Highest Number Of Positive Samples Research Articles

A superhydrophobic cone to facilitate the xenomonitoring of filarial parasites, malaria, and trypanosomes using mosquito excreta/feces.

Background: Molecular xenomonitoring (MX), the testing of insect vectors for the presence of human pathogens, has the potential to provide a non-invasive and cost-effective method for monitoring the prevalence of disease within a community. Current MX methods require the capture and processing of large numbers of mosquitoes, particularly in areas of low endemicity, increasing the time, cost and labour required. Screening the excreta/feces (E/F) released from mosquitoes, rather than whole carcasses, improves the throughput by removing the need to discriminate vector species since non-vectors release ingested pathogens in E/F. It also enables larger numbers of mosquitoes to be processed per pool. However, this new screening approach requires a method of efficiently collecting E/F. Methods: We developed a cone with a superhydrophobic surface to allow for the efficient collection of E/F. Using mosquitoes exposed to either Plasmodium falciparum, Brugia malayi or Trypanosoma brucei brucei, we tested the performance of the superhydrophobic cone alongside two other collection methods. Results: All collection methods enabled the detection of DNA from the three parasites. Using the superhydrophobic cone to deposit E/F into a small tube provided the highest number of positive samples (16 out of 18) and facilitated detection of parasite DNA in E/F from individual mosquitoes. Further tests showed that following a simple washing step, the cone can be reused multiple times, further improving its cost-effectiveness. Conclusions: Incorporating the superhydrophobic cone into mosquito traps or holding containers could provide a simple and efficient method for collecting E/F. Where this is not possible, swabbing the container or using the washing method facilitates the detection of the three parasites used in this study.

Chronic experimental genital leptospirosis with autochthonous Leptospira santarosai strains of serogroup Sejroe

Although ruminants may be experimental infected by different leptospiral strains, the majority of them determine acute infection. In contrast, members of serogroup Sejroe seems to be prevalent on chronic infection. Despite the importance of naturally occurring chronic genital infection, the parasite-host long-term relationship in genital tract remains to be elucidated. Few studies succeeded on reproducing the chronic genital infection it on experimental conditions, in this context, this paper aimed to assess the chronic experimental genital leptospirosis with autochthonous Leptospira santarosai strains of serogroup Sejroe. The animals were randomly divided into three experimental groups. Each group was inoculated with a different strain of the serogroup Sejroe (FV237, FV52 and U81). On the follow-up of the infection, blood, urine and vaginal fluid collection were performed, as well as the daily follow-up of the animals clinic, and on 30, 60 and 90 days post-infection (p.i.) was performed to collect uterine tissue, uterine lavage and follicular aspirate. There were no clinical signs or haematological alterations in any of the animals, and positive MAT was detected from day 4 p.i. The group inoculated with the FV237 strain showed the highest number of positive samples in the genital tract. The chronic model of leptospiral genital infection could be successfully established, using autochthonous strains that are not often studied. It is of extreme value in order to a better understanding on the pathophysiology of the genital leptospiral infection on ruminants and consequent reproductive disease.

Detection of Hepatitis E Antibodies in Kazakhstan: A Pilot Study.

IntroductionHepatitis E virus exposure is associated with sporadic cases of acute hepatitis and outbreaks in many countries worldwide. It is particularly dangerous for pregnant women, in whom the mortality rate is high. There are no previously published data reporting circulation of this virus in Kazakhstan.MethodsWe tested blood samples for IgG anti-hepatitis E virus antibodies in 199 Kazakh participants; of these 119 were workers at the EXPO 2017 building site in Astana, 35 were volunteers who got tested at the Astana City Hall on the World Hepatitis Day 2017, and 45 were volunteers who presented for screening at the Hepatogastroenterology Outpatient Clinic of the Republican Diagnostic Center, University Medical Center.Results11 (5.5%) individuals were positive for IgG anti-HEV antibodies, with a higher seroprevalence in males (7; 6.8%) vs females (4; 4.5%). The highest number of positive samples was in the 32–46 years age group.ConclusionsThis pilot study suggests that Hepatitis E virus has been circulating in Kazakhstan. Studies are needed to determine whether it continues to be present, which viral genotypes are involved and what are the best methodologies for preventing its spread.

Virological Situation in Poland in the 2016/2017 Epidemic Season Based on Sentinel Data.

