Role of extracellular matrix and prolactin in functional differentiation of bovine BME-UV1 mammary epithelial cells

Interactions between extracellular matrix (ECM) and epithelial cells are necessary for proper organisation and function of the epithelium. In the present study we show that bovine mammary epithelial cell line BME-UV1 cultured on ECM components, commercially available as Matrigel, constitutes a good model for studying mechanisms controlling functional differentiation of the bovine mammary gland. In contact with Matrigel BME-UV1 cells induce apicobasal polarity, and within 16 days form three dimensional (3D) acinar structures with a centrally localized hollow lumen, which structurally resemble mammary alveoli present in the functionally active mammary gland. We have shown that the 3D culture system enables a high expression and proper localisation of integrin receptors and tight junction proteins in BME-UV1 cells to be induced. This effect was not obtained in cells grown in the classical 2D culture system on plastic. Moreover, ECM highly stimulated the synthesis of one of the major milk proteins, beta-casein, even in the absence of prolactin. Our results show that contact with ECM plays an important role in the lactogenic activity of bovine MECs, however, prolactin is necessary for the efficient secretion of milk proteins.     

Haematological analysis of calves with bovine neonatal pancytopenia

Blood was obtained from 61 neonatal Holstein calves originating from a farm in Germany with a high incidence of bovine neonatal pancytopenia (BNP). In order to detect alterations that might be related to BNP, selected haematological analytes were determined. Haematological examinations demonstrated alterations in at least two of the three cell lineages in 10 calves (16.39 per cent). Six animals (9.84 per cent) developed a bleeding disorder indicative of BNP at approximately two weeks of age. None of these animals showed alterations in complete blood cell count at sampling in the first week of life. In weeks when calves with BNP were born, an increase in the number of apparently healthy calves demonstrating decreases in blood cell counts was observed.     

Serum and bronchoalveolar lavage fluid endothelin-1 concentrations as diagnostic biomarkers of canine idiopathic pulmonary fibrosis

Background: Diagnosis of canine idiopathic pulmonary fibrosis (IPF) is challenging. Endothelin‐1 (ET1) is a biomarker of IPF in humans, but whether ET1 can detect and differentiate IPF from other canine respiratory diseases is unknown. Objective: To evaluate whether measurement of the concentration of ET1 in serum and bronchoalveolar lavage fluid (BALF) can be used to distinguish canine IPF from chronic bronchitis (CB) and eosinophilic bronchopneumopathy (EBP). Animals: Twelve dogs with IPF, 10 dogs with CB, 6 dogs with EBP, 13 privately owned healthy West Highland White Terriers (WHWT), and 9 healthy Beagle dogs. Methods: Prospective, case control study. ET1 concentration was determined by ELISA in serum and in BALF. Results: No significant difference in serum ET1 concentration was detected between healthy Beagle dogs and WHWT. Serum ET1 concentration was higher in dogs with IPF (median interquartile range; 2.32 pg/mL, 2.05–3.38) than healthy Beagle dogs (1.28, 1.07–1.53; P < .001), healthy WHWT (1.56, 1.25–1.85; P < .001), dogs with EBP (0.94 0.68–1.01; P = .001), and dogs with CB (1.54 0.74–1.82; P = .005). BALF ET1 concentration was below the detection limit in healthy WHWT and in dogs with CB, whereas it was measurable in all dogs with IPF. A cut‐off serum concentration of 1.8 pg/mL had a sensitivity of 100% and a specificity of 81.2% for detection of IPF, with an area under the receiver operating characteristic curve of 0.818. Conclusions and Clinical Importance: Serum ET1 can differentiate dogs with IPF from dogs with EBP or CB. ET1 can be detected in BALF of dogs with IPF.     

Additivity of effects from dietary copper and zinc on growth performance and fecal microbiota of pigs after weaning

