Apparent prevalence of and risk factors for infection with Ostertagia ostertagi, Fasciola hepatica and Dictyocaulus viviparus in Swiss dairy herds.

Infections with helminth parasites can negatively affect performance of dairy cows. Knowledge on infection intensity, spatial distributions and risk factors are key to develop targeted treatment strategies. Canada and most EU countries have conducted large investigations, but respective data for Switzerland were missing. We now performed a bulk tank milk serosurvey for Ostertagia ostertagi, Fasciola hepatica, and Dictyocaulus viviparus on a total of 1036 voluntarily participating dairy herds that were sampled at confinement periods, i.e. in winter 2014/15 or 2015/16, respectively. All samples were analyzed with commercial ELISAs for antibodies (AB) against O. ostertagi and F. hepatica, and those of the first sampling period additionally with an in-house ELISA for AB against D. viviparus. Testing for the latter parasite was not done in the second year of the study, as the sampling period might have missed infections due to the short lived nature of specific antibodies. The possible influence of geographic, climatic, and farm management variables on AB levels were assessed for each parasite using scanning cluster and multiple regression analysis. Overall seroprevalence for O. ostertagi was 95.5% (95% C.I.: 94.0-96.6), with a mean optical density ratio (ODR) of 0.83, for F. hepatica 41.3% (95% C.I.: 38.3-44.4), and for D. viviparus 2.9% (95% C.I.: 1.6-4.7). There were no significant differences between the two sampling periods. For all parasites, significant geographic clusters of higher AB levels could be established. Furthermore, AB levels against all three parasites were positively correlated with each other, indicating either cross-reactions or co-infections. For O. ostertagi, herd size and percentage of pasture in the ration were positively correlated with AB levels. For F. hepatica, altitude above sea level (a.s.l.) positively, and milk production per cow and year was negatively correlated with AB levels. This work provides baseline data for further studies performing in-depth risk factor analysis and investigating management as well as targeted treatment options to control the parasites.

Comparison of methods to evaluate the energy supply in dairy herds.

Different methods to evaluate the nutritional and metabolic status in dairy herds are available. The aim of this study was to evaluate three different methods to assess the energy status of the herd: 1) the calculation of the energy balance of the herd with a new method based on the Swiss standard calculations, 2) the "efficiency of the feeding management" based on milk yield and milk protein content (%) obtained from DHI data, and 3) the percentage of cows per herd with an average milk yield (16-25 kg/day), but a low milk protein (< 3.2 %). The study included data from 47 farms enrolled in the herd reproduction survey program of the University of Berne, with a total of 653 cows included in one winter period. A correlation could only be seen between methods two and three. It is concluded that it is insufficient only to calculate the ration as an estimation of the energy status of the herd. A rough calculation of the ration is necessary to have an idea on the energy density. But the calculated ration seems to be considerably different from the amount of energy and nutrients of the diet finally absorbed by the cow. As the ingested ration is relevant for milk production and animal health, the relationship between milk yield and milk protein content as well as the percentage of cows with milk protein < 3.2% are considered to provide good information about the efficiency of the feeding management in dairy herds.

Expression of interleukin 4, interleukin 4 splice variants and interferon gamma mRNA in calves experimentally infected with Fasciola hepatica.

Interleukin 4 (IL-4) is expected to play a dominant role in the development of T helper (Th) 2 cells. Th2 immune responses with expression of relatively large amounts of interleukin 4 (IL-4) but little interferon gamma (IFN-gamma) are characteristic for chronic helminth infections. But no information is available about IL4 expression during early Fasciola hepatica (F. hepatica) infections in cattle. Therefore, we investigated F. hepatica specific IL-4 and IFN-gamma mRNA expression in peripheral blood mononuclear cells (PBMCs) from calves experimentally infected with F. hepatica. Cells were collected prior to infection and on post-inoculation days (PIDs) 10, 28 and 70. Interestingly, PBMCs responded to stimulation with F. hepatica secretory-excretory products (FhSEP) already on PID 10 and expressed high amounts of IL-4 but not of IFN-gamma mRNA suggesting that F. hepatica induced a Th2 biased early immune response which was not restricted to the site of infection. Later in infection IL-4 mRNA expression decreased whereas IFN-gamma mRNA expression increased slightly. Isolated lymph node cells (LNCs) stimulated with FhSEP and, even more importantly, non-stimulated LN tissue samples indicated highly polarized Th2 type immune responses in the draining (hepatic) lymph node, but not in the retropharyngeal lymph node. During preliminary experiments, two splice variants of bovine IL-4 mRNA, boIL-4delta2 and boIL-4delta3, were detected. Since a human IL-4delta2 was assumed to act as competitive inhibitor of IL-4, it was important to know whether expression of these splice variants of bovine IL-4 have a regulatory function during an immune response to infection with F. hepatica. Indeed, IL-4 splice variants could be detected in a number of samples, but quantitative analysis did not yield any clue to their function. Therefore, the significance of bovine IL-4 splice variants remains to be determined.

