Antimicrobial activity of a bovine myeloid antimicrobial peptide (BMAP‐28) against methicillin‐susceptible and methicillin‐resistant Staphylococcus aureus

A bovine myeloid antimicrobial peptide (BMAP‐28) is a member of the cathelicidin family which is included in the innate immune system of mammals. Recently, there have been many studies about antimicrobial peptides. This study aims to clarify whether BMAP‐28 has bactericidal activity against methicillin‐resistant Staphylococcus aureus (MRSA) and compares its activity against methicillin‐susceptible S. aureus (MSSA) and MRSA. We found that the peptide was effective in killing MRSA (minimal inhibitory concentration (MIC) range; 5–20 µg/mL). It was also revealed that MSSA (MIC range; 1.25–20 µg/mL) had two levels of susceptibility to BMAP‐28. We also examined the effect of BMAP‐28 on bacterial shape to visually show its activity. After exposure to the peptide, both MSSA and MRSA cells showed the morphological changes on their surfaces. Our results indicate that BMAP‐28 is a promising candidate for medicine against drug‐resistant bacteria.     

A bovine myeloid antimicrobial peptide (BMAP‐28) kills methicillin‐resistant Staphylococcus aureus but promotes adherence of the bacteria

The cathelicidin family is one of the several families of antimicrobial peptides (AMPs). A bovine myeloid antimicrobial peptide (BMAP‐28) belongs to this family. Recently, the emergence of drug‐resistant bacteria such as methicillin‐resistant Staphylococcus aureus (MRSA) has become a big problem. AMPs are expected to be leading compounds of new antibiotics against drug‐resistant bacteria. In this study, we focused on the activity of BMAP‐28 against bacterial cell surfaces. First, we observed morphological change of MRSA caused by BMAP‐28 using a scanning probe microscope. We also studied activities of BMAP‐28 against adherence of S. aureus to fibronectin, collagen type I, collagen type IV. We confirmed whether BMAP‐28 can bind to lipoteichoic acid (LTA) of S. aureus. BMAP‐28 was indicated as damaging the cell surface of MRSA. In a particular range of concentrations, BMAP‐28 promoted adherence of S. aureus against fibronectin and collagens. It was revealed that BMAP‐28 and LTA of S. aureus bound with each other. Our study showed the potential of BMAP‐28 which can damage MRSA and interact with LTA of S. aureus but promote its adherence in some concentrations. This study provides new points of which to take notice when we use AMPs as medicines.     

Molecular characterization of community‐associated methicillin‐resistant Staphylococcus aureus from pet dogs

Community‐associated methicillin‐resistant Staphylococcus aureus (MRSA) is a serious public health concern and in Australia, one that disproportionately affects Aboriginal people. Paralleling MRSA in human medicine, methicillin‐resistant S. pseudintermedius (MRSP) is an increasingly prevalent pathogen in veterinary medicine. We aimed to characterize the carriage of MRSA and MRSP in dogs and cats from predominantly Aboriginal communities in a very remote region of New South Wales (NSW), Australia. Pets (303 dogs and 80 cats) were recruited from six communities in western NSW. Three swabs were collected from each animal (anterior nares, oropharynx and perineum) and from skin lesions or wounds (if present) and cultured on selective media for methicillin‐resistant staphylococci. Human host‐adapted community‐associated MRSA representing four multilocus sequence types (ST1‐IV, ST5‐IV, ST72‐IV, ST93‐IV) were isolated from eight dogs (prevalence 2.6%, 95% confidence interval 1.3%–5.1%). Two ST5‐IV isolates from a single dog were phenotypically trimethoprim‐resistant, harbouring trimethoprim‐resistant gene dfrG within the SCCmec type IVo mobile genetic element. MRSA was not isolated from any cats and MRSP was not isolated from any dogs or cats. This study estimated a high prevalence of human host‐adapted community‐associated MRSA carriage in dogs despite an absence of MRSP. This suggests MRSA carried by dogs in remote NSW originate from human hosts. The cycle of transmission between people, dogs and common environmental sources warrants further investigation. To our knowledge, this is the first report of trimethoprim‐resistant ST5‐IV in eastern Australia and the first report of trimethoprim‐resistant ST5‐IV from a dog.     

