The Effects on Offspring of Diabetes Mellitus Induced by Chemical Agents in Rats with Advanced Pregnancy

ABSTRACT Background: Gestational diabetes mellitus is an increasingly frequent metabolic disorder that is important for both baby and mother. New studies on the development and treatment of the disease are required. Objective: To investigate the effects on offspring's survival and the biochemical values of diabetes mellitus, induced by different doses of two chemical agents among 35 rats with advanced pregnancy. Methods: The rats were randomly divided into five groups, with the rats in Group 1 as the control group. Alloxan was administered intraperitoneally at doses of 40 and 60 mg/kg in Groups 2 and 3, respectively. Streptozotocin was injected intraperitoneally at doses of 40 and 60 mg/kg in Groups 4 and 5, respectively. Deliveries were monitored, and offspring numbers, survival rates and congenital anomalies were recorded. At the end of the study, blood was drawn from one female offspring in each group; glucose, total protein, albumin, triglyceride, cholesterol, calcium and phosphorus levels were measured, and inter-group comparisons were made. Diabetic agents administered at various doses prolonged the duration of pregnancy. Results: Offspring's deaths were most frequent in the alloxan groups. The number of offspring mortalities in the streptozotocin group was higher than that of the control group, but lower than that of the alloxan group. No differences in glucose, total protein, albumin, triglyceride, cholesterol, calcium and phosphorus levels were observed between the groups. These results indicate that the female offspring, born from rats with gestational diabetes mellitus induced by different chemicals, were only clinically affected. No effect of the type of chemicals on the results was found. Conclusion: The use of streptozotocin in the studies on female offspring born from rats with gestational diabetes mellitus is recommended.

Influence of intra-mammary ozone administration on udder health in herds with contagious mastitis in the context of management practices.

This study investigates the effectiveness of intra-mammary ozone administration in the dry period and at the time of delivery for preventing against mastitis in herds with contagious mastitis. The cows were divided into five groups with 10 cows in each. Group 1 was administered an ozone-containing foam preparation via the teat canal into four udder quarters for 5 seconds at the beginning of the dry period; Group 2 was administered ozone at the beginning of the dry period and at the time of delivery; Group 3 was administered ozone at the time of delivery; Group 4 was administered a dry period udder preparation at the beginning of the dry period; and Group 5 was administered only teat seal at the beginning of the dry period. No statistically significant difference was found between the cows with regard to the SCC values at the beginning of the dry period and at the time of delivery (in cows without clinical mastitis, n=25). The SCC values were reported to decrease when the values at the beginning of the dry period and at the time of delivery were compared. All cows except two in Group 1 were detected to have clinical mastitis according to the frequency of microbial isolation in milk at the time of delivery. In conclusion, intra-mammary ozone administration did not prevent mastitis in the dry period or at the time of delivery in herds with contagious mastitis; moreover, it was determined to increase the rate of clinical mastitis in the postpartum period.

Effect of melatonin on torsion and reperfusion induced pathogenesis of rat uterus.

We investigated the use of melatonin to improve fertility and reduce uterine damage caused by torsion of the uterus in pregnant rats. We used 35 pregnant rats at gestational age 18 days. The animals were randomized into five groups. Group 1 was anesthetized only. Group 2 was subjected to experimental uterine torsion of 360° and the torsion was corrected after 6 h. Group 3 was subjected to uterine torsion of 360°, the torsion was corrected after 6 h and melatonin was administered at the time of correction. Group 4 rats were subjected to 360º uterine torsion and melatonin was administered 6 h later at the time of correction. Group 5 was administered melatonin followed by uterine torsion of 360 degrees followed by correction of torsion 6 h later. Samples were obtained from the uterine horns on the day 1 postpartum. We used Bax, Bcl-2 and caspase 3 staining to measure apoptosis in the uterine tissues. The mRNA levels of Rho-associated, coiled-coil containing protein kinases 1 (ROCK1), homeobox D10 (Hox4 HoxD10), TLR4, NFκB1, caveolin 1 (Cav1) heat shock protein 90 alpha (cytosolic), class B member 1 (Hsp90ab1) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were determined using quantitative real-time polymerase chain reaction analysis (qRT-PCR). Bax, Bcl-2 and caspase 3 were detected using immunohistochemistry. No difference was observed among groups with respect to abortion, neonatal mortality or congenital abnormalities. Compared to the control group, the mRNA levels of Rock1, Hox4, TLR4, NFκB1, Cav1 and Hsp90 genes were decreased significantly in the study groups; the decrease was greater in groups 3 and 4, which were treated with melatonin. The greatest amount of Bax staining was found in group 1 and the least amount of Bcl-2 staining was found in groups 4 and 5; the greatest amount of caspase 3 staining was found in group 2. Our findings indicate that melatonin reduced uterine torsion related tissue damage and that its application during torsion was more effective than application following removal of torsion.

