• 제목/요약/키워드: Subclinical mastitis

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Evaluation of microbiological, cellular and risk factors associated with subclinical mastitis in female buffaloes

  • de Oliveira Moura, Emmanuella;do Nascimento Rangel, Adriano Henrique;de Melo, Maria Celeste Nunes;Borba, Luiz Henrique Fernandes;de Lima, Dorgival Morais Junior;Novaes, Luciano Patto;Urbano, Stela Antas;de Andrade Neto, Julio Cesar
    • Asian-Australasian Journal of Animal Sciences
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    • 제30권9호
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    • pp.1340-1349
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    • 2017
  • Objective: This study aimed to evaluate the microbiological and cellular milk profile for the diagnosis of subclinical mastitis in female buffaloes and to assess risk factors for predisposition of the disease. Methods: Analyses were carried out by standard plate count (SPC), identification of species and antibiotic resistance, somatic cell count (SCC), electrical electrical conductivity of milk (ECM), and lactoferrin content in milk. Teat cups were swabbed to evaluate risk factors, observing hyperkeratosis, milking vacuum pressure and cleanliness of the site. Hence, 30 female buffaloes were randomly selected (15 from a group in early lactation and 15 in late lactation). Results: The most common bacteria in the microbiological examination were Staphylococcus spp., Streptococcus spp. and Corynebacterium sp. In the antibiotic sensitivity test, 10 (58.82%) of the 17 antibiotics tested were sensitive to all isolates, and resistant bacteria were Streptococcus uberis, Streptococcus dysgalactiae, Streptococcus haemolyticus, and Escherichia coli. It was observed that positive samples in the microbiological examination showed total bacterial count between $9.10{\times}10^3$ to $6.94{\times}10^6$ colony forming units/mL, SCC between 42,000 to 4,320,000 cells/mL and ECM ranging from 1.85 to 7.40 mS/cm. It was also found that the teat cups had high microbial counts indicating poor hygiene, and even faults in the cleanliness of the animals' waiting room were observed. It is concluded that values of SCC above 537,000 cells/mL and ECM above 3.0 mS/mL are indications of mammary gland infection for this herd; however, the association of these values with a microbiological analysis is necessary to more accurately evaluate the health status of mammary glands with subclinical mastitis. Conclusion: Through phenotypic characterization of bacteria involved in the samples, the genera Staphylococcus spp., Streptococcus spp., and Corynebacterimum bovis were the most prevalent in this study. Faults in environment and equipment hygienization are factors that are directly associated with mastitis.

전기전도도측정에 의한 유우준임상형 유방염의 진단에 관한 연구 2. 전기전도도치의 변동요인 (Studios on the Diagnosis of Subclinical Mastitis in Dairy Cows by the Measurement of the Electrical Conductivity 2. Factors Influencing Electrical Conductivity Value)

  • 강병규;신종봉
    • 한국임상수의학회지
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    • 제2권1호
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    • pp.115-120
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    • 1985
  • To probe the subclinical mastitis in a herd of cows in Chonnam district, the electrical conductivity(EC) of 825 foremilk samples were measured for 2 years. Normal (n=487) and mastitic(n=110) foremilk samples were classified by the California mastitis test (CMT) and direct somatic cell count(DSCC) and investigated the relations between the changes of the EC value and the calving history, age, days of postpartum, estrus and causative organism isolated. Obtained results are summarized as follows. 1. In the normal foremilk samples, positive correlation, though not significant, was found between the EC value and calving history (r=0.573) and age (r=0.247). 2. In the normal foremilk samples, the EC values were lowered at 30~120 days of postpartum through the whole lactation period and revealed a tendencies to higher values following the day of postpartum increased untill to the drying off (r=0.823), and the days of postpartum was recognized as one of a influencing factor on the EC value(p<0.05). 3. In the mastitic foremilk samples, significant correlation between EC value and resazurin reduction test (RRT) were observed (r=0.904, p<0.05). 4. In the mastitic foremilk samples, EC values were obtained in the E. coli infection as 63.9mM-NaCl, in the Streptococcus spp. infection as 60.5mM-NaCl and in the Staphylococcus spp. infection as 57.0mM-NaCl. 5. At day 0 of estrus, the mean EC values of normal and mastitic foremilk samples were 41.2mM-NaCl and 68.3mM-NaCl respectively and the EC value of day 0 of estrus was higher than that of days before and after estrus.

