Iranian Journal of Veterinary Research
Volume:9 Issue: 1, Winter 2008

This study was designed to examine the effects of retinoic acid adding to cumulus and/or fibroblast cells monolayer on the development of mouse early embryos. One-cell mouse embryos were obtained from NMRI mice after superovulation by an intraperitoneal injection of 5 IU equine chorionic gonadotrophin (eCG) followed 48 hrs later by 5 IU human chorionic gonadotrophin (hCG). Mouse embryonic fibroblasts (MEF) were obtained from mouse fetuses and cumulus cells (CC) were prepared from mouse cumulus-oocyte complexes (COCs). To produce monolayer of cumulus and fibroblast cells, 1.0 × 105 cells/ml were plated into culture dishes in 100 µl droplets. The collected mouse embryos were cultured randomly into six different conditions, being supplemented (experiment, Exp) or not (control, Con) with 0.28 µg/ml of retinol acetate methyl-β-cyclodextrin (RA) for 96 hrs at 37°C in 5% CO2 in air, including: (1) culture media only (Con 1); (2) culture media plus RA (Exp 1); (3) co-culture with CC (Con 2); (4) co-culture with CC plus RA (Exp 2); (5) co-culture with MEF (Con 3) and (6) co-culture with MEF plus RA (Exp 3). The culture medium was Alpha Modification of Minimum Essential Medium Eagle (α-MEM) + 10% fetal bovine serum (FBS) with 100 IU/ml penicillin and 100 µg/ml streptomycin. The proportions of embryos passing the two-cell block were significantly higher in the MEF (Con 3) group compared to the other treatment groups (P<0.05). The percentage of the two-cell passed embryos developing to the blastocyst stage was significantly higher in the co-culture groups than that of the culture medium alone (P<0.05). After 96 hrs in culture, the rate of blastocyst stage for both groups of CC co-culture treatment (Con 2 and Exp 2) was identical but, adding RA into the MEF co-culture (Exp 3) resulted significantly lower in vitro development than that of the Con 3 group (29.2% vs. 57.7%, P<0.05). These results suggest that supplementation of co-culture groups with RA could not affect the embryos passing the block and developing to the blastocyst stage, although the presence of RA into the culture medium alone may improve passing the critical two-cell stage. Also, in vitro addition of RA to cells without receptors for retinol during long term co-culture may result early embryonic growth retardation.

Sergentomyia sintoni is the natural vector of Sauroleishmania species of lizards. This sandfly is abundance in and around the burrows of great gerbils. S. sintoni was collected from peridomestic animal shelters, inside and around houses and also from the nearby burrows of the gerbil reservoir hosts, Rhombomys opimus, in several provinces of Iran. Mitochondrial Cytochrome b (Cyt b) of sandflies, which is a maternally-inherited gene marker was used to characterize different haplotypes and populations of this sandfly. The analyses were based on the last 717 bp of the Cyt b gene followed by 20 bp of intergenic spacer and the transfer RNA ser (TCN) gene, i.e. the 737 bp fragment (without primers) amplified with the primers CB1-SE and CB-R06. The ITS-rDNA gene was also used to find Leishmania infections in S. sintoni. Cyt b 5´ fragment sequences were obtained from 22 S. sintoni, Cyt b 3´ fragment sequences were also obtained from 22 and Cyt b Long fragment sequences were obtained from 19 S. sintoni. By using nested PCR of ITS-rDNA gene, at least two species of L. major and L. gerbilli s.l. were found in S. sintoni. It needs further studies for considering the vectorial role of S. sintoni in ZCL foci, it’s important role in maintaining L. major infections in the great gerbil and transmitting Leishmania species to people and among the reservoir hosts.

Bovine brucellosis is a zoonotic disease distributed worldwide and characterized by abortion and reduced fertility in cows. Since brucellosis eradication programme in Iran uses vaccination, test, slaughter and quarantine as control measures, it is essential to distinguish vaccine strains from strains that cause infections among vaccinated cattle herds. We developed and evaluated a multiplex polymerase chain reaction (PCR) assay to identify and differentiate the vaccine strains from wild field isolates, including all Brucella types usually found in cattle in Iran. Two pair primers were used to amplify eri and wbo regions of DNA sequences those strain-specific targets for B. abortus S19 and RB51 vaccine strains. This multiplex PCR method evaluated with DNA from reference strains, two vaccine strains and 29 field strains of Brucella. The results showed that the multiplex PCR can differentiate Brucella isolates into three categories: strain 19 (S19), strain RB51 and field strains. This PCR assay was successfully used, compared with traditional method to differentiate of S19 and RB51 from field Brucella isolates.