Clinical, microbiological, and molecular characterization of methicillin-resistant Staphylococcus aureus infections of cats

OBJECTIVE: To compare clinical information obtained from medical records of cats with methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible S aureus (MSSA) infections, evaluate antibiograms of MRSA and MSSA for multiple-drug resistance (MDR), and characterize the strain type and staphylococcal chromosome cassette (SCC)mec type of each MRSA. SAMPLE POPULATION: 70 S aureus isolates obtained from 46 cats. PROCEDURES: Clinical information obtained from medical records, including signalment, clinical signs, histologic examination of affected tissues, and outcomes, was compared between the 2 groups. Composite antibiograms of MRSA and MSSA were compared statistically. The MRSA strains were characterized by use of pulsed-field gel electrophoresis and SCCmec typing. RESULTS: No statistical differences in signalment or subjective differences in clinical signs or outcomes were detected between groups with MRSA or MSSA infection. Significant differences in antimicrobial resistance were detected, with MRSA having complete resistance to fluoroquinolone and macrolide antimicrobials, whereas MSSA maintained a high frequency of susceptibility. Seven pulsed-field patterns were observed in 15 MRSA strains; all but 1 were highly related. All MRSA isolates contained a type II SCCmec element. CONCLUSIONS AND CLINICAL RELEVANCE: Because MDR cannot be predicted in staphylococcal infections in cats on the basis of clinical signalment, culture and susceptibility testing are recommended whenever initial empirical treatment is unsuccessful. Molecular characterization of MRSA strains suggests that there has been reverse-zoonotic transmission from humans. IMPACT FOR HUMAN MEDICINE: The SCCmec type II element is typically associated with nosocomial MRSA infections of people. Cats may serve as reservoirs for MRSA infections in humans.     

饲用羽毛肽粉营养成分分析

采用国标及通用的方法,对饲用羽毛肽粉中各营养成分进行分析,并与其它饲料蛋白源进行比较。结果表明,饲用羽毛肽粉粗蛋白、粗脂肪、糖分和粗灰分、盐分含量分别为85.05%、7.9%、1%7、.76%、0.9%。饲用羽毛肽粉含有18种氨基酸,总氨基酸、必需氨基酸和呈味氨基酸分别占总量的70.47%、27.78%、38.70%,总氨基酸占蛋白质的82.86%,必需氨基酸和呈味氨基酸分别占氨基酸总量的39.42%和54.92%。饲用羽毛肽粉作为鱼粉替代品的限制性氨基酸为赖氨酸(Lys)、组氨酸(His)、色氨酸(Trp)、蛋氨酸+胱氨酸(Met+Cys)。饲用羽毛肽粉含14种脂肪酸,不饱和脂肪酸、单不饱和脂肪酸、多不饱和脂肪酸、ω-3不饱和脂肪酸分别占脂肪酸总量的25%、15%、10%、2%,EPA占脂肪酸含量的2%,未发现含有DHA。饲用羽毛肽粉含有丰富的营养成分,但其作为水产动物饲料蛋白源在氨基酸平衡及不饱和脂肪酸含量较鱼粉差,可通过调整营养配方,改善其应用价值。     

Prevalence and mechanism of antimicrobial resistance in Staphylococcus aureus isolates from diseased cattle, swine and chickens in Japan

