Comparison between intrauterine application of an antibiotic and an herbal product to treat clinical endometritis in dairy cattle - A randomized multicentre field study.

Uterine diseases are main indications for antibiotic use in dairy cows. To test a non-antibiotic treatment option, we compared the effect of an intrauterine cephapirin (Metricure®; cefapirin benzathin 500 mg per dosis; CEPH) with an intrauterine applied herbal product (25 ml of EucaComp® PlantaVet containing alcoholic extracts of Calendula officinalis L., Mellissa officinalis L., Origanum majorana L. and Eucalyptus globulus Labill. (EUC)) on the clinical cure of endometritis. Examinations of 816 cows between 21 and 35 days after calving were performed and cases of clinical endometritis (n = 169) were included. Diagnosis based on a scoring system for vaginal discharge. Study animals were randomly assigned to one of two treatment groups and treated immediately. After excluding animals with incomplete datasets, 136 cows (EUC: n = 61; CEPH: n = 75) remained for the final analysis. In total, 64% (EUC: 61%, CEPH: 67%) of analysed endometritis cases were considered as clinically cured 14 ± 2 days after the first treatment, 15% stayed uncured after the application of a maximum of two consecutive treatments, leading to an overall clinical cure rate of 85% (EUC: 82%, CEPH: 88%). No statistically difference in clinical cure rates could be observed between both treatment groups nor 14 ± 2 days after the first treatment (p = 0.956) neither regarding the overall cure rate (p = 0.923). In conclusion, the clinical cure of dairy cows' endometritis after the intrauterine application of the herbal product was non-inferior to the intrauterine application of the antibiotic cephapirin. These results could contribute to reduce the antimicrobial use in the daily veterinary routine treatment of endometritis.

Two Years after Coxiella burnetii Detection: Pathogen Shedding and Phase-Specific Antibody Response in Three Dairy Goat Herds.

The infection dynamics of Coxiella (C.) burnetii were investigated in three dairy goat herds (A, B, and C) 2 years after the first pathogen detection. A total of 28 and 29 goats from herds A and B, and 35 goats from herd C, were examined. Sera were analyzed on three sampling dates using phase-specific serology. Pathogen shedding was assessed using post-partum vaginal swabs and monthly bulk tank milk (BTM) samples. Dust samples from a barn and milking parlor were also collected monthly. These samples were analyzed with PCR (target IS1111). In herd A, individual animals tested seropositive, while vaginal swabs, BTM, and most dust samples tested negative. Herds B and C exhibited high IgG phase I activity, indicating a past infection. In herd B, approximately two-thirds of the goats shed C. burnetii with vaginal mucus, and irregular positive results were obtained from BTM. Herd C had two positive goats based on vaginal swabs, and BTM tested positive once. Dust samples from herds B and C contained C. burnetii DNA, with higher quantities typically found in samples from the milking parlor. This study highlights the different infection dynamics in three unvaccinated dairy goat herds and the potential use of dust samples as a supportive tool to detect C. burnetii at the herd level.

Immunization against Gonadotropin-Releasing Hormone in Female Beef Calves to Avoid Pregnancy at Time of Slaughter.

Precocious puberty in beef heifers can result in unwanted pregnancies due to accidental breeding by farm bulls. Inbreeding, premature calving followed by dystocia and a high stillbirth rate or slaughtering of pregnant heifers are the consequences of this behaviour. The aim of the study was to postpone puberty by using Improvac®, an anti-GnRH vaccine. Therefore, n = 25 calves were twice vaccinated, once at the age of 5 and then at 6.5 months. n = 24 calves served as unvaccinated case controls. The onset of puberty was assigned if progesterone analysis in the blood exceeded 1 ng/mL. Progesterone values were excluded if the corresponding serum cortisol levels were ≥60 nmol/L. Our target was met, as in the vaccinated group none of the calves exceeded a progesterone value >1 ng/mL until the scheduled age of slaughter at 11 months and only 12.5% of the animals exceeded a progesterone value of 1 ng/mL over the whole measuring period (>400 days) compared with 56.5% of the calves in the control group. In conclusion, the favourable results from our study using the vaccine Improvac® represent an animal-friendly, non-invasive and reliable way to avoid early pregnancy in heifers as well as the slaughter of pregnant cattle.

