Search results for: cryptosporidium hominis
Commenced in January 2007
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Edition: International
Paper Count: 32

Search results for: cryptosporidium hominis

32 In Silico Screening, Identification and Validation of Cryptosporidium hominis Hypothetical Protein and Virtual Screening of Inhibitors as Therapeutics

Authors: Arpit Kumar Shrivastava, Subrat Kumar, Rajani Kanta Mohapatra, Priyadarshi Soumyaranjan Sahu

Abstract:

Computational approaches to predict structure, function and other biological characteristics of proteins are becoming more common in comparison to the traditional methods in drug discovery. Cryptosporidiosis is a major zoonotic diarrheal disease particularly in children, which is caused primarily by Cryptosporidium hominis and Cryptosporidium parvum. Currently, there are no vaccines for cryptosporidiosis and recommended drugs are not effective. With the availability of complete genome sequence of C. hominis, new targets have been recognized for the development of effective and better drugs and/or vaccines. We identified a unique hypothetical epitopic protein in C. hominis genome through BLASTP analysis. A 3D model of the hypothetical protein was generated using I-Tasser server through threading methodology. The quality of the model was validated through Ramachandran plot by PROCHECK server. The functional annotation of the hypothetical protein through DALI server revealed structural similarity with human Transportin 3. Phylogenetic analysis for this hypothetical protein also showed C. hominis hypothetical protein (CUV04613) was the closely related to human transportin 3 protein. The 3D protein model is further subjected to virtual screening study with inhibitors from the Zinc Database by using Dock Blaster software. Docking study reported N-(3-chlorobenzyl) ethane-1,2-diamine as the best inhibitor in terms of docking score. Docking analysis elucidated that Leu 525, Ile 526, Glu 528, Glu 529 are critical residues for ligand–receptor interactions. The molecular dynamic simulation was done to access the reliability of the binding pose of inhibitor and protein complex using GROMACS software at 10ns time point. Trajectories were analyzed at each 2.5 ns time interval, among which, H-bond with LEU-525 and GLY- 530 are significantly present in MD trajectories. Furthermore, antigenic determinants of the protein were determined with the help of DNA Star software. Our study findings showed a great potential in order to provide insights in the development of new drug(s) or vaccine(s) for control as well as prevention of cryptosporidiosis among humans and animals.

Keywords: cryptosporidium hominis, hypothetical protein, molecular docking, molecular dynamics simulation

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31 Prevalence of Cryptosporidium spp. in Free-Living Wild Birds by Using Carbol Fuchsin Staining Methods in Konya, Turkey

Authors: Nermin Isik

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Cryptosporidiosis is one of the most common parasitic infection in domesticated, caged, wild birds. Cryptosporidium sp. has been reported in over 30 avian species worldwide. Cryptosporidium meleagridis, Cryptosporidium baileyi and Cryptosporidium galli are recognised avian species of Cryptosporidium. This study was carried out to determine the prevalance of Cryptosporidium sp. in wild birds in Konya province, Turkey. Faecal samples were collected from 65 wild birds including 52 Podicipedidae (Podiceps cristatus), 11 Rallidae (Fulicia Atra), 2 Anitadia (Aytha ferina). Faecal samples were stained with Modified Ziehl-Neelsen staining technigue, they were examined under light microscope for the presence of Cryptosporidium sp. oocyts. Among the 65 faecal samples, 11 (16.9%) were found to be infected with Cryptosporidium sp. oocysts. The results of this study indicate that wild birds may play an important role in the epidemiology of Cryptosporidium. In conclusion, Cryptosporidiosis has suggested zoonotic potential and thus warrant further attention. In addition, biological and genetic studies are required to provide more information on Cryptosporidiosis.

Keywords: Cryptosporidium sp, wild birds, Konya, Turkey

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30 Epidemiology, Clinical, Immune, and Molecular Profiles of Microsporidiosis and Cryptosporidiosis among HIV/AIDS patients

