Performance of three small-scale wastewater treatment plants. A challenge for possible re use.

The study focused on the assessment of the performance of three WWTPs in Greece by the estimation of the microbiological and chemical quality of influent and effluent sewage. Physicochemical parameters were recorded (temperature, pH, COD, BOD, suspended solids, conductivity), and meteorological data were collected (air temperature, rain). Microbiological parameters were analyzed (Escherichia coli, total coliforms, bacteriophages, Salmonella, human adenoviruses, Candida, Pseudallescheria boydii, helminths, parasites Cryptosporidium ssp., and Giardia spp.). Statistically significant correlations among the various aforementioned parameters were investigated, in an attempt to propose appropriate processing performance indicators. Furthermore, the study aimed to assess current joint ministerial decision (JMD) on wastewater reuse, for irrigation purposes; to evaluate its practicability and its potential for public health protection. In the vast majority, outlet samples from all three studied WWTPs were not appropriate for irrigation reuse purposes based on BOD50 and suspended solids limit values, set by the current JMD, for both limited and unrestricted irrigation applications. Reductions for E. coli, total coliforms, and bacteriophages were found to range between 2-3, 1.5-2.5, and 2-4 log10 values, respectively. Salmonella spp. was detected in outlet sewage samples from Patra (PAT), Arachova (ARH), and Livadeia (LEV), at 23% (3/13), 33% (4/12), and 38% (5/13), respectively. Molds were detected at 92.3% (12/13), 100% (13/13), and 91.6% (11/12), respectively, while Candida was found at 85% (11/13), 67% (8/12), and 46% (6/13). A high prevalence of Pseudallescheria boydii, in outlet samples from all studied WWTPs is an important public health issue, which underlines the need for further studies on this emerging fungal pathogen in wastewater reuse applications. Pseudallescheria boydii was found at 85% (11/13), 67% (8/12), and 46% (6/13), respectively. Helminths were found in both inlet and outlet samples, of all studied WWTPs, at 100%. Human adenoviruses, were detected at high percentages in outlet samples at 76.9% (10/13), 92.3% (12/13), 84.6% (11/13), respectively, while no influence of UV irradiation was recorded on the entry and exit loads of human adenoviruses. No influence of meteorological parameters was found on the microbiological and chemical parameters, with the exception of a weak positive correlation between environmental temperature and bacteriophages. A moderate positive correlation was found between BOD and suspended solids, bacteriophages, and total coliforms, bacteriophages and E. coli, and bacteriophages and adenoviruses. A significant positive correlation was found between total coliforms and E. coli, COD and BOD, and suspended solids and COD. No correlations were proved between human pathogens and bacterial indicator parameters. Collectively, our findings underlined the unsuitability of the current JMD on wastewater reuse in Greece, or public health protection. The study is expected to support the development of a public health risk assessment model based on quantitative risk assessment on the use of treated wastewater for irrigation purposes in Greece.

Computational and functional analysis of the putative SH2 domain in Janus Kinases.

Src homology 2 (SH2) domains interact in a highly specific manner with phosphorylated tyrosine residues on other signaling molecules. Protein tyrosine kinases (PTK) frequently contain SH2 domains, which often control signaling specificity. The Janus Kinases (JAKs) are a family of PTKs involved in signal transduction pathways mediated by various cytokines. Initial characterization of JAKs showed no identifiable SH2 domain. However, we have found substantial evidence supporting the existence of an SH2 domain in JAKs through the use of various web-based computational analysis programs. Predictive secondary and tertiary structures recognize an SH2 domain in JAKs. In addition, a three-dimensional homology model was constructed using the SH2 domains of Src tyrosine kinase and Syp tyrosine phosphatase as templates. These results, in conjunction with preliminary binding studies showing interactions with tyrosine phosphorylated proteins in activated splenocytes, suggest a functional role for this domain in JAKs.

Molecular cloning of a chicken JAK homolog from activated T cells.

A cDNA encoding a JAK-related protein was isolated from a chicken Tcell library by screening with a PCR-generated probe that corresponds to a conserved region in the kinase domain. Sequence analysis reveals an ORF of 3318nt, encoding a protein with a calculated molecular weight of 123000. Chicken JAK (cJAK) contains a double catalytic domain that is characteristic of the JAK family of tyrosine kinases. Compared with mammalian JAKs, the kinase domain shows 70% sequence identity with the corresponding region of the mammalian JAKs. Overall, cJAK shows approximately 59% amino acid identity with mammalian JAK3s, and 52% amino acid identity with mammalian JAK2s. cJAK is expressed predominantly in thymus and spleen, with lower levels in kidney, thyroid and liver. cJAK is also expressed at low levels in unstimulated splenic Tcells, whereas mRNA levels are increased after activation of the Tcells with Con A. The sequence analysis and pattern of expression suggests that this is an avian homolog of JAK3.