Evaluation of an alternative treatment protocol by aglepristone to induce parturition in ewes with an experimental model of early pregnancy toxemia.

A new protocol with aglepristone to induce parturition in ewes with pregnancy toxemia has been reported in the present manuscript. Four experimental groups were defined: Group AG5 (n = 10), Group DEX (n = 10), Group NC (n = 5) and Group PT (n = 5) in which ewes were injected twice with 10 mg/kg of aglepristone and 5 ml dexamethasone in first two groups, respectively; whereas negative control and pregnancy toxemia groups received no treatment for parturition induction. Different clinical parameters associated with parturition in ewes and their newborns were investigated. Blood hematology and biochemical measurements were carried out both in ewes and lambs. Blood pH values of lambs were recorded during the study. The injection time-lambing time, injection time-vaginal discharge intervals, placental expulsion periods, placental weight and vaginal delivery interval between lambs, hematological and biochemical results were not statistically different among the groups (p > 0,05). Increased NEFA and ß-HBA concentrations accompanied the disease and all ewes in AG, DEX and PT Groups developed clinical pregnancy toxemia (NEFA; P = 0,009) and ß-HBA; (P = 0,039). The differences in rectal body temperature of lambs were not significant (p > 0,05), whereas birth weight was found statistically significant among groups (p < 0,05). Blood pH, biochemical and hematologic measurements of lambs had also significant differences depending on different time points. Parturition pathology by means of incomplete cervical dilatation was severely observed in DEX Group. The results of this study show that aglepristone application in pregnancy toxemia to induce parturition could precisely control lambing time without any side effects in either mothers or lambs. Apart from these, it could be speculated that dexamethasone seems to induce parturition causing crucial pathologies, which results in important and risky changes in newborns' life. Incomplete cervical dilatation and continued ineffective uterine contractions could be a major factor of newborn losses because of placental separation and cessation of blood supply.

Effect of gestational diabetes mellitus on testis and pancreatic tissues of male offspring.

The purpose of this study was to determine the effect of gestational diabetes mellitus (GDM) on some reproductive characteristics, testicular and pancreatic oxidative status and pancreatic endocrine receptor densities of male offspring at post-pubertal stage. A total of 36 1-day-old Wistar Albino male offspring including 12 pups of nontreated mothers (control group), 14 pups of 40 mg/kg STZ-injected mothers (STZ-40 group) and 10 pups of 60 mg/kg STZ-injected mothers (STZ-60 group) were used. The offspring were euthanised on post-natal day 60, their blood, reproductive organs and pancreatic tissues were obtained and examined. When compared with the control group, there was a significant decrease in body and absolute reproductive organ weights, serum testosterone level, testicular and pancreatic catalase activities, pancreatic glutathione level, epididymal sperm concentration of both STZ-40 and STZ-60 groups as well as in testicular glutathione level of only STZ-60 group. Significant increases were determined in testicular and pancreatic malondialdehyde level and glutathione peroxidase activity in both groups and in fasting serum glucose of only STZ-60 group in comparison with the control group. Although some histopathological damages were observed in testes of both STZ-40 and STZ-60 groups, there were no detectable differences between the groups in density of insulin, glucagon and somatostatin receptors in pancreas. In conclusion, GDM has negative effects on reproductive efficiency and testicular-pancreatic tissue oxidant/antioxidant balance of male offspring at post-pubertal stage.