Evaluation of biochemical methods for estimating the sodium intake of dairy cows.

Samples of blood and urine were taken from 334 dairy cows in 29 herds, and the concentrations of sodium and potassium were measured in the serum and urine. The herds were split into five groups according to the amount of supplementary salt they were given: three groups given 10 to 20, 30 to 50 or 70 to 100 g salt per day, and two groups fed an ad libitum supply, given either in bowls or in the form of salt blocks, which were replaced either regularly or irregularly. The groups which received 70 to 100 g of salt daily or were supplied regularly ad libitum had significantly higher urinary sodium concentrations than the other groups. The group receiving 10 to 20 g a day had a significantly higher ratio of potassium:sodium in their urine than all the other groups, in which the ratio decreased as the level of supplementary salt increased. There were wider differences between the groups in terms of the urinary potassium:sodium ratio than in terms of the urinary sodium concentration but less variability within each group.

Influence of activation and germination on high pressure inactivation of ascospores of the mould Eurotium repens.

We investigated heat activation and germination of Eurotium repens ascospores to follow high pressure inactivation. Activation energy and entropy values strengthen the idea of protein denaturation as the underlying mechanism of heat activation. Preceding activation, germination or a combination of both affected high pressure inactivation in different ways. Activation followed immediately by high pressure treatment led to the most efficient improvement in inactivation. However, a pause after activation caused a partial re-establishment of the spores' stability and less efficient high pressure inactivation. Germination stabilized the spores against high pressure. A combined treatment of activation and germination led to an initially fast inactivation, but compared to high pressure treatment of only activated spores the time course of inactivation was slowed down.

Comparison of thelotomy and theloscopic triangulation for the treatment of distal teat obstructions in dairy cows--a retrospective study (1994-1998).

The medical records of 56 cows suffering from teat obstructions in the area of the rosette of Fürstenberg were reviewed; 22 were treated by thelotomy and 34 by theloscopic triangulation and the long-term results were reviewed by telephone conversations with the owners up to four years after the surgery. The milk flow at the fourth milking session after surgery was significantly better in the cows treated by theloscopy and, as a result, fewer of these cows required the teat sphinder to be cut. The cows treated by theloscopy also stayed in hospital for a significantly shorter period and suffered fewer episodes of mastitis.

The course of selected bone resorption marker concentrations in response to short-term hypocalcemia experimentally induced with disodium EDTA infusions in dairy cows.

The collagen metabolites hydroxyproline (HYP), deoxypyridinoline (DPD), and the carboxyterminal telopeptide of type I collagen (ICTP) are suitable markers for bone resorption in humans and several animal species. The purpose of this study was to describe the course of bone resorption markers during short-term hypocalcemia induced with disodium ethylenediaminetetraacetic acid (Na2EDTA) and to investigate whether bone resorption is increased in dairy cows under these conditions. EDTA infusions have been used as a model for periparturient paresis in dairy cows and to estimate the calcium mobilization rate from body reserves in ruminants. In this study, hypocalcemia was induced by means of a 5% Na2EDTA infusion (0.55 mg/kg/min Na2EDTA for 5 h = total dose of 100.6 g). Two experiments were conducted: (1) Six 4-11 years-old Brown Swiss cows were infused intravenously with EDTA for 5 h. Blood and urine samples were taken repeatedly from 1 day before until 10 days after infusion. (2) Towards the end of the lactation, the experiment was repeated with the same animals after a 14-day-period of feeding a low calcium diet (26 g/animal per day). The EDTA-infusion induced hypocalcemia and hypophosphatemia. The HYP-, DPD- and ICTP-concentration remained mainly unaffected during both infusions. Only DPD showed an increase during infusion and HYP an increase 2 days after the infusion. In conclusion, the EDTA infusion had little effect on the concentrations of the measured bone markers, which may be due to the fact that the serum calcium pool was refilled by increased absorption of Ca via the gastrointestinal tract. From these results, it can be concluded that bone resorption was not influenced by EDTA infusion.

