Importance of estrogens and estrogen-active compounds for udder health in cattle. A review

High oestrogen concentrations in blood or high intake of oestrogen-active compounds with forage can be associated with an enhanced occurrence of udder diseases. Mean somatic cell count (MSCC) can increase and milk yield can decrease. Subclinically infected udder quarters can develop clinical mastitis and the rate of new infections can be high. This review describes concentrations of oestrogens in peripheral blood plasma in cattle and occurrence of oestrogen-active compounds in forage. Relationships between oestrogens or oestrogen-active compounds and udder health are presented. The possible mechanisms of enhanced susceptibility of the udder to infection under the influence of oestrogens are discussed.     

Antimicrobial resistance in staphylococci from canine pyoderma: a prospective study of first-time and recurrent cases in Sweden

In a prospective study involving eight veterinary clinics during 1995 and 1996, samples from first-time and recurrent cases of canine pyoderma were collected by a needle technique. Three hundred and ninety-four staphylococci were isolated and their susceptibility to various antimicrobial drugs was assessed by a microdilution technique. Resistance to macrolides, lincosamides, fusidic add, tetracycline and streptomycin was significantly more common in isolates from the recurrent cases than from the first-time cases; 20 per cent of the isolates from the first-time cases were resistant to three or more of the antimicrobials tested, compared with 45 per cent of those from the recurrent cases. Coresistance between macrolide-lincosamides, tetracyclines and streptomycin was common. No resistance to penicillinase-stable beta-lactams was observed. A comparison with earlier studies indicated that there had been a marked increase in resistance during the previous five years.     

Carbonic anhydrase in the utero-vaginal junction of immature and mature ostriches

1. Sperm storage tubules in the ostrich start to develop at an early stage of oviductal growth. Concurrently, membrane-bound carbonic anhydrase was found in the cells of the storage tubules. 2. In mature ostriches the utero-vaginal junction averaged 11.5+/-2.1 cm in length and primary mucosal folds were extremely long and slender. Membrane-bound carbonic anhydrase was present in the cells of the sperm storage tubules. In the non-ciliated cells of the surface epithelium both membrane-bound and cytoplasmic activity was detected. 3. The possible role of carbonic anhydrase in the stimulation/inhibition of sperm motility by altering the pH was discussed.     

Genetic variation in sows' maternal behaviour, recorded under field conditions

The present study presents heritabilities for several maternal behavioural traits of sows recorded under field conditions (by the farmer) in Norwegian (N) and Finnish (SF) nucleus herds. Based on questionnaires to the farmers, maternal behaviour was scored on a scale from 1 to 7 for 11 traits in the Norwegian study and 5 of the same traits in the Finnish study. Based on the full relationship matrixes, significant heritabilities were found for the same traits in the two countries' breeding herds. The highest heritabilities were found for the sow's reaction to piglet screaming when handled (h²=0.16 (N), h²=0.12 (SF)), how often the sow showed fear during routine management (h²=0.14 (N), h²=0.17 (SF)), aggression towards humans (h²=0.11 (N)) and protests by the sow when moved to the farrowing pen (h²=0.00 (N) but h²=0.22(SF)). Except for the last trait, the results were very consistent across countries. The study shows that field recording of maternal behaviour with questionnaires is one way of detecting genetic variation between sows and seems to work in a large scale, under field conditions. Some of the estimated heritabilities were higher than corresponding heritabilities based on behavioural tests presented in previous studies. Sows' reaction to piglets screaming when handled, sows' fear of humans and sows' aggressive behaviour towards humans are possible candidate traits for selection.     

Macrophages in bovine term placenta: An ultrastructural and molecular study

Retention of foetal membranes (RFM) is a major reproductive disorder in dairy cows. An appropriate immune response is important for a physiological expulsion of the foetal membranes at parturition. Our study aims to provide a deeper insight into characteristics of foetal and maternal macrophages in bovine term placenta. We used transmission electron microscopy (TEM), immunohistochemistry and semi-quantitative RT-PCR to provide a deeper insight into characteristics of foetal and maternal macrophages in bovine term placenta. Semi-quantitative RT-PCR was used to define macrophage polarization in foetal and maternal compartments of normal term placenta. Gene expression of factors involved in M1 polarization [interferon regulatory factor-5 (IRF5), interleukin (IL)-12A, IL12B] and in M2 polarization (IL10) were studied. Ultrastructurally, foetal macrophages showed an irregular shape and large vacuoles, whereas the maternal macrophages were spindle shaped. By immunohistochemistry, macrophages were identified by a strong staining with the lysosomal marker Lysosome-associated membrane glycoprotein 1 (LAMP-1), while myofibroblast in the maternal stroma was positive for alpha-smooth muscle actin. We used the LAMP-1 marker to compare the density of foetal stromal macrophages in placentas of cows with RFM and in controls, but no statistically significant difference was observed. RT-PCR showed a higher expression of all studied genes in the maternal compartment of the placenta and generally a higher expression of M1-, compared to M2-associated genes. Our results indicated that at parturition placental macrophages predominantly show the pro-inflammatory M1 polarization. The higher expression of all the target genes in the maternal compartment may denote that maternal macrophages in bovine term placenta are more frequent than foetal macrophages.     