Authors: Roger WUMBA

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The objective of this study was to determine the prevalence of intestinal parasites, with special emphasis on microsporidia and Cryptosporidium, as well as their association with human immunodeficiency virus (HIV) symptoms, risk factors, and other digestive parasites. We also wish to determine the molecular biology definitions of the species and genotypes of microsporidia and Cryptosporidium in HIV patients. In this cross-sectional study, carried out in Kinshasa, Democratic Republic of the Congo, stool samples were collected from 242 HIV patients (87 men and 155 women) with referred symptoms and risk factors for opportunistic intestinal parasites. The analysis of feces specimen were performed using Ziehl–Neelsen stainings, real-time polymerase chain reaction (PCR), immunofluorescence indirect monoclonal antibody, nested PCR-restriction fragment length polymorphism, and PCR amplification and sequencing. Odds ratio (OR) and 95% confidence intervals were used to quantify the risk. Of the 242 HIV patients, 7.8%, 0.4%, 5.4%, 0.4%, 2%, 10.6%, and 2.8% had Enterocytozoon bieneusi, Encephalitozoon intestinalis, Cryptosporidium spp., Isospora belli, pathogenic intestinal protozoa, nonpathogenic intestinal protozoa, and helminths, respectively. We found five genotypes of E. bieneusi: two older, NIA1 and D, and three new, KIN1, KIN2, and KIN3. Only 0.4% and 1.6% had Cryptosporidium parvum and Cryptosporidium hominis, respectively. Of the patients, 36.4%, 34.3%, 31%, and 39% had asthenia, diarrhea, a CD4 count of ,100 cells/mm³, and no antiretroviral therapy (ART), respectively. The majority of those with opportunistic intestinal parasites and C. hominis, and all with C. parvum and new E. bieneusi genotypes, had diarrhea, low CD4+ counts of ,100 cells/mm³, and no ART. There was a significant association between Entamoeba coli, Kaposi sarcoma, herpes zoster, chronic diarrhea, and asthenia, and the presence of 28 cases with opportunistic intestinal parasites. Rural areas, public toilets, and exposure to farm pigs were the univariate risk factors present in the 28 cases with opportunistic intestinal parasites. In logistic regression analysis, a CD4 count of ,100 cells/mm³ (OR = 4.60; 95% CI 1.70–12.20; P = 0.002), no ART (OR = 5.00; 95% CI 1.90–13.20; P , 0.001), and exposure to surface water (OR = 2.90; 95% CI 1.01–8.40; P = 0.048) were identified as the significant and independent determinants for the presence of opportunistic intestinal parasites. E. bieneusi and Cryptosporidium are becoming more prevalent in Kinshasa, Congo. Based on the findings, we recommend epidemiology surveillance and prevention by means of hygiene, the emphasis of sensitive PCR methods, and treating opportunistic intestinal parasites that may be acquired through fecal–oral transmission, surface water, normal immunity, rural area-based person–person and animal–human nfection, and transmission of HIV. Therapy, including ART and treatment with fumagillin, is needed.

Keywords: diarrhea, enterocytozoon bieneusi, cryptosporidium hominis, cryptosporidium parvum, risk factors, africans

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29 Importance of Cryptosporidiosis in Dairy Calves

Authors: Mohammad Asadpour

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Cryptosporidium spp. is zoonotic pathogens transmissible from a variety of animals to humans and is a considerable public health concern. Calves have been identified in numerous reports as a major source of environmental contamination with this pathogen. Parasite has a different species that are the cases of zoonotic disease in immunodeficient people and neonatal calves. Cryptosporidium oocysts are extremely resistant to chlorine and other physical cases that commonly used in drinking water. Reproduction of resistant oocytes is a way for this monoxenous parasite to remain in the environment. Cryptosporidium parvum is the most important species that has human and cattle genotypes. Cryptosporidium is one of the most important causes of diarrhea in neonatal calves and also, one of the four causes of diarrhea symptoms in pre-weaned calves. Because of the incompetent immune system in calves, Cryptosporidium infection is the cause of a lot of problems in raising farms.

Keywords: Cryptosporidium spp, dairy calves, importance, veterinary medicine

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28 Identification and Molecular Characterization of Cryptosporidium Spp. in Pre-Wean Dairy Calves in Mashhad, Northeastern of Iran

Authors: Mohammad Asadpour, Gholamreza Razmi, Gholamreza Mohammadi, Abolghasem Naghibi

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Cryptosporidium Spp., protozoan parasites of the phylum Apicomplexa, have a wide spectrum of hosts including humans, domestic animals and wild mammals, birds, reptiles, amphibians and fish. Dairy cattle have been identified in numerous reports as a major source of environmental contamination with this pathogen. In this study, a Polymerase Chain Reaction (PCR), Restriction Fragment Length Polymorphism (RFLP) analysis of the Small-Subunit (SSU) rRNA gene was used to detect and identify Cryptosporidium Spp. in 300 fecal specimens from 1 to 30 days pre-wean calves in 10 farms in Mashhad, Iran. Eighty five (28.3%) and forty five (15%) of the specimens were positive for Cryptosporidium by microscopic and PCR examination respectively. Restriction digestion of the PCR products by VSPI and Ssp1 restriction enzymes and analysis of sequence data revealed the presence of C. parvum, bovine genotype in all isolates. Our findings suggest that cattle can be a source of Cryptosporidial infections for humans and animals in Mashhad area. This is the first published description of Cryptosporidium sub genotyping in Mashhad.