Effect of different doses of prostaglandin E2 on intrauterine pressure and uterine motility during diestrus in experimental cows.

Studies in human medicine proved the important role of prostaglandin E2, which stimulates uterine contractions in vivo and in vitro and has been extensively used to ripen the cervix around labor. We wanted to demonstrate that increasing the dosage of prostaglandin E2 (1.25 mg, 2.5 mg, 5 mg and 10 mg) provokes an increase in intrauterine pressure and uterine motility in cattle. Five healthy, lactating dairy cows were used as experimental animals for this study. Intrauterine pressure was recorded during the diestrus phase (1 recording per cow and diestrus phase) by means of a transcervically placed intraluminal pressure microtransducer. Physiologic uterine motility was recorded for 30 min, then placebo or one of the prostaglandin E2- dosages was administered through an indwelling catheter in the jugular vein, followed by a 2-h recording period (eight 15-min periods). Area under the curve (AUC), mean amplitude, frequency of pressure waves and intrauterine pressure were analyzed. Furthermore, we recorded protocols for monitoring heart and respiratory rates and side effects at 9 given examination times. Significant differences were found for the AUC, the mean amplitude and the intrauterine pressure (P < or = 0.05), whereas the number of pressure waves per 15 min did not differ significantly among treatments. Peak values for AUC, mean amplitude and intrauterine pressure were found during the first 15 min after administration of 10 mg of prostaglandin E2. Dose-effect curves showed that the 2.5 mg dosage provided the optimal ratio between myometrial stimulation and undesirable side-effects.

Biochemical markers of bone formation and resorption around parturition and during lactation in dairy cows with high and low standard milk yields.

Substantial changes occur in skeletal metabolism during lactation. These dynamic changes are monitored with biochemical bone markers. The goal of the present study was to follow these changes in lactating cows and to investigate whether cows with a higher milk yield have a higher mobilization rate of calcium from bone. Hydroxyproline, deoxypyridinoline, pyridinoline, and the carboxyterminal telopeptide of type I collagen (ICTP) were chosen as markers for bone resorption, whereas osteocalcin was used as a bone formation marker. Urine and blood samples were collected from cows with a mean standard milk yield of 4900 and 6500 kg, respectively, 14 d before, and 14 d, 1 mo, 1.5 mo, and monthly after parturition. Urinary hydroxyproline, deoxypyridinoline, and pyridinoline concentrations increased with time, but no differences between the two groups were evident. Concentrations of 1,25-dihydroxy vitamin D and ICTP of the two groups showed an increase 14 d after parturition. Furthermore, using multivariate regression models with age and milk yield as covariates, ICTP concentrations were higher in the group with a higher milk yield. In contrast, osteocalcin concentrations decreased 14 d after parturition and returned to prepartum values 1 mo after parturition. The increase of ICTP concentrations in both groups indicates that bone was substantially resorbed. At the same time, probably less Ca was embedded in bone, as indicated by the decrease of the osteocalcin concentrations. In conclusion, cows showed increased bone resorption around parturition, and cows with higher milk yield mobilize calcium more actively from bone than cows with lower milk yield.

Effect of sodium butyric acid, sodium valerianic acid, and osmolarity on contractility of specimens of intestinal wall obtained from the cecum and spiral colon of healthy cows.

