• Asian-Australasian Journal of Animal Sciences
• /
• v.21 no.1
• /
• pp.124-130
• /
• 2008

Streptococcus (Strep.) agalactiae is one of the major pathogens of bovine mastitis and is the main cause of subclinical infection. This study attempted to develop a rapid PCR diagnosis procedure using milk samples collected on filter paper disks. Chromatographic filter paper was employed as the preservation media and kept at room temperature for one to four weeks. The revival rate of Strep. agalactiae kept on dried filter paper disks was affected by the pretreatment preservation time. The revival test suggested that not all the bacteria in artificially contaminated milk samples on the filter paper disks could be recovered. After that, a PCR based on the 16-23S intergenic spacer region of Strep agalactiae was performed. The results distinguished the strep. agalactiae from major pathogens of bovine mastitis at a $2{\times}10^2$ colony forming units (CFU)/ml level, which showed similar sensitivity to the results from liquid milk samples. The results also showed that milk samples collected on filter paper disks could be kept at room temperature for one to four weeks with little negative effect on sensitivity and specificity. The field test showed that the diagnostic sensitivity and specificity was 96.15% and 98.60%, respectively. In conclusion, the protocol will provide a rapid and economic procedure for the detection of bovine mastitis.

• Korean Journal of Veterinary Research
• /
• v.47 no.1
• /
• pp.67-75
• /
• 2007

Environmental mastitis has increased particularly in well-managed or low somatic cell countherds that have successfully controled contagious pathogens. Major pathogens of environmental mastitisare Escherichia coli (E. coli) and Streptococcus uberis. The present study was conducted to investigate1,865 quaters of 241 Korean dairy farms from 2001 to 2004. Prevalence of major gram-negative bacteriaisolated from mastitis milk were E. coli (22.7%) and Enterobacter spp. (16.3%) in coliforms and Pseudomoassp. (10.3%) and Serratia spp. (7.9%) in non-coliforms. The results on antibiotic susceptibility by agardifusion test against these pathogens were 86.7% in piperaciliin, 94.6% in cefepime, 85.5% in amikacin,87.7% in gentamicin and so on. In contrast, the susceptibility against ampicillin (41.9%), cephalothin (9.9%),streptomycin (39.9%) and tetracycline (46.7%) appeared to be below 50%. Gram-negative bacteria showed(96.8%). Acording to year, distribution of high $256{\sim}64{\mu}g/ml$ on cephalothin get increased, but the othersare diferent. These findings demonstrate that major gram-negative bacteria were E. coli and Enterobacterspp. isolates, and often encountered the diverse antibiotic resistant patterns.

• Korean Journal of Veterinary Research
• /
• v.29 no.3
• /
• pp.383-391
• /
• 1989

This study was carried out to diagnostic test for bovine mastitis by the determination of adenosine triphosphate (ATP) based on firefly bioluminescence. The results obtained are follow; 1. The infection rate of bovine mastitis investigated with 521 cows in 47 dairy farms were found to be 3.6% of clinical form and 44.1% of subclinical form according to the degree of infection. 2. The light yield produced in firefly bioluminescence system was proportional to the concentration of ATP giving stright line within the range of 100PM~1uM. 3. When the number of somatic cell in milk was determined by the ATP assay and compared with three conventional methods such Fossomatic, California mastatic test (CMT), and rolling ball viscometer (RBV), it was shown that r=0.92 for Fossomatic, 0.63 for CMT and 0.7 for RBV. 4. The microorganisms causing mastitis were isolated Staphylococcus sp. (53.3%), Streptococcus sp. (17.9%), Micrococcus sp. (13.5%), Gram negative bacilli (6.3%), Gram positive bacilli (5.5%) and Yeast-like fungi (5.4%). 5. The endogeneous ATP levels of bacteria in a raw milk determined by the firefly bioluminescence system and compared with the results of the conventional methods. The correlation was 0.88 for raw milk.

• Korean Journal of Veterinary Service
• /
• v.30 no.1
• /
• pp.115-123
• /
• 2007

The therapeutic effect of honeybee venom collected using be venom collector on bovine mastitis was investigated. Mastitic cows from four farms were selected in the Yang-pyeong areas. Chronic mastitic cows were injected with the various concentrations of honey-bee venom per day. There was a significant difference in the reduction rates of somatic cell counts (SCC) according to treatment concentration and method of bee venom. The milk SCC were significantly decreased in all concentrations of bee venom 3 days after treatment. The reduction rates of SCC in treatment of 3, 6, 12 and 24mg honeybee venom were 20, 43, 63.3 and 65.8% respectively. Honeybee venom treatment consisted of two methods, a syringeful and a Bovivet Spenstift. The treatment with Bovivet Spenstift was more effective in the reduction rates of SCC compared with the syringeful. Thirty two out of 53 quarters were cured by Bovivet Spenstift with 12mg bee venom per day for 14 days. The venom cure rates of bovine mastitis by Escherichia coli, Stapylococcus aureus, Gram positive bacteria and Gram negative bacteria were 33.3, 75, 75 and 43.8% respectively. These results suggested that bee venom treatment (by Bovivet Spenstift with 12mg) might be effective for treatment of bovine mastitis.

