Pepper mild mottle virus: A plant pathogen with a greater purpose in (waste)water treatment development and public health management.

An enteric virus surrogate and reliable domestic wastewater tracer is needed to manage microbial quality of food and water as (waste)water reuse becomes more prevalent in response to population growth, urbanization, and climate change. Pepper mild mottle virus (PMMoV), a plant pathogen found at high concentrations in domestic wastewater, is a promising surrogate for enteric viruses that has been incorporated into over 29 water- and food-related microbial quality and technology investigations around the world. This review consolidates the available literature from across disciplines to provide guidance on the utility of PMMoV as either an enteric virus surrogate and/or domestic wastewater marker in various situations. Synthesis of the available research supports PMMoV as a useful enteric virus process indicator since its high concentrations in source water allow for identifying the extent of virus log-reductions in field, pilot, and full-scale (waste)water treatment systems. PMMoV reduction levels during many forms of wastewater treatment were less than or equal to the reduction of other viruses, suggesting this virus can serve as an enteric virus surrogate when evaluating new treatment technologies. PMMoV excels as an index virus for enteric viruses in environmental waters exposed to untreated domestic wastewater because it was detected more frequently and in higher concentrations than other human viruses in groundwater (72.2%) and surface waters (freshwater, 94.5% and coastal, 72.2%), with pathogen co-detection rates as high as 72.3%. Additionally, PMMoV is an important microbial source tracking marker, most appropriately associated with untreated domestic wastewater, where its pooled-specificity is 90% and pooled-sensitivity is 100%, as opposed to human feces where its pooled-sensitivity is only 11.3%. A limited number of studies have also suggested that PMMoV may be a useful index virus for enteric viruses in monitoring the microbial quality of fresh produce and shellfish, but further research is needed on these topics. Finally, future work is needed to fill in knowledge gaps regarding PMMoV's global specificity and sensitivity.

Microbial source tracking in shellfish harvesting waters in the Gulf of Nicoya, Costa Rica.

Current microbial water quality monitoring is generally limited to culture-based measurements of fecal indicator bacteria (FIB). Given the many possible sources of fecal pollution within a watershed and extra-intestinal FIB reservoirs, it is important to determine source(s) of fecal pollution as a means to improve water quality and protect public health. The principal objective of this investigation was to characterize the microbial water quality of shellfish harvesting areas in the Gulf of Nicoya, Costa Rica during 2015. In order to achieve this objective, the specificity and sensitivity of 11 existing microbial source tracking (MST) PCR assays, associated with cows (BacCow), dogs (BacCan, DogBac), domestic wastewater (PMMoV), general avian (GFD), gulls (Gull2), horses (HorseBac, HoF), humans (HF183, HPyV), and pigs (PF), were evaluated using domestic wastewater and animal fecal samples collected from the region. The sensitivity of animal-associated assays ranged from 13 to 100%, while assay specificity ranged from 38 to 100%. The specificity of pepper mild mottle virus (PMMoV) and human polyomavirus (HPyV) was 100% for domestic wastewater, as compared to 94% specificity of the HF183 Bacteroidales marker. PMMoV was identified as a useful domestic wastewater-associated marker, with concentrations as high as 1.1 × 105 copies/ml and 100% sensitivity and specificity. Monthly surface water samples collected from four shellfish harvesting areas were analyzed using culture-based methods for Escherichia coli as well as molecular methods for FIB and a suite of MST markers, which were selected for their specificity in the region. While culturable E. coli results suggested possible fecal pollution during the monitoring period, the absence of human/domestic wastewater-associated markers and low FIB concentrations determined using molecular methods indicated sufficient microbial water quality for shellfish harvesting. This is the first study to our knowledge to test the performance of MST markers in Costa Rica as well as in Central America. Given the lack of wastewater treatment and the presence of secondary sources of FIB, this study highlights the importance of an MST toolbox approach to characterize water quality in tropical regions. Furthermore, it confirms and extends the geographic range of PMMoV as an effective tool for monitoring domestic wastewater pollution.

Faecal pollution along the southeastern coast of Florida and insight into the use of pepper mild mottle virus as an indicator.

