Assessment of different diagnostic methods for the identification of subclinical endometritis in dairy cows with pathological puerperium and their reliability to conceive

The aim of study was to evaluate different methods for subclinical endometritis (SCE) diagnoses and their accuracy to predict pregnancy. The study was performed on 51 postpartum cross breed dairy cows with pathological puerperium. Artificial insemination (AI) was done in spontaneous estrus. Endometrium samples were taken by cytotape (CT), cervical mucus (CM) samples by Metricheck and smears for crystallization degree were obtained at AI day. Blood serum samples for progesterone (P4) concentration determination were taken at AI day and 17 days later. Cows were examined by ultrasound 35 days after AI to confirm pregnancy. Percent (%) of polymorphnuclear leukocytes (PMNs) taken by CT were expressed by ROC curve (with cut-off point set at 1%). Out of the total of 51 cows involved in the study, 13.73% true positive and 41.17% true negative cows were found (54.90%). The content of CM sampled with Metricheck was altered in 11 cows (21.57%), and one of them was pregnant (1.96%). An average macroscopic CM value for pregnant cows was 0.06±0.24 vs. 0.39±0.79 for non-pregnant (p<0.05). An average value of CM crystallization in pregnant cows was 2.50±1.04 vs. 1.85±1.15 in non-pregnant (p<0.05). P4 concentration was not statistically different at AI day, but was statistically significantly higher 17 days later in pregnant cows (p = 0.002). CT alterations may be a reliable diagnostic tool for SCE diagnosis, while macroscopic CM alterations may be used for CE detection. P4 determination is beneficial in pregnancy diagnosis while CM crystallization degree is helpful in determination of best time for AI.

Effect of Retained Placenta And Clinical Mastitis On Reproduction Parameters, Immune Response And Steroidogenic Receptors Gene Expression In Postpartum Crossbred Crossbreed Dairy Cows

The purpose of this study was to evaluate the effect of retained placenta (RP) and clinical mastitis (CM) on the reproductive efficiency of crossbred dairy cows during the postpartum period and to investigate the effect of them in some innate immune system indicators. For this, in the first experiment a total of 232 crossbred dairy cows were evaluated and divided into three groups as: healthy control, RP and CM. The incidence of RP and CM was evaluated after parturition until 30 days postpartum (DPP) and reproductive rates were measured. In experiment 2, 30 crossbred dairy cows were divided into three groups as in Experiment 1. Between 40 and 50 DPP, clinical, gynecological examination and endometrial cytobrush were performed to evaluate subclinical endometritis and gene expression of interleukin- 1β (IL-1β), interleukin-6 (IL-6), chemokine ligand -5 (CCL5), estrogen α (ESR1) and progesterone (PGR) receptors by qRT-PCR analysis. In Experiment 1, the conception rate at 1st AI was lower in RP and CM groups and pregnancy rate at 150 days decreased in cows with CM. Calving-to-1st AI interval and days open were shorter in healthy cows. In Experiment 2 the occurrence of subclinical endometritis was 26.7% and was higher in cows with RP and CM. The expression of IL-1β was increased in RP and CM groups, while IL-6 was less expressed in RP group. The chemokine CCL5, ESR1 and PGR were similar between groups. In conclusion, cows with RP and CM had their reproductive efficiency negatively affected and had they initial pro-inflammatory response improved by the increase of IL-β, while, IL-6 was lower in animals with RP probably because it was evaluated later in the postpartum.

Addition of Clinoptilolite In The Diet Reduces Uterine PMN Leukocytes And Open Days In Multiparous Lactating Dairy Cows Managed In A Mountain Tropical Pasture-Based System