The objective of this study was to compare different antigen preparations to produce monoclonal antibodies against bovine leukaemia virus gp51SU. The four antigen preparations for immunization of BALB/c mice were: CL: BLV-FLK cell lysate, UF: a fraction of CL (between 30 and 100 kDa), WVP: whole virus particles and SP: with ion exchange chromatography, gp51SU was semipurified. A total of nine successful fusions were performed which resulted in production of 23 monoclonal antibodies (mAbs) specific against gp51SU. The highest ratio of specific hybridoma colonies in each fusion was with SP preparation. Based on the reactivity of the mAbs in Western blotting, mAbs were classified into four groups: anti-gp51SU (23 mAbs), anti-gp30TM (8 mAbs), anti-Pr72 (5 mAbs) and antibodies against other viral proteins (7 mAbs). Some of the anti-gp51SU mAbs reacted with more than one band in Western blotting, suggesting that these colonies recognized not only gp51SU but also its precursors.

Cystic hydatid disease (hydatidosis) is one of the most important zoonosis that is caused by the larval stage of Echinococcus granulosus. As its diagnosis by clinical symptoms alone is difficult and confusing, serologic diagnostic techniques are used to confirm the disease. These techniques can also be used for epidemiologic studies. The present study was performed with a commercial human enzyme-linked immunosorbent assay (ELISA) kit for the diagnosis of hydatidosis in sera collected from sheep with hydatidosis. Sera were collected from 68 cases of hydatidosis proven by inspection of hydatid-infested livers and lungs of the sheep slaughtered in Mashhad abattoir and also from 11 healthy cases. Sera samples were examined by ELISA kit. The results showed that out of 68 cases of hydatidosis in sheep, 67 samples had positive absorbance. Also from 11 healthy samples, 9 had negative absorbance value. The sensitivity and specificity of the test were 99 and 82%, respectively. Therefore, it can be concluded that it is possible to use human ELISA kit for the diagnosis of hydatidosis in sheep.

The morphology of retinal photoreceptor layer was studied in continuous light-exposed and dark-adapted domestic male cats (Felis catus). The eyes of 12 healthy adult cats (4 in continuous light-exposed group, 4 in continuous dark-adapted group, and 4 in control group) were routinely fixed and studied by electron microscope. Results showed that the general structure of photoreceptor layer in this animal was the same as other species; rods were elongated and slender cells. Cone photoreceptors were shorter and stouter than rod photoreceptors. Cats exposed to continuous light for 24 hrs showed increased numbers of melanosome in retinal epithelial layer. The outer segments of rods and cones were long. Vacuoles increased the extracellular space and some condensed nuclei were observed in the inner segments. Continuous dark-adapted group showed a few melanosomes. In this group, the extracellular space, large swelling and condensed nuclei were more than those in other groups.

Citrullus colocynthis (CCT) is used in traditional medicine to inhibit the implantation of embryos. The objective of this study was to determine the number of embryos per pregnancy and the mortality rate in pregnant mice. 115 vaginal plug-positive mice were divided into 4 groups. The animals were given 30, 60 and 120 mg/kg hydroalcoholic extract of CCT until 17th day of gestation. Control group was fed with solvent. At the day 17, the animals were sacrificed and the number of pregnant mice and embryos per pregnancy were counted. We found that while in 30 mg/kg group the mean number of embryos per pregnancy was around 10, no embryo was found in other groups. Furthermore, 3 out of 30 mice in 30 mg/kg group died, while in 60 mg/kg and 120 mg/kg groups the number of death was 7 and 14, respectively. In conclusion, CCT reduces the number of embryos and increases the mortality rate in pregnant mice in a dose-dependent manner.

Testicles were isolated from dromedary camels in a local slaughterhouse at breeding and non-breeding seasons. Sperms were recovered from different parts of the epididymis (caput, corpus and cauda) and stained separately on slide glasses by eosin nigrosin staining method and dried by a hair dryer and carried to the laboratory. In the lab, slides were observed for evaluation of the proportion of live sperms and the proportion of sperms with cytoplasmic droplets under a light microscope. The proportions of live sperm cells were 76.8, 86.9 and 88.8% for caput, corpus and cauda epididymis, respectively. In the left testicle these values were 85.3, 83.1 and 88.4 for caput, corpus and cauda epididymis, respectively. No significant difference was also observed in the live sperm cells obtained from right and left testicles. The proportions of live sperm cells were 83, 90 and 86% in breeding and 80, 82 and 90.5% in non-breeding seasons for caput, corpus and cauda epididymis, respectively, which were not significantly different. The proportions of live sperms with protoplasmic droplets were 66, 70 and 74% in breeding and 73, 70 and 82% in non-breeding seasons for caput, corpus and cauda epididymis, respectively, which were not significantly different. The proportions of live sperms with protoplasmic droplets were significantly different neither among right and left testicles nor in different parts of epididymis. We concluded that sperm cells could be obtained from every part of the epididymis.

