Evaluation of highly sensitive diagnostic tools for the detection of P. falciparum in pregnant women attending antenatal care visits in Colombia.

BACKGROUND: In low transmission settings early diagnosis is the main strategy to reduce adverse outcomes of malaria in pregnancy; however, microscopy and rapid diagnostic tests (RDTs) are inadequate for detecting low-density infections. We studied the performance of the highly sensitive-RDT (hsRDT) and the loop mediated isothermal DNA amplification (LAMP) for the detection of P. falciparum in pregnant women. METHODS: A cross-sectional study was conducted in two malaria-endemic municipalities in Colombia. We screened pregnant women in the context of an antenatal care program in health facilities and evaluated five tests (microscopy, conventional RDT, hsRDT, LAMP and nested polymerase chain reaction-PCR) for the detection of P. falciparum in peripheral blood, using a quantitative reverse transcription PCR (qRT-PCR) as the reference standard. Diagnostic performance of hsRDT and LAMP were compared with routine testing. RESULTS: The prevalence of P. falciparum was 4.5% by qRT-PCR, half of those infections were subpatent. The sensitivity of the hsRDT (64.1%) was slightly better compared to microscopy and cRDT (59 and 53.8% respectively). LAMP had the highest sensitivity (89.7%) for detecting P. falciparum and the ability to detect very low-density infections (minimum parasite density detected 0.08 p/µL). CONCLUSIONS: There is an underestimation of Plasmodium spp. infections by tests routinely used in pregnant women attending antenatal care visits. LAMP methodology can be successfully implemented at local hospitals in malaria-endemic areas. The relevance of detecting and treating this sub-patent P. falciparum infections in pregnant women should be evaluated. TRIAL REGISTRATION: ClinicalTrials.gov, Identifier: NCT03172221 , Date of registration: May 29, 2017.

One Health collaborations for zoonotic disease control in Ethiopia.

Zoonotic diseases require a One Health approach for successful control and elimination due to the nature of their transmission between animals and humans. One Health recognises that the health of humans, animals, and the environment are all interconnected. Ethiopia has committed itself to controlling five prioritised zoonotic diseases (rabies, anthrax, brucellosis, leptospirosis and echinococcosis), using a One Health approach. The National One Health Steering Committee (NOHSC) provides a framework for national stakeholders to address gaps in multisectoral communication, coordination and collaboration. In addition, the NOHSC oversees the formation of several specialised disease-focused groups, referred to as 'Technical Working Groups' (TWGs). These TWGs are responsible for developing disease prevention and control strategies, as well as implementing disease-focused public health activities and providing recommendations to the NOHSC. Ethiopia's success using the One Health approach and its efficient control of zoonotic diseases will depend on the commitment of all member Ministries to support the NOHSC and TWGs, as well as to build capacity in Ethiopia's workforce and laboratories, a task supported by its many international partners.

Les zoonoses étant par nature des maladies transmissibles entre les animaux et l'homme, l'approche Une seule santé est la seule qui permette de les contrôler efficacement en vue de les éliminer. Le concept Une seule santé repose sur la prise en compte de l'interconnexion entre la santé humaine, celle des animaux et celle de l'environnement. L'Éthiopie s'est fixé pour objectif de lutter contre cinq maladies zoonotiques classées comme prioritaires (rage, fièvre charbonneuse, brucellose, leptospirose et échinococcose) en suivant une approche Une seule santé. Le comité de pilotage national Une seule santé (NOHSC) apporte un cadre permettant aux parties prenantes du pays de résoudre les problèmes de communication, de coordination et de collaboration intersectorielles. En outre, le NOHSC supervise la création de plusieurs groupes de travail techniques dédiés à des maladies spécifiques. Ces groupes de travail sont chargés d'élaborer des stratégies de prévention et de contrôle, de mettre en oeuvre des activités de santé publique axées sur ces maladies et de formuler des recommandations à l'intention du NOHSC. La réussite des efforts déployés par l'Éthiopie pour appliquer les principes Une seule santé et l'efficacité de la lutte contre les maladies zoonotiques dépendront de l'engagement des ministères concernés à soutenir le NOHSC et les groupes de travail techniques et à renforcer les capacités des ressources humaines et des laboratoires éthiopiens, tâche qui bénéficie de l'appui de nombreux partenaires internationaux.