Antimicrobial administration is essential for the control and treatment of diseases in animals, but the emergence and prevalence of antimicrobial-resistant Staphylococcus aureus is a significant concern during animal production. Here we investigated the antimicrobial susceptibility of S. aureus from diseased food-producing animals and molecularly characterized the methicillin-resistant and fluoroquinolone-resistant isolates. A total of 290 S. aureus isolates obtained from cattle (n=246), swine (n=16), and chickens (n=28) between 2003 and 2009 were examined for antimicrobial susceptibility against 9 antimicrobials using an agar dilution method. Resistance to penicillin (PC) was most frequently found (24.8%), followed by oxytetracycline (OTC, 10.0%), dihydrostreptomycin (4.1%), erythromycin (EM, 3.1%), enrofloxacin (ERFX, 2.1%), and kanamycin (1.7%). The PC resistance rate was significantly higher in swine than in cattle (P<0.01) and chickens (P<0.01). The resistance rates to OTC, EM and ERFX were significantly higher in swine and chickens than in cattle (P<0.05). Methicillin-resistant S. aureus (MRSA) was recovered from milk derived from a cow with mastitis in 2003; sequence type 8, SCCmec type IV and spa type t024. In the six ERFX-resistant strains isolated after 2003, amino acid substitutions in ParC with/without GyrA were detected. As the prevalence of MRSA and FQ-resistant S. aureus in the animals should be noticed, continuous monitoring is necessary to control resistance to clinically important antimicrobials in S. aureus from food-producing animals.     

Methicillin‐resistant Staphylococcus aureus in Resident Animals of a Long‐term Care Facility

Animals provide benefits to elderly and chronically ill people by decreasing loneliness, increasing social interactions, and improving mental health. As a result, many hospitals and long‐term care facilities allow family pets to visit ill or convalescing patients or support animal‐assisted therapy programs. These include programs that have resident animals in long‐term care facilities. Despite the benefits, there are concerns about disease transmission between pets and patients. Antibiotic‐resistant bacteria, such as methicillin‐resistant Staphylococcus aureus (MRSA), are a recognized problem in healthcare settings leading to refractory infections and potentially life‐threatening illnesses. MRSA has been isolated from numerous animal species, yet few studies are available on the carriage of this pathogen in animals residing in long‐term care facilities. Our objective was to characterize MRSA carriage among resident animals in a long‐term care facility. Methods: To document MRSA colonization, nasal swabs from 12 resident animals (one dogs and 11 cats) of a long‐term care facility were collected weekly for 8 weeks. Staphylococcus isolates were characterized by antimicrobial susceptibility and MRSA isolates were further characterized by pulsed‐field gel electrophoresis (PFGE). PFGE isolate patterns were compared with an existing database of MRSA isolate patterns at the Minnesota Department of Health. Results: Two of 11 cats were colonized with MRSA. MRSA was recovered from five of eight weekly samples in one cat and two of eight weekly samples in the other cat. All isolates were classified as USA100 (healthcare‐associated strains). Discussion: Long‐term care resident animals may acquire MRSA. Clonally related strains were identified over the 8‐week sampling period. It is unclear if pets serve as an on‐going source of infection to their human companions in long‐term care facilities.     

Insulin‐independent actions of glucagon‐like peptide‐1 in wethers

Insulin‐independent actions of glucagon‐like peptide‐1 (GLP‐1) are not yet clear in ruminants. Four Suffolk mature wethers (60.0 ± 6.7 kg body weight (BW)) were intravenously infused with insulin (0.5 mU/kg BW/min; from 0 to 90 min) and GLP‐1 (0.5 μg/kg BW/min; from 60 to 150 min) with both hormones co‐administered from 60 to 90 min, in a repeated‐measure design under euglycemic clamp for 150 min, to investigate whether GLP‐1 has insulin‐independent actions. Jugular blood samples were taken at 15‐min intervals for plasma hormones and metabolites analysis. Compared to baseline concentrations (at 0 min), insulin infusion decreased (P < 0.05) plasma concentrations of glucagon, non‐esterified fatty acids (NEFA), lactate, nonessential amino acids (NEAA), branched‐chain amino acids (BCAA), total amino acids (TAA) and urea nitrogen (UN). Insulin plus GLP‐1 infusion induced a greater increase (P < 0.05) in plasma concentrations of insulin and triglyceride (TG), but decreased (P < 0.05) glucagon, total cholesterol (T‐Cho), NEAA and UN plasma concentrations. GLP‐1 infusion increased (P < 0.05) NEFA, β‐hydroxybutyrate and TG, but decreased (P < 0.05) glucagon, T‐Cho, NEAA, BCAA and UN plasma concentrations. In conclusion, GLP‐1 exerts extrapancreatic roles in ruminants not only insulin‐independent but probably, in contrast to non‐ruminants, antagonistic to insulin effects.     