The influenza Sentinel surveillance system is a source of valuable data about the dynamics of epidemic seasons in Poland. During the epidemic season 2016/2017, more than 1,000 samples were examined, of which 48% were positive for influenza diagnosis. The predominance of influenza A/H3N2/ was confirmed. Influenza B viruses were noted in only 1% of the samples tested. After the analysis in age groups, the highest number of positive samples was observed in the group of 26-44years. Infections caused by influenza-like viruses were confirmed only in 3% of cases. The Sentinel surveillance system makes it possible to evaluate the spread of the influenza virus in each epidemic season.

Role of Immunohistochemistry in the Diagnosis of Leptospirosis in Neotropical Primates

Background: Leptospirosis is considered the most widespread zoonosis worldwide, occurring more frequently in tropical and developing regions. The aim of the present study was to detect the presence of Leptospira spp. in different primate tissues, using immunohistochemical (IHC) assays, taking advantage of the considerable number of necropsies compatible with a diagnosis of leptospirosis in neotropical primates at the Animal Pathology Laboratory (APL) of the University of Passo Fundo (UPF) in the northern region of Rio Grande do Sul.Materials, Methods & Results: Paraffin-embedded primate tissue samples were selected from necropsy examinations and subjected to IHC. The streptavidin-biotin-peroxidase method was used with diaminobenzidine chromogen (DAB) to verify immunostaining. Of the101 primates tested for Leptospira spp., 51.48% were positive; taining was distributed between lung (76.92%), liver (44.23%), and kidney (32.69%) tissue. Analysis of the combined anatomopathological verification data of the studied organs revealed a high frequency of lesions commonly observed in the tissues of animals exposed to the pathogen. For complementary diagnosis, an anti-Leptospira spp. antibody test was performed in primates at the UPF-Zoo, from which a population of the necropsied animals originated. The microscopic agglutination test (MAT) was utilized, which demonstrated 90.47% positivity in 21 individuals; sejroe and panama were the most frequent serovars.Discussion: Different intensities of tissue immunostaining were observed. Areas of fragmented or diffuse staining were considered to indicate equal positivity to that indicated by areas of staining with preserved morphology. Of 52 Leptospirapositive primates, most presented some degree of staining in lung samples, which shows a high level of involvement for this organ in primate leptospirosis. Conventional pathological diagnostic methods do not allow fort issue antigen recognition, thus making the IHC technique important to facilitate conclusive antigen sample verification. In the liver, leptospires were detected mostly between the sinusoids, hepatocytes, and Kupffer cells. In kidney tissues, staining indicated small agglomerates in the tubular lumen, interstitium, and glomeruli. All these forms of presentation have been previously reported. Considering that we detected the highest number of positive samples in lung tissue, followed by those from liver and kidney tissue, we argue that the IHC technique, when applied to samples of these three tissues, decreases the chance of false negatives. Anatomopathological studies of primate leptospirosis are scarce. In dogs, renal lesions are characterized by the necrosis and degeneration of tubular epithelium, cellular debris, and hyaline cylinders. In the liver, hepatocyte cord dissociation and biliary pigment accumulation within the canaliculi and hepatocellular necrosis are observed. These findings are similar to those from our study. In the lung, diffuse alveolar lesions are reported, with hemorrhage and edema, in addition to capillaritis. The high frequency of Leptospira-positive animals determined by serological examination was consistent with the IHC findings, thus confirming the pathogen’s high prevalence in neotropical primate populations in the studied region. Serological surveys on primate populations have already been carried out and have revealed frequency and serovar variations between regions. Immunohistochemical examination allows the detection of leptospires in various tissues and should be used based on the characteristics of the investigated case.

Applicability of FLOTAC® technique in recovering equine strongyle larvae in the pasture: A comparison study

The FLOTAC® technique represents a highly sensitive method for the isolation of oocysts, eggs, and larvae of parasites in faeces. This assay could be used for detecting free-living stages of nematodes in the pasture but no attempt has been assessed so far. Therefore, the performance of FLOTAC® technique for isolating infective larvae of nematodes in the environment was investigated and compared with the spontaneous sedimentation (SST) and centrifugal sedimentation (CST) techniques. The study was conducted in a horse farm located in northeastern Brazil, where the occurrence of strongyle larvae had been previously reported. Pasture samplings were collected monthly from January to May 2016 in a 376 m2 crop area harvested with the Guinea grass Panicum cultivar Massai. The recovery of third-stage larvae (L3) was performed using the FLOTAC®, SST and CST techniques. Values of Cohen's kappa coefficient, sensitivity, specificity, positive and negative predictive values, and accuracy of each technique were assessed. Although strongyle larvae were evenly detected, with the FLOTAC® technique yielded the highest number of positive samples (i.e., 41%, 41/100, p < .0001). The main parasites isolated belonged to the Cyathostominae and Strongylinae subfamilies. Based on these results, the FLOTAC® technique should be considered as practical and safe method for the isolation of nematode larvae in the pasture, thus opening a new potential use for this tool in the field.