Toda labor eficaz de control y eliminación de las enfermedades zoonóticas, por la propia naturaleza de su transmisión entre animales y personas, pasa por abordar estas patologías desde los planteamientos de Una sola salud, noción esta que parte del reconocimiento de que salud humana, animal y ambiental están siempre interconectadas. Etiopía está embarcada en el innegociable empeño de combatir cinco enfermedades zoonóticas consideradas prioritarias (rabia, carbunco bacteridiano, brucelosis, leptospirosis y equinococosis) trabajando desde la óptica de Una sola salud. El Comité Directivo Nacional de Una sola salud proporciona a los interlocutores del país un marco de referencia que sirve para subsanar las lagunas existentes en cuanto a comunicación, coordinación y colaboración entre los diversos sectores. Ese órgano, además, supervisa la formación de varios grupos especializados y centrados en una u otra enfermedad, denominados grupos de trabajo técnicos, que tienen por cometido elaborar estrategias de prevención y control de una enfermedad concreta, llevar adelante acciones de salud pública dirigidas contra ella y formular recomendaciones para el Comité Directivo. El éxito de Etiopía a la hora de aplicar los postulados de Una sola salud y de combatir eficazmente las enfermedades zoonóticas dependerá del nivel de compromiso con que todos los ministerios copartícipes presten apoyo al Comité Directivo y los grupos de trabajo técnicos y ayuden a instaurar en el país un tejido lo bastante solvente de laboratorios y recursos humanos, empresa esta en la que Etiopía cuenta con el respaldo de sus numerosos asociados internacionales.

A decade of improvement: New York State fluid milk quality.

The microbiological and sensory qualities of New York State (NYS) fluid milk products were assessed as part of an ongoing fluid milk quality program. Commercially packaged pasteurized fluid milk samples were collected twice a year over the 10-yr period from 2001 to 2010 from 14 NYS dairy processing facilities and analyzed at the Milk Quality Improvement Program (MQIP) laboratory. Each sample was tested throughout refrigerated storage (6°C) on day initial, 7, 10, and 14 for standard plate count (SPC), coliform count (CC), and sensory quality. Over the 10-yr period, the percentage of samples with bacterial numbers below the Pasteurized Milk Ordinance (PMO) limit of 20,000 cfu/mL at d 14 postprocessing ranged from a low of 21.1% in 2002 to a high of 48.6% in 2010. Percent samples positive for coliforms during that same period ranged from a high of 26.6% in 2002 to a low of 7.5% in 2007. Mean d 14 sensory scores ranged from a low of 6.0 in 2002 to a high of 7.3 in 2007. Samples contaminated with coliforms after pasteurization have significantly higher SPC counts and significantly lower sensory scores on d 14 of shelf-life than those not contaminated with coliforms. Product factors such as fat level were not significantly associated with SPC, CC, or sensory quality of the product, whereas the factor processing plant significantly affected overall product quality. This study demonstrates that overall fluid milk quality in NYS, as determined by microbiological and sensory analyses, has improved over the last decade, and identifies some challenges that remain.

Evaluation of various selective media for the detection of Pseudomonas species in pasteurized milk.

Pseudomonas spp. are common gram-negative, post-pasteurization contaminants that contribute to spoilage of pasteurized dairy products. This study evaluated 5 common selective media for detecting Pseudomonas spp. in pasteurized milk. The performance of each selective medium for recovering 12 different Pseudomonas isolates (selected to represent a diversity of pasteurized milk isolates) was compared with that of standard plate count agar pour plates. Pseudomonas isolates showed varying abilities to produce colonies on different selective media. For 2 of 12 isolates, a 48-h incubation time was required for colony formation on any of the media tested. Violet red bile agar and coliform Petrifilm (3M, St. Paul, MN) were less effective than standard plate count agar pour plates at recovering Pseudomonas, regardless of incubation time, and MacConkey agar showed poor detection efficiency compared with SPCP after a 48-h incubation (R(2) = 0.26). Therefore, the use of violet red bile agar, MacConkey agar, or coliform Petrifilm may not be sufficient for detecting common Pseudomonas spp. in milk. The methods showing the highest detection efficiencies were crystal violet tetrazolium agar (CVTA) pour plates (R(2) = 0.95) and CVTA plates inoculated by spiral plating (R(2) = 0.89) incubated at 32 °C for 48 h. Overall, plating milk samples on CVTA followed by a 48-h incubation at 32 °C was the most effective selective method for recovering a diversity of Pseudomonas spp. from milk.