A review of the characteristics and treatment of methicillin‐resistant Staphylococcus aureus (MRSA) in the horse and a case series of MRSA infection in four horses

Methicillin‐resistant Staphylococcus aureus (MRSA) is an emerging cause of serious bacterial infection in the horse, with an increasing number of cases reported over the last decade. MRSA, along with other commensal staphylococcal species, can reside on the mucosa of several sites in the horse, particularly the nose. Nasal carriage of MRSA appears rare amongst horses in the community, although a higher prevalence has been found in hospitalised horses. MRSA infections can involve a variety of body sites, but most commonly encountered are soft tissue infections of either traumatic or surgical wounds. MRSA strain types isolated from horses are typically multidrug‐resistant and usually differ from those recovered from humans and other small animal species. Treatment of infection can be prolonged and is dependent on timely, accurate diagnosis and on appropriate therapy; often guided by antimicrobial susceptibility testing. The purpose of this review is to provide clinically relevant information for the equine practitioner and, for illustration, the diagnosis, treatment and outcome of 4 clinical cases of MRSA infection in horses is discussed.     

Antimicrobial activity of cathelicidins BMAP28, SMAP28, SMAP29, and PMAP23 against Pasteurella multocida is more broad-spectrum than host species specific

The antimicrobial activity of linear, cationic alpha-helical peptides from cattle (BMAP28), sheep (SMAP28 and SMAP29), and pigs (PMAP23) were assessed to determine if activity was selective for Pasteurella multocida from a particular animal species or broad-spectrum against all P. multocida tested. The antimicrobial activities of synthetic peptides were determined for P. multocida isolated from cattle (10 isolates), sheep (10 isolates), and pigs (10 isolates) in a broth microdilution assay. All thirty isolates of P. multocida were susceptible to BMAP28 (MICs and MBCs, 1.0-1.9 microM); SMAP28 and SMAP29 (MICs and MBCs, 0.2-0.7 microM); and PMAP23 (MICs and MBCs, 4.3 to > or = 6.8 microM). Overall, the results of this study suggest that synthesized cathelicidins from cattle, sheep, and pigs had broad-spectrum antimicrobial activity against all P. multocida.     

Comparison of three markers for the determination of bacterial protein in terminal ileal digesta in the growing pig

The aim of the study was to compare three methods commonly used to determine the concentrations of bacterial protein in digesta collected from the terminal ileum of growing pigs that had been fed a casein‐based diet. The amounts of bacterial protein in terminal ileal digesta were determined using three different markers: 2.6‐diaminopimelic acid (DAPA) and the d ‐amino acids, d ‐aspartic acid (d ‐Asp) and d ‐glutamic acid (d ‐Glu). The effectiveness of each marker was compared against a control based on physical fractionation by centrifugation. The total bacterial protein concentrations derived from the markers d ‐Asp and d ‐Glu were significantly different (p = 0.05) to those calculated from DAPA and the control, but there was no difference between DAPA and the control. The percentage of bacterial nitrogen ranged from 40% to 52% dependent on the marker used. Bacterial protein expressed as a percentage of the total protein, ranged from 48% to 62%, a substantial proportion of which (12–28%) was derived from lysed bacterial cells. Statistical correlations between the estimation methods were low. Such poor correlation between the markers may be the result of random errors such as variance in the epimerization of the two d ‐amino acids during protein hydrolysis. DAPA was accepted as a reliable marker for determining microbial protein in ileal digesta.     