Glycoprotein B genotype correlates with cell tropism in vivo of human cytomegalovirus infection.

Human cytomegalovirus (HCMV) strains can be classified into four genotypes of the glycoprotein B (gB). In a previous study, the gB genotype 1 was found more frequently in bone marrow transplant recipients with nonfatal HCMV infection than in patients who died from HCMV disease [Fries et al. (1994): Journal of Infectious Diseases 169:769-774]. The distribution and cell tropism of different gB types in vivo were investigated. The gB type of HCMV was determined in blood or urine specimen from 76 organ and 47 bone marrow transplant recipients using PCR and restriction fragment length polymorphism (RFLP). The leukocyte populations (polymorphonuclear leukocytes, monocytes, T lymphocytes, non-T lymphocytes) of 20 viremic patients were purified by a fluorescence-activated cell sorter (FACS) and examined for HCMV infection by PCR. Sequence analysis of four randomly selected strains showed that gB types were similar to published sequences and no atypical gB types were found. Within the compartments blood and urine, the gB types were almost equally distributed, whereas the gB type 1, in contrast to gB types 2 and 3, did not infect T lymphocytes in vivo. These data show that the gB type correlates with viral tropism in vivo and thus provides further evidence that the gB variation may indeed influence the virulence of HCMV.

Prevalence of antibodies to Borrelia burgdorferi and tick-borne encephalitis virus in an endemic region in southern Germany.

With the intention to evaluate the frequency of asymptomatic infections with TBE virus and B. burgdorferi clinical data and serum specimens were collected from 393 individuals living in an area endemic for both agents (Freiburg, southern Germany). Sera were examined by ELISA. Borderline and positive results were checked by immunoblotting. Only specific antibodies detected by immunoblotting (B. burgdorferi: 22 kDa, 31 kDa, 34 kDa, 39 kDa, 83 kDa; TBE: glycoprotein E) were assessed as positive findings. Specific antibodies to B. burgdorferi were detected in 17/105 individuals with possible symptoms of borreliosis (16%) and in 36/288 individuals without current or previous symptoms of borreliosis (12.5%). Antibody to TBE virus was demonstrated in 34/361 individuals (9.4%) without clinical symptoms of TBE or vaccination against TBE. Thirty individuals had been immunised against TBE (10.6%) and two had clinical TBE one year ago. Antibodies against both agents were detected only in 1.5% of all subjects. Considering the low seroprevalence, antibody screening is not recommended prior to TBE vaccination.

Killing activity of microwaves in milk.

The killing activity of microwaves of 2450 MHz frequency and 600 W power on Pseudomonas aeruginosa, Escherichia coli, Enterobacter sakazakii, Klebsiella pneumoniae, Staphylococcus aureus, Candida albicans, Mycobacterium terrae and poliomyelitis vaccine-virus suspended in five infant formula preparations was investigated. The samples were brought to the boil (85-100 s depending on milk type). They had reached average temperatures of 82-93 degrees C at this point. Most of the vegetative organisms were killed. In those samples where growth was still detectable after microwave treatment, a significant reduction in viable micro-organisms (at least 5000-fold) was noted. We conclude that microwave beating to the boil is a convenient and fast method to reduce microbial contamination of infant feeds. However, care should be taken to ensure that milk is adequately cooled to the required temperature before it is fed to an infant.

Congenital rubella syndrome despite repeated vaccination of the mother: a coincidence of vaccine failure with failure to vaccinate.

Congenital rubella syndrome occurred in a boy born to a mother who had been properly vaccinated three times before conception, but developed only low titers of rubella antibody. The non-vaccinated father most likely infected the mother on a home visit during the third to fifth weeks of gestation. He served in the army and fell ill during an outbreak of a rubella-like disease in his military unit. The mother subsequently developed a slight rash, but rubella IgM antibodies were lacking and rubella infection was not suspected. This incidence demonstrates the necessity of vaccinating all children, including boys and young men, in order to reduce the number of infections and prevent further cases of congenital rubella syndrome.

Detection of human cytomegalovirus in urine by DNA-DNA and RNA-DNA hybridization.

A diagnostic hybridization assay for detecting human cytomegalovirus (HCMV) DNA in urine specimens was developed by using cloned viral DNA and in vitro-synthesized RNA probes. Both probes detected 3 pg of homologous DNA and hybridized with DNA of HCMV but not with other viral or human cellular DNA tested. In 95 urine specimens simultaneously tested by cell culture, the sensitivity of hybridization was at least 83%, and the specificity was at least 92%. This assay will be useful for rapid viral diagnosis with wide clinical applications such as screening of immunocompromised patients and quantitation of viral shedding in patients with primary or reactivated HCMV infection who may be receiving antiviral therapy.

Amplified direct immunofluorescence (AMDI) for detection of Epstein-Barr virus nuclear antigen.

