Antimicrobial susceptibility of Staphylococcus spp. isolated from mammary parenchymas of slaughtered dairy cows

The high prevalence of the great resistance to antimicrobials shown by the aetiological agents of infectious bovine mastitis, often leads to chronic recurrent mastitis, one of the main causes of early culling of dairy cows. The purpose of this study was to determine the in vitro susceptibility pattern of Staphylococcus spp. isolated from mammary parenchymas of slaughtered dairy cows, to different anti-microbials. A total of 45 Staphylococcus spp. strains [33 coagulase-negative Staphylococcus (CNS) and 12 Staphylococcus aureus (SA)] were used. These strains were tested with 12 different antimicrobials by the Kirby and Bauer standardized disc diffusion method; 84.44% and 86.66% of the 45 strains were resistant to ampicillin and penicillin, respectively. The highest sensitivity was to cephalothin (84.44%), gentamicin (80%) and to sulphazotrin (77.77%). CNS showed higher resistance (P < 0.05) than SA. The in vitro susceptibility pattern of Staphylococcus spp. strains isolated from mammary parenchymas studied in the present investigation was similar to that observed in recent studies of in vitro and in vivo susceptibility patterns of Staphylococcus spp. isolated from cases of bovine mastitis.     

In vitro model of parathyroid hormone-related protein secretion from mammary cells isolated from lactating cows

Parathyroid hormone-related protein (PTHrP) is produced by the lactating mammary gland and is present in milk in a biologically active form. The goal of this investigation was to determine if cells cultured from the lactating mammary glands of cows would secrete PTHrP in vitro. Mammary acini were isolated from lactating cows at 1–6 wk after calving, and fresh or cryopreserved mammary acini were cultured for 14 d on Type I collagen. Cultures on thick layers of collagen (2.5 mm) were detached and allowed to contract on Day 6. PTHrP production was measured by N-terminal radioimmunoassay and bioassay (increased cAMP levels in ROS 17/2.8 osteoblast-like cells). The mammary cells reached confluence at Day 6. PTHrP production was low at Day 2 (<0.5 ng/ml) but increased to peak production (2–4 ng/ml) at approximately Day 6 and remained constant until Day 14. Immunoreactive and bioactive PTHrP levels in the culture medium correlated well. The cultures produced lactoferrin (2,000–2,300 ng/ml) and α_s1-casein (14–19 ng/ml). Prolactin stimulated PTHrP production approximately 50% on Days 6–14. PTHrP production was increased approximately 100% by treatment with epidermal growth factor (10 ng/ml) for 2 d. Morphologic evaluation of cultures on thick, contracted collagen at Day 14 revealed an inner layer of mammary epithelial cells overlying myoepithelial cells and an outer layer of collagen containing stromal cells. Immunohistochemistry demonstrated positive staining for PTHrP and cytokeratin in both mammary epithelial and myoepithelial cells and a-smooth muscle actin in myoepithelial cells. These data demonstrated that cryopreserved mammary tissue from lactating cows could be cultured in vitro and secreted PTHrP in a regulated manner. This in vitro model will be useful to investigate the function and regulation of PTHrP in the lactating mammary gland.     

Enzymatic activity of yeasts isolated from the inflamed mammary secretion in dairy cows

The aim of the study was to evaluate enzymatic activities of yeasts isolated from inflammatory mammary secretion. The yeasts isolated from cows with clinical and sub-clinical mastitis (134 strains) included: Candida krusei (62 strains), Candida kefyr (48 strains), Candida lusitaniae (17 strains) and Candida famata (7 strains). The API ZYM system was used containing substrates to assess 19 hydrolytic enzymes. Substantial differences in the number and activity of hydrolyses were demonstrated in individual species. In Candida krusei, acid phosphatase showed the highest activity (4.36 points), in Candida kefyr and Candida lusitaniae--leucine arylamidase (4.93 and 4.25 points, respectively), in Candida famata--alpha-glucosidase (4.75 points). No activity of trypsin, chymotrypsin, alpha-galactosidase, beta-glucuronidase, alpha-mannosidase or alpha-fucosidase was observed in any of the yeasts examined.     