Genome‐wide association study for fatty acid composition in Japanese Black cattle

Fatty acid composition is one of the important traits in beef. The aim of this study was to identify candidate genomic regions for fatty acid composition by genome‐wide association study with 50 K single nucleotide polymorphism (SNP) array in Japanese Black cattle. A total of 461 individuals and 40 657 SNPs were used in this study. We applied genome‐wide rapid association using mixed model and regression (GRAMMAR) and genomic control approaches to estimate the associations between genotypes and fatty acid composition. In addition, two SNPs in fatty acid synthase (FASN) (T1952A) and stearoyl‐CoA desaturase (SCD) (V293A) genes were also genotyped. Association analysis revealed that 30 significant SNPs for several fatty acids (C14:0, C14:1, C16:1 and C18:1) were located in the BTA19 FASN gene located within this region but the FASN mutation had no significant effect on any traits. We also detected one significant SNP for C18:1 on BTA23 and two SNPs for C16:0 on BTA25. The region around 17 Mb on BTA26 harbored two significant SNPs for C14:1 and SNP in SCD in this region showed the strongest association with C14:1. This study demonstrated novel candidate regions in BTA19, 23 and 25 for fatty acid composition.     

Expression analyses of candidate genes related to meat quality traits in squabs from two breeds of meat‐type pigeon

In this study, meat quality traits were compared between squabs from two pigeon breeds: one Chinese indigenous breed, the Shiqi (SQ) meat‐type pigeon, and an imported breed, the white king (WK) meat‐type pigeon. Breed differences were detected in the content of intramuscular fat (IMF) in the breast muscle. SQ squabs had significantly higher IMF content than the WK birds. The shear force value (an objective measure of meat tenderness) of SQ birds was also relatively lower than that of the WK squabs. Further analysis of fatty acids profile revealed that SQ squabs exhibited significant advantage in the synthesis of polyunsaturated fatty acids, while WK squabs were significantly higher in the sum of monounsaturated fatty acids. Breast muscle in the SQ squabs was also significantly higher in the ratio of polyunsaturated fatty acids to saturated fatty acids, as well as the sum of omega 6 fatty acids. Variability of expression levels of functional genes in relation to fat accumulation and meat tenderness was analysed by qRT‐PCR. Gene expression analyses showed that the hepatic expression of LPL (lipoprotein lipase), FABP4 (fatty acid‐binding protein 4), and CAPN2 (calpain‐2) were significantly higher in the SQ squabs. In the breast muscle tissue, the FABP3 (fatty acid‐binding protein 3) and CAPN2mRNA abundance was significantly higher in SQ squabs. Our results suggested that these differentially expressed genes might be candidate genes used in the programmes of targeted selection for squabs with higher IMF content, tender meat, and more favourable fatty acids composition.     

Characterization of the cathelicidin cluster in the Japanese quail (Coturnix japonica)

The Japanese quail has several advantages as a low‐fat meat bird with high immunity against diseases. Cathelicidins (CATHs) are antimicrobial peptides that play an important role in innate immunity. The aim of this study was to characterize the CATH cluster in the Japanese quail (Coturnix japonica). The Japanese quail CATH (CjCATH) cluster, contains four CATH genes, as in the chicken. The coding sequences of CjCATHs exhibited >85.3% identity to chicken CATHs. The predicted amino acid sequences of the four CjCATH genes contained the cathelin‐like domain characteristic of CATH proteins. Polymorphisms were detected in the open reading frames (ORFs) of all CjCATH sequences. Two amino acid substitutions were observed in the antimicrobial region of the mature peptide of CjCATH2, and predicted to influence peptide function. CjCATH1 is expressed in lung, heart, bone marrow and bursa of Fabricius (BF). CjCATH2 is expressed in bone marrow. CjCATH3 is expressed in lung, heart, bone marrow, BF, tongue and duodenum. CjCATHB1 is expressed in bone marrow and BF. This study is the first to characterize CATH genes in the Japanese quail, and identifies novel antimicrobial peptide sequences belonging to the cathelicidin family, which may play a role in immunity in this species.     