A superhydrophobic cone to facilitate the xenomonitoring of filarial parasites, malaria, and trypanosomes using mosquito excreta/feces

Background: Molecular xenomonitoring (MX), the testing of insect vectors for the presence of human pathogens, has the potential to provide a non-invasive and cost-effective method for monitoring the prevalence of disease within a community. Current MX methods require the capture and processing of large numbers of mosquitoes, particularly in areas of low endemicity, increasing the time, cost and labour required. Screening the excreta/feces (E/F) released from mosquitoes, rather than whole carcasses, improves the throughput by removing the need to discriminate vector species since non-vectors release ingested pathogens in E/F. It also enables larger numbers of mosquitoes to be processed per pool. However, this new screening approach requires a method of efficiently collecting E/F. Methods: We developed a cone with a superhydrophobic surface to allow for the efficient collection of E/F. Using mosquitoes exposed to either Plasmodium falciparum, Brugia malayi or Trypanosoma brucei brucei, we tested the performance of the superhydrophobic cone alongside two other collection methods. Results: All collection methods enabled the detection of DNA from the three parasites. Using the superhydrophobic cone to deposit E/F into a small tube provided the highest number of positive samples (16 out of 18) and facilitated detection of parasite DNA in E/F from individual mosquitoes. Further tests showed that following a simple washing step, the cone can be reused multiple times, further improving its cost-effectiveness. Conclusions: Incorporating the superhydrophobic cone into mosquito traps or holding containers could provide a simple and efficient method for collecting E/F. Where this is not possible, swabbing the container or using the washing method facilitates the detection of the three parasites used in this study.

A superhydrophobic cone to facilitate the xenomonitoring of filarial parasites, malaria, and trypanosomes using mosquito excreta/feces.

Background: Molecular xenomonitoring (MX), the testing of insect vectors for the presence of human pathogens, has the potential to provide a non-invasive and cost-effective method for monitoring the prevalence of disease within a community. Current MX methods require the capture and processing of large numbers of mosquitoes, particularly in areas of low endemicity, increasing the time, cost and labour required. Screening the excreta/feces (E/F) released from mosquitoes, rather than whole carcasses, improves the throughput by removing the need to discriminate vector species since non-vectors release ingested pathogens in E/F. It also enables larger numbers of mosquitoes to be processed per pool. However, this new screening approach requires a method of efficiently collecting E/F. Methods: We developed a cone with a superhydrophobic surface to allow for the efficient collection of E/F. Using mosquitoes exposed to either Plasmodium falciparum, Brugia malayi or Trypanosoma bruceibrucei, we tested the performance of the superhydrophobic cone alongside two other collection methods. Results: All collection methods enabled the detection of DNA from the three parasites. Using the superhydrophobic cone to deposit E/F into a small tube provided the highest number of positive samples (16 out of 18) and facilitated detection of parasite DNA in E/F from individual mosquitoes. Further tests showed that following a simple washing step, the cone can be reused multiple times, further improving its cost-effectiveness. Conclusions: Incorporating the superhydrophobic cone into mosquito traps or holding containers could provide a simple and efficient method for collecting E/F. Where this is not possible, swabbing the container or using the washing method facilitates the detection of the three parasites used in this study.