When cheese gets the blues: Pseudomonas fluorescens as the causative agent of cheese spoilage.

A bacterial contamination of fresh, low-acid cheese that resulted in production of a blue fluorescent pigment on the surface of the cheese was determined to be caused by Pseudomonas fluorescens biovar IV, a gram-negative bacteria that produces a blue, nondiffusible pigment as well as the soluble pigment pyoverdin, which fluoresces under UV light. Ten isolates collected from contaminated cheese and environmental samples were initially identified as P. fluorescens using 16S rDNA sequencing, but only 8 of the isolates produced blue pigment and fluoresced under UV light when re-inoculated onto fresh, low-acid cheese. The Biolog Metabolic Fingerprint system (Biolog Inc., Hayward, CA) and the Analytical Profile Index (BioMerieux Vitek Inc., Hazelwood, MO) for nonenteric gram-negative species as well as EcoRI ribotyping did not differentiate between the isolates that produced blue color and those that did not. Pulsed field gel electrophoresis with the enzyme XbaI was able to distinguish between the isolates that produced pigment and those that did not and allowed for identification of a specific environmental site (i.e., an overhead cheese vat agitator system) as the likely source of product contamination.

Results from raw milk microbiological tests do not predict the shelf-life performance of commercially pasteurized fluid milk.

Analytical tools that accurately predict the performance of raw milk following its manufacture into commercial food products are of economic interest to the dairy industry. To evaluate the ability of currently applied raw milk microbiological tests to predict the quality of commercially pasteurized fluid milk products, samples of raw milk and 2% fat pasteurized milk were obtained from 4 New York State fluid milk processors for a 1-yr period. Raw milk samples were examined using a variety of tests commonly applied to raw milk, including somatic cell count, standard plate count, psychrotrophic bacteria count, ropy milk test, coliform count, preliminary incubation count, laboratory pasteurization count, and spore pasteurization count. Differential and selective media were used to identify groups of bacteria present in raw milk. Pasteurized milk samples were held at 6°C for 21 d and evaluated for standard plate count, coliform count, and sensory quality throughout shelf-life. Bacterial isolates from select raw and pasteurized milk tests were identified using 16S ribosomal DNA sequencing. Linear regression analysis of raw milk test results versus results reflecting pasteurized milk quality consistently showed low R(2) values (<0.45); the majority of R(2) values were <0.25, indicating small relationship between the results from the raw milk tests and results from tests used to evaluate pasteurized milk quality. Our findings suggest the need for new raw milk tests that measure the specific biological barriers that limit shelf-life and quality of fluid milk products.

Tracking spore-forming bacterial contaminants in fluid milk-processing systems.

The presence of psychrotolerant Bacillus species and related spore formers (e.g., Paenibacillus spp.) in milk has emerged as a key biological obstacle in extending the shelf life of high-temperature, short-time pasteurized fluid milk beyond 14 d. A recently developed rpoB DNA sequence-based subtyping method was applied to characterize spoilage bacteria present in raw milk supplies for 2 processing plants, and to assess transmission of these organisms into pasteurized products. Thirty-nine raw milk samples and 11 pasteurized product samples were collected to represent the processing continuum from incoming truck loads of raw milk to packaged products. Milk samples were held at 6 degrees C for up to 16 d and plated for bacterial enumeration at various times throughout storage. Among the 88 bacterial isolates characterized, a total of 31 rpoB allelic types representing Bacillus and Paenibacillus spp. were identified, including 5 allelic types found in both raw milk and finished product samples. The presence of the same bacterial subtypes in raw and commercially pasteurized milk samples suggests that the raw milk supply represents an important source of these spoilage bacteria. Extension of the shelf life of high-temperature, short-time pasteurized fluid milk products will require elimination of these organisms from milk-processing systems.