OBJECTIVE: To compare the effect of various concentrations of sodium butyric acid and sodium valerianic acid, as well as various osmolarities, on contractility of ex-vivo intestinal wall specimens obtained from the cecum and spiral colon of each of several healthy cows. SAMPLE POPULATION: Full-thickness preparations of intestinal wall, dissected parallel to the longitudinal smooth muscle layers, harvested from freshly slaughtered healthy cows. PROCEDURE: Specimens of intestinal wall were incubated for 5 minutes with various concentrations of sodium butyric acid and sodium valerianic acid as well as various osmolar concentrations of NaCl, using a crossover design. Isometric contractions were induced 7 times with carbachol (CH; 5 X 10(-6) mol/L). Contractility was defined as the maximum amplitude of contraction and the amplitude of contraction 2 minutes after addition of CH. RESULTS: Repeated addition of CH did not result in a significant effect on contractility of specimens from the cecum and spiral colon. Contractility after addition of CH was not significantly affected by prior incubation with various concentrations of sodium butyric acid or sodium valerianic acid or after an increase of osmolarity. Maximum amplitude of contraction was significantly higher in specimens from the spiral colon, compared with specimens from the cecum. CONCLUSIONS: Increases in concentrations of sodium butyric acid or sodium valerianic acid and increases in osmolarity did not inhibit contractility of intestinal wall specimens from the cecum and spiral colon of a group of healthy cows.

Allergen-specific IgE levels against crude mould and storage mite extracts and recombinant mould allergens in sera from horses affected with chronic bronchitis.

Immunoglobulin E antibody (IgE) levels against four recombinant (r) mould allergens (r-Aspergillus fumigatus [rAsp f] 7, 8 and 9; r-Alternaria alternata 1 [rAlta1]) and crude mould (Aspergillus fumigatus, Alternaria alternata, Penicillium notatum) and storage mite extracts were determined by ELISA in sera from 24 pulmonary sound control horses and 26 horses suffering from chronic bronchitis/bronchiolitis (CB), also called chronic obstructive pulmonary disease (COPD). Serum IgG and IgA titres were also determined against Aspergillus fumigatus extract and rAsp f 8.IgE against the crude extracts could be measured in all sera, but there was no significant difference between CB-affected and control horses. In contrast, only 8-30% of the horses, depending on the r-allergen tested, had detectable IgE levels in serum against the r-allergens. Horses with CB had significantly more often detectable IgE levels than controls against rAlt a 1 (10/26 and 3/24, respectively, p=0. 054), rAsp f 7 (13/26 and 2/24, respectively, p<0.01) and rAsp f 8 (11/26 and 1/24, respectively, p<0.01). Only four horses (three CB-affected and one healthy, p0.05) had detectable IgE levels against rAsp f 9. Furthermore, CB-affected horses were often sensitised against two or more r-allergens (13/26 of the CB-affected horses) while only one of the 24 healthy horses had positive IgE levels against more than one r-allergens. Similarly to IgE levels, no significant differences between CB-affected and healthy horses were found for IgG titres against the Aspergillus fumigatus extract. However, horses with CB had significantly higher serum IgG titres against rAsp f 8 than healthy controls (median=28 versus 10 relative ELISA units [REU], p<0.01). Additionally, horses with detectable IgE titres against rAsp f 8 had significantly higher IgG titres against this r-allergen than horses with undetectable IgE titres (median IgG titres=46 and 13 REU, respectively; p<0.01). For serum IgA titres, neither differences between healthy and CB-affected animals nor correlations between IgA and IgG or IgE titres could be found. These results show that horses suffering from CB are more often sensitised to some Aspergillus fumigatus and Alternaria alternata allergens than control horses and that they are partly sensitised to the same fungal proteins as mould-allergic human patients. Furthermore, this study shows that r-allergens allow a much more sensitive determination of specific serum antibody levels by ELISA than crude mould extracts.

Interferon-gamma and interleukin-4 mRNA expression by peripheral blood mononuclear cells from pregnant and non-pregnant cattle seropositive for bovine viral diarrhea virus.