The effect of ephedrine at 0 (E0C0, n = 10), 8 (E8C0, n = 10) and 10 (E10C0, n = 10) mg per kg metabolic body weight or mixture of ephedrine/caffeine, at doses of 8 mg ephedrine/80 mg caffeine (E8C80, n = 10), or 10 mg ephedrine/100 mg caffeine (E10C100, n = 7), per kg metabolic body weight on body composition of feedlot Mehraban ram lambs (8-month-old) was studied. The lambs were fed for 95 days with a fattening ration ad lib., and ephedrine and ephedrine/caffeine, dissolved in distilled water, were drenched daily. The control sheep (E0C0) were drenched with distilled water only. Ephedrine/caffeine mixture caused a significant decrease in weight and daily gain. Dressing percentage was not affected by treatment, but carcass depreciation (shrinkage) was significantly reduced in E10C100 treatment (1.3% vs. 2.0 in control lambs). The mixture significantly increased the crude protein (in dry matter) but decreased dry matter and fat contents of the carcass meat. Internal fat (absolute values and as a percentage of slaughter weight) was significantly higher in the control sheep as compared with other groups. Serum glucose concentration was significantly lower in the control than in other groups. Serum cholesterol levels increased in groups receiving the ephedrine (E8C0 and E10C0) compared with the control, but caffeine returned their values to the control levels. Total serum protein level increased slightly but significantly in E8C0 and E10C100 groups, and serum total lipid and triacylglycerol levels did not change significantly. The results showed that oral administration of ephedrine/caffeine altered the body composition of Mehraban fat-tailed rams, and that feeding of 10 mg ephedrine and 100 mg caffeine per kg metabolic body weight (E10C100) was most effective in changing the body composition.

Studies on the pathogenesis of Babesia ovis infection following blood transfusion of infected blood to sheep with intact spleen and splenectomised sheep showed that all animals developed fever concurrent with a parasitaemia that were occurred within 2-4 days post-inoculation (dpi), clinical signs of disease were severe and included varying degrees of anorexia, listlessness, anaemia, moderate jaundice and haemoglobinuria. In intact animals, the hyperthermia returned to normal on the fourth day after the peak pyrexia and parasitaemia was eliminated within the course of the disease in four cases. However, other cases which had severe clinical signs of the disease were died. The parasitaemia reached a maximum of 7% in splenectomised sheep 7-8 dpi; in animals with intact spleen, the parasitaemia was much lower and reached to a maximum of 1%. In both of the infected groups, the red cell counts, haematocrit and haemoglobin concentration fell soon after the appearance of parasitaemia, reaching their lowest levels simultaneously with the peak parasitaemia. The total leukocyte counts were significantly decreased. The total serum bilirubin levels of the infected group rose above the normal and peaked on 14-16 dpi; the rise in AST, BUN and creatinine levels were slight. The kidneys and lungs were the organs most severely affected by experimental infection with B. ovis. Acute alveolar oedema and infiltration of neutrophils and macrophages in interstitial tissue were present, acute diffuse proliferative glomerulitis, congestion and stasis in glomerular capillaries and acute tubular necrosis were also present.

A study was undertaken in southern peninsular state of India, the Tamil Nadu State, to assess the farmers’ “Willingness to pay” (WTP) for receiving annual health care services to their dairy animals. The districts of the state were categorized as “Livestock developed” (LD) and “Livestock under developed” (LUD) based on initial base line developed. Contingent valuation (CV) approach was used to study the farmers’ maximum WTP value for two types of health care services: (a) providing health care services at government veterinary centres (in-centre) and (b) extending health care services at farmers’ doorsteps (farm gate). A payment card (PC) format was used to assess the farmers’ maximum WTP for receiving health care services to cows and buffaloes. The Maximum Likelihood Interval technique was used on interval midpoints. Overall mean WTP value for annual health care services in cows was INR 202.34 for in-centre services, while it was INR 261.66 for home services. Similarly, overall mean WTP value for annual health care services in buffaloes was INR 135.78 for in-centre services and INR 186.20 for farm gate services. The mean stated WTP values for both in-centre and at home services in the LD districts were highest as compared to LUD districts, leaving a scope for increased cost recovery. These WTP estimates exhibited the scope of cost recovery measure that can be implemented in lieu of free services extended currently, besides presenting a clue for designing a “vet-claim” policy in line with “medi-claim” policy for humans.