Evaluation of surimi, fat and water content in a low/no added pork sausage formulation using response surface methodology.

Response surface methodology (RSM) was employed for simultaneous analysis of the effects of added surimi (0-40%), fat (5-30%) and water (10-35%), on the physical, textural and sensory characteristics of fresh breakfast pork sausages. Experimental design allowed for evaluation of potential interactive effects between these ingredients. Sausages were evaluated for texture, colour, water holding capacity (WHC) and sensory attributes. Three optimum recipes, R1 (25.3% surimi, 22.2% fat, 12.7% water, 25.3% pork), R2 (12.2% surimi, 5.5% fat, 38.7% water, 33.2% pork) and R3 (25.3% surimi, 6.3% fat, 28.5% water, 25.3% pork), were determined and these were evaluated against a full-fat commercial control (R4). Force values of R1 were not significantly different to R4, however, force values for R2 and R3 were lower (P<0.001). No significant differences were observed between R1, R3 and R4 for visual colour or sensory acceptability scores throughout the study, whereas scores for R2 were lower. Sensory analysis indicated that R2 had lower scores for texture (P<0.01), chewiness (P<0.01), acceptability (P<0.01), flavour (P<0.05) and preference (P<0.01). Results from this study suggest that it is possible to successfully replace pork meat with functional fish proteins in the manufacture of sausage type products.

Gaps in the childhood malaria burden in Africa: cerebral malaria, neurological sequelae, anemia, respiratory distress, hypoglycemia, and complications of pregnancy.

Evaluations of the African childhood malaria burden do not fully quantify the contributions of cerebral malaria (CM), CM-associated neurological sequelae, malarial anemia, respiratory distress, hypoglycemia, and pregnancy-related complications. We estimated the impact of these malaria manifestations on members of the African population < 5 years old. Calculations were based on an extensive literature review that used National Library of Medicine search engines, other bibliographic sources, and demographic data. In sub-Saharan Africa, CM annually affects 575,000 children < 5 years of age and 110,000 (approximately 19% case fatality rate [CFR]) die. Childhood survivor, of CM experience developmental and behavioral impairments: each year, 9,000-19,000 children (> 2% of survivors of CM) < 5 years of age in Africa experience neurological complications lasting > 6 months. Severe malarial anemia heavily burdens hospitals with rising admission and CFRs and with treatments that are complicated by limited and sometimes contaminated blood supplies. Severe malarial anemia occurs 1.42-5.66 million times annually and kills 190,000-974,000 (> 13% CFR) children < 5 years of age annually. Respiratory distress, hypoglycemia, and overlapping clinical manifestations cause 1.12-1.99 million cases and > 225,000 (> 18% CFR) additional deaths among African children with malaria. Maternal, placental, or fetal malaria infection during pregnancy adversely affects development and survival of fetuses and newborns through low birth weight (LBW), maternal anemia, and possibly abortion and stillbirth. Between 167,000 and 967,000 cases of malaria-associated LBW occur yearly; malaria-induced LBW kills 62,000-363,000 (> 38% CFR) newborns each year. All the gaps in the burden comprise 0.4-1.7 million deaths annually, > 50% of which are due to severe malarial anemia. These malaria-induced medical problems constitute major clinical, public health, and research challenges in that they may contribute to more than double the mortality than is generally acknowledged.

Identification and characterization of elevated microbial counts in bulk tank raw milk.

