Laboratory diagnosis of paraneoplastic pemphigus.

BACKGROUND: Paraneoplastic pemphigus (PNP) is a multiorgan disease characterized by antibodies against plakins, desmogleins and the α2-macroglobulin-like-1 (A2ML1) protein, in association with an underlying neoplasm. Accurate diagnosis relies on the demonstration of these autoantibodies in serum. OBJECTIVES: To evaluate the value of different laboratory techniques in the serological diagnosis of PNP. METHODS: We performed immunoblotting, envoplakin (EP) enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence (IIF) on rat bladder, radioactive immunoprecipitation and a nonradioactive combined immunoprecipitation-immunoblot assay. Additional assays included BP180 ELISA and BP230 ELISA. We included the sera of 19 patients with PNP and 40 control subjects. RESULTS: The sensitivities were 63% for anti-EP ELISA, 74% for rat bladder IIF, 89% for immunoblotting, 95% for radioactive immunoprecipitation and 100% for nonradioactive immunoprecipitation. Specificities ranged from 86% to 100%. The BP180 and BP230 ELISAs had low sensitivity and specificity for PNP. The combination of rat bladder IIF and immunoblot showed 100% sensitivity and specificity. The analysis of sequential PNP sera showed that antibody titres may decrease over time, possibly resulting in negative outcomes for EP ELISA and rat bladder IIF studies. CONCLUSIONS: The detection of autoantibodies against EP and periplakin, or A2ML1 by immunoprecipitation is most sensitive for PNP. The combination of rat bladder IIF and immunoblotting is equally sensitive and highly specific, and represents an alternative valuable and relatively easy approach for the serological diagnosis of PNP.

[The clinical effectiveness of a post milking teat disinfection method with a foaming iodophor teat dip]. / Untersuchungen zur klinischen Wirksamkeit eines Zitzendesinfektionsverfahrens nach dem Melken mit einem schaumbildenden jodophoren Dippmittel.

The effect of postmilking teat dipping with a foaming iodophor agent on incidence of intramammary infections (IMI), incidence of clinical mastitis, somatic cell count and the characteristics of udder tissue and teat was investigated in a positively controlled field study. Two groups of animals were compared. Teats were dipped with a foaming iodophor in the treatment group (TG, 122 animals) while teats in the control group (CG, 121 animals) were dipped with a conventional iodophor teat dip with the same iodine content. A bacteriological examination of quarter milk samples divided the study period in two parts. The incidence of new IMI did not differ between the groups (1st part of trial: TG vs. CG: 6.84% vs. 9.16%, 2nd part of trial: 7.78% vs. 7.82%). There were no differences between the treatment groups regarding incidence of clinical mastitis. We detected 0.64 clinical cases per 100 days in the treatment group vs. 0.50 in the control group. The development of SCC was comparable in both groups. Teat skin and teat duct conditions showed variation during the study period. Clinical efficacy of postmilking teat disinfection with a foaming iodophor was comparable to the treatment with a conventional iodophor product.

Separation of metal ions by capillary electrophoresis--diversity, advantages, and drawbacks of detection methods.

Capillary electrophoresis (CE) has been applied to metal-ion analysis during the last 10 years. To improve sensitivity and selectivity different modes of detection have been adapted or developed. The selection of commercially available detection systems for metal-ion analysis is still primarily limited to UV-Vis detection, although other commercial systems, e.g. fluorescence, conductivity, or interfaces for coupling to mass spectrometry (MS) or inductively coupled plasma mass spectrometry (ICP-MS) are becoming available. High demands are made on any detector used in CE, because the analytical signal has to be extracted from less than 1 nL of sample, which corresponds to a total amount of < or = 10(-12) to 10(-15) mol analyte. This paper compares currently available and recently developed detection methods for CE as applied to the analysis of metal ions. Commercially available techniques, for example UV-Vis, fluorescence, or mass spectrometry, and other new detection methods including electrochemistry, radioactivity, and XRF, are discussed and future trends are anticipated.

Efficacy of a herd-specific vaccine against Staphylococcus aureus to prevent post-partum mastitis in dairy heifers.

A placebo-controlled field study was performed to evaluate the effect of a herd-specific vaccine against Staphylococcus aureus on intramammary infection (IMI), somatic cell count (SCC) and clinical mastitis. Three hundred and twenty-one heifers were assigned to two groups. Heifers in the vaccination group (n = 164) were vaccinated twice, i.e. 5 and 2 weeks before their expected calving date. Animals of the control group (n = 157) received the same treatment with a placebo containing no bacterial antigen. Quarter milk samples were collected immediately after parturition prior to the first machine milking, 3-4 weeks after calving and before the onset of treatment in animals with signs of clinical mastitis during the first 3 months after calving. For comparison of SCC the data from the monthly milk test records were evaluated. The prevalence of S. aureus in quarter milk samples taken at calving and 3-4 weeks post-partum did not differ significantly between the vaccine and control group. Incidence of clinical mastitis during the first 3 months after calving and the prevalence of S. aureus in quarter milk samples taken before the onset of treatment did not differ significantly between the groups. The SCC was lower in vaccinated than in control heifers. However, the difference was only significant on the third milk test day. Regarding prevalence of IMI with S. aureus and incidence of clinical mastitis the use of a herd-specific vaccine against S. aureus did not prove to be efficient on this farm.