AIMS: To identify faecal pollution along the southeastern Florida coast and determine the performance of a reverse transcription-quantitative polymerase chain reaction (RT-qPCR) method for pepper mild mottle virus (PMMoV). METHODS AND RESULTS: In 2014, bimonthly surface water samples were collected from inlets, exposed to runoff and septic seepage, and coastal sites, exposed to ocean outfalls. Analysis of culturable enterococci and a suite of microbial source tracking (MST) markers (BacHum, CowM2, DogBact, HF183, HPyV, PMMoV) revealed faecal pollution, primarily of human origin, at all sites. Since PMMoV was detected more frequently than other MST markers, the process limits of quantification (undiluted to 10-2 dilution) and detection (10-2 dilution) for the RT-qPCR method were determined by seeding untreated wastewater into the coastal waters. Simulated quantitative microbial risk assessment, employing human norovirus as a reference pathogen, calculated a 0·286 median risk of gastrointestinal illness associated with the PMMoV limit of detection. CONCLUSIONS: All sites met the U.S. EPA recreational water criteria, despite detection of domestic wastewater-associated MST markers. PMMoV correlated only with human-associated MST markers. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated that PMMoV is an important domestic wastewater-associated marker that should be included in the MST toolbox; therefore, future studies should thoroughly investigate the health risks associated with its detection and quantification in environmental waters.

Reduction of nutrients, microbes, and personal care products in domestic wastewater by a benchtop electrocoagulation unit.

To preserve environmental and human health, improved treatment processes are needed to reduce nutrients, microbes, and emerging chemical contaminants from domestic wastewater prior to discharge into the environment. Electrocoagulation (EC) treatment is increasingly used to treat industrial wastewater; however, this technology has not yet been thoroughly assessed for its potential to reduce concentrations of nutrients, a variety of microbial surrogates, and personal care products found in domestic wastewater. This investigation's objective was to determine the efficiency of a benchtop EC unit with aluminum sacrificial electrodes to reduce concentrations of the aforementioned biological and chemical pollutants from raw and tertiary-treated domestic wastewater. EC treatment resulted in significant reductions (p < 0.05, α = 0.05) in phosphate, all microbial surrogates, and several personal care products from raw and tertiary-treated domestic wastewater. When wastewater was augmented with microbial surrogates representing bacterial, viral, and protozoan pathogens to measure the extent of reduction, EC treatment resulted in up to 7-log10 reduction of microbial surrogates. Future pilot and full-scale investigations are needed to optimize EC treatment for the following: reducing nitrogen species, personal care products, and energy consumption; elucidating the mechanisms behind microbial reductions; and performing life cycle analyses to determine the appropriateness of implementation.

A case study of enteric virus removal and insights into the associated risk of water reuse for two wastewater treatment pond systems in Bolivia.

Wastewater treatment ponds (WTP) are one of the most widespread treatment technologies in the world; however, the mechanisms and extent of enteric virus removal in these systems are poorly understood. Two WTP systems in Bolivia, with similar overall hydraulic retention times but different first stages of treatment, were analyzed for enteric virus removal. One system consisted of a facultative pond followed by two maturation ponds (three-pond system) and the other consisted of an upflow anaerobic sludge blanket (UASB) reactor followed by two maturation (polishing) ponds (UASB-pond system). Quantitative polymerase chain reaction with reverse transcription (RT-qPCR) was used to measure concentrations of norovirus, rotavirus, and pepper mild mottle virus, while cell culture methods were used to measure concentrations of culturable enteroviruses (EV). Limited virus removal was observed with RT-qPCR in either system; however, the three-pond system removed culturable EV with greater efficiency than the UASB-pond system. The majority of viruses were not associated with particles and only a small proportion was associated with particles larger than 180 µm; thus, it is unlikely that sedimentation is a major mechanism of virus removal. High concentrations of viruses were associated with particles between 0.45 and 180 µm in the UASB reactor effluent, but not in the facultative pond effluent. The association of viruses with this size class of particles may explain why only minimal virus removal was observed in the UASB-pond system. Quantitative microbial risk assessment of the treated effluent for reuse for restricted irrigation indicated that the three-pond system effluent requires an additional 1- to 2-log10 reduction of viruses to achieve the WHO health target of <10(-4) disability-adjusted life years (DALYs) lost per person per year; however, the UASB-pond system effluent may require an additional 2.5- to 4.5-log10 reduction of viruses.