Somatic SNPs of the BRCA2 gene at the fragments encoding RAD51 binding sites of canine mammary tumors.

Mammary tumors are the most common tumor type both in women and in female dogs. In women, heritable breast cancers have been linked mutations in the breast cancer susceptibility gene BRCA2 and it contains eight BRC repeats in exon 11 that bind to RAD51. In this study, we investigated the sequence variations of BRC1-BRC8 and C-terminus of canine BRCA2 gene. From a total of 64 canine patients with mammary tumors, 31 mammary tumors with benign and malign carcinomas and the 3 normal mammary glands were used for the study. In this study, 19 SNPs of exon 11 of BRCA2 in canine mammary tumors were detected for the first time. The c.2383A>C (T1425P) SNP was found to be the most probable disease-associated nsSNP. Our findings suggest that T1425P variation in BRC3 to be the most probable disease-associated nsSNP and may affect RAD51 binding strength.

Parturition induction in ewes by a progesterone receptor blocker, aglepristone, and subsequent neonatal survival: Preliminary results.

The clinical effects of aglepristone treatment to induce parturition in ewes and their newborns were reported. Three experimental groups were defined: group AG5 (n = 5), group AG10 (n = 5), and group CG (n = 5) in which ewes were injected twice with 5, 10 mg/kg of aglepristone, and saline solution of ewes, respectively. Different parameters associated with parturition in ewes and their newborns were investigated. Serum progesterone, oxytocin, and free and conjugated total estrogens were measured after treatments until parturition. No statistical difference was found from first aglepristone administration to onset of lambing between AG5 and AG10 (23.90 ± 6.20, 40.00 ± 6.71 hours). Parturition induction in two groups shortened the gestational length significantly compared with the control group (P = 0.003). Dystocia was observed in two ewes in group AG10. The placental weight showed statistically significant difference only between the AG10 and CG (P = 0.039), but no difference was observed in the placental expulsion period between the groups. Decrease in food consumption 24 to 36 hours after parturition in all ewes and skin necrosis in an ewe in group AG5 were observed. Progesterone concentration was significantly lower in AG5 than that in ewes in group AG10 and CG (P < 0.05). No difference was observed in concentrations of free total estrogens and oxytocin between groups. The body temperature of lambs was significantly different between AG10 and CG groups both right after (P = 0.011) and 12 hours after parturition (P = 0.014). The lambs in CG had the highest mean birth weight (4.29 ± 0.28 kg), which was significantly different from the induced groups. No significant difference of blood pH and blood gases values between groups was identified both at birth and 12 hours after parturition for lambs. Significant differences could clearly be observed in total protein and blood urea nitrogen and total protein findings 12 hours after parturition (P < 0.05), whereas no difference was found in blood glucose, albumin, inorganic phosphor, triglyceride, or total cholesterol parameters. The results of this study show that the administration of aglepristone to induce parturition can precisely control lambing time without any side effects in either mothers or lambs.

Relationship between California Mastitis Test score and ultrasonographic teat measurements in dairy cows.

OBJECTIVE: The majority of published studies about mastitis are related to the control and prevention of mastitis, with particular emphasis on eliminating predisposition factors. The objective of the current study was to determine the role of teat morphology as an important factor in the aetiology of mastitis. DESIGN: Ultrasonographic measurements were taken from 190 teats from 100 dairy cows of different breeds. PROCEDURE: Mastitis in cows was diagnosed by the California Mastitis Test (CMT) and microbiological tests. The data were evaluated in the light of the clinical history of the animals. RESULTS: Main effects of breed on teat diameter at the position of the Furstenberg rosette (FTD) and teat cistern diameter (CD), that of age on FTD and overall teat diameter (OTD), and that of CMT score on CD and OTD were significant (P < 0.05, P < 0.01). Number of lactations, pregnancy, the lactation period and the udder lobe (rear, front) were not found to have a significant effect on any of the measurements. Results showed that CD was significantly smaller (1.44 +/- 0.04 cm) (P < 0.01) in CMT-positive udder lobes than that in CMT-negative lobes. No difference was detected in canal length, CD, teat wall thickness, OTD or FTD between the CMT-positive and -negative lobes. CONCLUSION: The occurrence of mastitis could be related to specific ultrasonographic teat measurements (e.g. CD, OTD and FTD) and these may be important in the breeding of cows with a predisposition to mastitis, as well in the evaluation of in-herd cows in terms of udder/teat deformities.