A study of the dynamics of digital dermatitis in 742 lactating dairy cows

Digital dermatitis (DD) is a contagious disease of cattle affecting the skin of the claw. The disease presents with a range of severities and can be associated with lameness. Information about the disease dynamics of DD is scarce. Parity and lactation stage have been identified as risk factors for DD and studies have also indicated that not all cows are equal regarding their risk of recurrent disease and prospects for cure from DD. The aim of this study was to investigate host heterogeneity to DD and to identify disease patterns of DD and factors associated with the development and resolution of lesions. In three commercial dairy herds, 742 lactating cows were observed for DD lesions weekly for 11 or 12 weeks. The effects of parity, lactation stage and duration of preceding episode on the hazard of transitions between healthy and lesion states were analysed using a multilevel multistate discrete-time model. One or more DD lesions were observed in 460 cows and lesions were observed in 2426 out of 10,585 observations. In total, 1755 uncensored episodes with DD lesions were observed. Early lactation was associated with a reduced risk of developing lesions compared with mid and late lactation. Lesions that developed in late lactation had a greater likelihood of resolution compared with lesions that developed during early lactation. There was a reduced risk of lesions developing in parity 3 compared with parity 1 cows, but an increased risk of lesions developing in parity 2 compared with parity 1 cows. In the present study, the mean duration of uncensored DD episodes was 1.7 weeks indicating that the duration of the majority of DD lesions might be shorter than the 42 days reported previously. The transitions between disease states suggest that DD is a dynamic disease, and that the early stage lesions are more transient than expected from previous studies. We conclude that studies with shorter observation intervals than one week are needed to fully understand and describe the individual and group dynamics of DD.     

Fractional excretion of electrolytes in lactating dairy cows.

Samples of serum and urine were obtained simultaneously from 56 healthy lactating cows to determine ranges of fractional excretion (FE) of calcium (Ca), phosphate (PO4), magnesium (Mg), sodium (Na), potassium (K), and chloride (Cl). Samples were obtained at 3 stages of lactation: period 1 = 1 to 7 days, 2 = 83 to 112 days, and 3 = 175 to 197 days. The FE of electrolytes were significantly different among periods 1, 2, and 3 for Ca (P less than 0.001), PO4 (P less than 0.025) and Mg (P less than 0.025), but were not significantly different for Na, K, and Cl. Least squares mean FE of Ca was lowest in period 1 and not significantly different for periods 2 and 3, whereas mean FE values for PO4 and Mg were highest in period 2 and not significantly different for periods 1 and 3. The mean FE values of Na, K, and Cl did not change with stage of lactation. Age and category of milk production (high, medium, and low) did not influence the FE values of the electrolytes.     

Pharmacokinetics of sulphadiazine-trimethoprim in lactating dairy cows

Five Finnish Ayrshire cows in mid or end-lactation were treated with 40 mg sulphadiazine/kg and 8 mg trimethoprim/kg using intravenous (i.v.), intramuscular (i.m.) and subcutaneous (s.c.) routes. Elimination of sulphadiazine was not affected by the route of administration (median t1/2 4.4-5.0 h) while elimination of trimethoprim was strongly limited by slow absorption from the injection site after s.c. and i.m. administration (median for apparent t1/2 21-25 h) compared to that after i.v. administration (median t1/2 1.2 h; p < 0.05). The median bioavailability of trimethoprim was also decreased, being 37% and 55% after s.c. and i.m. administration, respectively. When i.v. administration was used, trimethoprim concentration exceeded 0.1 mg/l in milk between 0.15-8 h while sulphadiazine concentrations above 2 mg/l were maintained from 0.5-2 h to 8 h. After s.c. and i.m. administration sulphadiazine in milk behaved similar to that after i.v. administration, while trimethoprim time-concentration curves were flat and trimethoprim concentrations were around 0.1 mg/l for an extended period of time (8-12 h). Median Cmax values in milk were only 0.07 mg/l and 0.10 mg/l for s.c. and i.m. administrations, respectively. After s.c. administration, 4 out of 5 cows showed signs of pain. After i.m. administration, 2 of the cows showed clear signs of pain and one had some local tenderness at the site of injection.     