Audit of birth defects in 34,109 deliveries in a tertiary referral center.

The objective of the study is to determine the proportion and different types of birth defects among the children born in Hospital Kuala Lumpur. A cross-sectional study was conducted for a period of 18 months where all consecutively born infants, dead or alive were included. There were total of 34,109 births recorded during this period. The proportion of birth defects in Hospital Kuala Lumpur was 3.1% (n = 1056). The commonest involved were the hematology system, (157.7 per 10,000 births), the central nervous system, genitourinary system and chromosomal anomalies. The proportion was significantly higher in males and in the Chinese (p < 0.001). The commonest abnormalities are Glucose 6 Phosphate Deficiency (157.7/10000), Down's syndrome (12.6/10000), thalassaemia (8.8/10000), cleft lip and/or palate (7.6/10000) and anencephaly (7.3/10000). Neural tube defect is common and ranked second after G6PD deficiency. There is a need for a birth defect registry to assess the extent of the problem in Malaysia.

Delivery of macrosomic babies: management and outcomes of 330 cases.

This study reviews the deliveries of macrosomic babies and their outcomes. A total of 330 macrosomic (birth weight > or =4 kg) cases were studied retrospectively from July 1999 to December 1999 in the Maternity Hospital of Kuala Lumpur. The variables studied included induction of labour, mode of delivery and the incidence of maternal and perinatal complications. Three hundred and thirty macrosomic infants were delivered during the period of study. Vaginal delivery was achived in 56% of the study cases. The percentage of vaginal delivery was higher among those who had induction of labour (63%) compared to the group without induction of labour (50%). Vaginal delivery was planned in 267 mothers and of these 69% achieved vaginal delivery. Twelve per cent of the macrosomic infants were delivered by elective caesarean section. Shoulder dystocia occurred in 4.9% of vaginal deliveries. Eighty-eight neonates were admitted to the special care nursery unit and 57% of these infants were delivered by elective caesarean section. Perineal trauma occurred in 26% of vaginal deliveries. Post-partum haemorrhage occurred in 32% of caesarean deliveries compared to 4% in vaginal deliveries. Two cases of stillbirths were documented but no maternal death occurred during the period of study. Vaginal delivery is the most frequent mode of delivery for a fetus weighing in excess of 4 kg and vaginal delivery should be attempted in the absence of contraindications, because vaginal delivery has less maternal morbidity compared to caesarean delivery. However, shoulder dystocia remains a significant complication of vaginal delivery for macrosomic fetuses.

An immunoscintigraphic evaluation of the engineered human monoclonal antibody (hCTMO1) for use in the treatment of ovarian carcinoma.

OBJECTIVE: To assess the safety and targeting ability of the engineered human antibody (hCTMO1) in women with ovarian carcinoma. DESIGN: The monoclonal antibody labelled with Indium-111 was administered to women with suspected primary or recurrent ovarian carcinoma six days pre-operatively. The first group of women was given a dose of 0.1 mg per kg body weight of radiolabelled antibody. A second group of women received 1 mg per kg body weight and finally a third group was given 1 mg per kg body weight of unlabelled antibody followed one hour later by 0.1 mg per kg body weight of radiolabelled antibody. All the women were then imaged using a gamma camera one hour and up to 96 hours after injection. PARTICIPANTS: Fourty-four women in whom there was a high suspicion of primary ovarian carcinoma on the basis of ultrasound or CT imaging and serum CA125 and those in whom there was a suspicion of recurrent ovarian carcinoma after being treated for histologically confirmed carcinoma. SETTING: The Queen's Medical Centre, Nottingham and University Hospital Vrije Universiteit, Amsterdam, The Netherlands. RESULTS: At the low dose of antibody the sensitivity for detection of ovarian carcinoma was 70%. After increasing the dose of antibody and also after pre-dosing with unlabelled antibody the sensitivity increased to 100%, but there was a large number of false positive results at the higher dose, and therefore the specificity was low. The liver and bone marrow were the organs with the highest activities. CONCLUSION: The genetically engineered antibody hCTMO1 is safe for use in women. This antibody effectively targets ovarian carcinoma and has greater potential as a vector for therapeutic use than as a diagnostic agent.