This study was aimed to assess the effect of adding clinoptilolite in the diet on uterine health and reproductive performance in multiparous lactating dairy cows managed in a tropical pasture-based system above 2500 meters of altitude. Seventy-seven multiparous Holstein crossbred cows were allocated randomly into two groups: clinoptilolite supplemented cows (CLG, n = 42) and non-supplemented cows as control (CG, n = 35). Cows from CLG were supplemented with clinoptilolite from 30 days (50 g/cow/day) before to 60 days after calving (200 g/cow/day). Data were analyzed by general linear model, least squares means and chi-square test of SAS. In CLG cows, percentages of uterine PMN leukocytes (P<0.0001) and proportion of subclinical endometritis (P=0.0187) were lower than in CG. The interval calving to first corpus luteum was shorter (P=0.0759) in CLG than CG, and calving to first service interval was similar between treatments. Cows from CLG became pregnant 35 days earlier than CG cows (P=0.0224). In conclusion, daily addition of clinoptilolite in the diet decreased the proportion of cows with subclinical endometritis and shortened the interval from calving to conception in lactating dairy cows.

Clinical morphological reasoning of the diagnosis of subclinical cows endometritis

The project presents the results of the clinical and morphological substantiation of the diagnosis of endometritis in cows. The scientific and production part of the experiment was carried out by the method of selection of analog groups, taking into account the reproductive age and live weight, according to generally accepted methods. Two groups of animals were formed. Group one - cows with subclinical endometritis, group two - with no signs of this pathology, which is morphological control. Of these, seven animals were healthy and thirteenth with signs of endometritis. Of these, pathology in an acute form was detected in three animals, in a latent form in ten animals. The formation of a group of animals with a normal course of the postpartum period was carried out on the basis of techniques, including examination of the external genitalia and rectal examinations. General clinical indicators were also taken into account: temperature, pulse and respiration. The studies were carried out on the basis of the Department of Anatomy and Histology of Animals named after Professor A.F. Klimov FGBOU VO MGAVMiB - MBA named after K.I. Scriabin, as well as in ZAO Povadino breeding plant. Hematological studies were carried out on the basis of the Chance Bio laboratory. A comprehensive methodological approach was used in the work, including clinical examination of animals, macroscopic morphometry, anatomical preparation, morphological blood tests, statistical analysis of the obtained digital data. The normative morphological parameters of the uterus in black-white Holsteinized cows were established as the baseline for assessing the nature of structural changes in the organ with subclinical endometritis. Revealed pathognomonic morphological signs of the uterus in subclinical endometritis, expressed in an increase in its cavity and horns, the accumulation of exudate in them, in the swelling and dullness of the endometrium. In the course of the conducted hematological studies, the manifestation of an inflammatory reaction was noted, expressed in an increase in the number of leukocytes, stab and segmented neutrophils and monocytes. The article presents the clinical and morphological substantiation of the diagnosis of subclinical endometritis, based on a comprehensive assessment of changes in the morphometric parameters of the uterus and hematological changes occurring in the body.

Determination of thiol/disulphide homeostasis as a new indicator of oxidative stress in dairy cows with subclinical endometritis

The objective of this study was to determine thiol/disulphide homeostasis (TDH) in infertile cows with subclinical endometritis (SCE). Endometrial cytological samples were collected using a cytobrush to diagnose SCE in 36 infertile cows. According to the results of the cytology examination, those with acute endometritis were classified as Group I (n = 20) and those with chronic endometritis were classified as Group II (n = 16). A control group was formed of heifers as Group III (n = 20). Blood samples were taken from each group on the day of diagnosis (day 0) to analyse TDH. In the cytology examination, both the Giemsa method and immunocytochemical staining were applied to determine chronic inflammation and activity status. In 55.55% (20/36) of the infertile cows with cytological endometritis, the inflammation was determined to be active, and in 44.44% (16/36) it had become chronic. The native thiol and total thiol levels were found to be statistically significantly lower in the acute (206.54 ± 8.30 μmol/L; 227.11 ± 9.30 μmol/L) and chronic SCE cases (225.15 ± 11.89 μmol/L; 247.96 ± 10.80 μmol/L) compared to the heathy control group (308.47 ± 13.59 μmol/L; 336.83 ± 15.5 μmol/L respectively) (P<0.001). Disulphide levels, disulphide/total thiol, native thiol/total thiol and disulphide/native thiol ratios were similar in all the groups (P>0.05). The diagnostic accuracy of native thiol, which can be used in the diagnosis of SCE, was 92.8%, that of total thiol was 89.3% and that of disulphide was 64.3% according to the ROC curve analysis. These results demonstrate that TDH is a reliable and sensitive indicator of oxidative stress in cow SCE, and that abnormal TDH might play a role in SCE pathogenic mechanisms. This is the first study to evaluate thiol/disulphide homeostasis in dairy cows with SCE as a new indicator of oxidative stress.