Causes of Abortions in South American Camelids in Switzerland-Cases and Questionnaire.

Over the last decade, South American camelids (SAC) have gained increasing popularity in Switzerland. They are used for several purposes such as fiber and meat production, as companion or guard animals and for trekking activities. The purpose of this study was to investigate the frequency and reasons for pregnancy loss and perinatal death in SAC herds. Within the scope of this study, early embryonic losses could not be identified, as pregnancy examinations by ultrasonography are not performed routinely. Aborted and stillborn fetuses were collected, necropsied and analyzed for infectious abortifacients. A nationwide survey among breeders was carried out. During a 1.5-year period, only eight cases of aborted or stillborn alpaca and llama (out of a population of 6550 animals) were reported by the breeders, and their causes were subsequently analyzed. In half of the cases, Coxiella burnetii was identified in the fetoplacental material. Abortions and stillbirths were reported to be rare in Swiss herds. As a conclusion, recording of embryonic losses through ultrasound training of veterinarians should be impaired and breeders motivated to have abortions and perinatal mortality examined. Special focus should be laid on C. burnetii due to its zoonotic risk.

Evaluation of an investigative model in dairy herds with high calf perinatal mortality rates in Switzerland.

The objective of this study was to evaluate an investigative model which encompassed the risk factors, incidence, timing and causes of perinatal mortality (PM) (0-48 h) on high risk dairy farms (PM of >5% in the previous year) in Switzerland. This pilot-study was carried out on 47 predominantly Holstein PM calves from 21 dairy farms, between September 2016 and January 2018. Gross pathological examinations of calves and placentae as well as histopathological examinations of internal organs and placental tissue were performed. Further investigations included microbiological examinations: broad-spectrum bacterial and fungal culture, detection of Chlamydia abortus, Coxiella burnetii, pathogenic Leptospira spp. and Neospora caninum by real-time PCR (qPCR) and of bovine viral diarrhoea virus (BVDV) by Ag-ELISA. Maternal blood samples were used for serology of bovine herpesvirus 1 (BHV-1), Brucella abortus, Chlamydia abortus, Coxiella burnetii and nine pathogenic leptospiral serovars and the evaluation of trace element status. A questionnaire was completed with the farmer, which included general farm characteristics and case-related data. Inbreeding coefficients (IC) were calculated for pure-bred matings. At the farm-level, the PM rate was 10.0% (5.3-28.2%) and at the cow-level, 11.5%. These values, from high-risk farms, were approximately five-times higher than the contemporary national bovine PM rate (2.3%) in Switzerland. The risk factors associated with these high PM rates were the self-selection of high risk herds, the high proportion of primiparae in these herds (45%) and the evidence of widespread pathogenic infections on these farms (exposure: 67% of herds, 53% of dams; infection: 57% of herds, 45% of calves). The majority (68.1%) of calves died intrapartum. The most commonly diagnosed initiating/ultimate cause of death (UCOD) was infection (34%) of which Coxiella burnetii was the most frequently detected pathogen, by antigen. The most frequently diagnosed proximate cause of death (PCOD) was asphyxia (44.7%), though multiple PCOD was also common (21.3%). This study was the first detailed investigation of bovine PM in Switzerland. Infectious causes were diagnosed more frequently than expected. While the findings from these high PM Swiss herds may have limited external validity, the investigative model adopted and the detailed research methodologies employed can be replicated and re-evaluated, respectively, in future studies on PM internationally.

Therapy of retained fetal membranes in cattle: Comparison of two treatment protocols.

This prospective study was conducted to compare two treatment protocols for retained foetal membranes (RFM) in cattle. The routine therapy consisting of intrauterine tetracycline boluses was compared with the administration of ozone foam, both intrauterine therapies without attempts to manually remove the placenta. Cows undergoing caesarean section, suffering from uterine torsion or prolapse of the uterus, and other illnesses than RFM on the first day post-partum were excluded. Values for baseline variables as age, breed, yearly milk yield, parity, pregnancy duration, calving season and number of routine intrauterine treatments were not different between the groups. Number of days with rectal temperature >39.7 °C within the first 10 days in milk (DIM) were greater in the ozone group. Number of escape therapies were recorded when there was imposing of this therapy as a result of cows having a fever (based on rectal temperature >39.7 °C) and a significant reduced food intake. Escape therapies consisted of parenteral administration of tetracyclines and NSAIDS during the first 10 DIM. Number of escape therapies, however, was not different between the groups. Results from use of a logistic regression model with fever as a dichotomous variable indicated there was no effect of breed, studbook, type of stall, pasture, business and no effect of age or milk yield. In a second assessment, cows were evaluated until 200 DIM. Results from a survival analysis indicated group (Te/Oz) did not affect rate of culling from the herd.