Urinary pseudouridine excretion in lactating dairy cows

Introduction   A number of systems based on metabolizable protein, such as that adopted in the UK (A gricultural and F ood R esearch C ouncil 1992) have been developed to improve the accuracy of protein rationing for ruminants. Quantification of microbial protein synthesis in the rumen is a fundamental requirement of all such systems. In the UK system, microbial protein supply is predicted from an estimate of fermentable metabolizable energy intake, using a correction for the effects of level of feeding on the energetic efficiency of microbial protein synthesis. Use of such an approach is however subject to considerable error due to large variations in the energetic efficiency of microbial protein synthesis (A gricultural R esearch C ouncil 1984). Consequently there is an urgent requirement for an on-farm diagnostic marker of microbial protein supply as a basis for adjusting diets to maximize efficiency of dietary nitrogen utilization by dairy cows (D ewhurst et al. 1996). Urinary purine derivative excretion has been proposed as a noninvasive index of microbial protein supply in ruminant animals (T opps and E lliot 1965). Use of this microbial marker is based on the assumption that purines entering the duodenum are essentially microbial in origin (M c A llan 1982), and that following metabolism, their derivatives are quantitatively recovered in the urine (C hen et al. 1990; V erbic et al. 1990). Purine metabolites excreted in ruminant urine are primarily derived from the metabolism of absorbed purines, but as a consequence of tissue adenosine triphosphate and nucleic acid turnover, a proportion of purine bases are not salvaged and re-utilized, but enter catabolic pathways, constituting an endogenous loss.     

Pharmacokinetics of sodium cephapirin in lactating dairy cows

Sodium cephapirin was administered (10 mg/kg of body weight, IM) at 8-hour intervals in 4 consecutive doses to each of 6 lactating dairy cows. Blood, normal milk, mastitic milk, urine, and endometrial tissue samples were collected serially. Mean peak cephapirin concentrations in serum were 13.3 micrograms/ml 10 minutes after the 1st injection and were 15.8 micrograms/ml 20 minutes after the 4th injection (post[initial]injection hour [PIH] 24.33). The overall elimination rate constant value was 0.66/h and plasma clearance was 760 ml/h/kg. Mean peak cephapirin concentration in normal milk was 0.11 microgram/ml at PIH 2 and mean peak cephapirin concentration in mastitic milk was 0.18 microgram/ml at PIH 4. Cephapirin was not detected in the endometrium. The highest concentration of cephapirin in urine was 452 micrograms/ml, 2 hours after the 4th dose (PIH 26).     

Characterization of erythromycin-resistant methylase genes from multiple antibiotic resistant Staphylococcus spp isolated from milk samples of lactating cows

OBJECTIVE: To isolate and characterize erythromycin-resistant methylase genes in multiple-antibiotic resistant staphylococci isolated from milk samples. ANIMALS: 300 lactating cows. PROCEDURE: 23 erythromycin-resistant staphylococci were isolated from milk samples of 300 lactating cows. The prevalence of erythromycin-resistant methylase (erm) genes, ermC and ermA genes, and the multicomponent macrolide efflux pump in staphylococci msrA genes were identified and characterized by use of multiplex polymerase chain reaction (PCR), Southern hybridization, restricted fragment length polymorphism (RFLP) analysis, and dot-blot hybridization. RESULTS: Biochemical characterization indicated that 3 of 23 (13%) isolates were coagulase-positive Staphylococcus aureus, and the rest were coagulase-negative. Multiplex PCR resulted in amplification of a 520-base pair (bp) region of the ermC gene from the cell lysates of a strain of S simulans M-21 and S sciuri M-28. The ermC gene in both isolates was found on a 3-kilobase plasmid. The ermA gene was found on the chromosome of 21 isolates, and 6 RFLP patterns were observed. None of the isolates harbored the msrA gene. CONCLUSIONS: Erythromycin-resistant Staphylococcus spp isolated from milk samples of lactating cows may serve as reservoirs of erm genes homologous to those described in human isolates. However, the chromosomal insert patterns and prevalence of these genes, the sizes of plasmids harboring the genes, and the number of inserts of the genes (copy number) may differ from that of human isolates.     