Use of GnRH antagonists in ovarian remnant syndrome experimentally induced in rats.

PURPOSE: The objective of this study was to demonstrate the efficacy of cetrorelix, a GnRH antagonist, in rats with experimentally induced ovarian remnant syndrome. METHODS: 25 Wistar female rats at seven to eight weeks of age and weighing 200-250 g were used. The rats were randomly divided into five groups: the first group was used as a control group; the second and third groups underwent a sham operation; and the fourth and fifth groups underwent bilateral hemiovariectomy. At the first proestrus detected by vaginal cytology from postoperative day 2, the animals in groups 1, 2 and 5 received placebo and the animals in groups 3 and 4 received cetrorelix subcutaneously. In the study, the Kruskal-Wallis analysis of variance was used for comparison of the results of vaginal irrigation, histopathblogical examination, and of blood FSH and LH values, and the Mann Whitney U-test was used for determination of the differences between the groups. RESULTS: It was determined that according to vaginal cytology results, estrus-like cytological changes disappeared in a shorter time and according to histopathology results, the number of follicles were fewer in the ovarian remnant syndrome-induced and cetrorelix-injected group 4 (p < 0.05), but there was no difference between the groups for FSH and LH concentrations. CONCLUSIONS: Ovarian remnant syndrome is a complication of bilateral ovariohysterectomy. In cases with this syndrome, certain treatment is possible with re-operation. However, it may not always be possible to perform an operation, or even if operated, it is difficult to determine the place of the residual ovarian tissue. In this study, it was determined that the use of cetrorelix as a GnRH antagonist in rats with ovarian remnant syndrome reduced the duration of estrogenic affect.

Lack of correlation between the electrical conductivity of milk and the blood progesterone levels in cows.

A study was conducted to investigate the possibility of measuring the electrical conductivity (EC) of cow's milk as a practical and cost-effective means of determining oestrus in cows. Prostaglandin injections were used to synchronise the oestrous cycle of 15 cows. From the onset of oestrus, milk and blood samples were collected from each cow every 2nd day over a period of 22 days. A portable, commercially available instrument was used to measure the EC of the milk. The blood progesterone levels were determined by means of an enzyme immuno-assay method. No correlation was found between the EC of milk and the blood progesterone levels of the cows tested. Electrical conductivity measurements of milk are of no value in determining the onset of oestrus in cows.

Evaluation of relationship between repeat breeding and Fasciola hepatica and hydatid cyst infections in cows in Elazig district of eastern Turkey.

Sera samples from 89 dairy cows with repeat breeding and 111 healthy pregnant dairy cows (controls) from Elazig province of eastern Turkey were tested for presence of Fasciola hepatica and Hydatid cyst antibodies by use of an indirect enzyme linked immunosorbent assay (indirect-ELISA). One hundred and twenty one (60.5%) and 93 (46.5%) cows were found to be positive to F. hepatica and Hydatid cyst antibodies, respectively. Fifty two of 89 cows (58.4%) with a history of repeat breeding were seropositive to F. hepatica, and 43 of 89 (48.3%) were seropositive to Hydatid cyst antibodies. Whereas, seropositivity rates were 62.1% (69/11) for F. hepatica and 45% (50/111) for Hydatid cyst in healthy pregnant cows (control group). There were no statistically relationship between infected and control group (P>0.05). In conclusion, we could not detect any relationship between repeat breeding and F. hepatica and Hydatid cyst infections in cattle.