Prediction of calving time in dairy cattle.

This prospective study was carried out to predict the calving time in primiparous (n=11) and multiparous (n=22) Holstein-Friesian cows using the combination of data obtained from the RumiWatch noseband-sensor and 3D-accelerometer. The animals included in the study were fitted with the RumiWatch noseband-sensor and 3D-accelerometer at least 10days before the expected calving day. The calving event was defined as the time of the first appearance of the calves' feet outside the vulva, and this moment was determined by farm staff and/or confirmed by video monitor. As primiparous and multiparous cows behaved differently, two models including data of noseband-sensors and 3D-accelerometers were used to predict the calving time in each group. Lying bouts (LB) increased and rumination chews (RC) decreased similarly in both groups; besides that, boluses (B) decreased and other activities (OA) increased significantly in multiparous and primiparous cows, respectively. The sensitivity (Se) and specificity (Sp) for prediction of the onset of calving within the next 3h were determined with the logistic regression and ROC analysis (Se=88.9%, 85% and Sp=93.3%, 74% for multiparous and primiparous cows, respectively). This pilot study revealed that the RumiWatch system is a useful tool to predict calving time under farm conditions.

Immunization against GnRF in adult cattle: a prospective field study.

BACKGROUND: Suppression of cyclic activity in cattle is often desired in alpine farming and for feedlot cattle, not intended for breeding. A cattle specific anti-GnRF vaccine (Bopriva™) is registered for use in heifers and bulls in different countries. In adult cows vaccinated with Bopriva™, the median period until recurrence of class III follicles was 78 days from the day of the 2nd vaccination and reversibility could be proven, as out of 11 experimental cows 10 cows became pregnant at first, and one cow at second insemination. In the present study, 76 healthy, cyclic Eringer heifers and cows were vaccinated twice with Bopriva™ 3-7 weeks apart, to prevent estrus during alpine pasturing. Blood samples were taken for progesterone and GnRF antibody titer analysis on the day of inclusion (7-9 d before the first vaccination) and at the first vaccination. At the same time, gynaecological examinations were performed. When estrus occurred in the course of the alpine pasturing season, a gynaecological examination was done including analysis of a blood sample (progesterone, anti-GnRF antibody titer). Cows were followed for fertility out to 26 months post second vaccination. RESULTS: Median duration of estrus suppression was 191 days after the second vaccination (when the 2 vaccinations were given 28-35 days apart). From n = 13 cows showing signs of estrus on the alpine pasture, n = 7 could not be confirmed in estrus (serum progesterone value >2 ng/ml, no class III follicles seen using ultrasonography). Median duration between second vaccination and next calving was 496 days (25%/75% quartiles: 478/532 days). CONCLUSION: Bopriva™ induced a reliable and reversible suppression of estrus for more than 3 months in over 90% of the cows.

Simplicimonas-like DNA in vaginal swabs of cows and heifers cross-reacting in the real-time PCR for T. foetus.