Effect of teat dipping with a germicide barrier teat dip in late gestation on intramammary infection and clinical mastitis during the first 5 days post-partum in primiparous cows.

The effect of teat dipping with a barrier teat dip prior to parturition on intramammary infection (IMI) and clinical mastitis during the first 5 days post-partum was investigated in a split udder trial in 149 Holstein-Frisian heifers. Their left front and right hind quarters were dipped three times weekly (i.e. Monday, Wednesday and Friday) with a barrier teat dip containing 0.1% polyvidon iodine from day 260 of gestation until parturition. The opposite quarters (right front and left hind quarter) served as untreated control. Bacteria were isolated from 52.2% of quarter milk samples collected immediately after parturition prior to first machine milking. Staphylococcus aureus and coagulase-negative staphylococci (CNS) were predominantly found in the samples (29.2 and 35.6% of the positive samples, respectively). At parturition 6.7% of the heifers showed signs of clinical mastitis and another 27.5% developed signs of clinical mastitis during the first five days of lactation. No significant differences were found between treated and control quarters regarding IMI and incidence of clinical mastitis. Teat dipping prior to parturition in primigravid dairy heifers did not improve udder health in this trial.

[Effect of puerperal mastitis in primiparous cows on milk production, cell count and culling]. / Einfluss von peripartalen Mastitiden bei Erstkalbinnen auf Milchleistung, Zellgehalt und Abgangsrate.

A retrospective cohort study was performed to investigate the effect of clinical mastitis in heifers during the first week post partum (p.p.) on subsequent health and productivity. Primiparous cows that had calved between August 15th, 1996 and August 14th, 1997 (n = 1389) were included in the study. Milk samples were collected from each quarter post partum prior to first milking. The predominant group of bacteria found were Staphylococcus spp. (69%) followed by Streptococcus spp.(12%) and E. coli (5%). Intramammary infections at calving increased the risk of clinical mastitis within the first week p.p. The cow incidence of mastitis until day 7 p. p. was 38%. Cows were classified into three groups. Group 1: Animals with clinical mastitis prior to calving (n = 35); Group 2: Animals with clinical mastitis between calving and 7 days p. p. (n = 503); Group 3: Animals without mastitis until 7 days p. p. (n = 851). Mastitis prior to parturition and mastitis within the first week p.p. increased the risk of further cases of mastitis and culling during the first 45 days of lactation. Milk yield of Group 2 was lower and somatic cell counts were higher than in Group 3.

[Individual animal risk factors for Prototheca mastitis in cattle]. / Tierindividuelle Risikofaktoren für die Protothekenmastitis des Rindes.

Risk factors for mastitis caused by Prototheca zopfii were determined in a retrospective case control study. 248 cows culled for protothecal mastitis in a large dairy herd were included in the study. The diseased cows were compared to their non affected herdmates in respect to age, stage of lactation, production and milk composition. The effect of previous cases of mastitis during the same lactation or in previous lactations and the effect of antibiotic pretreatment were determined in a split udder analysis comparing left quarters to right quarters. Age and mastitis history especially antimicrobial pretreatment were important risks factors for developing protothecal mastitis. Risk of protothecal mastitis increased with parity. Quarters with a history of clinical mastitis during the same or during previous lactations were at higher risk than other quarters (OR 1.98, p < 0.01 and 1.44, p = 0.06, respectively). Pretreatment with antibiotics increased the risk of protothecal mastitis (OR 3.19, p < 0.001). Milk yield was lower in cows that developed protothecal mastitis. Somatic cell counts were higher than in control cows. Incidence of protothecal mastitis peaked in the second month of lactation.

Transient isotachophoretic-electrophoretic separations of lanthanides with indirect laser-induced fluorescence detection.

Indirect laser-induced fluorescence was used for the detection of several lanthanide species separated by capillary electrophoresis. Quinine sulfate was the fluorescent component of the background electrolyte, and α-hydroxyisobutyric acid was added as a complexing agent to enable the separation of analyte ions that have similar mobilities. The UV lines (333-364 nm) of an argon ion laser were used as the excitation source with a diode array detector for monitoring the fluorescent emission at 442 nm. Electrokinetic injections and transient isotachophoresis were implemented to stack the analyte ions into more concentrated zones. On-line preconcentration factors were determined to be ∼700 and resulted in limits of detection for La(3+), Ce(3+), Pr(3+), Nd(3+), Sm(3+), and Eu(3+) in the low-ppb range (6-11 nM).

Analysis of fission products using capillary electrophoresis with on-line radioactivity detection.

Capillary electrophoresis has been used to separate metal ions characteristically associated with nuclear fission. Indirect UV absorbance and on-line radioactivity detection were used simultaneously to monitor the analytes. The radioactivity detector consists of conical plastic scintillating material with the capillary passing through the center to provide a 4π detection geometry. The wide end of the cone is optically coupled to a photomultiplier tube. Transient isotachophoretic techniques were employed to stack large volumes of samples which had low specific activities. Radioactivity detection of (152)Eu and (137)Cs was achieved at the nanocurie level for 80-100 nL injections. The detector is approximately 80% efficient, enabling samples resident in the detector window for 0.1 min to be reliably assayed. The separation of (137)Cs and (137m)Ba isotopes, which are in secular equilibrium, was modeled to demonstrate the effects of the rapid decay of (137m)Ba.