Effects of stocking density on oxidative stress status and mammary gland permeability in early lactating dairy cows

The current study was conducted to investigate the effect of stocking density (SD) on oxidative stress status and mammary gland permeability in early lactating dairy cows. Thirty‐two dairy cows were allocated into 16 blocks, basing on parity, previous milk yield, and body weight, and were then randomly assigned into one of the two treatments as follows: 75% (75SD) and 100% (100SD) SD. The cows were fed with same diet throughout the 8‐week experimental period. The milk yield and milk sample were collected on two consecutive days during the 8‐week experimental period weekly. Plasma samples were collected on fourth and eighth experimental weeks. Raw, energy‐corrected, and 4% fat‐corrected milk yield were significantly higher in 75SD‐cows than that of 100SD‐animals, respectively. The milk somatic cell count was lower in 75SD‐cows than that of 100SD‐animals. The levels of Na⁺, Na⁺/K⁺, bovine serum albumin and plasmin were lower in 75SD‐cows than those of 100SD‐cattle, respectively. The 75SD‐cows had reduced insulin and insulin/glucagon levels but higher prolactin and growth hormone concentrations, compared with those of 100SD‐animals, respectively. In conclusion, compared with low SD‐animals, early lactating cows with higher SD had higher oxidative stress status, which further led to a greater mammary gland permeability.     

Blood flow to the distal part of the teat (mammary papilla) of lactating dairy cows

Relative blood flow to tissues of the distal part of the teat (mammary papilla) in 5 lactating dairy cows was determined, using 15-microns nonradioactive carbonized microspheres. Highest relative blood flow occurred in the epithelial region (zone A) of the teat canal (papillary duct) which comprised both teat canal epithelium and papillated portion of the stratum papillare. Blood flow to these tissues was more than 4 times greater than blood flow to equivalent tissues of the mucosal rosette. The high rate of metabolic activity indicated by this relatively high blood flow may be required to support secretion of antimicrobial substances and continual synthesis of epithelial cells to replace those lost during milking.     

菊芋粕对泌乳奶牛的营养价值评定

选用6头2～3胎泌乳中期健康荷斯坦奶牛,采用3×3有重复拉丁方试验设计,通过消化试验就菊芋粕对泌乳奶牛的营养价值进行评定。试验设3个日粮处理,处理1为基础日粮,处理2为菊芋粕替代15%基础日粮(干物质基础),处理3为菊芋粕替代30%基础日粮(干物质基础)。结果表明,菊芋粕风干样品中干物质、粗蛋白、粗脂肪、粗纤维、中性洗涤纤维、酸性洗涤纤维、粗灰分、钙、磷和产奶净能分别为92.43%,12.98%,0.81%,12.67%,25.93%,21.44%,3.92%,0.27%,0.20%和 5.18 MJ/kg;菊芋粕对泌乳奶牛干物质消化率、粗蛋白消化率、中性洗涤纤维消化率、酸性洗涤纤维消化率分别为61.78%,73.21%,51.65%,61.14%,消化能和产奶净能分别为10.13和5.18 MJ/kg;3个处理组试牛产奶量和乳成分无显著差异(P>0.05),表明菊芋粕是一种营养价值略低但养分消化率较高的能量饲料。     

Stability of plasma metabolites and hormones in lactating dairy cows

Plasma concentration stability of glucose, free fatty acids, ketone bodies, growth hormone, insulin were determined in lactating dairy cows. Concentrations of these metabolites and hormones were measured during a 36- to 48-hour period in 3 normal, mature dairy cows in the 2nd month of lactation. Samples were taken at 30-minute intervals; also, intensive sampling (every 10 minutes) was done at varying times in relation to feeding and milking. Of the 5 components measured, glucose concentration was the most stable, easiest to assay, and most reliable for use as a diagnostic aid in assessing metabolic carbohydrate disturbances in dairy cattle.     