Cows on an alpine pasture were presented with severe signs of vaginitis. To rule out infection with Tritrichomonas foetus, vaginal swabs were taken and real-time PCR based on detection via fluorescence resonance energy transfer (FRET) probes and targeting the first internal transcribed spacer (ITS-1) of nuclear ribosomal DNA (rDNA) was performed. PCR was positive in 25 of totally 34 assessed cows. However, the melting profiles of the probes targeting the diagnostic PCR products differed from the T. foetus positive control. Subsequent sequencing of the amplicons revealed 91% identity to Simplicimonas sp. sequences deposited in GenBank™. Furthermore, there was no clear association between positive PCR result and presence of vaginitis. To investigate the distribution of this Simplicimonas-like organism in cows, more herds grazing on the same alpine pastures as well as unrelated cows were tested. In total, 133 cows and 16 heifers were sampled, 53 cows and 6 heifers even twice. Vaginitis was evident in 43 cows and 4 heifers. All-over-positivity of PCR was 44%, including nine tests performed on heifers. Melting peak analysis indicated Simplicimonas-like organisms in all positive samples. Culture attempts in bovine InPouch ™ TF failed. No association between a positive PCR result and the presence of vaginitis was found. This is, to the best of our knowledge, the first report on Simplicimonas-like DNA in vaginal swabs of female cattle. Our data suggest that when testing vaginal swabs of cattle by means of T. foetus PCR, false positive reactions due to Simplicimonas-like organisms may occur.

Comparison of the effect of a CIDR-Select Synch versus a long-term CIDR based AI protocol on reproductive performance in multiparous dairy cows in Swiss dairy farms.

BACKGROUND: Synchronization programs have become standard in the dairy industry in many countries. In Switzerland, these programs are not routinely used for groups of cows, but predominantly as a therapy for individual problem cows. The objective of this study was to compare the effect of a CIDR-Select Synch and a 12-d CIDR protocol on the pregnancy rate in healthy, multiparous dairy cows in Swiss dairy farms. METHODS: Cows (N = 508) were randomly assigned to CIDR-Select Synch (N = 262) or 12-d CIDR (N = 246) protocols. Cows in the CIDR-Select Synch group received a CIDR and 2.5 ml of buserelin i.m. on d 0. On d 7, the CIDR insert was removed and 5 ml of dinoprost was administered i.m.. Cows in the 12-d CIDR group received the CIDR on d 0 and it was removed on d 12 (the routine CIDR protocol in Swiss dairies). On d 0 a milk sample for progesterone analysis was taken. Cows were inseminated upon observed estrus. Pregnancy was determined at or more than 35 days after artificial insemination. As a first step, the two groups were compared as to indication for treatment, breed, stud book, stall, pasture, and farmer's business using chi square tests or Fisher's exact test. Furthermore, groups were compared as to age, DIM, number of AI's, number of cows per farm, and yearly milk yield per cow using nonparametric ANOVA. A multiple logistic model was used to relate the success of the protocols to all of the available factors; in particular treatment (CIDR-Select Synch/12-d CIDR), milk progesterone value, age, DIM, previous treatment of the uterus, previous gynecological treatment, and number of preceding inseminations. RESULTS: The pregnancy rate was higher in cows following the CIDR-Select Synch compared to the 12-d CIDR protocol (50.4% vs. 22.4%; P < 0.0001). CONCLUSION: The CIDR-Select Synch protocol may be highly recommended for multiparous dairy cows. The reduced time span of the progesterone insert decreased the number of days open, improved the pregnancy rate compared to the 12-d CIDR protocol and the cows did not to have to be handled more often.

Alopecia areata in Eringer cows.

Alopecia areata is a hair loss disorder in humans, dogs and horses with a suspected autoimmune aetiology targeting anagen hair follicles. Alopecia areata is only sporadically reported in cows. Recently, we observed several cases of suspected alopecia areata in Eringer cows. The aim of this study was to confirm the presumptive diagnosis of alopecia areata and to define the clinical phenotype and histopathological patterns, including characterization of the infiltrating inflammatory cells. Twenty Eringer cows with alopecia and 11 Eringer cows without skin problems were included in this study. Affected cows had either generalized or multifocal alopecia or hypotrichosis. The tail, forehead and distal extremities were usually spared. Punch biopsies were obtained from the centre and margin of alopecic lesions and normal haired skin. Histological examination revealed several alterations in anagen hair bulbs. These included peri- and intrabulbar lymphocytic infiltration, peribulbar fibrosis, degenerate matrix cells with clumped melanosomes and pigmentary incontinence. Mild lymphocytic infiltrative mural folliculitis was seen in the inferior segment and isthmus of the hair follicles. Hair shafts were often unpigmented and dysplastic. The large majority of infiltrating lymphocytes were CD3(+) T cells, whereas only occasional CD20(+) lymphocytes were present in the peribulbar infiltrate. Our findings confirm the diagnosis of T-cell-mediated alopecia areata in these cows. Alopecia areata appears to occur with increased frequency in the Eringer breed, but distinct predisposing factors could not be identified.