Diagnostic Efficiency of Different Serological Tests and Real time PCR for Detecting Brucella Infection in Camels' Sera

px; > <Evaluation of the real-time PCR, rose bengal test (RBT), competitive ELISA, andcomplement fixation test (CFT) was done on 335 camels sera. Real-time PCR,classified 335 camel serum samples to 268 (80%) as positive and 67 (20%) asnegative. Real-time PCR, using species specific primers, distinguished 94/104 serumsamples due to B. abortus, 4/104 samples due to B. melitensis and 6/104 due tomixed infection. The results of serological tests revealed that modified mRBT75using 75 µl of serum, detected the highest number of positive samples 271 (80.9%),while 262 (78.2%), 257 (76.7%), 253 (75.5%) and 245 (73.1%) samples were foundto be positive for brucellosis using CFT, cELISA, mRBT50, and RBT25, respectively.Compared to other serological tests, the CFT proved to have the best results in thecriteria of test validations, namely; specificity (88%), PPV (96.9%), NPV (80.8%), PLR(7.9), NLR (0.06) and DOR (133.8). The Kappa (K) statistic agreements valuesbetween real-time PCR and rose bengal (RBT25), modified (mRBT50), (mRBT75),cELISA and CFT was 0.562 (± 0.053), 0.613 (± 0.052), 0.725 (± 0.048), 0.710 (± 0.047)and 0.801 (± 0.041), respectively. The authors recommend the use of real-time PCRon camel sera to confirm the disease.

The effect of temperature and chlorine residual on the presence of Legionella spp. in water systems of public and tourist facilities

Introduction: Legionella bacteria are ubiquitous microorganisms mostly found in artificial water environments, especially those which produce aerosol, such as swimming pools, saunas, and spas. Development of Legionella depends on several factors, including water temperature and chlorine. The aim of this study was to evaluate the occurrence of Legionella spp. to the temperature and free residual chlorine in tap water, water from fountains, swimming pools, cooling and heating systems.&#x0D; Methods: We collected 238 samples of water taken from different places and analyzed the presence of Legionella spp. by BAS EN ISO 11731-2:2009 - Water quality – Detection and enumeration of Legionella - Part 2: A direct membrane filtration method for waters with low bacterial counts, as samples were the waters with low expectancy of bacterial contamination compared to the temperature and free residual chlorine. The X2 test was used to show statistical significance.&#x0D; Results: Legionella spp. was detected in 18.62% of tap water and in 8.82% samples of water taken from fountains, swimming pools, and cooling and heating systems. The highest number of positive samples were detected in waters with the temperature higher than 20°C and lower than 50°C. The highest number of positive samples were reported by the concentration of free residual chlorine lower than 0.2 mg/l. The X2 test showed a statistically significant difference between positive and negative results for the presence of Legionella spp. among three groups of water samples.&#x0D; Conclusion: The research has shown a connection between environmental factors and the presence of Legionella spp. in the water systems of public and tourist facilities.

Comparative performance of isolation methods using Preston broth, Bolton broth and their modifications for the detection of Campylobacter spp. from naturally contaminated fresh and frozen raw poultry meat

The performance of different isolation methods was evaluated for the detection of Campylobacter from naturally contaminated raw poultry meat. Therefore, fresh and frozen poultry meat samples were analysed using the standard procedure (ISO 10272-1:2006), enrichment in Preston broth, and enrichment in modified Bolton broth (supplemented with (i) potassium clavulanate (C-BB), (ii) triclosan (T-BB), (iii) polymyxin B (P-BB)). The enrichment cultures were streaked onto both modified charcoal cefoperazone deoxycholate agar (mCCDA) and RAPID'Campylobacter agar (RCA). Moreover, direct plating on mCCDA and RCA was performed to quantify Campylobacter. In total, 33 out of 59 fresh retail meat samples (55.9%) were Campylobacter positive. For both fresh and frozen poultry meat samples, enrichment in Bolton broth (ISO 10272-1:2006) resulted in a higher number of positive samples than enrichment in Preston broth. Supplementation of Bolton broth with potassium clavulanate (C-BB) and triclosan (T-BB) enhanced the Campylobacter recovery from fresh poultry meat compared to non-supplemented Bolton broth, although the use of C-BB was less applicable than T-BB for Campylobacter recovery from frozen samples. Additionally, the use of RCA resulted in a higher isolation rate compared to mCCDA. The present study demonstrates the impact of culture medium on the recovery of Campylobacter from fresh and frozen naturally contaminated poultry meat samples and can support laboratories in choosing the most appropriate culturing method to detect Campylobacter.