The bacterial composition of bulk tank milk from 13 farms was examined over a 2-wk period to characterize sudden elevations in the total bacterial count referred to as "spikes." Bulk tank milk samples collected at each pick-up were analyzed for standard plate count, Petrifilm aerobic count, somatic cell count, gram-negative organisms, and streptococci. Twenty standard plate count spikes were observed: 12 associated with streptococci, 4 associated with gram-negative organisms, 2 associated with streptococci and gram-negative organisms, and 2 that were not definitively characterized. Spikes ranged from 14,000 to 600,000 cfu/ml. Streptococcus uberis was isolated as the predominant organism from 11 spikes, and Escherichia coli was isolated from 4 spikes. Statistical analysis of total bacterial counts indicated a high correlation (r = 0.94) between standard plate counts and Petrifilm aerobic count. Regression analysis of standard plate counts and Petrifilm aerobic counts yielded the equation log10 (standard plate count) = 0.73 + 0.85log10 (Petrifilm aerobic count), indicating that the correlation, although strong, is not one to one. In a related pilot study, triplicate bulk tank milk samples were collected and analyzed for total bacterial count and presumptive streptococcus, gram-negative, and staphylococcus counts. Two-way ANOVA of these triplicate data indicated a lack of significant variation among the triplicate samples, suggesting that one sample can reliably gauge the microbial status of the entire bulk tank.

Genetic and biological variation in equine infectious anemia virus Rev correlates with variable stages of clinical disease in an experimentally infected pony.

Genetic and biological variation in the regulatory protein Rev of equine infectious anemia virus (EIAV) were examined throughout a clinically dynamic disease course of an experimentally infected pony. Following infection with the virulent EIAV(Wyo), the pony underwent a variable disease course, including an acute fever episode at 12 days postinfection (DPI), multiple recurrent fever episodes until 135 DPI, a prolonged subclinical period, and two late fever episodes. Viral RNA was isolated from the inoculum and sequential sera samples, and the rev exon 2/gp45 overlapping ORFs were amplified, cloned, and sequenced. Novel variants were found throughout infection, and genetic analyses indicated that both the Rev and gp45 ORFs were under selective pressure. The Rev variant predominant in the inoculum, R1, remained predominant during the early periods following infection (until 35 DPI); however, R1 was replaced by new predominant variants during the recurrent fever period (67-135 DPI). R1 reemerged as the predominant variant during the afebrile period, but a new predominant variant, R93, was associated with the late fever episodes. Rev variants predominant during recurrent febrile and late-febrile periods had significantly higher Rev-mediated nuclear export activity than the variants predominant during the acute and afebrile periods. Statistical correlation was found between Rev activity and different stages of clinical disease. Together, these results suggest that genetic and biological variation in rev may be a contributing factor in EIAV disease progression.

Fluid milk vitamin fortification compliance in New York State.

Current US regulations, as specified in the Pasteurized Milk Ordinance, require vitamin A fortification of all reduced fat fluid milk products. The addition of vitamin D is optional in all fluid products. Acceptable vitamin concentrations in fortified milks are 2000 to 3000 International units per quart for vitamin A and 400 to 600 International units per quart for vitamin D. Vitamin A and D levels were analyzed in fortified milk products collected over a 4-yr period in New York State. Samples of whole fat, 2% fat, 1% fat, and nonfat milks were collected twice per year from up to 31 dairy processing plants. For vitamin A, 44.5% of 516 samples were in compliance with current regulations, and 47.7% of 648 samples were within the acceptable range for vitamin D. Most milk samples that were out of compliance were underfortified.

Allogeneic bone marrow transplantation for low-grade lymphoma.

Advanced low-grade lymphomas are usually incurable with conventional-dose chemotherapy. It is uncertain whether cures are possible with high-dose therapy and bone marrow transplant from a human leukocyte antigen (HLA)-identical sibling. We sought to determine the outcome of HLA-identical sibling bone marrow transplants in advanced low-grade lymphoma in an observational study of 113 patients conducted at 50 centers participating in the International Bone Marrow Transplant Registry (IBMTR). The median patient age was 38 years (range, 15 to 61). Eighty percent had stage IV disease at the time of transplantation. The median number of prior chemotherapy regimens was two (range, 0 to 5). Thirty-eight percent had refractory disease and 29% a Karnofsky performance score (KPS) less than 80%. All patients underwent allogeneic bone marrow transplantation from a HLA-identical sibling donor. The conditioning regimen included total-body irradiation (TBI) in 82% of patients; cyclosporine was used for graft-versus-host disease prophylaxis in 74%. Survival, disease-free survival, recurrence rate, treatment-related mortality, and causes of death were determined. Three-year probabilities of recurrence, survival, and disease-free survival were 16% (95% confidence interval [CI], 9% to 27%), 49% (95% CI, 39% to 60%), and 49% (95% CI, 39% to 59%), respectively. Higher survival was associated with pretransplant KPS >/=90%, chemotherapy-sensitive disease, use of a TBI-containing conditioning regimen, and age less than 40 years. We conclude that high-dose therapy followed by transplantation from a HLA-identical sibling leads to prolonged survival in some patients with advanced low-grade lymphoma. Most mortality is treatment-related, and recurrences are rare.