早期断奶对藏西北绒山羊背最长肌氨基酸和脂肪酸含量的影响

[目的] 研究早期断奶对藏西北绒山羊背最长肌中氨基酸和脂肪酸含量的影响。[方法] 将18只健康羔羊按照体重相近原则随机分为试验组和对照组,试验组在28日龄（D28）实施早期断奶,对照组在60日龄（D60）自然断奶,羔羊断奶后饲喂相同的代乳料至75日龄;从两组随机选取5只羔羊进行屠宰采样,对其背最长肌中氨基酸和脂肪酸指标进行测定分析。[结果] 在藏西北绒山羊背最长肌中检测出17种氨基酸,包括赖氨酸、亮氨酸等7种必需氨基酸,天冬氨酸、精氨酸等10种非必需氨基酸。两组断奶羔羊肉中总氨基酸含量、必需氨基酸含量、非必需氨基酸含量均无显著差异（P>0.05）,28日龄断奶组羔羊肉中EAA/TAA、EAA/NEAA比值显著（P<0.05）高于60日龄断奶羔羊。共检测出29种饱和脂肪酸、不饱和脂肪酸,以棕榈酸和油酸含量最高,早期断奶对羔羊肉中饱和脂肪酸含量、单不饱和脂肪酸含量以及多不饱和脂肪酸含量影响不显著（P>0.05）。[结论] 藏西北绒山羊羔羊肉中所含氨基酸和脂肪酸种类多、含量丰富,实施早期断奶、及早补充颗粒料对肉中脂肪酸和氨基酸含量无影响,且可提高EAA/TAA、EAA/NEAA比值,使其更符合FAO/WHO氨基酸评分模式;该试验可为藏西北绒山羊羔羊实施早期断奶、提高母羊的繁殖率提供参考。     

Effects of a DHA‐rich unextracted microalgae as a dietary supplement on performance,carcass traits and meat fatty acid profile in growing‐finishing pigs

Two 125‐day experiments of the same design were conducted to evaluate the effects of a heterotrophically grown microalgae (AURA) containing docosahexaenoic acid (DHA) on pig performance, carcass traits and the fatty acid composition of lean and adipose tissue. In each experiment, 144 Hypor pigs were blocked by sex, allocated to three treatment groups, and fed 0, 0.25% or 0.50% AURA in isonutritive, isocaloric diets. Pigs were weighed on days 0, 28, 56, 84 and 112. Feed and water intakes were measured every 28 days. Pigs were slaughtered on day 125. Data from the two studies were analysed as a single data set. Performance and carcass traits did not differ between treatments. Both microalgae treatment levels enriched (p < .05) Longissimus lumborum (LL) and backfat in DHA and improved (p < .05) their ratios of n‐6 to n‐3 fatty acids.     

Characterization and expression of non‐polymorphic liver expressed antimicrobial peptide 2: LEAP‐2 in the Japanese quail,Coturnix japonica