Studying the Elimination Pattern of Caprine Arthritis Encephalitis Virus in the Milk of Infected Females

Background: Small ruminants can be infected by lentiviruses, such as Maedi-Visna Virus (MVV) and Caprine ArthritisEncephalitis Virus (CAEV). The main route of transmission is via ingestion of contaminated colostrum and milk although vertical transmission can occur. Recently, several studies for molecular detection of CAEV in milk, using conventional PCR and real-time PCR are being carried out. Considering the elimination of CAEV through the milk of infected animals and the importance of this virus in the goat production, the aim of this study was to evaluate the elimination pattern of CAEV in milk, evaluating the frequency and the concentration eliminated during the lactation.Materials, Methods &amp; Results: A cohort of four negative females for CAEV was inseminated with semen experimentally infected with CAEV-Cork strain. They were located in stalls at the Hospital of Ruminants from School of Veterinary Medicine and Animal Science from University of São Paulo, Brazil. Goats received coast-cross hay, pellet feeding, mineral salt and water ad libitum. All females were observed every day during pregnancy. After lambing, kids received warm bovine colostrum and bovine milk powder during two months. Forty milk samples were collected at five-day interval during two months. A mixture of five milliliters from each teat was obtained and cDNA extraction was performed using DNA Mini Kit. Initially, real-time PCR was performed using an endogenous control for research of the constitutive gene (12S) for goats. Using positive samples in the first reaction, another reaction was performed using specific primers for lentiviruses based on the gag gene (conserved in retroviruses). In order to compare the results, nested-PCR was performed. After realtime PCR, cDNA was detected in samples from one female, corresponding to the day of calving, 14th, 20th, 25th, 35th and 40th day postpartum (15%; 6/40). The absence of amplified cDNA in thirty days postpartum, as well as in the final twenty days of lactation, was observed. Sample corresponding to the 7th day postpartum was not obtained. The virus concentration throughout lactation grew up until forty days postpartum. After this period, there was no cDNA amplification. In Nested PCR, positive results were detected in samples corresponding to the day of calving, 15th days, 20th days and 30th days postpartum, only.Discussion: cDNA was detected in samples from one positive female, during forty days postpartum, but not on the 30th. On the other hand, amplified cDNA was observed on 30th day by nested-PCR. In this case, a false negative result was observed after real-time PCR, probably because sample corresponding to 30th days may not have been properly homogenized, so that the fraction used in real-time PCR was not representative. A higher number of positive samples were expected due to the higher sensitivity of the technique used. The low viral concentration in the milk due to high antibody titers, for example, leaded to a small number of cells containing the agent, reducing the possibility of detection. cDNA was not detected in any sample from three infected females. A possible false-positive serological reaction or the very low viral concentration in milk samples could explain the negative results, although some animals might be infected by a strain that could not be recognized by PCR.

Prevalence of potentially zoonotic gastrointestinal parasites in canine faeces in Ibadan, Nigeria.

Humans can get infected through direct or indirect contact with infective stages of zoonotic parasites shed to the environment through dog faeces. This study was designed to investigate the presence of gastrointestinal parasites present in dog faeces shed on the street of Ibadan metropolis, one of the largest cities in Africa. Twenty-three locations were randomly selected using grid-sampling method. A total of 203 faecal samples collected from the streets of selected areas were processed for detection of helminth eggs and protozoan oocysts using flotation technique. Eggs/oocysts per gram of faeces was counted using modified McMaster technique. The prevalence of gastrointestinal parasites was 43.3% (88/203). Single and multiple infections were 69 (78.4%) and 19 (21.6%) respectively. The parasites detected were Ancylostoma sp. 24.6% (50/88) Isospora sp. 14.2% (29/88), Toxocara sp. 9.8% (20/88), Uncinaria sp. 2.5% (5/88) and Strongyloides sp, 3.9% (8/88). Ancylostoma sp. (320 × 102 epg) and Uncinaria sp. (5 × 102 epg) had the highest and least intensity respectively. Streets within residential areas having markets had the highest number of positive samples. All the genera of parasites detected in this study have zoonotic potential. The high prevalence of zoonotic parasites detected in dog faeces from Ibadan metropolis showed that infected stray dogs roam the streets and constitute potential risk to human health. This study suggests the need for enforcement of laws restraining roaming or straying dogs and proper veterinary care of dogs. None declared.