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Development of a Rapid PCR Test for Identification of Streptococcus agalactiae in Milk Samples Collected on Filter Paper Disks

  • Wu, Jiusheng;Liu, Yuehuan;Hu, Songhua;Zhou, Jiyong
    • Asian-Australasian Journal of Animal Sciences
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    • 제21권1호
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    • pp.124-130
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    • 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.

Bioluminescence 반응에 의한 ATP 측정을 이용한 젖소 유방염 진단에 관한 연구 (Diagnostic test for bovine mastitis by the determination of ATP based on firefly bioluminescence)

  • 김태종;김종배;이승배;전용수
    • 대한수의학회지
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    • 제29권3호
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    • pp.383-391
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    • 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.

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젖소 유방염 유즙에서 분리한 Streptococcus uberis의 항생제 감수성 및 유전학적 특성 (Antimicrobial susceptibility and genetic characteristics of Streptococcus uberis isolated from bovine mastitis milk)

  • 이길;강현미;정충일;문진산
    • 대한수의학회지
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    • 제47권1호
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    • pp.33-41
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    • 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.

Genome-wide DNA methylation pattern in a mouse model reveals two novel genes associated with Staphylococcus aureus mastitis

  • Wang, Di;Wei, Yiyuan;Shi, Liangyu;Khan, Muhammad Zahoor;Fan, Lijun;Wang, Yachun;Yu, Ying
    • Asian-Australasian Journal of Animal Sciences
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    • 제33권2호
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    • pp.203-211
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    • 2020
  • Objective: Staphylococcus aureus (S. aureus) is one of the major microorganisms responsible for subclinical mastitis in dairy cattle. The present study was designed with the aim to explore the DNA methylation patterns using the Fluorescence-labeled methylation-sensitive amplified polymorphism (F-MSAP) techniques in a S. aureus-infected mouse model. Methods: A total of 12 out-bred Institute of Cancer Research female mice ranging from 12 to 13 weeks-old were selected to construct a mastitis model. F-MSAP analysis was carried out to detect fluctuations of DNA methylation between control group and S. aureus mastitis group. Results: Visible changes were observed in white cell counts in milk, percentage of granulocytes, percentage of lymphocytes, CD4+/CD8+ ratio (CD4+/CD8+), and histopathology of mice pre- and post-challenge with S. aureus. These findings showed the suitability of the S. aureus-infected mouse model. A total of 369 fragments was amplified from udder tissue samples from the two groups (S. aureus-infected mastitis group and control group) using eight pairs of selective primers. Results indicated that the methylation level of mastitis mouse group was higher than that in the control group. In addition, NCK-associated protein 5 (Nckap5) and transposon MTD were identified to be differentially methylated through secondary polymerase chain reaction and sequencing in the mastitis group. These observations might play an important role in the development of S. aureus mastitis. Conclusion: Collectively, our study suggests that the methylation modification in Nckap5 and transposon MTD might be considered as epigenetic markers in resistance to S. aureus-infected mastitis and provided a new insight into S. aureus mastitis research in dairy industry and public health.

