An improved screening technique for isolation of pseudomonas pseudomallei from clinical specimens

doi:10.3109/00313027909061954

Pathology

Volume 11, Issue 2, 1979, Pages 293-297

https://doi.org/10.3109/00313027909061954 Get rights and content

Summary

A selective medium consisting of trypticase soy agar with 4% glycerol, 5 mg/l crystal violet, 50 mg/l neutral red and 4 mg/l of gentamicin was devised for isolation of Pseudomonas pseudomallei from clinical specimens. Absorption of neutral red was found to be suitable for differentiating this organism from other bacteria, while gentamicin was effective in selecting Ps. pseudomallei from organisms commonly found in clinical material. The medium was more suitable for screening clinical specimens than MacConkey's agar with colistin-S because it was more selective and allowed multiple specimens to be inoculated on a single plate. Eight thousand clinical specimens from an area endemic for melioidosis were screened on the selective medium. This resulted in the recovery of 8 isolates of Ps. pseudomallei that would not have been detected using routine culture media alone.

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Gentamicin and tobramycin—an in vitro comparison using 1400 clinical isolates
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Selection and rapid identification of Pseudomonas pseudomallei from other Gram-negative bacteria
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Inhibition of swarming of proteus by sodium tetradecyl sulphate, p-phenethyl alcohol and p-nitrophenylglycerol
Appl. Microbiol
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There are more references available in the full text version of this article.

Cited by (162)