Four experiments were conducted to determine the interactive effects of pharmacological amounts of Zn from ZnO and Cu from organic (Cu-AA complex; Cu-AA) or inorganic (CuSO(4)) sources on growth performance of weanling pigs. The Cu was fed for 4 (Exp. 1) or 6 (Exp. 2, 3, and 4) wk after weaning, and Zn was fed for 4 (Exp. 1) or 2 (Exp. 2, 3, and 4) wk after weaning. Treatments were replicated with 7 pens of 5 or 6 pigs per pen (19.0 ± 1.4 d of age and 5.8 ± 0.4 kg of BW, Exp. 1), 12 pens of 21 pigs per pen (about 21 d of age and 5.3 kg of BW, Exp. 2), 5 pens of 4 pigs per pen (20.3 ± 0.5 d of age and 7.0 ± 0.5 kg of BW, Exp. 3), and 16 pens of 21 pigs per pen (about 21 d of age and 5.7 kg of BW, Exp. 4). In Exp. 1 and 2, Cu-AA (0 vs. 100 mg/kg of Cu) and ZnO (0 vs. 3,000 mg/kg of Zn) were used in a 2 × 2 factorial arrangement. Only Exp. 1 used in-feed antibiotic (165 mg of oxytetracycline and 116 mg of neomycin per kilogram feed), and Exp. 2 was conducted at a commercial farm. In Exp. 3, sources of Cu (none; CuSO(4) at 250 mg/kg of Cu; and Cu-AA at 100 mg/kg of Cu) and ZnO (0 vs. 3,000 mg/kg of Zn) were used in a 3 × 2 factorial arrangement. In Exp. 4, treatments were no additional Cu, CuSO(4) at 315 mg/kg of Cu, or Cu-AA at 100 mg/kg of Cu to a diet supplemented with 3,000 mg/kg of Zn from ZnO and in-feed antibiotic (55 mg of carbadox per kilogram of feed). In Exp. 1 and 2, both Zn and Cu-AA improved (P < 0.001 to P = 0.03) ADG and ADFI. No interactions were observed, except in wk 1 of Exp. 2, where Zn increased the G:F only in the absence of Cu-AA (Cu-AA × Zn, P = 0.04). A naturally occurring colibacillosis diarrhea outbreak occurred during this experiment. The ZnO addition reduced (P < 0.001) the number of pigs removed and pig-days on antibiotic therapy. In Exp 3, ADFI in wk 2 was improved by Zn and Cu (P < 0.001 and P = 0.09, respectively) with no interactions. In wk 1, G:F was reduced by ZnO only in the absence of Cu (Cu × Zn, P = 0.03). Feeding Zn decreased fecal microbiota diversity in the presence of CuSO(4) but increased it in the presence of Cu-AA (Cu source × Zn, P = 0.06). In Exp. 4, Cu supplementation improved the overall ADG (P = 0.002) and G:F (P < 0.001). The CuSO(4) effect on G:F was greater (P < 0.001) than the Cu-AA effect. Our results indicate that pharmacological amounts of ZnO and Cu (Cu-AA or CuSO(4)) are additive in promoting growth of pigs after weaning.     

The role of European starlings in the spread of coccidia within concentrated animal feeding operations

To investigate the relationship between European starlings and bovine coccidiosis we collected samples from European starlings, cattle feed bunks, cattle water troughs, and cattle feces within concentrated animal feeding operations (CAFOs). These samples were screened for coccidia spp. to investigate (i) the prevalence of coccidia in starlings using CAFOs; (ii) if there is a relationship between bovine coccidiosis and starling numbers; (iii) if coccidia contamination of cattle feed and water is related to the number of starlings observed on CAFOs. Coccidia belonging to the genus Eimeria were detected in cattle feces and one water sample but no Eimeria spp. were detected in European starlings or cattle feed. However, many European starling samples were positive for Isospora. Starling use of CAFOs did not appear to be associated with coccidia spp. shedding by cattle and there was no correlation between starling numbers and contamination of cattle feed and water, suggesting that starling do not contribute to the amplification and spread of Eimeria in CAFOs.     

Epidemiology of bovine anaplasmosis in dairy herds from Costa Rica

Bovine anaplasmosis is endemic and occurs in almost all areas of livestock production of Costa Rica. The aim of this study was to determine the seroprevalence and risk factors of anaplasmosis in dairy farms of Costa Rica by the recombinant truncated MSP-5 (rMSP-5) enzyme-linked immunosorbent assay (ELISA). Serum samples were obtained from 733 cattle from 20 commercial dairy herds of Costa Rica. The overall seroprevalence was 37.2% and herd seroprevalence ranged from 20.0 to 72.0%. The age-specific seroprevalence was 49.3% in young and 33.4% in adult animals. The main risk factors associated with seroprevalence were season of occurrence of clinical cases (rainy season) (OR=22.8), presence of tabanids (OR=9.5) and stable flies (OR=6.2), stable flies control measures (OR=3.2), non-use of ear tattoos (OR=2.8), interval of veterinary visit (≤ 60 days) (OR=2.7), altitude of the farms (<800 masl) (OR=2.6) and age (<2 years) (OR=1.8). The results indicated that exposure of cattle to Anaplasma marginale is common in dairy herds of Costa Rica and endemic instability situation probably is due to inadequate vector control.     

The expression of interferon-stimulated gene 15 in equine endometrium

Establishment of pregnancy is critically dependent upon a precisely orchestrated embryo-maternal interaction leading to a receptive uterine environment. The up-regulation of the interferon-stimulated protein 15 kDa (ISG15) during pregnancy has been described in various species and has been hypothesized to be part of the molecular repertoire that makes the uterus receptive to conceptus development. In the current study, the expression of ISG15 and enzymes involved in ISG15ylation was examined at the mRNA and protein level in equine endometrium at Day 14 of the luteal phase and at Day 14 and 50 of pregnancy. ISG15 mRNA showed a 2.63-fold higher expression at Day 14 of pregnancy when compared to Day 14 of the cycle, while mRNA abundance at Day 50 of pregnancy was unchanged compared to Day 14 of the cycle. Upon Western blot analysis using anti-ISG15 antibody, several higher molecular weight bands could be observed, representing proteins conjugated to ISG15. No free ISG15 could be detected. The pattern of ISG15 reactive proteins differed from those observed in non-uterine samples. Upon immunohistochemistry, ISG15 reactive proteins located primarily to luminal and glandular epithelial cells, while stromal cells showed weaker staining. In conclusion, the expression of ISG15-conjugated proteins in equine endometrium did not differ between cyclic and pregnant 14 days after ovulation and Day 50 of pregnancy. It is hypothesized that the unique subset of ISG15ylated proteins expressed in endometrial tissue contributes to normal cellular function and that, unlike other species, the modification of ISG15-conjugated proteins is not an active contributor to conceptus-maternal interaction in the mare.