Keywords: cryptosporidium, genotype, dairy calves, 18S rRNA, Mashhad

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27 Detection of Cryptosporidium Oocysts by Acid-Fast Staining Method and PCR in Surface Water from Tehran, Iran

Authors: Mohamad Mohsen Homayouni, Niloofar Taghipour, Ahmad Reza Memar, Niloofar Khalaji, Hamed Kiani, Seyyed Javad Seyyed Tabaei

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Background and Objective: Cryptosporidium is a coccidian protozoan parasite; its oocysts in surface water are a global health problem. Due to the low number of parasites in the water resources and the lack of laboratory culture, rapid and sensitive method for detection of the organism in the water resources is necessarily required. We applied modified acid-fast staining and PCR for the detection of the Cryptosporidium spp. and analysed the genotypes in 55 samples collected from surface water. Methods: Over a period of nine months, 55 surface water samples were collected from the five rivers in Tehran, Iran. The samples were filtered by using cellulose acetate membrane filters. By acid fast method, initial identification of Cryptosporidium oocyst were carried out on surface water samples. Then, nested PCR assay was designed for the specific amplification and analysed the genotypes. Results: Modified Ziehl-Neelsen method revealed 5–20 Cryptosporidium oocysts detected per 10 Liter. Five out of the 55 (9.09%) surface water samples were found positive for Cryptosporidium spp. by Ziehl-Neelsen test and seven (12.7%) were found positive by nested PCR. The staining results were consistent with PCR. Seven Cryptosporidium PCR products were successfully sequenced and five gp60 subtypes were detected. Our finding of gp60 gene revealed that all of the positive isolates were Cryptosporidium parvum and belonged to subtype families IIa and IId. Conclusion: Our investigations were showed that collection of water samples were contaminated by Cryptosporidium, with potential hazards for the significant health problem. This study provides the first report on detection and genotyping of Cryptosporidium species from surface water samples in Iran, and its result confirmed the low clinical incidence of this parasite on the community.

Keywords: Cryptosporidium spp., membrane filtration, subtype, surface water, Iran

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26 A Survey on Frequency of Cryptosporidiosis and Giardiasis in Horses in Ahvaz South-West of Iran

Authors: Ali R. Ghadrdan-Mashhadi, Hosein Hamidi-Nejat, Parisa Alizadehnia

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Cryptosporidia and Giardia are protozoan parasites that have worldwide distribution and infect a variety of animals. Although, the infection to these parasites rarely caused to illness in horses, but some veterinarian recorded the clinical signs (such as diarrhea and malabsorbtion) especially in foals. In present study, the frequency of Cryptosporidiosis and Giardiasis in horses in Ahvaz investigated. The feces samples were taken from 100 horses that keep in seven horse breeding clubs, during spring and summer. The ages of horses were from 1 month to 27 years old. Fecal samples were stained by modified Ziehl-Neelsen and Tri-chrome methods. Results were analyzed with Chi-square Test and Fisher’s exact test. The results showed that the rate of infection to Cryptosporidium and Giardia were 18% and 40%, respectively. There weren't significant differences between infection to Cryptosporidium and Giardia with sex, age and fecal constancy. Although, the rate of infection to Cryptosporidium in studied horses is very similar to other studies but it seems, the rate of infection to Giardia is high in compare to other studies were done in the other countries.