Detection of the ethanol consumption markers ethyl glucuronide and ethyl sulfate in urine samples from inmates of two German prisons

Abstinence from ethanol is necessary in various situations. Among these are jail terms. Nevertheless, it is a matter of fact that ethanol is illegally produced and ingested in prisons. So far, data regarding drug prevalence in jail have mainly been collected by questionnaires. To get an objective database for the prevalence of ethanol consumption in jail, a cross-sectional study was performed. Inmates of two German prisons (Offenburg and Freiburg) were asked to give a urine sample at an unknown and random point of time. Participation was voluntary and did lead to neither negative consequences nor benefits. All samples were anonymized. Using the consumption markers ethyl glucuronide (EtG) and ethyl sulfate (EtS), the urine samples were tested for previous ethanol consumption. Analyses were performed by a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. In total 676 male inmates participated in this study. The participation rate was 70-75% of all permanent inmates in Offenburg and 30.6% in Freiburg. Ten of the 555 (1.8%) samples from Offenburg and 1 of the 121 (0.8%) samples from Freiburg were positive for ethanol consumption markers with concentrations ranging from trace amounts to 1400 ng/mL for EtG and up to 510 ng/mL for EtS, respectively. The number of participants in this study was rather high, so that the results represent a good cross section, at least for Offenburg, the jail with the higher number of positive samples.

Presence of pathogenic Vibrio species in fresh mussels harvested in the southern Rias of Galicia (NW Spain)

Spain is the third mussel producer worldwide, and the main supplier to the European market, coming 98% of this production from Galicia. Pathogenic species from the genus Vibrio pose a considerable public health threat as the causative agents of both sporadic and epidemic human infections. Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus are the most significant pathogens within this genus. The aims of this study were to investigate the prevalence of human pathogenic Vibrio species in Galician mussels from different geographical areas, and determine their pathogenic profiles by qPCR as rapid diagnostic method. Out of the 101 samples, 72% of the V. parahaemolyticus positive samples, carried at least one virulence factor (tdh and/or trh), being trh was the most prevalent (37%). The highest number of positive samples was found during October and September. During the period of study neither Vibrio vulnificus nor V. cholerae were detected. After enrichment, positive samples were tested for antibiotic resistance genes (ermB and tetQ) by qPCR. No resistance was detected against tetracycline. On the contrary, 52% of the samples were positive for ermB, and out of these, 78% also had, either one or both, V. parahaemolyticus virulence factors.

Study The Some Aspects of The Immune Response For Pregnant Women Infected with T.Gondii and Determine The Genotyping of This Parasite

Objectives: This study aims to find out the immune response (antibodies and cytokines) for women with disease T.Gondii and correlation between them calibrated and the effects on the patients of the difference age groups. Methodology: Were collected (200) samples of blood and (30) samples of umbilical cord of women and pregnant women with disease T.Gondii and revisions to hospitals in Najaf and laboratories for the purposes of abortion or treatment of disease, while the number of women samples control amounted to 30 samples . Pregnant women and control were divided into three age groups, namely, (15-25), (26-36) and (37-47) respectively. The study involved collecting samples and tested the first test by latex and then examined by the technique of adsorption-linked immunosorbent assay (ELISA) for the detection of antibodies (IgG and IGM). The study reported that technique is more important than the ELISA test for latex which was (P = 0.007). Results: The results of this study include following : the difference between the age groups and abortion rate in women was age groups (15-25) and (26-36) is the highest at 40% and 51%, respectively, and compared with the pregnant women and the disease infected T.Gondii. The study pointed to the existence of a significant difference in the proportion of antibodies in different age groups when compared with the control group. Adding to the cytokines. The study showed that there is a relationship between genes Toxo B1, ITS-1, Tg-8, Tg-9, SAG-1 and SAG-2 by examining the DNA taken of the blood and placenta of women with T.Gondii. And recorded the highest number of positive samples (52) in the sample ITS-1 of the rest of the other genes. Statistical relationship was the most important in the diagnosis of the parasite (P = 0.001). Conclusion: The correlation between of the cytokines and toxoplasmosis with according of abortion. Recommendations: To detect genotypes that lead to spontaneous abortion

Nucleotide Correlations Between Rotavirus C Isolates in Clinical Samples from Outbreaks and in Sewage Samples.

