Application of the Limulus amoebocyte lysate test to the detection of Gram-negative bacterial endotoxins in normal and mastitic milk

Hartman, Ivo; Gil, Ziv; Saran, A.

doi:10.1016/s0034-5288(18)33435-0

Cited by 17 publications

(7 citation statements)

References 4 publications

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“…Other workers have used the gelation-based LAL test for milk monitoring (Hartman et al 1976;Svensson and Hahn-Hagerdal 1987;May et al 1989). In the case of milk, the threshold of a positive reaction in the gelation test was 10 3± 10 4 total viable bacteria per millilitre of milk sample.…”

Section: Resultsmentioning

confidence: 99%

“…In the case of milk, the threshold of a positive reaction in the gelation test was 10 3± 10 4 total viable bacteria per millilitre of milk sample. Hartman et al (1976) utilized the gelation-based LAL test to discriminate mastitic milk due to Gram-negative mastitis from mastitic milk from predominantly Gram-positive mastitis by testing serially diluted samples of milk with the LAL gelation test and assigning a cut-off dilution as indicative of the condition. Similarly, the assay presented in the current work is based on dilutions; therefore, it is semiquantitative for diagnosing process deviations.…”

Section: Resultsmentioning

confidence: 99%

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Monitoring the microbial contamination of beef carcass tissue with a rapid chromogenic Limulus amoebocyte lysate endpoint assay

Siragusa

Kang

Cutter³

2000

Lett Appl Microbiol

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A chromogenic Limulus amoebocyte lysate (LAL) endpoint assay was found to be an accurate and rapid means of gauging levels of beef carcass microbial contamination within 10 min. The assay demonstrated a high correlation with the total mesophilic bacterial and coliform surface populations from inoculated beef carcass surface tissues. This assay was tested on a set of actual beef carcass surface samples (n = 121) demonstrating the utility of the chromogenic LAL test as a means of monitoring carcass microbial contamination in a near real‐time fashion. Classifying the chromogenic LAL results into four contamination groups was found to be a sound means of utilizing the resultant chromogenic LAL data for detecting carcasses with high levels of microbial contamination. For beef carcass testing, this assay can be used with no instrumentation other than the required 37 °C incubator and, as an option, a microplate reader.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Monitoring the microbial contamination of beef carcass tissue with a rapid chromogenic Limulus amoebocyte lysate endpoint assay

Siragusa

Kang

Cutter³

2000

Lett Appl Microbiol

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“…In peracute cases of toxic mastitis, severe dehydration develops as a consequence of toxic shock, and this can be rapidly followed by coma and death (Eberhart 1984, Radostits and others 1994); such cases can become moribund before any gross physical changes occur in the milk from the affected quarter (Radostits and others 1994). A confirmatory diagnosis requires the demonstration of toxins, a laborious and expensive procedure (Hartman and others 1976, Waage and others 1994).…”

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confidence: 99%

Clinical and laboratory findings in cases of toxic mastitis in cows in Northern Ireland

Menzies¹,

McBride²,

McDowell³

et al. 2000

Veterinary Record

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This paper describes the clinical and laboratory findings from 264 cases of toxic mastitis in cows in Northern Ireland between October 1995 and May 1997. Nearly all the cases occurred during the winter housing period, with 84 per cent occurring between November and March inclusive, and 30 per cent in March. Sixty per cent of the cases occurred within one month of calving, and 29 per cent within four days of calving. The most common clinical signs were lethargy (92 per cent), discoloured milk (90 per cent), anorexia (72 per cent), tachypnoea (23 per cent), diarrhoea (23 per cent), recumbency (18 per cent) and staggering (15 per cent). Severe pyrexia (18 per cent) and clinical dehydration (44 per cent) were relatively common findings. Pure growths of Escherichia coli were isolated from 50 per cent of the milk samples, but 11 per cent yielded no bacterial growth. In vitro sensitivity tests indicated that enrofloxacin was effective against 98 per cent of the bacteria isolated, and framycetin and amoxycillin/clavulanic acid against 91 per cent. Abnormally high blood urea levels were observed in 31 per cent of cases, high blood creatinine levels in 42 per cent, and severe leucopenia in 56 per cent. Of the cases which were followed up, 14 per cent died, 21 per cent were culled early and a further 22 per cent lost milk production from the affected quarter.

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“…Virtually all of these have reported ng/mL LoDs, for endotoxin, but none are serogroup-specific. Researchers have used heat [46,102], chemical treatment with chloroform [103], acids [104,105], alkali [106,107], or ether [108] to improve sensitivity with some success when using heat or chemical extraction of the endotoxin [46,109]. However, the results show poor reproducibility between researchers (Table 1).…”

Section: Limulus Amoebocyte Lysate Assay and The Rabbit Pyrogen Testmentioning

confidence: 99%

Detection Methods for Lipopolysaccharides: Past and Present

Stromberg¹,

Mendez²,

Mukundan³

2017

≪i>Escherichia Coli

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Lipopolysaccharide (LPS) is the primary component of the outer membrane of Gram-negative bacteria. LPS aids in protecting bacterial cells, and also defines the unique serogroups used to classify bacteria. Additionally, LPS is an endotoxin and the primary stimulator of innate immune cells in mammals, making it an ideal candidate for early detection of pathogens. However, the majority of methods for detection of LPS focus on detection of the endotoxic component of the molecule, lipid A. Since lipid A is largely conserved among bacterial species and serogroups, these detection approaches are highly nonspecific. Thus, the importance of identifying the O-polysaccharide antigenic portion of LPS, which confers serogroup specificity, has received a great deal of attention in recent years. However, methods that are highly selective to the O-antigens are typically less sensitive than those that target the endotoxin. Here we present a history and comparison of the sensitivity of these methods and their value for detecting bacteria in a variety of different sample types.

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Application of the Limulus amoebocyte lysate test to the detection of Gram-negative bacterial endotoxins in normal and mastitic milk

Cited by 17 publications

References 4 publications

Monitoring the microbial contamination of beef carcass tissue with a rapid chromogenic Limulus amoebocyte lysate endpoint assay

Monitoring the microbial contamination of beef carcass tissue with a rapid chromogenic Limulus amoebocyte lysate endpoint assay

Clinical and laboratory findings in cases of toxic mastitis in cows in Northern Ireland

Detection Methods for Lipopolysaccharides: Past and Present

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