Povezanost između polimorfizma gena CXCR1 i rizika od endometritisa u krava holštajnske pasmine

Endometritis is one of the reproductive diseases that can cause disturbances of postpartum uterine health in cattle. Therefore, identification of resistant genotypes to endometritis is essential. The objective of this study was to evaluate the association between the single nucleotide polymorphism in the interleukin-8 receptor-α (CXCR1) gene and the possibility of endometritis in Holstein dairy cows. For this purpose, blood samples were collected from multiparous dairy cows with a history of clinical endometritis (n = 30), and normal, healthy cows as the control group (n = 10). Deoxyribonucleic acid (DNA) was isolated from a blood sample. To determine genotype, the polymerase chain reaction-restriction fragment length polymorphism technique (PCR-RFLP) was used. The results indicated the presence of a different proportion of polymorphisms (G > C) in the CXCR1 gene in cows with clinical endometritis, compared to the control group. Statistical analyses showed that there is a significant correlation between the incidence of the disease and the CXCR1 genotype in nucleotide position 956. The incidence rate of clinical endometritis was associated with the CXCR1.956 genotype; cows with genotype GC had a higher incidence of clinical endometritis compared with cows with the GG genotype. Overall, the results showed that CXCR1 polymorphism could be a useful marker for identification of resistant genotypes to endometritis in Holstein dairy cows.Endometritis je reproduktivna bolest koja može poremetiti poslijeporođajno zdravlje maternice u goveda, stoga je važna identifikacija genotipova otpornih na tu bolest. Cilj ovoga istraživanja bio je procijeniti povezanost između polimorfizma pojedinačnog nukleotida na genu interleukin-8 receptoru-α (CXCR1) i vjerojatnosti pojave endometritisa u holštajnskih mliječnih krava. U tu su svrhu prikupljeni uzorci krvi mliječnih krava multipara s kliničkim endometritisom u anamnezi (n = 30) i zdravih krava kao kontrolne skupine (n = 10). Iz uzoraka krvi izolirana je DNA. Genotip je određivan metodom lančane reakcije polimeraze – polimorfizmom dužine restrikcijskih fragmenata (PCR-RFLP). Rezultati upućuju na prisutnost različitih omjera polimorfizama (G > C) gena CXCR1 u krava s kliničkim endometritisom u usporedbi s kontrolnom skupinom. Statistička analiza pokazala je znakovitu korelaciju između incidencije bolesti i genotipa CXCR1 u poziciji nukleotida 956. Incidencija kliničkog endometritisa bila je povezana s genotipom CXCR1.956; krave s genotipom GC imale su veću incidenciju kliničkog endometritisa u usporedbi s kravama s genotipom GG. Općenito, rezultati pokazuju da bi polimorfizam CXCR1 mogao biti koristan biljeg za identifikaciju genotipova otpornih na endometritis u holštajnskih mliječnih krava

Evaluation of Possible Effects of Hyoscine in Xylazine-Induced Fetal Death in Pregnant Rats

Although xylazine is widely used in domestic animals as a sedative, analgesic, and muscle relaxant, its side effects on the uterus prevent its utilization in pregnant animals or in embryo transfer. Although the effects of xylazine on increasing uterine contractions have been confirmed, no reliable report of fetal death due to xylazine administration has been published. Hyoscine is an anticholinergic medication that has antimuscarinic and antispasmodic effects in the uterine tissue of pregnant cattle during in vitro studies, therefore, we investigated if administration of xylazine in the last third of pregnancy could increase fetal death and if hyoscine could prevent its adverse effects. Twenty adult female rats, after mating with four adult male rats and confirming pregnancy, were randomly divided into two equal control and treatment groups. On the 18th day of pregnancy, the number of fetuses per rat was determined using ultrasonography. Rats in the treatment group received hyoscine (1 mg/kg, intraperitoneally) for 3 days. Subsequently, all rats were administered xylazine (10 mg/kg, intraperitoneally) for 3 days. On the 21st day of pregnancy, the number of living and dead fetuses was counted after laparotomy. Also, the weight and dimensions of the fetuses were measured. The results showed that although more fetuses lost their lives in the treatment group compared to the control group, the statistical difference in the percentage of fetal mortality in the two groups was not significant (p 0.05). In addition, the comparison of the mean weight, body length, and body width of living and dead fetuses in both groups showed that there was no statistically significant difference between these groups (p 0.05). It could be concluded that maternal xylazine intake in rats could cause about 18-25% of fetal mortality. However, the use of hyoscine to prevent fetal death induced by xylazine is not recommended