Keywords: Ahvaz, cryptosporidium, giardia, horse

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25 Determination of Some Etiologic Agents in Calves with Diarrhea

Authors: Nermin Isik, Ozlem Derinbay Ekici, Oguzhan Avci

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The aim of this study was to determination of role infection in neonatal calves in Central Anatolian, Turkey. A total 300 fecal samples were collected from diarrheic neonatal calves, aged between 0–90 days from Konya, Karaman, and Aksaray from January to April 2014. Fecal specimens from calves with clinically diarrheic symptoms were examined for the presence of Bovine Coronavirus, Bovine Rotavirus, Cryptosporidium sp., and E. coli by commercially available capture direct enzyme linked immunosorbent assay (ELISA) kit and Modified Ziehl Neelsen method (MZN). Calves were grouped according to their age as follows: 1-14, 15-29, and 30-90 days. Cryptosporidium sp. infection was detected in 52.8%, 58.8%, and 39.2% by ELISA and 33.9%, 47%, 26.7% by MZN in the respective age groups. The seroprevalance of Rotavirus (12.5 %, 40 %, 12.5 %), Coronavirus (2.5%, 0%, 3.5%) and E. coli (5%, 4.7%, 8.9%) infections were determined according to the age groups respectively. Cryptosporidium sp. was the most detected enteropathogen (52 %) of calves and coronavirus was the least detected (2 %). The detection rate of the mixed enfection was 12.3%. In conclusion, it must be evaluated by mix infections in calves with diarrhea. These results will provide an important contribution against the factors that cause diarrhea

Keywords: cryptosporidium sp., bovine coronavirus, bovine rotavirus, E.coli, calve, ELISA

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24 Prevalence of Mycoplasma hominis and Ureaplasma urealyticum as Causative Agents of Non-Gonococcal Urethritis in Men and Determination of Anti-Bacterial Resistance Rates

Authors: Recep Keşli, Cengiz Demir, Onur Türkyılmaz

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Objective: The aim of this study was to determine the prevalence of Mycoplasma hominis and Ureaplasma urealyticum as the causative agents in men with non-gonococcal urethtritis, and anti-bacterial resistance rates. Methods: The Study was carried out in the two Medical Microbiology Laboratories belonging to: Konya Education and Research Hospital and ANS Practice and Research Hospital, Afyon Kocatepe University, between January 2012 and December 2015. Urethral samples were obtained from patients by using a swab. Mycoplasma hominis and Ureaplasma urealyticum were detected by using Mycoplasma IST-2 kit (bio-Mérieux, Marcy l'Étoile, France). Neisseria gonorrhoea was excluded by Gram staining and culture methods. Results: Of all the one hundred and eighty-eight male patients with urethritis, forty M. hominis and forty two U. urealyticum were detected. Resistance rates of M. hominis strains against to doxycycline, ofloxacin, erythromycin, tetracycline, ciprofloxacin, azithromycin, clarithromycin, and pristinamycin were found as 5 %, 65 %, 25 %, 5 %, 80 %, 20 %, 20 %, 20 %, 5 %, respectively. Resistance rates of U. urealyticum strains against to doxycycline, ofloxacin, erythromycin, tetracycline, ciprofloxacin, azithromycin, clarithromycin, and pristinamycin were found as 4.7 %, 66.6 %, 23.8 %, 4.75 %, 81 %, 19 %, 19 %, 4.7 % respectively. No resistance was detected against to josamycin, for both the strains. Conclusions: It was concluded that; ciprofloxacin and ofloxacin had the weakest; josamycin, doxycycline, and tetracycline had the strongest in vitro anti-bacterial activity, for treatment of the NGU. So josamycin, doxycycline, and tetracycline should be preferred as the first choice of anti-bacterial agents, for treatment of the patients with non-gonococcal male urethritis.

Keywords: antimicrobial resistance, Mycoplasma hominis, non-gonococcal urethritis, Ureaplasma urealyticum

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23 Study on the Presence of Protozoal Coinfections among Patients with Pneumocystis jirovecii Pneumonia in Bulgaria

Authors: Nina Tsvetkova, Rumen Harizanov, Aleksandra Ivanova, Iskra Rainova, Nina Yancheva-Petrova, Dimitar Strashimirov, Raina Enikova, Mihaela Videnova, Eleonora Kaneva, Iskren Kaftandjiev, Viktoria Levterova, Ivan Simeonovski, Nikolay Yanev, Georgi Hinkov