Spontaneous myometrial contractility in cows suffering from endometritis-Influence of localisation, smooth muscle layer and cycle phase. An in vitro study.

Contractility of the healthy bovine myometrium depends on the reproductive state. Furthermore, contractility is influenced by localisation and the direction of smooth muscle strips. However, little is known about the contractile behaviour of the uterus when affected by endometritis. In our study, myometrial specimens from the larger horn (near the corpus and near the tip) in cows suffering from endometritis in estrus (n=8) or diestrus (n=8) were collected after slaughter. Two strips were prepared from each region corresponding to the circular and the longitudinal muscle layers, respectively. The spontaneous contractility of these strips was recorded in an organ bath. To analyse the results, the 2.5h recordings were divided into five periods of 30 min each. The variables area under curve (AUC) and maximal (A(max)) and minimal amplitude (A(min)) were calculated separately for each period, and the results were analysed using a non-parametric model regarding the influence of cycle phase (estrus vs. diestrus), region (corpus vs. tip) and muscle layer (circular vs. longitudinal). The values of both AUC and A(max) increased significantly over time. Muscle layer had a significant effect on AUC (corpus, tip) and A(max) (tip): the values of circular layers were increased compared to longitudinal layers. Dividing the data into subgroups allowed us to analyse them additionally according to muscle layer: In longitudinal layers, A(max) was increased at the corpus as compared with the tip. In this model, the factor cycle phase did not produce any significant difference in spontaneous myometrial activity. However, data of all variables showed non-significant higher values in estrus than in diestrus samples.

In vitro effects of bethanechol on abomasal and duodenal smooth muscle preparations from dairy cows with left displacement of the abomasum and from healthy dairy cows.

This study investigated the effects of bethanechol (BeCh) on abomasal and duodenal smooth muscle preparations from dairy cows with left displacement of the abomasum (LDA) and from healthy dairy cows, and determined the role of muscarinic acetylcholine receptor subtypes 2 and 3 (M(2) and M(3)) in mediating contraction. Concentration-response curves for BeCh, with or without prior incubation with an M(2) antagonist (AF-DX 116) or an M(3) antagonist (4-DAMP), were established and evaluated. BeCh induced a significant, concentration-dependant increase in the contractility variables for all locations in both groups of cows. The inhibiting effect of 4-DAMP was stronger than that of AF-DX 116, which suggested that contractions were mediated by M(3) and to a lesser extent by M(2). The basal tone of abomasal smooth muscle was reduced in cows with LDA, which indicated hypotonia. The use of BeCh as a prokinetic drug in cows with gastrointestinal motility disorder warrants further investigation.

Effects of a single administration of prostaglandin F2alpha, or a combination of prostaglandin F2alpha and prostaglandin E2, or placebo on fertility variables in dairy cows 3-5 weeks post partum, a randomized, double-blind clinical trial.

BACKGROUND: Delayed uterine involution has negative effects on the fertility of cows; use of prostaglandin F2alpha alone as a single treatment has not been shown to consistently improve fertility. Combined administration of PGF2alpha and PGE2 increased uterine pressure in healthy cows. We hypothesized, that the combination of both prostaglandins would accelerate uterine involution and have, therefore, a positive effect on fertility variables. In commercial dairy farming, the benefit of a single post partum combined prostaglandin treatment should be demonstrated. METHODS: 383 cows from commercial dairy farms were included in this study. Uterine size and secretion were evaluated at treatment 21-35 days post partum and 14 days later. Cows were randomly allocated to one of three treatment groups: PGF2alpha and PGE2, PGF2alpha or placebo. For every animal participating in the study, the following reproduction variables were recorded: Interval from calving to first insemination, days open, number of artificial inseminations (AI) to conception; subsequent treatment of uterus, subsequent treatment of ovaries. Plasma progesterone level at time of treatment was used as a covariable. For continuous measurements, analysis of variance was performed. Fisher's exact test for categorical non-ordered data and exact Kruskal-Wallis test for ordered data were used; pairwise group comparisons with Bonferroni adjustment of significance level were performed. RESULTS: There was no significant difference among treatment groups in uterine size. Furthermore, there was no significant difference among treatments concerning days open, number of AI, and subsequent treatment of uterus and ovaries. Days from calving to first insemination tended to be shorter for cows with low progesterone level given PGF2alpha and PGE2 in combination than for the placebo-group (P = 0.024). CONCLUSION: The results of this study indicate that the administration of PGF2alpha or a combination of PGF2alpha and PGE2 21 to 35 days post partum had no beneficial effect upon measured fertility variables. The exception was a tendency for a shorter interval from calving to first insemination after administration of the combination of PGF2alpha and PGE2, as compared to the placebo group. Further research should be done in herds with reduced fertility and/or an increased incidence of postpartum vaginal discharge.