Escherichia coli O157:H7 is an important human pathogen causing haemorrhagic colitis, haemolytic-uraemic syndrom and thrombotic thrombocytopenic purpura. In this study, 100 ground beef samples were collected randomly from beef markets in June 2004. For isolation of the bacteria, samples were firstly enriched in modified trypticase soy broth, followed by plating onto sorbitol MacConkey agar supplemented with cefixime and potassium tellurite. Consequently, the suspected non-sorbitol fermenting (NSF) colonies were confirmed by biochemical tests and employed for polymerase chain reaction (PCR) assay, using primers specific for O157 and H7 antigens gene. In this study, 7 NSF E. coli colonies were isolated; in PCR assay only one of them confirmed as E. coli O157:H7. The PCR assay employed in this study may be a possible alternative to immunological assays which detects somatic and flagellar antigens.

Human fascioliasis due to unknown species and animal fascioliasis caused by both or one of Fasciola spp. are commonly seen in Iran. To compare electrophoretic patterns of somatic and excretory-secretory antigens of F. hepatica and F. gigantica by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), the adult flukes were collected from infected slaughtered bovine livers. E/S and somatic antigens were prepared by incubation and homogenizing of adult flukes, respectively. The antigens were electrophoresed using SDS-PAGE. Following SDS-PAGE, E/S proteins of F. hepatica and F. gigantica were characterized by the presence of 6 common major peptide bands with molecular weights of 15, 16, 20, 24, 33 and 42 kDa. Differences between F. hepatica and F. gigantica somatic proteins were noticed. F. gigantica had 11 major protein bands with molecular weights of 18, 22, 24, 33, 36, 42, 46, 57, 60, 62 and 68 kDa, whereas F. hepatica had proteins characterized by 8 distinct bands with molecular weights of 18, 22, 24, 33, 36, 42, 46 and 62 kDa.

A 4-year-old female native goat with the history of inappetence and no defecation was referred to the Department of Clinical Sciences, School of Veterinary Medicine of Shiraz, Shiraz, southern Iran. During exploratory laparotomy, pyloric obstruction and displacement of the abomasum to the left side was observed. Obstruction of pylorus was due to a ball-shaped phytobezoar. The goat was followed and had a good condition one month after the operation. It was concluded that the displacement of the abomasum to the left side has been occurred subsequent to the abomasal obstruction.

Members of the genus Gongylonema are nematodes that commonly infect ruminants, particularly sheep and goats. An 11-year-old donkey mare was referred to the Veterinary Teaching Hospital, School of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, northeastern Iran, with a history of acute colitis. At post-mortem examination, there were several white to pink zigzag tracks in the mucosa of the oesophagus and the stomach which contained long white worms. Microscopic examination of the parasite revealed that it was Gongylonema pulchrum.

A 4-year-old female miniature pincher with a 40-day history of weight loss, lethargy and vomiting was referred to Small Animal Hospital, Faculty of Veterinary Medicine, University of Tehran. The case also had severe jaundice, chronic diarrhoea and steatorrhoea. The efforts for saving the life of the case were not successful and finally the animal died of hypovolaemic shock and electrolyte imbalance. At necropsy, the body was cachectic, oedematous and severely icteric. A haemorrhagic ascitic fluid, without fibrin and clot was also noticed in the abdominal cavity. Firm tumour masses originated from pancreas were seeded to peritoneum with multiple attachments to duodenum. There were also metastasis to regional lymph nodes and liver. Based on histopathologic characteristics of the tumour, the mass was diagnosed as relatively well-differentiated exocrine pancreatic adenocarcinoma.

This study was conducted to determine the use of three-dimensional ultrasonography (3DU) in ocular imaging of the dog and measuring of its optical axis. 12 healthy mixed-breed dogs including 6 males and 6 females were studied. 3DU of the eyes were done using a 5–12 MHz linear trapezoid transducer. 3DU of the eyes were evaluated and the normal optical long axis through a line between the cornea and the optic disc in three-dimensional images were measured. In 3D images, vitreous body, anterior chamber, and lens cortex and its nucleus had a distinct anechogenic to hypoechogenic pattern. Details of the eyes compartments were better observed by rotating the images in all possible angles and planes using 3D facilities. Anterior and posterior lens capsule and the optic disk were hyperechogenic. The mean ± SD optical axis was 20.7 ±0.9 mm in males and 26.3 ± 0.6 mm in females (P<0.05). No significant difference existed between the measurements made in left and right eyes. We found marked advantages in image acquisition for interpretation of all aspects of the ocular structures.