were imaged in the sagittal plane by ultrasound. Length, maximal height of the cranial and caudal poles, area and circumference of the left and right adrenal glands and also the internal diameter of the abdominal aorta were measured. Analysis showed that there is a positive correlation between the adrenal gland dimensions and weight and aortic diameter. The ratio of adrenal gland dimensions and the aortic diameter was calculated, but this ratio showed a great amount of variability in tandem with a significant correlation to the weight; Meaning that the ratio of the adrenal gland dimensions to the aortic diameter was not a reliable index for evaluating the size of the normal adrenal gland in animals with different weights. Therefore, we used the weight along with adrenal measurements in a linear regression model, and then, we were able to estimate the adrenal gland dimensions in different weights. Knowing the relationship of normal adrenal dimensions with the indices such as weight, age, sex and diameter of abdominal aorta can help the clinicians to diagnose the adrenal gland diseases.     

Glutathione content of in vivo and in vitro matured canine oocytes collected from different reproductive stages

Glutathione (GSH) concentrations of oocytes are considered as an important marker of the cytoplasmic maturation. The present study was designed to compare GSH concentrations of in vivo and in vitro matured canine oocytes. In vivo matured oocytes were collected 72 hr after ovulation by flushing fallopian tubes after laparotomy. Ovaries were collected from bitches with different reproductive stages, and collected oocytes were divided into 2 groups according to the size viz. < 120 microm and > 120 microm in diameter and cultured for 72 hr in Tissue Culture Medium-199 supplemented with 10% FBS, 2.2 mg/ml sodium bicarbonate, 2.0 microg/ml estrogen, 0.5 microg/ml FSH, 0.03 IU/ml hCG, and 1% penicillin-streptomycin solution in the presence or absence of 50 microM beta-mercaptoethanol. GSH concentrations were determined by the dithionitrobenzoic acid-glutathione disulfide (DTNB-GSSG) reductase recycling assay. GSH concentrations of immature canine oocytes were 2.9 and 3.8, 3.5 and 6.8, and 3.1 and 6.5 pM/oocyte for < 120 microm and > 120 microm in diameter oocyte groups at anestrous, follicular and luteal stage, respectively (P<0.05). In vivo matured oocytes had significantly higher GSH concentrations compared with in vitro matured oocytes. The GSH content was 19.2 pM/oocyte for in vivo matured oocytes, while 4.1 to 8.1 and 5.7 to 13.2 pM/oocyte for in vitro matured oocytes cultured in the absence or presence of beta-mercaptoethanol, respectively (P<0.05). Presence of beta-mercaptoethanol increased GSH synthesis in canine oocytes cultured in vitro, and oocytes collected from follicular and luteal stage was superior to anestrus oocytes.     

Molecular typing of Iranian field isolates Mycoplasma synoviae and their differentiation from the live commercial vaccine strain MS-H using vlhA gene

1. The single-copy domain of the N-terminal region of the vlhA gene of Mycoplasma synoviae was sequenced, analysed and verified and used to type Iranian field isolates of M. synoviae and the MS-H live vaccine strain. In addition, a restriction fragment length polymorphism (RFLP) method was developed to differentiate between field isolates of Iranian and MS-H vaccine strains.

2. All sequences were analysed and aligned; the percentage similarity of the DNA was calculated and dendrograms were constructed. Based on single nucleotide polymorphism (SNP) that existed in all field isolates in Iran, the PCR-RFLP method allowed the differentiation of all M. synoviae field isolates from the vaccine strain.

3. Using phylogenetic analysis, the isolates were assigned to 8 unique genotypes and, within each group, DNA had a high level of similarity.

4. DNA sequence analysis and PCR-RFLP of the amplicon based on percent similarity and evolutionary relationship appeared to be useful tools for strain differentiation whether M. synoviae clinical isolates from infected chickens were derived from the vaccine strain or wild-type strains.

5. This study confirms the potential value of strain typing for epidemiological purposes and suggests that phylogenetic studies are essential to understand the true relationships between strains.     

Livestock feed resources availability,feeding practices,and nutrient balances in high Himalaya (Gurez) valley of Kashmir

Tropical Animal Health and Production - Assessment of the ruminant livestock feed sources availability, existing feeding practices, and nutrient balances was embarked at different zones of Gurez...