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The Pneumocystis jirovecii (P. jirovecii) and protozoan of the genera Acanthamoeba, Cryptosporidium, and Toxoplasma gondii are opportunistic pathogens that can cause life-threatening infections in immunocompromised patients. Aim of the study was to evaluate the coinfection rate with opportunistic protozoal agents among Bulgarian patients diagnosed with P. jirovecii pneumonia. Thirty-eight pulmonary samples were collected from 38 patients (28 HIV-infected) with P. jirovecii infection. P. jirovecii DNA was detected by real-time PCR targeting the large mitochondrial subunit ribosomal RNA gene. Acanthamoeba was determined by genus-specific conventional PCR assay. Real-time PCR for the detection of a Toxoplasma gondii and Cryptosporidium DNA fragment was used. Pneumocystis DNA was detected in all 38 specimens; 28 (73.7%) were from HIV-infected patients. Three (10,7%) of them were co-infected with T. gondii and 1 (3.6%) with Cryptosporidium. In the group of non-HIV-infected (n=10), Cryptosporidium DNA was detected in an infant (10%). Acanthamoeba DNA was not found in the tested samples. The current study showed a relatively low rate of coinfections of Cryptosporidium spp./T. gondii and P. jirovecii in the Bulgarian patients studied.

Keywords: coinfection, opportunistic protozoal agents, Pneumocystis jirovecii, pulmonary infections

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22 In Vitro Antimycoplasmal Activity of Peganum harmala on Mycoplasma hominis Tunisian Strains

Authors: Nadine khadraoui, Rym Essid, Olfa Tabbene, Imen Chniba, Safa Boujemaa, Selim Jallouli, Nadia Fares, Behija Mlik, Boutheina Ben Abdelmoumen Mardassi

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Background and aim: Mycoplasma hominis is an opportunistic pathogen that can cause various gynecological infections such cervicitis, infertility, and, less frequently, extra-genital infections. Previous studies on the antimicrobial susceptibility of Mycoplasma hominis Tunisian strains have highlighted a significant resistance, even multi-resistance, to the most used antibiotic in the therapy of consequential infections. To address this concern, the present study aimed for the alternative of phytotherapy. Peganum harmala seed extract was tested as an antibacterial agent against multidrug-resistant M.hominis clinical strains. Material and Methods: Peganum harmala plant was collected from Ain Sebaa, Tabarka, North West region of Tunisia in April 2018, air-dried, grounded and extracted by different solvents.The crude methanolic extract was further partitioned with n-HEX, DCM, EtOAC and n-BuOl. Antibacterial activity was evaluated against M. hominis ATCC 23114 and 20 M. hominis clinical strains.The antimycoplasmal activity was tested by the microdilution method, and MIC values were determined. Phytochemical analysis and hemolytic activity on human erythrocytes were also performed. The active fraction was then subjected to purification, and the chemical identification of the active compound was investigated. Results: Among the tested fractions, the n-BuOH extract was the most active fraction since it exhibited an inhibitory effect against M. hominis ATCC 23114 and 80% of the tested clinical strains with MIC between 125 and 1000 µg/ml. The phytochemical analysis of the n-BuOH revealed its metabolic abundance in polyphenols, flavonoids and condensed tannin with levels of 257.37 mg AGE/g, 172.27 mg EC/g and 58.27 mg EC/g, respectively. In addition, P. harmala n-BuOH extract exhibited potent bactericidal activity against all M. hominis isolates with CMB values ranging between 125 and 4000 µg/ml. Further, the active fraction exhibited weak cytotoxicity effect at active concentrations when tested on human erythrocytes. The active compound was identified by gas chromatography–mass spectrometry as an indole alkaloid harmaline. Conclusion: In summary, Peganum harmala extract demonstrated an interesting anti-mycoplasmal activity against M. hominis Tunisian strains. Therefore, it could be considered as a potential candidate for the treatment of consequential infections. However, further studies are necessary to evaluate its mechanism of action in mycoplasmas.

Keywords: mycoplasma hominis, peganum harmala, antibioresistance, phytotherapy, phytochemical analysis

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21 Cryptosporidium Parvum oocytic Antigen Induced a Pro-Inflammatory DC Phenotype

Authors: Connick K, Lalor R, Murphy A, O’Neill S. M., Rabab S. Zalat, Eman E. El Shanawany