Microbiological and chemical quality of raw milk in New York State.

Between December 1993 and January 1996, samples of raw milk from bulk tanks were collected by licensed milk haulers at the time of pick-up from 855 randomly selected farms in New York State (representing approximately 10% of all dairy farms in the state). The milk was examined for microbial and chemical qualities. Bacterial numbers were determined by standard plate count, laboratory-pasteurized count, coliform count, heat-resistant spore-forming psychrotroph count, aerobic spore count (mesophilic), rapid psychrotrophic count, and preliminary incubation count. The frequency distributions for these counts are presented. Paired correlation analyses between the microbiological parameters showed low correlations between test results; no correlation coefficients were > 0.8. The four highest positive correlation coefficients were found between standard plate count and rapid psychrotrophic count (0.7685), rapid psychrotrophic count and preliminary incubation count (0.6648), standard plate count and preliminary incubation count (0.5800), and aerobic spore count and laboratory-pasteurized count (0.5393). All other correlation coefficients were < 0.5. Milk freezing points and acid degree values were determined for all samples. Frequency distributions for these results are also presented.

Evaluation of adenosine triphosphate-bioluminescence hygiene monitoring for trouble-shooting fluid milk shelf-life problems.

An ATP-bioluminescence hygiene monitoring system was used to evaluate food contact surfaces in four fluid milk plants experiencing shelf-life problems. Postpasteurization surfaces, including gaskets, pipe fittings, valves, filler parts, and hand-washed items, were evaluated. Swab results, measured in relative light units proportional to total recovered ATP, were compared with results from the standard method of microbiological swab contact for adjacent sites of equal area. Microbiological procedures included standard plate count, coliform count, and Gram-negative bacteria count. Standard plate counts were < 1, 1 to 50, and > 50 cfu in 65, 22, and 13% of swabbed sites of < 100 RLU (relative light units); in 9, 36, and 55% of sites of 100 to 150 RLU; and in 22, 18, and 60% of sites of > 50 RLU, respectively. Thirteen sites were found with standard plate counts > 10,000 cfu per site and identified with the hygiene monitoring system (> 150 RLU). Gram-negative bacteria were the dominant bacterial type in a majority of these samples. Gram-negative bacteria were detected in a total of 22 sites tested; mean counts were 2100 cfu per site for Gram-negative bacteria and 20 cfu per site for coliform bacteria. Although limited to use on accessible sites, the hygiene monitoring system proved to be an effective, rapid tool for identifying the possible sources of postpasteurization contamination in the fluid milk plants evaluated.

The person behind the eponym: Wilhelm Frederick von Ludwig (1790-1865).

In 1836, Wilhelm Frederick von Ludwig described a fast-spreading, nearly always fatal infection involving the connective tissues of the neck and floor of the mouth. Named after this Stuttgart physician, the condition has been known since as "Ludwig's angina". This biographical sketch highlights the life and times of the man behind the eponym, who was lauded for his surgical prowess at the age of 19, went on to become president of the Württemberg Medical Association, and whose name and the condition he described continue to be recognized today.

Multiple eruptive dermatofibromas in an HIV-positive man.

Multiple dermatofibromas (DFs) may occur in association with altered immunity, including systemic lupus erythematosus and iatrogenic immunosuppression. We report a case of multiple eruptive DFs which occurred in a patient positive for human immunodeficiency virus (HIV). The association of eruptive DFs and HIV infection has not been previously reported. The mechanism for the development of DFs in the setting of immune disturbance remains unclear. In the setting of HIV infection, DFs may clinically mimic Kaposi's sarcoma.