Comparison of α1-Antitrypsin, α1-Acid Glycoprotein, Fibrinogen and NOx as Indicator of Subclinical Mastitis in Riverine Buffalo (Bubalus bubalis)

  • Guha, Anirban;Guha, Ruby;Gera, Sandeep
    • Asian-Australasian Journal of Animal Sciences
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    • 제26권6호
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    • pp.788-794
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    • 2013
  • Mastitis set apart as clinical and sub clinical is a disease complex of dairy cattle, with sub clinical being the most important economically. Of late, laboratories showed interest in developing biochemical markers to diagnose sub clinical mastitis (SCM) in herds. Many workers reported noteworthy alternation of acute phase proteins (APPs) and nitric oxide, (measured as nitrate+nitrite = NOx) in milk due to intra-mammary inflammation. But, the literature on validation of these parameters as indicators of SCM, particularly in riverine milch buffalo (Bubalus bubalis) milk is inadequate. Hence, the present study focused on comparing several APPs viz. ${\alpha}_1$-anti trypsin, ${\alpha}_1$-acid glycoprotein, fibrinogen and NOx as indicators of SCM in buffalo milk. These components in milk were estimated using standardized analytical protocols. Somatic cell count (SCC) was done microscopically. Microbial culture was done on 5% ovine blood agar. Of the 776 buffaloes (3,096 quarters) sampled, only 347 buffaloes comprising 496 quarters were found positive for SCM i.e. milk culture showed growth in blood agar with $SCC{\geq}2{\times}10^5$ cells/ml of milk. The cultural examination revealed Gram positive bacteria as the most prevalent etiological agent. It was observed that ${\alpha}_1$-anti trypsin and NOx had a highly significant (p<0.01) increase in SCM milk, whereas, the increase of ${\alpha}_1$-acid glycoprotein in infected milk was significant (p<0.05). Fibrinogen was below detection level in both healthy and SCM milk. The percent sensitivity, specificity and accuracy, predictive values and likelihood ratios were calculated taking bacterial culture examination and $SCC{\geq}2{\times}10^5$ cells/ml of milk as the benchmark. Udder profile correlation coefficient was also used. Allowing for statistical and epidemiological analysis, it was concluded that ${\alpha}_1$-anti trypsin indicates SCM irrespective of etiology, whereas ${\alpha}_1$-acid glycoprotein better diagnosed SCM caused by gram positive bacteria. NOx did not prove to be a good indicator of SCM. It is recommended measuring both ${\alpha}_1$-anti trypsin and ${\alpha}_1$-acid glycoprotein in milk to diagnose SCM in buffalo irrespective of etiology.

Detection and Identification of $\beta$-lactamase, Enterotoxin and Other Exotoxins Genes of Staphylococcus aureus by PCR

  • Yoon, Y.H.;Kim, K.I.
    • Asian-Australasian Journal of Animal Sciences
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    • 제16권3호
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    • pp.425-429
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    • 2003
  • Staphylococcus aureus is a major pathogen for cattle, causing various forms of subclinical and clinical mastitis and could be a causative agent of food poisoning, it produces various superantigenic exotoxins which have a great public health significance. A total of 72 S. aureus clinical isolates from dairy farms located in Kyunggi Province Korea were examined for the species identification by biochemical method, and for the detection of $\beta$-lactamase, enterotoxin and other exotoxins genes by PCR. The results of species identification by biochemical method agreed with those of PCR done with species specific primer STA-AU. $\beta$-lactamase is an enzyme closely associated with the resistance to antibiotic penicillin, which is an important means of treatment of mastitis, all the isolates were positive for the presence of genes encoding $\beta$-lactamase, which were reproduced in penicillin susceptibility disc assay. Six types of toxin genes, Staphylococcal enterotoxin (SE)A, SEB, SEC, SEE, toxic shock syndrome toxin (TSST-1) and exfoliative toxin A (ET A) were detected in 72 isolates by PCR associated genotypic method in this study, none of the isolates carried the genes for enterotoxin D (SED) and exfoliative toxin B (ETB). The occurrence rate of exotoxin genes rated as 12.5%, and the precision of the PCR identification results has been confirmed using the reference strains.