• Korean Journal of Veterinary Research
• /
• v.55 no.4
• /
• pp.253-257
• /
• 2015

Oregano (Origanum vulgare) is an aromatic herb belonging to the Lamiaceae family. This study evaluated the therapeutic effect of oregano essential oil (OEO) on clinical bovine mastitis caused by Staphylococcus (S.) aureus and/or Escherichia (E.) coli. Thirty-two quarters from 18 lactating cows with subclinical mastitis were selected from a dairy farm and were divided into 4 groups (8 quarters/group): negative control (CON), positive control (GEN) and two OEO-treated groups (OEO-1 and OEO-2). Each group was intramammarily treated with saline, 10 g/tube of gentamicin ointment, and single and double doses of 0.9 mL OEO ointment twice a day for three days, respectively. After the treatments, physical udder conditions were greatly improved in GEN and OEO groups. In CON group, somatic cell counts (SCCs), number of white blood cells (WBC) and bacteria were increased. However, in OEO groups, SCCs and number of WBC were significantly decreased and S. aureus and E. coli were not detected in milk as compared with those before the treatments as well as GEN group. These results suggest that OEO may be a useful alternative to antibiotics for the control of subclinical bovine mastitis caused by S. aureus and/or E. coli.

• Asian-Australasian Journal of Animal Sciences
• /
• v.25 no.3
• /
• pp.353-360
• /
• 2012

Mastitis is a highly morbid disease that requires detection at the subclinical stage. Tropical countries like India mainly depend on milch buffaloes for milk. The present study was conducted to investigate whether the trace minerals viz. copper (Cu), iron (Fe), zinc (Zn), cobalt (Co) and manganese (Mn) and enzyme activity of lactate dehydrogenase (LDH), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) in riverine buffalo milk can be used as an indicator of subclinical mastitis (SCM) with the aim of developing suitable diagnostic kit for SCM. Trace elements and enzyme activity in milk were estimated with Atomic absorption Spectrophotometer, GBC 932 plus and biochemical methods, respectively. Somatic cell count (SCC) was done microscopically. The cultural examination revealed Gram positive bacteria as the most prevalent etiological agent. A statistically significant (p<0.01) increase in SCC, Fe, Zn, Co and LDH occurred in SCM milk containing gram positive bacterial agents only. ALP was found to be elevated in milk infected by both gram positive and negative bacteria. The percent sensitivity, specificity and accuracy, predictive values and likelihood ratios were calculated taking bacterial culture examination and $SCC\geq2{\times}10^5$ cells/ml of milk as the benchmark. Only ALP and Zn, the former being superior, were found to be suitable for diagnosis of SCM irrespective of etiological agents. LDH, Co and Fe can be introduced in the screening programs where Gram positive bacteria are omnipresent. It is recommended that both ALP and Zn be measured together in milk to diagnose buffalo SCM, irrespective of etiology.

• Korean Journal of Veterinary Research
• /
• v.24 no.1
• /
• pp.91-98
• /
• 1984

A total of 466 foremilk from dairy farms in Chonnam district was examined for the subclinical mastitis over a period of one year, using a method of the electrical conductivities(EC); absolute conductivity(AC) and differential conductivity(DC) and quarter difference value(QD), in relation to the California mastitis test(CMT) and the direct somatic cell count(DSCC). The compatibility and efficiency rating between the EC values and the other screening tests was conducted. Obtained results are summarized as follows. 1. A linear relationship was found between the EC values and the CMT scores and direct somatic cell counts and it was found that electrical conductivity measurements were comparable with other screening tests for diagnosing animals with mastitis. 2. Compatibilities between the EC and CMT were 70.4% in AC, 74.6% in DC and 70.7% in QD, and that of the EC and DSCC were 53.0% in AC, 63.1% in DC and 53.2% in QD. On the other hand, relative efficiency ratings of Postle's equation between EC and CMT were 37.3% in AC, 26.5% in DC and 13.6% in QD, and that of the EC and DSCC were 33.1% in AC, 20.2% in DC and 11.9% in QD. 3. In the foremilk samples collected from damaged quarters determined by EC, the false positive rate wart higher than the false negative rate, and consequently tests of EC produced lower compatibility or efficiency rating scores. These tendencies suggested that any factors other than the mastitic condition influencing the EC values might be existed.