Influence of Escherichia coli and Arcanobacterium pyogenes isolated from bovine puerperal uteri on phenotypic and functional properties of neutrophils

When cows develop endometritis after birth, Escherichia coli and Arcanobacterium pyogenes are usually the most prominent bacteria present in bovine uterine lochial secretions. A. pyogenes alone is rarely found in the course of a disturbed puerperium. This was confirmed in this study, since average and high-grade uterine contaminations were always associated with the presence of both bacteria. The contamination grade was positively correlated with uterine polymorphonuclear granulocyte (PMN) numbers and negatively correlated with blood PMN numbers. Whether E. coli and A. pyogenes affect the phenotype and function of bovine PMN in a similar or differential way was subject to in vitro studies. PMN were tested in the presence of washed bacterial fragments or culture supernatants taken as a source for soluble and/or secreted bacterial products. Fragments and soluble products differed only quantitatively in their effects on PMN. Usually, long-time exposure (24h) of PMN to fragments induced the strongest effects. Accelerated death of granulocytes was only moderately induced by both E. coli and A. pyogenes products. Both E. coli and A. pyogenes products induced the enhanced expression of a membrane molecule detected by mAb IL-A110 and of CD11b. Expression of other surface structures remained largely unchanged (MHC class I, CD11c). Functional parameters of PMN (phagocytosis; generation of reactive oxygen species, ROS; antibody-independent cellular cytotoxicity, AICC) generally declined after pre-incubation for 24h with products of E. coli or A. pyogenes. Interestingly, soluble products of A. pyogenes stimulated the phagocytosis of PMN. However, co-incubation with E. coli products abrogated this stimulatory effect. The results supply evidence for similar modes of action of the gram-negative E. coli and the gram-positive A. pyogenes on bovine PMN. Alterations in PMN function and phenotype are mainly triggered by direct contact between bacterial fragments and PMN. Inhibition experiments with polymyxin B demonstrated that E. coli-mediated effects were not solely due to the action of lipopolysaccharide. The dominant functional depression of neutrophils by E. coli products strengthens the suggestion that the earlier appearance of E. coli in the uterus may support the co-infection of this organ by A. pyogenes at later times.     

Characterization of leukocytotoxic and superantigen-like factors produced by Staphylococcus aureus isolates from milk of cows with mastitis.

Staphylococcus aureus is a major pathogen for cattle, causing various forms of subclinical and clinical mastitis. Two groups of virulence factors (leukotoxins and superantigens) are supposed to play an important role in the initiation and/or the exacerbation of this disease. In order to detect all known and putative members of leukotoxins and SAgs (superantigens), we tested secreted factors of different S. aureus isolates in flow cytometry-based assays.Isolates were sampled from 68 cows of different farms and cultured for 24h in vitro. Supernatants were then coincubated with purified polymorphonuclear granulocytes (PMN) or combinations of blood mononuclear cells (MNC) and PMN. Viable PMN and MNC were determined by quantitative flow cytometry. In addition, we recorded the proliferation-inducing potential of isolate supernatants for bovine MNC. Based on these criteria, the supernatants of S. aureus isolates fell in three groups. The first group (n=32), termed LT-SNs (leukotoxin-containing supernatants), killed purified granulocytes (neutrophils and eosinophils) in vitro. The second group of supernatants (n=20), termed SAg-SN (superantigen-containing supernatants), induced activation and proliferation of mononuclear cells (MNC) and, only in the presence of MNC, resulted in a selective depletion of neutrophils after 24h in vitro. The third group of supernatants (n=16) contained neither LTs or SAgs. Functionally, SAg-SNs behaved like purified staphylococcal enterotoxin A (SEA) or SEB tested in parallel. The absence of SAg-like activity in LT-SNs was confirmed by heat treatment of LT-SNs, which destroyed the leukocytotoxic activity, but did not reveal any MNC-activating potential. This study, therefore, suggests, that pathogenic S. aureus isolates either produce leukotoxins or superantigens and that both groups of virulence factors can easily be differentiated by the functional assays described.The prevalence of leukotoxin- or superantigen-producing isolates was comparable among cattle with subclinical (LT=41%; SAg=30.8%) mastitis. The higher frequency of LT-producing isolates in cases of clinical mastitis (LT=55.2%; SAg=27.6%) was not significant. At least, these findings argue against the dominant role of superantigens or leukotoxins in S. aureus-induced bovine mastitis.     

Tentative Avenanthramide‐Modifying Enzyme in Oats

A decrease of the concentration of the synthetic avenanthramide N‐(4′‐hydroxy‐(E)‐cinnamoyl)‐5‐hydroxyanthranilic acid in a buffered slurry of milled oat groats (Avena sativa L.) was temperature and pH‐dependent, with a maximum rate at 30°C and pH 9. The reaction was inhibited in the presence of 2‐mercaptoethanol, acetic acid and at high temperature; suggestive of enzymatically catalyzed nature. Among eight different synthetic avenanthramides tested, the tentative enzyme had highest affinity for avenanthramides comprising caffeic and p‐coumaric acids and lowest for those comprising sinapic and ferulic acids. The activity was found in samples from several oat cultivars and was equally pronounced in both bran and endosperm flour of oats. Steeping of oat grains did not influence the reaction.