A conserved active site PenA β-lactamase Ambler motif specific for Burkholderia pseudomallei/B. mallei is likely responsible for intrinsic amoxicillin-clavulanic acid sensitivity and facilitates a simple diagnostic PCR assay for melioidosis
2023, International Journal of Antimicrobial Agents
Burkholderia pseudomallei is a soil- and water-dwelling Gram-negative bacterium that causes melioidosis in humans and animals. Amoxicillin-clavulanic acid (AMC) susceptibility has been hailed as an integral part of the screening algorithm for identification of B. pseudomallei, but the molecular basis for the inherent AMC susceptibility of this bacterium remains undefined. This study showed that B. pseudomallei (and the closely-related B. mallei) wild-type strains are the only Burkholderia spp. that contain a ⁷⁰STSK⁷³ PenA Ambler motif. This motif was present in >99.5% of 1820 analysed B. pseudomallei strains and 100% of 83 analysed B. mallei strains, and is proposed as the likely cause for their inherent AMC sensitivity. The authors developed a polymerase chain reaction (PCR) assay that specifically amplifies the penA ⁷⁰ST(S/F)K⁷³-containing region from B. pseudomallei and B. mallei, but not from the remaining B. pseudomallei complex species or the ⁷⁰STFK⁷³ region from the closely-related penB of B. cepacia complex species. The abundance and purity of the 193-bp PCR fragment from putative B. pseudomallei isolates from clinical and environmental samples is likely sufficient for reliable confirmation of the presence of B. pseudomallei. The PCR assay is designed to be especially suited for use in resource-constrained areas. While not further explored in this study, the assay may allow diagnosis of putative B. mallei in culture isolates from animal and human samples.
Miscellaneous Bacterial Infections
2022, Greene's Infectious Diseases of the Dog and Cat, Fifth Edition
Dermatophilosis
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  Cause: Dermatophilus congolensis, family Dermatophilaceae.
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  First Described: 1915 in cattle from the Belgian Congo (Van Saceghem).¹
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  Affected Hosts: Skin commensals on cattle, horses, small ruminants. Secondary hosts: humans, dogs, cats, platypus.
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  Geographic Distribution: Worldwide, but infections are more common in humid environments.
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  Primary Mode of Transmission: May be acquired by dogs and cats through contact with affected primary hosts or from contaminated fomites or soil.
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  Major Clinical Signs: Exudative dermatitis (dogs), deep soft-tissue abscesses with fistulation (cats).
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  Differential Diagnoses: Dogs: superficial pyoderma, mycobacteriosis, allergic dermatoses, immune-mediated dermatoses, drug eruptions, thermal or caustic skin damage. Cats: mycobacteriosis, nocardiosis, fungal infections, cutaneous neoplasia.
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  Human Health Significance: Humans often develop infection as a result of contact with colonized or infected animals (especially horses, cattle, and elephants), although infections are very rare. No cases have been reported in association with contact with a dog or cat.
Melioidosis and Glanders
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  Causes: Burkholderia pseudomallei (melioidosis) and Burkholderia mallei (glanders/farcy), gram-negative, aerobic, rod-shaped bacteria that belong to the family Burkholderiaceae.
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  First Described: Burkholderia pseudomallei was first described by Whitmore and Krishnaswami in 1912² in Burma. Glanders (B. mallei) was reported in ancient Greece by Hippocrates, c.450 BC; 100 years later Aristotle named the disease “meles.” The organism was isolated for the first time by Loeffler and Schuetz in 1882.³
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  Affected Hosts: Burkholderia pseudomallei: Humans, non-human primates, captive marine mammals (e.g., dolphins, whales, sea lions), birds, reptiles, sheep, goats, cattle, alpaca, pigs, horses, mules, cats, dogs, and other wildlife (kangaroos, meerkats). Burkholderia mallei: Donkeys, mules, horses, camels, humans, cats, dogs, large carnivores (wolves, bears, lions, tigers, leopards, jackals, and hyenas).
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  Geographic Distribution: Burkholderia pseudomallei: Tropics and subtropics. Burkholderia mallei: South America (Brazil), northern Africa, China, Iraq, Pakistan, India, Mongolia, Turkey, and United Arab Emirates.
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  Primary Mode of Transmission: Burkholderia pseudomallei: Cutaneous inoculation via breaches in the integument; inhalation/aspiration of aerosols or muddy water, respectively, ingestion. Burkholderia mallei: Inhalation of aerosols, ingestion of contaminated water or feed, or via contact with fomites in solipeds, especially in situations where husbandry is substandard, and ingestion of infected meat in carnivores.