The acceptance of the fetal allograft by pregnant women and mice seems to be associated with a shift from a Th 1 dominated to a Th 2 dominated immune response to certain infectious agents. The goal of this study was to examine cytokine expression in peripheral blood mononuclear cells (PBMCs) from cattle immune to bovine viral diarrhea virus (BVDV) to determine whether pregnancy also has an influence on the type of immune response in this species. Forty-six heifers and cows between 14 months and 13 years of age were included in this study. Twenty-four were seropositive and 22 seronegative for BVDV. Eleven of the seropositive animals and 11 of the seronegative animals were in the eighth month of gestation, the remaining animals were virgin heifers. PBMC from these animals were analyzed for Interferon (IFN)-gamma and Interleukin (IL)-4 mRNA expression by real-time RT-PCR after stimulation with a non-cytopathic strain of BVDV. Additionally, an ELISA was performed to measure IFN-gamma in the supernatants of stimulated cell cultures. In BVDV seropositive animals, IFN-gamma mRNA levels were significantly higher than in BVDV seronegative animals and there was a significant positive correlation between the changes in IFN-gamma and IL-4 mRNA expression. There was, however, no significant difference in IFN-gamma and IL-4 mRNA levels between pregnant and non-pregnant animals. These results are inconsistent with BVDV inducing a Th1 or Th2 biased immune response. Furthermore, a shift in the cytokine pattern during bovine pregnancy was not evident.

Sulfidoleukotriene generation from peripheral blood leukocytes of horses affected with insect bite dermal hypersensitivity.

Sulfidoleukotrienes (sLT) generated in vitro after incubation of equine peripheral blood leukocytes (PBL) with different inducing agents were determined in 18 healthy and 16 insect bite dermal hypersensitivity (IDH)-affected horses. PBL from these 32 horses were stimulated with Concanavalin A, Parascaris equorum, Culicoides nubeculosus and Simulium extracts, and with a six-Grass mix. The cells of all but four horses generated sLT after incubation with Concanavalin A; these four horses did also not produce sLT with the other inducing agents. Of the 28 remaining horses (12 affected with IDH and 16 healthy), all but three generated sLT with the P. equorum extract. The six-Grass mix did not induce sLT production in any of the tested horses. sLT generation with Concanavalin A and Parascaris was statistically not different between IDH-affected and healthy horses. PBL of the diseased horses, however, produced significantly more sLT with the Culicoides (p < 0.01) and Simulium (p < 0.05) extracts than those of the healthy animals. Additionally, sLT generation with the Culicoides extract was measured at different times of the year in one IDH-affected animal and remained high even in winter, when the horse was asymptomatic. sLT and histamine release were determined in 10 horses in parallel. Positive correlations of 0.81 and 0.82 for Concanavalin A and Parascaris (p < 0.01 and p < 0.05, respectively), and of 0.95 and 0.94 for Culicoides and Simulium (p < 0.01) were found between sLT and histamine release. These results indicate that, alike in humans, sLT are released in vitro from equine basophils along with histamine in response to various stimuli and that immediate type hypersensitivity reactions to Culicoides and Simulium are often involved in the pathogenesis of IDH. Thus, sLT generation from equine basophils offers an in vitro diagnostic tool for IDH even in sensitised but asymptomatic horses.

[Physical examination of ruminants showing signs of abdominal pain]. / Klinische Untersuchung der Wiederkäuer beim Symptom "Kolik".

This paper presents a review of the pathogenesis of abdominal pain in ruminants, the complete clinical examination of animals with colic and the symptomatology of the different conditions producing abdominal pain. Further diagnostic procedures such as abdominal ultrasonography, explorative laparotomy and diagnostic laparoscopy are discussed.

[New aspects on the etiopathogenesis and treatment of cecal dilatation/-dislocation in cows--a review]. / Neue Erkenntnisse zur Atiopathogenese und Behandlung der Blinddarmdilatation/-torsion bei der Kuh--Eine Ubersicht.

This paper describes the latest results of research in the areas of etiopathogenesis and treatment of cecal dilatation/-dislocation (CDD) in cows. Similar etiopathogenic mechanisms of CDD and displacement of the abomasum, as has been hypothesized in the literature, were not confirmed. Spontaneous CDD is most likely the consequence of a dysfunction of the spiral colon. Bethanechol is an appropriate motility-modulating drug for medical and postsurgical treatment of spontaneous CDD.

[Epidemiology and risk factors of cecal dilatation/dislocation and abomasal displacement in dairy cows]. / Epidemiologie und Risikofaktoren der Blinddarmdilatation und Labmagenverlagerung bei der Milchkuh.