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Cryptosporidium parvum is an opportunistic intracellular parasite that causes mild to severe diarrhea in human and animal populations and is an important zoonotic disease globally. In immunocompromised hosts, infection Canbe life-threatening as no effective treatments are currently available to control infection. To increase our understanding of the mechanisms that play a role in host-parasite interactions at the level of the immune response, we investigated the effects of Cryptosporidium parvum antigen (CPA) on bone marrow-derived (DCS). Herein we examined cytokine secretion and cell surface marker expression on DCs exposed to CPA. We also measured cytokine production in CD4+ cells co-cultured with CPA primed DCs in the presence of anti-CD3. CPA induced a significant increase in the production of interleukin(IL)-12p40, IL-10, IL-6, and TNF-α by DCs and enhanced the expression of the cell surface markers TLR4, CD80, CD86, and MHC11. CPA primed DC co-cultured in the presence of anti-CD3 with CD4+ T-cells inhibited the secretion of Th2 associated cytokines, notably IL-5 and IL-13, with no effects on the secretions of interferon (IFN)-γ, IL-2, IL-17, and IL-10. These findings support studies in the literature that CPA can induce the full maturation of DCs that subsequently initiate Th1 immune responses critical to the resolution of C. parvum infection.

Keywords: cryptosporidium parvum, dendritic cells, IL-12 p70, cell surface marker

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20 Unraveling the Evolution of Mycoplasma Hominis Through Its Genome Sequence

Authors: Boutheina Ben Abdelmoumen Mardassi, Salim Chibani, Safa Boujemaa, Amaury Vaysse, Julien Guglielmini, Elhem Yacoub

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Background and aim: Mycoplasma hominis (MH) is a pathogenic bacterium belonging to the Mollicutes class. It causes a wide range of gynecological infections and infertility among adults. Recently, we have explored for the first time the phylodistribution of Tunisian M. hominis clinical strains using an expanded MLST. We have demonstrated their distinction into two pure lineages, which each corresponding to a specific pathotype: genital infections and infertility. The aim of this project is to gain further insight into the evolutionary dynamics and the specific genetic factors that distinguish MH pathotypes Methods: Whole genome sequencing of Mycoplasma hominis clinical strains was performed using illumina Miseq. Denovo assembly was performed using a publicly available in-house pipeline. We used prokka to annotate the genomes, panaroo to generate the gene presence matrix and Jolytree to establish the phylogenetic tree. We used treeWAS to identify genetic loci associated with the pathothype of interest from the presence matrix and phylogenetic tree. Results: Our results revealed a clear categorization of the 62 MH clinical strains into two distinct genetic lineages, with each corresponding to a specific pathotype.; gynecological infections and infertility[AV1] . Genome annotation showed that GC content is ranging between 26 and 27%, which is a known characteristic of Mycoplasma genome. Housekeeping genes belonging to the core genome are highly conserved among our strains. TreeWas identified 4 virulence genes associated with the pathotype gynecological infection. encoding for asparagine--tRNA ligase, restriction endonuclease subunit S, Eco47II restriction endonuclease, and transcription regulator XRE (involved in tolerance to oxidative stress). Five genes have been identified that have a statistical association with infertility, tow lipoprotein, one hypothetical protein, a glycosyl transferase involved in capsule synthesis, and pyruvate kinase involved in biofilm formation. All strains harbored an efflux pomp that belongs to the family of multidrug resistance ABC transporter, which confers resistance to a wide range of antibiotics. Indeed many adhesion factors and lipoproteins (p120, p120', p60, p80, Vaa) have been checked and confirmed in our strains with a relatively 99 % to 96 % conserved domain and hypervariable domain that represent 1 to 4 % of the reference sequence extracted from gene bank. Conclusion: In summary, this study led to the identification of specific genetic loci associated with distinct pathotypes in M hominis.

Keywords: mycoplasma hominis, infertility, gynecological infections, virulence genes, antibiotic resistance

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19 Simulation and Fabrication of Plasmonic Lens for Bacteria Detection

Authors: Sangwoo Oh, Jaewoo Kim, Dongmin Seo, Jaewon Park, Yongha Hwang, Sungkyu Seo

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Plasmonics has been regarded one of the most powerful bio-sensing modalities to evaluate bio-molecular interactions in real-time. However, most of the plasmonic sensing methods are based on labeling metallic nanoparticles, e.g. gold or silver, as optical modulation markers, which are non-recyclable and expensive. This plasmonic modulation can be usually achieved through various nano structures, e.g., nano-hole arrays. Among those structures, plasmonic lens has been regarded as a unique plasmonic structure due to its light focusing characteristics. In this study, we introduce a custom designed plasmonic lens array for bio-sensing, which was simulated by finite-difference-time-domain (FDTD) approach and fabricated by top-down approach. In our work, we performed the FDTD simulations of various plasmonic lens designs for bacteria sensor, i.e., Samonella and Hominis. We optimized the design parameters, i.e., radius, shape, and material, of the plasmonic lens. The simulation results showed the change in the peak intensity value with the introduction of each bacteria and antigen i.e., peak intensity 1.8711 a.u. with the introduction of antibody layer of thickness of 15nm. For Salmonella, the peak intensity changed from 1.8711 a.u. to 2.3654 a.u. and for Hominis, the peak intensity changed from 1.8711 a.u. to 3.2355 a.u. This significant shift in the intensity due to the interaction between bacteria and antigen showed a promising sensing capability of the plasmonic lens. With the batch processing and bulk production of this nano scale design, the cost of biological sensing can be significantly reduced, holding great promise in the fields of clinical diagnostics and bio-defense.