Messenger RNA Expression of Selected Factors at Different Sites of the Bovine Endometrium Associated With Uterine Health

Recent studies have elucidated the role of several pro-inflammatory factors as mediators of inflammatory processes in the bovine endometrium. Only few studies, however, have analyzed samples collected from different regions of the uterus of the same animal. In this study, we tested the hypothesis that on a molecular level, clinical endometritis is characterized by inflammatory responses spread over the entire endometrium. Furthermore, we assume that subclinical endometritis is described by an inflammation of local regions of the uterus. Therefore, the objective of this study was to assess the mRNA expression of uterus-associated pro-inflammatory factors at five pre-defined endometrial sites, i.e., corpus uteri, left horn base, left horn tip, right horn base, and right horn tip, in cows with clinical and subclinical endometritis and in healthy controls. We analyzed the mRNA expression of interleukin 1 alpha, interleukin 1 beta, C-X-C motif chemokine ligand 8, prostaglandin-endoperoxide synthase 2, protein tyrosine phosphatase receptor type C, carcinoembryonic antigen related cell adhesion molecule 1, and mucin 4 and 16. Based on vaginoscopy and endometrial cytology (≥ 5% polymorphonuclear neutrophils) between 28 to 34 days in milk, 18 Simmental cows were categorized in clinical endometritis group (n = 7), subclinical endometritis group (n = 4), and healthy group (n = 7). In general, the analyses revealed a great variation of mRNA expression between sites and animals. Differences were found between different uterine health statuses, but the variation between the sampling sites within the groups was not significant (P &gt; 0.05). This indicates that inflammatory processes at the end of the postpartum period can be regarded as multi-focal or spread throughout the uterus independent from the uterine health status.

Effect of Cefazolin on endometrial cytology and reagent test strips parameters in bovine endometritis

Endometritis is one of uterine disorders in dairy cows causing low fertility. This study explored effects of intrauterine (IU) infusion of Cefazolin on endometrial cytology and reagent test strips parameters (leukocyte esterase [LE] and pH) in bovine subclinical endometritis. A total of 90 cows at 49-56 days in milk (DIM) diagnosed for subclinical endometritis were divided randomlyinto three groups, group I (n = 35), treated with IU infusion of 2 grams Cefazolin (Zinol ®, Pharco) diluted with 50 ml saline; group II (n = 28), treated with IU infusion of 2 gramsCephalexin (Ceporex®, GlaxoSmithKline) diluted with 50 ml saline; group III (n = 27), cows kept as untreated control. Cytological examination and reagent test strips were performed in uterine material derived with the cotton swab technique before treatment program and repeated two times later, in 10 days interval. The cows were artificially inseminated at first oestrous after the end of treatment program and conception rates were evaluated. After the end of treatment program, polymorphonuclear cells (PMN) decreased significantly (P <0.05) in Cefazolin and Cephalexin groups (3.23% and 4.39% respectively) compared tocontrol (24.89%) group, also means of LE in these groups became significantly (P<0.05) lower. The Ph value decreased after treatment, this reduction was significant after the first dose of Cefazolin, while in Cephalexin reduction of pH became significant after second dose. Cefazolin had conception rate (77.14%) significantly (P<0.05) higher than cephalexin and control groups (57.14% and 25.93% respectively). Cefazolin and Cephalexin decrease uterine PMN, LE concentration. The pH value after treatment by Cefazolin was lower than in Cephalexin. Moreover, treatment with Cefazolin resulted in a significantly higher conception rate. Thus, Cefazolin is recommended for the treatment of subclinical endometritis in dairy cows.