Academic obstetrics and gynaecology in the UK.

An investigation into the current structure of academic obstetrics and gynaecology was performed by circular to 33 departments in the UK. All departments responded. Of the 217 clinical academic staff, 32% were funded by National Health Service trusts and charities. Between 1 January 1995 and 1 January 1997, 84 posts were advertised eliciting a total of 302 applicants. Of the 73 staff that resigned over the same period, only 11 remained in academic obstetrics and gynaecology. The most commonly listed concern affecting career choice was listed as pressures from research and clinical practice.

Cellular proliferation in the placenta in normal human pregnancy and pregnancy complicated by intrauterine growth restriction.

OBJECTIVE: To determine the incidence of cellular proliferation in the placenta throughout the three trimesters of normal pregnancy, and in the third trimester of pregnancy complicated by intrauterine growth restriction (IUGR). METHODS: Placental samples were obtained from 17 first-trimester pregnancies, 9 second-trimester pregnancies, 33 uncomplicated third-trimester pregnancies, and 21 third-trimester pregnancies complicated by IUGR. These samples were then stained by immunohistochemical technique, using the monoclonal antibody MIB-1. RESULTS: The incidences of cellular proliferation in the four groups were as follows: first trimester (n = 17): 11.8% of cells (8.51-17.04); second trimester (n = 9): 9.88% of cells (5.04-10.99); normal third trimester (n = 33): 3.15% of cells (2.07-3.7); IUGR third trimester (n = 21): 3.7% of cells (3.02-4.85). The decline in cellular proliferation throughout the three trimesters of pregnancy was statistically significant (P < .0001 Kruskall-Wallis test). The Spearman rank correlation for proliferative index against gestational age had a P value less than .0001 (Rho corrected for ties = -0.81). There was no statistically significant difference in the incidence of cellular proliferation between normal third-trimester and IUGR third-trimester samples. CONCLUSION: The incidence of cellular proliferation in the placenta declines as pregnancy progresses, a finding that agrees with previous work by others. The incidence of cellular proliferation was not altered in cases of IUGR.

Biodistribution of 111In-labelled engineered human antibody CTM01 (hCTM01) in ovarian cancer patients: influence of prior administration of unlabelled hCTM01.

mAb hCTM01 binds a carcinoma-associated antigen, the MUC1 gene product. The antigen is also present in the circulation, and administration of 111In-labelled hCTM01 results in the formation of immune complexes with enhanced accumulation in the liver. To avoid the unwanted effect of circulating radioactive immune complexes, a strategy to remove the circulating antigen was investigated using a split-dosage schedule. Eleven patients suspected of having ovarian carcinoma were injected with 1 mg/kg unlabelled hCTM01, 1 h before receiving 0.1 mg/kg 111In-labelled hCTM01 (100 M Bq). The amount of radioactivity was determined in resected tumour tissue, various normal tissues and blood samples obtained at laparotomy 6 days postinjection (p.i.). In all patients, the circulating antigen decreased to its nadir after the unlabelled antibody infusion and immune complex formation was demonstrated. Uptake in tumour deposits 6 days p.i. was 11.1 times higher than in normal tissues (P < 0.0001) and 5.9 times higher than in blood (P < 0.0001). 111In activity in liver tissue was comparable to 111In uptake in tumour tissue, and considerably lower than previously reported in patients not pretreated with unlabelled antibody. The split-dosing strategy would appear to be advantageous for use of hCTM01 as a specific carrier for the delivery of cytotoxic agents to patients with ovarian cancer.

Placental apoptosis in normal human pregnancy.