Keywords: plasmonic lens, FDTD, fabrication, bacteria sensor, salmonella, hominis

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18 Foodborne Outbreak Calendar: Application of Time Series Analysis

Authors: Ryan B. Simpson, Margaret A. Waskow, Aishwarya Venkat, Elena N. Naumova

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The Centers for Disease Control and Prevention (CDC) estimate that 31 known foodborne pathogens cause 9.4 million cases of these illnesses annually in US. Over 90% of these illnesses are associated with exposure to Campylobacter, Cryptosporidium, Cyclospora, Listeria, Salmonella, Shigella, Shiga-Toxin Producing E.Coli (STEC), Vibrio, and Yersinia. Contaminated products contain parasites typically causing an intestinal illness manifested by diarrhea, stomach cramping, nausea, weight loss, fatigue and may result in deaths in fragile populations. Since 1998, the National Outbreak Reporting System (NORS) has allowed for routine collection of suspected and laboratory-confirmed cases of food poisoning. While retrospective analyses have revealed common pathogen-specific seasonal patterns, little is known concerning the stability of those patterns over time and whether they can be used for preventative forecasting. The objective of this study is to construct a calendar of foodborne outbreaks of nine infections based on the peak timing of outbreak incidence in the US from 1996 to 2017. Reported cases were abstracted from FoodNet for Salmonella (135115), Campylobacter (121099), Shigella (48520), Cryptosporidium (21701), STEC (18022), Yersinia (3602), Vibrio (3000), Listeria (2543), and Cyclospora (758). Monthly counts were compiled for each agent, seasonal peak timing and peak intensity were estimated, and the stability of seasonal peaks and synchronization of infections was examined. Negative Binomial harmonic regression models with the delta-method were applied to derive confidence intervals for the peak timing for each year and overall study period estimates. Preliminary results indicate that five infections continue to lead as major causes of outbreaks, exhibiting steady upward trends with annual increases in cases ranging from 2.71% (95%CI: [2.38, 3.05]) in Campylobacter, 4.78% (95%CI: [4.14, 5.41]) in Salmonella, 7.09% (95%CI: [6.38, 7.82]) in E.Coli, 7.71% (95%CI: [6.94, 8.49]) in Cryptosporidium, and 8.67% (95%CI: [7.55, 9.80]) in Vibrio. Strong synchronization of summer outbreaks were observed, caused by Campylobacter, Vibrio, E.Coli and Salmonella, peaking at 7.57 ± 0.33, 7.84 ± 0.47, 7.85 ± 0.37, and 7.82 ± 0.14 calendar months, respectively, with the serial cross-correlation ranging 0.81-0.88 (p < 0.001). Over 21 years, Listeria and Cryptosporidium peaks (8.43 ± 0.77 and 8.52 ± 0.45 months, respectively) have a tendency to arrive 1-2 weeks earlier, while Vibrio peaks (7.8 ± 0.47) delay by 2-3 weeks. These findings will be incorporated in the forecast models to predict common paths of the spread, long-term trends, and the synchronization of outbreaks across etiological agents. The predictive modeling of foodborne outbreaks should consider long-term changes in seasonal timing, spatiotemporal trends, and sources of contamination.