Vaginal Fornix Discharge Cellularity and Its Leukocyte Esterase Activity for Diagnosis of Endometritis in Dairy Cows

The objective of the present study was to evaluate the application of some strip test markers (i.e., leukocyte esterase (LE) activity, protein, nitrate and pH) for diagnosis of endometritis in dairy cows using vaginal fornix discharge. Also, the total white blood cell count (t-WBC/l) of this secretion and degenerative changes of neutrophils in cervical cytology were used as alternative methods to predict progression of the endometritis severity. Holstein cows (n=215) between 30-40 days in milk (DIM) were included and examined. Giemsa-stained smear was prepared from cervical mucus. Cervical cytology test was considered as reference screening method for the detection of subclinical endometritis. The LE activity and t-WBC in the vaginal fornix discharge of subclinical endometritis cows were significantly higher than those from healthy cows. Sensitivity and specificity were 78% and 73% for LE10 activity (10 minutes after contacting with discharges) and 60% and 69% for t-WBC (cut off point=210 cells/l) for diagnosis of subclinical endometritis, respectively. There was a good agreement between LE10 activity, t-WBC and cervical cytology test with a Kappa coefficient of 0.4 and 0.42, respectively (P<0.0001). Total WBC count in discharge and degenerative neutrophils (DN) percentages increase simultaneously with the degree and severity of endometritis. There was a highly significant (P<0.01) correlation between t-WBC and some reagent strip test markers (LE activity, protein and nitrate) in clear discharge of studied cows. In conclusion, the present results suggest the LE activity and t-WBC in vaginal fornix discharge could be used as non-invasive reliable and valid methods for screening of subclinical endometritis in postpartum dairy herds

Characterization of single nucleotide polymorphism in the 5'-untranslated region (5'-UTR) of Lactoferrin gene and its association with reproductive parameters and uterine infection in dairy cattle

Uterine infection is one of the reproductive diseases that can have disturbing postpartumuterine health in cattle. Therefore, identification of resistant genotypes to uterine infection isimportant. Lactoferrin (LF) is one of the major antimicrobial compounds in the normal uterinedischarges of cows. We hypothesized that allelic diversity in LF gene may contribute tosusceptibility or resistance to uterine infection. We investigated the single nucleotidepolymorphism genes identified in the 5' untranslated region (5'-UTR, position +32) of the LF geneusing Allele-specific PCR method in cows with and without uterine infection. Blood samples werecollected from 89 multiparous Holstein dairy cows with a history of uterine infection (n = 51), andcows without disease as the control group (n= 38). The results indicated the presence of differentproportion of polymorphisms (G > C) in the 5'-UTR area of cows in the all groups. The results ofAllele specific PCR was in complete agreement with sequencing method. Statistical analysis did notshow any statistically significant correlation between disease and SNP in 5'-UTR. While, there was asignificant difference in the mean of reproductive parameters of cows without polymorphismcompare to those of with SNP in 5'-UTR. Cows with +32:CC genotype and +32:GC genotype (cowswith SNP in UTR) had lower average of services per conception and days open compared to cowswith the +32:GG genotypes. However, no significant difference in the calving to first service wasfound between these genotypes. Further studies will be required to determine critical SNPs in LFgene and status of the risk of uterine infection and embryo survival in cows