The aims of the study were to determine the prevalence of cecal dilatation/dislocation (CDD) and abomasal displacement (DA) in Switzerland, to identify risk factors for both diseases, and to compare directly their epidemiologic situation. The epidemiologic study included 158 cases of CDD and 149 cases of DA from the cases referred to both University Clinics of Berne and Zurich. The results showed that DA was associated with nutrition-related risk factors: use of minerals and sodium chloride, inadequate concentrate feeding and beginning of the feeding. Furthermore, breed was significantly associated with DA, but not with CDD. Finally, milk yield and pasture were also significantly included in the models. For CDD, nutrition-related risk factors were also found: pasture in summer, use of corn pellets and corn silage in winter. In the final model, protein concentrate was also included. Although both diseases were found at comparable frequencies, the results of this study indicate marked differences between the epidemiology of occurrence of CDD and DA. Therefore, the hypothesis of a common etiopathogenesis appears unlikely.

Abortion rate during the second half of gestation of cows undergoing surgery in lateral recumbency as compared with cows of a control population.

In a retrospective study, we compared the abortion rates of 59 cows in the second half of gestation, admitted to the Clinic for Food Animals and Horses (1993-1996) with a strictly focal problem (head region, udder and teats, locomotor system) requiring surgery in lateral recumbency, to the abortion rate of a control cow population. Before surgery, a uterus relaxant was given in 42 cases, and 17 cows were untreated as to pregnancy. Cows included in the reproduction health program of the Department of Fertility and Reproduction were used as a control population. Data of 1,736 cows pregnant in the second half of gestation per year were available for the same time period (1993-1996). Abortion rate was 5.08% for the cows undergoing surgery in lateral recumbency as compared with 5.18% for the cows of the control population. There was no significant difference between the two populations (P > 0.05). Thus, the risk for abortion in the cows in question is not elevated as compared with the control population.

Cow level sampling factors affecting analysis and interpretation of milk urea concentrations in 2 dairy herds.

The goals of this study were to determine the influence of the variations among udder quarters, the somatic cell count, the time of sampling during the day, sample conservation, and centrifugation on milk urea (UREA) concentrations, and to propose a sample collection procedure for herds that are not on a Dairy Herd Improvement (DHI) program. Forty cows from 2 herds with different feeding practices were randomly selected. The quarter sampled and the somatic cell count did not significantly influence UREA concentrations. Milk urea concentrations were highest in the morning. The diurnal pattern was not influenced by intrinsic factors like parity, days postpartum, or daily milk yield. The UREA concentrations were significantly higher after refrigeration for one week (mean UREA change = +0.41 +/- 0.24 mmol/L, P = 0.0001) and freezing for one month (mean UREA change = +1.52 +/- 1.25 mmol/L, P = 0.0001). Urea concentrations were slightly higher in lactoserum than in whole milk (mean UREA difference = +0.17 +/- 0.24 mmol/L, P = 0.0001). Although this study included only 2 herds and does not allow extrapolation, differences were found in the diurnal pattern of UREA in these 2 herds, which possibly reflect differences in feeding strategy. With consideration of these results, a 6-point sampling procedure for herds that are not on a DHI program is proposed.

Factors affecting milk urea nitrogen and protein concentrations in Quebec dairy cows.

Milk urea nitrogen (MUN), together with percentage milk protein (PROT), are increasingly used as indicators of the protein-energy balance and for monitoring nutrition and diagnosing feeding disorders. The goal of this study was to analyze the effects of parity, milk yield, days in lactation, somatic-cell count, and herd and feeding factors on MUN and PROT. In 10 dairy herds, one milk sample was taken from each of 418 cows, within +/- 2 days of the routine milk-test visit. We used a four-step multiple linear-regression model with backward elimination, including interactions between herd and the different factors. For both dependent variables, there were significant interactions with herd. Herd-specific models were markedly different; however, the daily amount of protein concentrates fed remained (and had a positive coefficient) in seven of 10 herd models for MUN. This factor is easy to record under field conditions and has to be considered in the evaluation of the ration by means of MUN and PROT. Overfeeding of rumen soluble protein can be easily diagnosed and corrected using MUN analyses. The relationships between MUN and PROT in respect of the factors parity, daily milk yield, and days postpartum also vary considerably among herds.