OBJECTIVES: The study aims were to conclusively demonstrate apoptosis in the human placenta and to quantify its incidence at different stages of pregnancy. STUDY DESIGN: Placental samples were obtained from 28 first-trimester pregnancies and 38 uncomplicated third-trimester pregnancies. Light microscopy, electron microscopy, and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate marker nick end-labeling staining were used to identify apoptosis. Light microscopy was used to quantify its incidence. RESULTS: Apoptosis has been conclusively demonstrated within placental tissue. Quantification of apoptosis (medians and interquartile ranges) was as follows: first trimester (n = 28), 0.07% of cells (0.05% to 0.14%); third trimester (n = 39), 0.14% of cells (0.09% to 0.20%). The incidence of apoptosis was significantly higher in the third trimester than in the first trimester (p < 0.01, Mann-Whitney U test). CONCLUSIONS: Placental apoptosis increases significantly as pregnancy progresses, suggesting that it may play a role in the normal development and aging of the placenta.

The effect of circulating antigen on the biodistribution of the engineered human antibody hCTM01 in a nude mice model.

Clinical studies are currently underway to assess the biodistribution and therapeutic potential of the genetically engineered human antibody hCTM01 directed against polymorphic epithelial mucin (PEM) in patients with ovarian carcinoma. The present study was undertaken to assess the effect of circulating PEM antigen on the biodistribution of the anti-PEM antibody in mice bearing MUC-1 transfected adenocarcinoma cell lines. Tumour xenografts were established from three cell lines: 413-BCR, which expressed antigen on the cell surface and also shed antigen into the circulation, E3P23, which expressed the antigen but did not shed into the circulation, and a negative control (410.4 MUCI). Groups of five mice were injected with 1.0 mg/kg antibody, imaged after 72 h and then sacrificed, followed by assay of tissue uptake. The results showed a clear difference in the tumour and liver uptake, with the non-secreting cell line showing almost twice the tumour uptake and approximately 20% of the liver uptake of the secreting cell line.

Increased placental apoptosis in intrauterine growth restriction.

OBJECTIVES: Our purpose was to investigate a possible role for apoptosis in the pathophysiologic mechanisms of intrauterine growth restriction. STUDY DESIGN: Placental samples were obtained from 43 uncomplicated third-trimester pregnancies and from 26 pregnancies complicated by intrauterine growth restriction. The definition used to identify cases of intrauterine growth restriction depended on three criteria: clinical evidence of suboptimal growth, ultrasonographic evidence of deviation from an appropriate growth percentile, and individualized birth weight ratios <10th percentile. Light microscopy was used to quantify the incidence of apoptosis. Electron microscopy and TUNEL (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling) staining were used to confirm the occurrence of apoptosis. RESULTS: Quantification of apoptosis (medians and interquartile ranges) resulted in the following values: normal third trimester (n = 43) 0.14% of cells (0.08% to 0.20%) and intrauterine growth restriction third trimester (n = 26) 0.24% of cells (0.16% to 0.29%). The incidence of apoptosis was significantly higher in placentas from pregnancies with intrauterine growth restriction compared with normal third-trimester placentas (p < 0.01, Mann Whitney U test). CONCLUSIONS: These results suggest that apoptosis may play a role in the pathophysiologic mechanisms of intrauterine growth restriction.

Biodistribution of (111)indium-labeled engineered human antibody CTMO1 in ovarian cancer patients: influence of protein dose.