An in vitro study on spontaneous myometrial contractility in the mare during estrus and diestrus.

Uterine smooth muscle specimens were collected from euthanatized mares in estrus and diestrus. Longitudinal and circular specimens were mounted in organ baths and the signals transcribed to a Grass polygraph. After equilibration time and 2 g preload, their physiologic isometric contractility was recorded for a continuous 2.0 h. Area under the curve, frequency and time occupied by contractions were studied. Differences between cycle phases, between muscle layers, and over the recorded time periods were statistically evaluated using linear mixed-effect models. In the mare, physiologic contractility of the uterus decreased significantly over time for all variables evaluated (time as covariate on a continuous scale). For area under the curve, there was a significant effect of muscle layer (longitudinal > circular). For frequency, higher values were recorded in estrus for circular smooth muscle layer, whereas higher values were seen in longitudinal smooth muscle layers during diestrus. In longitudinal layer and in diestrus, more time was occupied by contractions than in circular layer, and in estrus. This study is describing physiologic myometrial motility in the organ bath depending on cycle phase.

Spontaneous in vitro contractile activity of specimens from the abomasal wall of healthy cows and comparison among dairy breeds.

OBJECTIVE: To characterize and compare in vitro contractility patterns of sections of abomasal wall harvested from cattle of 3 dairy breeds. SAMPLE POPULATION: Longitudinal and circular smooth muscle preparations harvested from the antrum and body of the abomasum of 30 recently slaughtered Holstein-Friesian, Brown Swiss, and Simmental X Red Holstein cows. PROCEDURE: Spontaneous isometric contractions of specimens in tissue baths of modified Krebs solution were recorded during a 4-hour period. Maximal amplitude, frequency of contractions, and change of basal tension were used to characterize contractility. Statistical analyses were used to test for differences among time periods, among breeds, between specimen locations, and between fiber orientations. RESULTS: Myoactivity patterns of abomasal smooth muscle preparations are highly variable and differ on the basis of location and fiber orientation. Frequency of contractions differed significantly among time periods for longitudinally oriented specimens with decreasing frequencies of contractions over time. Maximal amplitude of the longitudinally oriented specimens from the antrum increased significantly, whereas maximal amplitude of the circularly oriented specimens from the antrum decreased significantly. Values did not differ significantly among breeds. CONCLUSIONS AND CLINICAL RELEVANCE: Patterns of spontaneous contractility of abomasal wall specimens are not homogeneous. During a 4-hour recording period, maximal amplitude and frequency of contractions of specimens varied significantly with respect to orientation and location; however, spontaneous contractile myoactivity did not differ significantly among breeds. Therefore, breed predisposition for displaced abomasum is not correlated with spontaneous activity of smooth muscle specimens.

An in vitro study on spontaneous myometrial contractility in the cow during estrus and diestrus.

Myometrial smooth muscle strips were collected from slaughtered cows in estrus and diestrus. Longitudinal and circular smooth muscle strips were mounted in organ baths and after equilibration time and 2g preload, their physiologic contractility was recorded for 3h. Area under the curve (AUC), mean amplitude (MA) and frequency of contractions (F) were studied. Differences between cycle phases, between muscle layers and over the recorded time period were statistically evaluated. In the cow, physiologic contractility patterns (measured as AUC and MA) of circular versus longitudinal myometrial strips are always different during the 3h recording. Significant differences between estrus versus diestrus are only found for circular layers, but not for longitudinal layers. Significant differences over time are only found for longitudinal layers.