日粮阴阳离子差在泌乳奶牛日粮中的应用

日粮阴阳离子差（DCAD）可调控家畜体内的酸碱平衡。近年来,由于其对泌乳奶牛生产性能的影响而越来越受到研究者的重视。文章就阴阳离子差的计算,阴阳离子差对泌乳奶牛干物质采食量、泌乳量和乳成分、瘤胃参数的影响,及对DCAD在我国泌乳奶牛日粮中的应用的展望作了综合阐述。     

Hypocalcaemia associated with high dose rates of zinc oxide to lactating dairy cows

Clinical hypocalcaemia was associated with single doses of three oi more days; supply of zinc oxide to dairy cows for facial eczema control. Experimental hand dosing of a single dose of zinc oxide (40 and 120 mg Zn/kg b wt) to Jersey cows caused a significant drop in serum calcium concentration at 24 h followed by a rise to concentrations higher than controls at subsequent samplings over four days. At the highest zinc dose rate, 78% of serum calcium values were equal to or lower than the lowest control value at 24 h post dosing. Serum magnesium concentrations had a reciprocal relationship to the calcium levels at all samplings. Serum zinc concentrations were most elevated over the 48 h post dosing, returning to normal levels at the fifth day. Following the highest zinc dose milk yields were reduced by approximately 1.5 kg milk/cow/day for 48 h. Similar results were obtained when cows grazed pasture on which large amounts of zinc oxide had been sprayed.     

Behavioural signs of oestrus during pregnancy in lactating dairy cows

Abstract

Two herds of approximately 50 dairy cows were observed for oestrous behaviour for 6 weeks, 12 times a day for 30 minutes. It appeared that 3.08% of the pregnant cows showed oestrous behaviour during pregnancy (EBP) in such an intensity that they would have been considered in oestrus. With a less rigid oestrus detection threshold, 10.8% of the pregnant cows would have been considered to be in oestrus. Animals showed EBP during all months of pregnancy, but most of the behavior was observed in the middle of the gestation period.     

Incidence and treatment of dermatitis digitalis in dairy cows

In dairy cows of the Black Pied Lowland, Bohemian Pied breeds and their crossbreds loosely in large cow houses VKK 900 on slatted floors, 24% incidence of dermatitis digitalis was recorded during the period of one year. Relations between the occurrence of this disease, age and efficiency of dairy cows, time of parturition and season of the year were studied. Therapeutical effects of several methods of treatment were evaluated and compared. No effect of age and efficiency of dairy cows, nor of the year season on the occurrence of this disease was observed. A significantly higher occurrence was proved in the period before and after parturition, when 80% of the total occurrence of digitalis dermatitis were diagnosed. During this period, however, the cows were housed in a stable with markedly worse environmental circumstance than those in production stable. 91% cases of dermatitis were diagnosed on the digits of pelvic limbs. Relapses were determined only in five dairy cows. There was no case of the disease occurring in calves reared in the prophylactorium of the calf house. Therapeutical results were best after repeated mass treatment of the digits of dairy cows in 5% formaldehyde baths. The results of this study point at a conclusion that probable pathogens of this disease are specific infection agents, or that there are more synergic pathogens. A significant pre-disposition factor are bad environmental circumstances.     

Hypocalcaemia associated with high dose rates of zinc oxide to lactating dairy cows

Clinical hypocalcaemia was associated with single doses of three oi more days’ supply of zinc oxide to dairy cows for facial eczema control. Experimental hand dosing of a single dose of zinc oxide (40 and 120 mg Zn/kg h wt) to Jersey cows caused a significant drop in serum calcium concentration at 24 h followed by a rise to concentrations higher than controls at subsequent samplings over four days. At the highest zinc dose rate, 78% of serum calcium values were equal to or lower than the lowest control value at 24 h post dosing. Serum magnesium concentrations had a reciprocal relationship to the calcium levels at all samplings. Serum zinc concentrations were most elevated over the 48 h post dosing, returning to normal levels at the fifth day. Following the highest zinc dose milk yields were reduced by approximately 1.5 kg milk/cow/day for 48 h. Similar results were obtained when cows grazed pasture on which large amounts of zinc oxide had been sprayed.