An Enzyme-Linked Immunosorbent Spot Assay Measuring Borrelia burgdorferi B31-Specific Interferon Gamma-Secreting T Cells Cannot Discriminate Active Lyme Neuroborreliosis from Past Lyme Borreliosis: a Prospective Study in the Netherlands.

Two-tier serology testing is most frequently used for the diagnosis of Lyme borreliosis (LB); however, a positive result is no proof of active disease. To establish a diagnosis of active LB, better diagnostics are needed. Tests investigating the cellular immune system are available, but studies evaluating the utility of these tests on well-defined patient populations are lacking. Therefore, we investigated the utility of an enzyme-linked immunosorbent spot (ELISpot) assay to diagnose active Lyme neuroborreliosis. Peripheral blood mononuclear cells (PBMCs) of various study groups were stimulated by using Borrelia burgdorferi strain B31 and various recombinant antigens, and subsequently, the number of Borrelia-specific interferon gamma (IFN-γ)-secreting T cells was measured. We included 33 active and 37 treated Lyme neuroborreliosis patients, 28 healthy individuals treated for an early manifestation of LB in the past, and 145 untreated healthy individuals. The median numbers of B. burgdorferi B31-specific IFN-γ-secreting T cells/2.5 × 105 PBMCs did not differ between active Lyme neuroborreliosis patients (6.0; interquartile range [IQR], 0.5 to 14.0), treated Lyme neuroborreliosis patients (4.5; IQR, 2.0 to 18.6), and treated healthy individuals (7.4; IQR, 2.3 to 14.9) (P = 1.000); however, the median number of B. burgdorferi B31-specific IFN-γ-secreting T cells/2.5 × 105 PBMCs among untreated healthy individuals was lower (2.0; IQR, 0.5 to 3.9) (P ≤ 0.016). We conclude that the Borrelia ELISpot assay, measuring the number of B. burgdorferi B31-specific IFN-γ-secreting T cells/2.5 × 105 PBMCs, correlates with exposure to the Borrelia bacterium but cannot be used for the diagnosis of active Lyme neuroborreliosis.

The influence of the molar ratio of cholesteryl hemisuccinate/dipalmitoylphosphatidylcholine on 'liposome' formation after lipid film hydration.

The morphology of the structures formed after hydration of lipid films of cholesteryl hemisuccinate/dipalmitoylphosphatidylcholine (CHEMS/DPPC) was investigated in low ionic strength solutions. The importance of addition of a charge inducing agent/geometrical structure such as CHEMS for the formation of stable vesicle dispersions upon hydration was demonstrated. The encapsulated volume measured for CHEMS/DPPC ratios below 1:50 was low. For a ratio of CHEMS/DPPC of 1:30 EM micrographs showed mainly small unilamellar vesicles, with particle sizes between 0.07 and 0.3 microns, together with a small number of much larger vesicles. For ratios of CHEMS/DPPC above 0.1 only unilamellar vesicles and no bilayer stacks were found. The results confirm the hypothesis by Hauser (Biochim. Biophys. Acta 772 (1984) 37-50), that the structures formed upon hydration of charged phospholipid films are unilamellar vesicles, while for neutral phospholipid films upon hydration bilayer stacks and multilamellar vesicles are formed. The effect of CHEMS on the liposome bilayer structure can be mainly ascribed to its charge inducing properties and presumably to a minor extent to its molecular geometry, or to a combination of both.

The relationships of salmonellae from infected broiler flocks, transport crates or processing plants to contamination of eviscerated carcases.

Three flocks raised for broiler or roaster performance tests were studied to determine the incidence and sources of salmonellae during the growing period, transport and processing and to relate these to contamination of processed carcasses. Day old chicks in two of the tests, (tests IV and V), were treated with a culture of intestinal anaerobes derived from mature chickens. The incidence of salmonellae during the growing period was too low to permit any conclusions about the efficacy of this culture in preventing Salmonella infection, but it had no adverse effect on flock performance. Carcasses from all three flocks were contaminated with salmonellae. Although the test IV flock was raised free of salmonellae, 46% of the carcasses tested from this flock were contaminated. The apparent source was the transport crates, 99% of which yielded salmonellae before the flock was loaded. In test V, 92% of the carcasses tested yielded salmonellae. The apparent sources were: flock infection (apparently originating from the parent flock), contaminated crates, spread during transport, and plant contamination. The flock of test VI was infected with Salmonella albany, and 54% of the carcasses tested were contaminated with this serovar. Carcasses of chicks infected early in life were more likely to be contaminated than those of chickens which contacted salmonellae later in the growing period.

Flock infection and transport as sources of salmonellae in broiler chickens and carcasses.

Cultural monitoring was used to determine the incidence and sources of salmonellae in a 4160-bird broiler flock raised on litter in 32 pens. Twenty-five of the pens remained apparently free of salmonellae during the 49-day growing period. Salmonella johannesburg, first detected in the meat meal component of the starter ration, was recovered from the litter of seven pens and from the intestines of dead or culled chicks from two pens. Salmonella alachua was also recovered from two of these pens. Culture of swabs collected from the plastic crates used to transport this flock for processing showed that 97/112 (86.6%) were contaminated with salmonellae (15 serovars) before the birds were loaded. The crate washer at the plant did not remove salmonellae from these crates: 97/132 (73.5%) crates sampled after washing yielded salmonellae. Eleven serovar were recovered, including S. johannesburg and S. alachua introduced by the infected flock. Twelve of 31 chickens (38.7%) collected when the birds were unloaded at the processing plant were intestinal carriers of S. johannesburg and/or S. alachua and 29 (93.5%) were external carriers. Salmonella johannesburg, S. alachua and four other serovars were isolated from the feathers of these birds. Eleven of 25 (44%) carcasses tested from this flock yielded salmonellae. Salmonella johannesburg or S. alachua, first isolated from the infected flock, were recovered from five carcasses and S. haardt and S. Typhimurium, first isolated from the transport crates were recovered from six carcasses.

Sources of salmonellae in an uninfected commercially-processed broiler flock.

Cultural monitoring was used to study the incidence and sources of salmonellae in a 4160 bird broiler flock during the growing period, transport and processing in a commercial plant. No salmonellae were isolated from any of 132 litter samples of 189 chickens cultured during the seven-week growing period, even though nest litter samples from four of the eight parent flocks yielded salmonellae and Salmonella worthington was isolated from the meat meal component of the grower ration. On arrival at the plant, 2/23 birds sampled carried S. infantis on their feathers, although intestinal cultures failed to yield salmonellae. Three of 18 processed carcasses samples yielded salmonellae (S. infantis, S. heidelberg, S. typhimurium var copenhagen). The most likely source of these salmonellae was the plastic transport crates, since 15/107 sampled before the birds were loaded yielded salmonellae (S. infantis, S. typhimurium). The crate washer at the plant did not reduce the incidence of Salmonella-contaminated crates, since 16/116 sampled after washing yielded salmonellae (S. infantis, S. typhimurium, S. heidelberg, S. schwarzengrund, S. albany).