Liver‐expressed antimicrobial peptide 2 (LEAP‐2) is a cationic peptide that plays an important role in innate immunity for host defense. The aim of this study was to characterize the LEAP‐2 gene in the Japanese quail (Coturnix japonica). Japanese quail LEAP‐2 (CjLEAP‐2) was identified from the Japanese quail draft genome database by a local BLAST analysis using chicken LEAP‐2 (GgLEAP‐2). The exon‐intron structure of CjLEAP‐2, analyzed from three quails, is composed of three exons, as is the chicken LEAP‐2 homolog (GgLEAP‐2). An analysis of the coding sequence revealed that CjLEAP‐2 is 231 bp long, like GgLEAP‐2, and 93% identical to GgLEAP‐2 at the nucleic acid level. The predicted amino acid sequence of CjLEAP‐2 contained the liver‐expressed antimicrobial peptide 2‐precursor domain and four cysteine residues characteristic of the LEAP‐2 protein. The amino acid sequence of the mature peptide of CjLEAP‐2 was 100% identical to that of GgLEAP‐2. We confirmed that CjLEAP‐2 was transcribed in at least seven tissues, including the digestive system. Additionally, the mature peptide region of CjLEAP‐2 exhibited no polymorphisms in 99 quails from six strains. Taken together, these findings indicate that CjLEAP‐2 is non‐polymorphic and therefore, it likely plays an important role in the innate immunity of quail as it does in chicken.     

Detection of methicillin‐resistant Staphylococcus aureus (MRSA) in the microbial flora from the conjunctiva of healthy donkeys from Sicily (Italy)

Objective To describe the bacterial flora present in the normal conjunctiva of donkeys from Sicily (Italy). Animals studied A total of 46 healthy donkeys housed in 3 locations within the territory of Palermo (Sicily, Italy) were studied. Donkeys ranged from 2 to 13 years of age, with a median age of 6 years. Procedures Forty‐six conjunctival swabs were obtained from both eyes of each animal, and specimens were cultured for aerobic bacteria. Furthermore, the antimicrobial activity of methicillin (1 μg) and oxacillin (5 μg) on Staphylococcus spp. isolates was evaluated, and a specific PCR assay, which allows the detection of mecA gene specific for methicillin‐resistant Staphylococcus aureus (MRSA) strains, was performed. Results Forty of 46 (86.9%) donkeys were positive for bacteria. Eighty bacterial isolates, representing 9 bacteria genera, were successfully cultured. The most frequently recovered bacterial genus was Staphylococcus (52/80 isolates; 65%). Several strains (20/80 isolates; 25%) belonging to the Enterobacteriaceae family were also isolated, among which the most frequently isolated genus was Enterobacter (eight isolates). Of the 52 Staphylococcus spp. isolates, 14 (26.9%) strains were oxacillin/methicillin resistant. The mecA gene was detected in 6/52 (11.5%) strains. Conclusions This study contributes to the knowledge about normal ocular flora and MRSA occurrence in donkey farms in Sicily.     

Effects of dietary n‐6/n‐3 fatty acid ratio on nutrient digestibility and blood metabolites of Hanwoo heifers

The objectives of this study were to investigate the effects of dietary n‐6/n‐3 fatty acid (FA) ratio on digestibility, blood metabolites and FA profile of Hanwoo heifers. Fifteen Hanwoo heifers (22 ± 3 months old; 357 ± 69.7 kg) were randomly assigned to three dietary treatments with n‐6/n‐3 FA ratios of 2.07, 5.18 and 7.37. The animals were housed individually in digestion crates and fed total mixed rations at 2.2% of body weight for 2 weeks of adaptation and 1 week of collection. Treatment effects on in vivo digestibility, plasma metabolite and fatty acid profiles, and in vitro ruminal fermentation and fatty acid profiles were examined. In vivo digestibility was not affected (P > 0.05) by dietary n‐6/n‐3 FA ratio. However, in vitro dry matter digestibility and concentrations of total volatile fatty acids and propionate decreased (P < 0.05) linearly with increasing n‐6/n‐3 FA ratio. Plasma insulin and progesterone increased linearly (P < 0.05), but linolenic acid and total n‐3 FA decreased linearly (P < 0.05) with increasing n‐6/n‐3 ratio. Increasing the dietary n‐6/n‐3 FA ratio can increase the n‐6/n‐3 FA ratio in plasma and ruminal fluid as well as plasma progesterone secretion.