Rotavirus C (RVC) is detected in both sporadic cases and outbreaks of gastroenteritis worldwide. However, the epidemic dynamics of RVC in populations remain poorly understood because the detection rate is low. In this study, raw sewage samples were collected from a wastewater treatment plant in Yokohama, Japan, over 5years, in 12-month period from September to August, to identify the RVC strains in these samples and compare them with the RVC strains circulating in the population. RVC strains were detected in 15 of the 118 raw sewage samples collected between 2007 and 2012. The highest number of positive samples detected per period (seven) was in 2008-2009. A fragment (225 nucleotides) of the VP7 gene of RVC from 14 sewage samples was sequenced. The nucleotide sequences of 11 strains were completely consistent with those of clinical strains identified in Yokohama. A phylogenetic analysis showed that 13 strains from the sewage samples clustered with several Yokohama outbreak strains and were closely related to the clinical strains (except sewage-derived strain Y11-SW0805-C). Our study demonstrates a correlation between clinical and sewage strains of RVC based on a genetic analysis, and shows that monitoring environmental samples is an effective way to study the strains circulating in a population, including in asymptomatic or mildly symptomatic patients, even when these infections are not detected in clinical samples. This is the first report of the surveillance of RVC in sewage samples in Yokohama, Japan, for molecular epidemiological analysis.

Detection of veterinary drug residues in surface waters collected nearby farming areas in Galicia, North of Spain

The occurrence of pharmaceuticals in the aquatic environment has become a matter of concern in the last decade due to potential risks posed to non-target organisms and the potential for unintended human exposure via food chain. This concern has been driven by a high detection frequency for drugs in environmental samples; these substances are produced in large quantities and are used in both veterinary and human medicine, leading to deposition and potential effects in the environment. However, few studies have focused on the presence of pharmaceuticals in rural areas associated with farming activities in comparison to urban areas. The aim of this study is to investigate the occurrence of pharmaceutically active compounds in surface waters collected from urban and rural areas in northwestern Spain. A monitoring study was conducted with 312 river water samples analysed by high-performance liquid chromatography coupled to tandem mass spectrometry. Positive detection of pharmaceuticals was made for 51 % of the samples. Decoquinate, sulfamethazine, sulfamethoxypyridazine and trimethoprim were the drugs most frequently detected, being present in more than 10 % of the samples. The sampling sites located downstream of the discharge points for wastewater treatment plants yielded the highest number of positive samples, 13 % of the positive samples were detected in these sites and 38 % of the samples collected near the collection point of a drinking water treatment plant were positive.

Development of a novel polymerase chain reaction-enzyme-linked immunosorbent assay for the diagnosis ofTrichophyton rubrumonychomycosis

The prevalence of onychomycosis has increased steadily in the past decade. An accurate diagnosis at the outset is important for successful and cost-effective treatment of patients. However, current diagnostic tests for onychomycosis are not rapid, sensitive or specific. To develop a microsatellite-based polymerase chain reaction (PCR)-enzyme-linked immunosorbent assay (MS-ELISA) for the detection of Trichophyton rubrum, which is the most common aetiological agent of onychomycosis. An archival set of 434 nail and skin specimens from 217 patients was included as the test sample in this study. We also compared MS-ELISA with an earlier published topoisomerase PCR-ELISA (TI-ELISA) using template DNA extracted by another method. The MS-ELISA detected the highest number of positive samples (69%) followed by direct microscopy (56%), TI-ELISA (44%) and fungal culture (30%). When an identical DNA extraction method was applied to 120 specimens, the MS-ELISA proved to be twice as sensitive as the TI-ELISA. We have optimized a target gene and DNA extraction method for rapid detection of T. rubrum onychomycosis.

Monitoring Current-Season Spread of Potato virus Y in Potato Fields Using ELISA and Real-Time RT-PCR.

The current-season spread of Potato virus Y (PVY) was investigated in New Brunswick, Canada, in 11 potato fields planted with six different cultivars in 2009 and 2010. In all, 100 plants selected from each field were monitored for current-season PVY infections using enzyme-linked immunosorbent assay (ELISA) and real-time reverse-transcription polymerase chain reaction (RT-PCR) assay. Average PVY incidence in fields increased from 0.6% in 2009 and 2% in 2010 in the leaves to 20.3% in 2009 and 21.9% in 2010 in the tubers at the time of harvest. In individual fields, PVY incidence in tubers reached as high as 37% in 2009 and 39% in 2010 at the time of harvest. Real-time RT-PCR assay detected more samples with PVY from leaves than did ELISA. A higher number of positive samples was also detected with real-time RT-PCR from growing tubers compared with the leaves collected from the same plant at the same sampling time. PVY incidence determined from the growing tubers showed a significant positive correlation with the PVY incidence of tubers after harvest. Preharvest testing provides another option to growers to either top-kill the crop immediately to secure the seed market when the PVY incidence is low or leave the tubers to develop further for table or processing purposes when incidence of PVY is high.