Low antigen concentrations could be identified in human cells by sequential application of (a) FITC-labeled human antibodies at high dilution, (b) rabbit antiserum to the hapten fluorescein isothiocyanate (FITC), and (c) FITC-anti-rabbit globulin. The high specificity of direct immunofluorescence (a) was not affected by the amplifying steps (b) and (c). Using this AMDI technique Epstein-Barr (EB) virus nuclear antigen (EBNA) could be specifically stained with human sera up to a dilution of 1 : 4000. Owing to the high dilutions applied, unwanted antibody reactivity in the FITC-labeled serum could be blocked by preincubating with unlabeled undiluted human sera. Thus EBNA was selectively stained in EB virus producer cells. Moreover, EBNA was specifically detected in human tumor biopsy material by the use of AMDI.

Simple mathematical deductions in the seroepidemiology of viral infections. I. Herpesvirus group (herpesvirus hominis, varicella-zoster virus, cytomegalovris, Epstein-Barr-Virus).

Large samples of nonselected persons collected in South-West Germnay were investigated for the prevalence of serum antibodies to the human Herpesviruses HSV, VZV, CMV and EBV. According to "catalytic models", which compare the infection spread to simple chemical reactions of molecules, a mathematical approximation of the serum surveys was performed. Through a new deduction of the exponential function y = k (l - e-r+) a simple way was found to estimate the annual attack rates in percent of the susceptible, seronegative people. While it was possible to represent the prevalence of serum antibodies to VZV by a continuous curve, a biphasic curve to the antibody prevalence rates in the epidemiology of the other Herpesviruses proved to be more adequate indicating changes in hormonal balance and social behaviour. The use of the epidemiologic parameters k and r for the characterization of the test sensitivity was examined for CMV. By evaluation of the mean antibody titres to CMV, HSV, and EBV throughout different age groups, information about the reactivation of the Herpesvirus diseases could be obtained.

Solid-phase enzyme immunoassay for immunoglobulin M antibodies to cytomegalovirus.

A sensitive, solid-phase enzyme immunoassay for the detection of immunoglobulin M antibodies to cytomegalovirus is described. The results of the enzyme immunoassay correlated well with those obtained by an indirect immunofluorescence method. Horseradish peroxidase proved to be a more sensitive label than alkaline phosphatase. Nonspecific reactions, occurring with commercially available cytomegalovirus antigens, could be avoided by using a nuclear antigen prepared from sonically disrupted nuclei of cytomegalovirus-infected cells.

IgM antibodies to cytomegalovirus during pregnancy.

IgM antibodies to cytomegalovirus (CMV) could be detected in about 7 per cent of 629 pregnant women whereas in 225 nonpregnant control women of similar age distribution only 2.6 per cent showed CMV IgM antibodies. Intrauterine CMV infections were almost exclusively detected among the CMV IgM positive gravides. The high incidence of CMV IgM antibodies in pregnant women can be possibly explained by an increased rate of CMV reactivations during pregnancy. We were able to show that during CMV reactivation an intrauterine infection might occur.

Sensitive method to detect non-complement-fixing antibodies to Epstein-Barr virus nuclear antigne.

The anti-complement immunofluorescence (ACIF) test was modified to detect non-complement-fixing antibodies to Epstein-Barr (EB) virus nuclear antigen (IBNA). These EBNA antibodies belong to the immunoglobulin classes IgA and IgG. In our method anti-human gamma-globulin (AHG) was bound to the EBNA antibodies before the complement was added. If only non-complement-fixing antibodies are present the complement can be fixed to the AHG. Within only a few weeks of the onset of infectious mononucleosis (IM) the non-complement-fixing EBNA antibodies reach high titers while the complement-fixing antibodies (detected by the ACIF test) are not yet present. Anti-EBNA-IgM antibodies were not found in the IgM fractions of sera taken at different stages of IM.

[Quantitative and virus specific serum IgM detection in newborns (author's transl)]. / Quantitativer und virusspezifischer Serum-IgM-Nachweis bei Neugeborenen

Serum IgM concentration was determined in umbilical cord blood samples from 1000 newborns. 34 cases presenting elevated IgM values (greater than or equal to 30 mg%) were tested for virus specific IgM antibodies. We could demonstrate such virus specific IgM antibodies against Cytomegalovirus (4), against Rubella, Influenza A and Influenza B (2 each) and against Coxsackie virus (1). The quantitation of IgM in blood samples of child bearing women showed no elevated values compared to those of non-pregnant women of similar age groups.

Rapid method to detect rubella immunoglobulin M and immunoglobulin A antibodies.

Immunoglobulin (Ig) G was removed from serum specimens by precipitation with gamma chain-specific anti-human IgG of rabbit origin. The remaining rubella virus-specific IgM (and IgA) antibodies were then detected by the rubella hemagglutination-inhibition test. This procedure has proven to be as reliable as estimations carried out with IgM fractions separated on a sucrose density gradient.