• Korean Journal of Veterinary Research
• /
• v.47 no.1
• /
• pp.33-41
• /
• 2007

Streptococcus spp. comprising Streptococcus (S.) uberis S. dysgalactiae strains is major causeof bovine mastitis from particularly well-managed or low somatic cell count herds that have successfullycontrolled contagious pathogens. In this study, antimicrobial susceptibility and genetic characteristics of S.uberis isolated from clinical or subclinical mastitis milk at 2003 were investigated. Eighty seven isolatesof Streptococus spp. were identified by the conventional biochemical methods. The antimicrobialsusceptibility by disk diffusion method was determined for 46 S. uberis, 11 S. bovis, 10 S. oralis, 6 S.uberis and 14 other Streptococcus spp.. Overall, the tested strains were susceptible to tetracycline (11.5%),amikacin (14.9%), streptomycin (16.1%), neomycin (26.4%), kanamycin (35.6%), gentamicin (65.2%),oxacillin (70.1%), ampicillin (75.9%), chloramphenicol (78.2%), and cephalothin (97.7%). Additionally, S.uberis strains were susceptible to pencillin G (97.8%), but resistant to erythromycin (76.0%) by minimalinhibitory concentration test. The multiple-drug resistance rate of isolated bacteria to 4 more thanamplification fingerprinting patterns amplifed with primer 8.6d showed that 3 to 8 number of distinguishableDNA fragments ranged from 180 bp to 1,20 bp. Thirty seven isolates of S. uberis strains were subtypedinto 8 distinct patterns. Each subtype revealed a typical pattern of antimicrobial susceptibilities. Thesefindings demonstrate that S. uberis isolates were mastitis pathogens of diverse serotypes, and oftenencountered the diverse resistant patterns.

• Journal of Veterinary Clinics
• /
• v.25 no.5
• /
• pp.346-354
• /
• 2008

The combined effects of EDTA-Tris and eighteen antimicrobial agents have been evaluated in eight clinically isolated methicillin-resistant bacteria (Staphylococcus aureus, Escherichia coli, Streptococcus uberis and Streptococcus agalactiae) from bovine mastitis. The antimicrobial activity was evaluated by measuring the minimal bactericidal concentration (MBC) for the antibiotics alone or in combination with EDTA-Tris in Mueller-Hilton broth and milk. Combined use of EDTA-Tris and antibiotics potentiated or antagonized activity of antibiotics against mastitic pathogens. Milk increased the antibiotic potency of erythromycin and spiramycin on S. aureus. Culture in milk changed patterns of EDTA-Tris combinational effects compared with that in standard Mueller-Hilton broth. Combined with EDTA-Tris in milk, synergic effects were observed in colistin, dihydrostreptomycin, kanamycin, erythromycin, gentamycin, oxytetracycline, streptomycin to E. coli, Str. uberis, and Str. agalactiae. However, significant antagonistic effects of milk on antibiotic susceptibility in combination with EDTA-Tris were noted in neomycin, streptomycin, penicillin, roxithromycin, and amoxicillin. This study indicates that combination therapy of EDTA-Tris with antibiotics in bovine mastitis should be used with caution because of the possible antagonistic effects of antibiotic combination with EDTA-Tris on mastitic pathogens. In addition, antibiotic susceptibility test in combination with EDTA-Tris in milk culture condition can be benefit in search of effective treatment regimen for some antibiotic-resistant bacteria of mastitis.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.14 no.2
• /
• pp.137-142
• /
• 2007

The antibacterial activity of Korean honey bee venom (KBV) was examined against seven major bacterial mastitis pathogens, Enterococcus faecium, Escherichia coli, Methicillin resistant Staphylococcus aureus (MRSA), Staphylococcus aureus, Streptococcus intermedius, Streptococcus oralis and Streptococcus uberis isolated from infected mammary quarters of cows. Seven bacterial mastitis pathogens were studied for antibacterial activity of the KBV by disc diffusion assay, minimal inhibitory concentrations (MIC) and bacterial count in milk samples. The KBV showed activity against Ent. faecium, E. coli, MRSA, Staph. aureus, Strep. intermedius. The order of susceptibility of the bacteria against the KBV was Staph. aureus>MRSA>E. coli>Strep. intermedius>Ent. faecium>Strep. oralis>Strep. uberis. The MIC against Staph. aureus, MRSA and E. coli were stronger effect as compared with standard drug. The effect of the KBV ($100{\mu}g\;ml^{-1}$) on the viability of Ent. faecium, E. coli, MRSA, Staph. aureus, Strep. intermedius, Strep. oralis and Strep. uberis in milk differed significantly with each other within 12 h incubation period. The results indicate that KBV has significant antibacterial effects against major bacterial mastitis bacteria, Ent. faecium, E.coli, MRSA, Staph. aureus, Strep. intermedius. Results of the study indicate the potential use of KBV as alternative to antibiotic therapy. Further investigations are needed though to confirm its efficacy and its effects on the animals.