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  Major Clinical Signs: Range from minor skin lesions, subacute to chronic pneumonia, to fulminant sepsis (B. pseudomallei).
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  Differential Diagnoses: Mycobacteriosis, neoplasia, other saprophytic gram-negative infections, for example, Chromobacterium violaceum.
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  Human Health Significance: Burkholderia pseudomallei is estimated to clinically affect approximately 165,000 people per year, 10% to 40% fatally. Both B. pseudomallei and B. mallei are classified as a Tier 1 Select biothreat agents by the United States CDC.
Chromobacteriosis
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  First Described: Schroeter 1872.⁴
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  Cause: Chromobacterium violaceum.
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  Affected Hosts: Humans, pigs, cattle, dogs, non-human primates, water buffalo, Macquarie turtle, red panda, horse.
- •
  Geographic Distribution: Worldwide but mostly tropical and subtropical regions.
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  Primary Mode of Transmission: Cutaneous inoculation after exposure of wounds to contaminated water or soil.
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  Major Clinical Signs: Localized cutaneous and regional lymph nodes, multi-systemic abscessation, pneumonia, and sepsis.
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  Differential Diagnoses: Burkholderia pseudomallei infection, and other causes of gram-negative sepsis.
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  Human Health Significance: Infections are acquired from the environment and are not transmitted between animals and humans. Infections in humans are often fatal.
Melioidosis: Case reports of confirmed Burkholderia pseudomallei in Saudi Arabia
2020, Journal of Infection and Public Health
Citation Excerpt :
B. pseudomallei can grow in, MacConkey, or cystine-lactose-electrolyte deficient agars. Using Ashdown’s medium, a gentamicin-containing liquid transport broth will increase the likelihood of successful culture from sputum, throat swabs [8]. Working on samples should be carried out in a biosafety level 2 practices equipped microbiology lab [9].
Melioidosis is an infectious disease of tropical climates. The disease is caused by the bacterium Burkholderia pseudomallei. Most cases are diagnosed in southeast Asia and northern Australia. Some imported cases diagnosed in returning tourists, soldiers, and immigrants from endemic areas. It caught much attention since the Centers for Disease Control and Prevention (CDC) designated B. pseudomallei as an agent for biological warfare and terrorism. We describe two cases of a 26-year-old Saudi woman who had fulminant sepsis soon after returning from Thailand & a 48-year-old woman with a long history of fever. B. pseudomallei was isolated from both patients blood cultures, and they had different consequences. A confirmed case of melioidosis was not reported before in Saudi Arabia.
Disruption of the Burkholderia pseudomallei two-component signal transduction system BbeR-BbeS leads to increased extracellular DNA secretion and altered biofilm formation
2020, Veterinary Microbiology
Citation Excerpt :
Modified pBluescript SK(–) and pBHR1 constructs were maintained in the E. coli strains SM10 λpir and S17-1 λpir, respectively. B. pseudomallei strains were routinely cultured in Lysogeny Broth (LB) containing 1 % w/v tryptone, 0.5 % w/v yeast extract, and 0.5 % w/v NaCl, 2 YT medium containing 1.6 % w/v tryptone, 1 % w/v yeast extract, and 0.5 % w/v NaCl, or Ashdown’s medium prepared as described previously (Ashdown, 1979). Solid media contained 1.5 % w/v bacteriological agar.
Two-component signal transduction systems (TCSTS) are abundant among prokaryotes and regulate important functions, including drug resistance and virulence. The Gram-negative bacterium Burkholderia pseudomallei, which causes the severe infectious disease melioidosis, encodes 136 putative TCSTS components. In silico analyses of these TCSTS indicated that the predicted BbeR-BbeS system (BPSL1036-BPSL1037) displayed significant amino acid sequence similarity to the Shigella flexneri virulence-associated OmpR-EnvZ osmoregulator. To assess the function of the B. pseudomallei BbeR-BbeS system, we constructed by allelic exchange a ΔbbeRS double mutant strain lacking both genes, and single ΔbbeR and ΔbbeS mutants. All three mutant strains caused disease in the BALB/c acute melioidosis model at the same rate as the wild-type strain, displayed unchanged swarming motility on semi-solid medium, and were unaffected for viability on high-osmolarity media. However, when cultured at 37 °C for at least 14 days, ΔbbeS and ΔbbeR colonies developed a distinct, hypermucoid morphology absent in similarly-cultured wild-type colonies. At both 30 °C and 37 °C, these hypermucoid strains produced wild-type levels of type I capsule but released increased quantities of extracellular DNA (eDNA). Upon static growth in liquid medium, all B. pseudomallei strains produced pellicle biofilms that contained DNA in close association with bacterial cells; however, the ΔbbeS and ΔbbeR strains produced increased biofilms with altered microscopic architecture compared to the wild-type. Unusually, while the ΔbbeS and ΔbbeR single-deletion mutants displayed clear phenotypes, the ΔbbeRS double-deletion mutant was indistinguishable from the wild-type strain. We propose that BbeR-BbeS indirectly affects eDNA secretion and biofilm formation through cross-talk with one or more other TCSTS.