Thirty-one patients suspected of having ovarian cancer received a single i.v. injection of radiolabeled (100 MBq (111)In) engineered human CTMO1 (hCTMO1) to investigate its potential as an internalizing drug carrier. hCTMO1 is a complementary-determining region-grafted human IgG4 monoclonal antibody recognizing an ovarian carcinoma-associated antigen, the MUC-1-gene product. The amount of radioactivity was determined in tumor tissue, various normal tissues, including liver biopsies, and blood samples obtained at laparotomy, 6 days after injection of either 0.1 or 1.0 mg hCTMO1/kg of body weight. Circulating antigen-15-3 was measurable in all patients before injection, and immune complex formation was already present at the end of infusion. In the 0.1 mg/kg group, most of the radioactivity was bound to immune complexes, whereas in the 1.0 mg/kg group, most was bound to IgG monomers. Increasing the hCTMO1 dose 10-fold did not influence the overall disappearance of (111)In from the blood, but the elimination half-life of (111)indium bound to immune complexes was increased 2-fold. Uptake in tumor tissue 6 days postinjection at the 0.1 mg/kg dose was 7.6 times higher (P = 0.0009) than in normal tissue and 2.5 times higher (P = 0.03) than in blood. At the 1.0 mg/kg dose, the uptake in tumor tissue was 14.0 times higher (P = 0.0003) than in normal tissue and 8.1 times higher (P = 0.0007) than in blood. Liver activity was substantial (23.7 +/- 10.5 and 18.3 +/- 6.7% of the injected dose/kg for the 0.1 and 1.0 mg/kg dose group, respectively). These results are superior to those found with other clinically tested anti-MUC-1 gene product antibodies. hCTMO1 seems to be a suitable carrier for cytotoxic agents in ovarian carcinoma patients; the better uptake results and tumor-to-blood ratios are obtained at the higher dose of 1.0 mg hCTMO1/kg body weight.

Changes in the PR interval--fetal heart rate relationship of the electrocardiogram during fetal compromise in chronically instrumented sheep.

OBJECTIVES: The evaluation of the changes in the relationship of the PR interval and fetal heart rate during prolonged fetal compromise in sheep at levels of acidosis comparable to those seen during human fetal compromise and to see whether these changes are potentially of use in the detection of fetal distress. STUDY DESIGN: A retrospective analysis of continuous fetal electrocardiogram recordings during graded fetal hypoxemia in 20 chronically cannulated fetal sheep was performed. Baseline recordings during normoxemia were compared with recordings during hypoxemia by use of Fisher's exact test and the Student t test. RESULTS: Sixteen of the 20 cases could be used for final analysis. Twelve showed a statistically significant change from a predominantly negative relationship between the PR interval and the fetal heart rate during normoxemia to a predominantly positive relationship during hypoxemia. Two cases showed an obvious trend in the same direction, which was statistically not significant. In two other cases no change in the relationship was observed. CONCLUSION: A changing relation between the PR interval and the fetal heart rate is of potential use in the detection of fetal compromise.

The potential impact of PR interval analysis of the fetal electrocardiogram (FECG) on intrapartum fetal monitoring.

A retrospective study was performed at the Queens Medical Centre, Nottingham, UK to evaluate the potential value of PR interval analysis of the FECG compared to conventional intrapartum assessment with fetal heart rate monitoring. Two-hundred sixty-five labours were selected for monitoring. Outcome was assessed by the number of fetal scalp blood samples (FBS) performed and the associated incidence of acidosis in the first stage of labour, the mode of delivery and whether or not this was expedited for fetal heart abnormality or an abnormal scalp pH. The condition of the fetus at delivery was assessed by arterial and venous blood acid-base status, Apgar score and the need for admission to the neonatal intensive care unit. Conventional electronic fetal heart rate monitoring (EFM) was used in all labours. The addition of PR interval assessment would potentially reduce the numbers of normal FBSs being carried out from 85.5% to 26.8% and the proportion of cases of missed acidosis at delivery from 8.5% to 4.5%. These results highlight the potential benefit of PR interval analysis in improving interpretation of the intrapartum cardiotocograph and need to be tested by prospective randomised controlled study.

The fetal electrocardiogram.

Advances in microprocessing technology have made fetal ECG analysis a feasible adjunct to fetal surveillance. Time interval and morphology changes of the FECG occur during fetal hypoxia. The use of these changes to detect a fetus at risk of intrapartum asphyxia awaits validation in terms of both future and ongoing clinical trials. Recognition of FECG changes during decelerations may improve the sensitivity of EFM. Antepartum FECG analysis has potential for the detection of a number of pathological fetal conditions, including intrauterine growth retardation, but remains hampered by low signal-to-noise ratios, rendering successful signal acquisition unreliable.