Melioidosis case series
2019, Clinical Infection in Practice
Citation Excerpt :
Gram-negative bacilli with bipolar staining may be identified from the sputum, pus or wound swab of an infected patient. Otherwise, the organism grows readily on a wide variety of culture media, and selectively on Ashdown's media.10,11 Suspected cases should be notified to the laboratory processing specimens; B. pseudomallei is a hazard group 3 pathogen, and prophylaxis for laboratory workers may be required depending on type of exposure and underlying risk factors for severe disease.12
Melioidosis is a potentially fatal infection caused by Burkholderia pseudomallei, a bacterium endemic to certain tropical and subtropical world regions. It most frequently enters the body through small abrasions on the feet or lower limbs when individuals walk barefoot, or in open footwear, through contaminated wet or muddy environments. We report on three female cases successfully treated for melioidosis within the Newcastle upon Tyne NHS Hospitals over a 12-month period; a 52-year-old with a mediastinal abscess, a 38-year-old with an intra-cerebral abscess, and a 69-year-old with pneumonia and bacteraemia. Two patients were diabetic, the latter with established microvascular complications, but the third patient had no immunosuppressive risk factor identified. All had recently travelled to Thailand, although one remained solely within the confines of her holiday resort and another did not leave the city of Pattaya. Clinicians must remain vigilant to the potential for imported cases of melioidosis into the UK in travellers returning from endemic areas, such as Southeast Asia.
Caprine humoral response to burkholderia pseudomallei antigens during acute melioidosis from aerosol exposure
2019, PLoS Neglected Tropical Diseases
Burkholderia pseudomallei causes melioidosis, a common source of pneumonia and sepsis in Southeast Asia and Northern Australia that results in high mortality rates. A caprine melioidosis model of aerosol infection that leads to a systemic infection has the potential to characterize the humoral immune response. This could help identify immunogenic proteins for new diagnostics and vaccine candidates. Outbred goats may more accurately mimic human infection, in contrast to the inbred mouse models used to date. B. pseudomallei infection was delivered as an intratracheal aerosol. Antigenic protein profiling was generated from the infecting strain MSHR511. Humoral immune responses were analyzed by ELISA and western blot, and the antigenic proteins were identified by mass spectrometry. Throughout the course of the infection the assay results demonstrated a much greater humoral response with IgG antibodies, in both breadth and quantity, compared to IgM antibodies. Pre-infection sera showed multiple immunogenic proteins already reactive for IgG (7–20) and IgM (0–12) in most of the goats despite no previous exposure to B. pseudomallei. After infection, the number of IgG reactive proteins showed a marked increase as the disease progressed. Early stage infection (day 7) showed immune reaction to chaperone proteins (GroEL, EF-Tu, and DnaK). These three proteins were detected in all serum samples after infection, with GroEL immunogenically dominant. Seven common reactive antigens were selected for further analysis using ELISA. The heat shock protein GroEL1 elicited the strongest goat antibody immune response compared to the other six antigens. Most of the six antigens showed the peak IgM reactivity at day 14, whereas the IgG reactivity increased further as the disease progressed. An overall MSHR511 proteomic comparison between the goat model and human sera showed that many immune reactive proteins are common between humans and goats with melioidosis.

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Pathology

Summary

References (8)

Gentamicin and tobramycin—an in vitro comparison using 1400 clinical isolates

Pathology

Cowan and Steel's Manual for the Identification of Medical Bacteria

Selection and rapid identification of Pseudomonas pseudomallei from other Gram-negative bacteria

Am. J. Clin. Pathol

Inhibition of swarming of proteus by sodium tetradecyl sulphate, p-phenethyl alcohol and p-nitrophenylglycerol

Appl. Microbiol

Cited by (162)

A conserved active site PenA β-lactamase Ambler motif specific for Burkholderia pseudomallei/B. mallei is likely responsible for intrinsic amoxicillin-clavulanic acid sensitivity and facilitates a simple diagnostic PCR assay for melioidosis

Miscellaneous Bacterial Infections

Melioidosis: Case reports of confirmed Burkholderia pseudomallei in Saudi Arabia

Disruption of the Burkholderia pseudomallei two-component signal transduction system BbeR-BbeS leads to increased extracellular DNA secretion and altered biofilm formation

Melioidosis case series

Caprine humoral response to burkholderia pseudomallei antigens during acute melioidosis from aerosol exposure