Keywords: foodborne outbreak, national outbreak reporting system, predictive modeling, seasonality

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17 Multiple Etiologies and Incidences of Co-Infections in Childhood Diarrhea in a Hospital Based Screening Study in Odisha, India

Authors: Arpit K. Shrivastava, Nirmal K. Mohakud, Subrat Kumar, Priyadarshi S. Sahu

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Acute diarrhea is one of the major causes of morbidity and mortality among children less than five years of age. Multiple etiologies have been implicated for infectious gastroenteritis causing acute diarrhea. In our study fecal samples (n=165) were collected from children (<5 years) presenting with symptoms of acute diarrhea. Samples were screened for viral, bacterial, and parasitic etiologies such as Rotavirus, Adenovirus, Diarrhoeagenic Escherichia coli (EPEC, EHEC, STEC, O157, O111), Shigella spp., Salmonella spp., Vibrio cholera, Cryptosporidium spp., and Giardia spp. The overall results from our study showed that 57% of children below 5 years of age with acute diarrhea were positive for at least one infectious etiology. Diarrhoeagenic Escherichia coli was detected to be the major etiological agent (29.09%) followed by Rotavirus (24.24%), Shigella (21.21%), Adenovirus (5.45%), Cryptosporidium (2.42%), and Giardia (0.60%). Among the different DEC strains, EPEC was detected significantly higher in <2 years children in comparison to >2 years age group (p =0.001). Concurrent infections with two or more pathogens were observed in 47 of 160 (28.48%) cases with a predominant incidence particularly in <2-year-old children (66.66%) compared to children of 2 to 5 years age group. Co-infection of Rotavirus with Shigella was the most frequent combination, which was detected in 17.94% cases, followed by Rotavirus with EPEC (15.38%) and Shigella with STEC (12.82%). Detection of multiple infectious etiologies and diagnosis of the right causative agent(s) can immensely help in better management of acute childhood diarrhea. In future more studies focusing on the detection of cases with concurrent infections must be carried out, as we believe that the etiological agents might be complementing each other’s strategies of pathogenesis resulting in severe diarrhea.

Keywords: children, co-infection, infectious diarrhea, Odisha

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16 The Use of Non-Parametric Bootstrap in Computing of Microbial Risk Assessment from Lettuce Consumption Irrigated with Contaminated Water by Sanitary Sewage in Infulene Valley

Authors: Mario Tauzene Afonso Matangue, Ivan Andres Sanchez Ortiz

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The Metropolitan area of Maputo (Mozambique Capital City) is located in semi-arid zone (800 mm annual rainfall) with 1101170 million inhabitants. On the west side, there are the flatlands of Infulene where the Mulauze River flows towards to the Indian Ocean, receiving at this site, the storm water contaminated with sanitary sewage from Maputo, transported through a concrete open channel. In Infulene, local communities grow salads crops such as tomato, onion, garlic, lettuce, and cabbage, which are then commercialized and consumed in several markets in Maputo City. Lettuce is the most daily consumed salad crop in different meals, generally in fast-foods, breakfasts, lunches, and dinners. However, the risk of infection by several pathogens due to the consumption of lettuce, using the Quantitative Microbial Risk Assessment (QMRA) tools, is still unknown since there are few studies or publications concerning to this matter in Mozambique. This work is aimed at determining the annual risk arising from the consumption of lettuce grown in Infulene valley, in Maputo, using QMRA tools. The exposure model was constructed upon the volume of contaminated water remaining in the lettuce leaves, the empirical relations between the number of pathogens and the indicator of microorganisms (E. coli), the consumption of lettuce (g) and reduction of pathogens (days). The reference pathogens were Vibrio cholerae, Cryptosporidium, norovirus, and Ascaris. The water quality samples (E. coli) were collected in the storm water channel from January 2016 to December 2018, comprising 65 samples, and the urban lettuce consumption data were collected through inquiry in Maputo Metropolis covering 350 persons. A non-parametric bootstrap was performed involving 10,000 iterations over the collected dataset, namely, water quality (E. coli) and lettuce consumption. The dose-response models were: Exponential for Cryptosporidium, Kummer Confluent hypergeomtric function (1F1) for Vibrio and Ascaris Gaussian hypergeometric function (2F1-(a,b;c;z) for norovirus. The annual infection risk estimates were performed using R 3.6.0 (CoreTeam) software by Monte Carlo (Latin hypercubes), a sampling technique involving 10,000 iterations. The annual infection risks values expressed by Median and the 95th percentile, per person per year (pppy) arising from the consumption of lettuce are as follows: Vibrio cholerae (1.00, 1.00), Cryptosporidium (3.91x10⁻³, 9.72x 10⁻³), nororvirus (5.22x10⁻¹, 9.99x10⁻¹) and Ascaris (2.59x10⁻¹, 9.65x10⁻¹). Thus, the consumption of the lettuce would result in greater risks than the tolerable levels ( < 10⁻³ pppy or 10⁻⁶ DALY) for all pathogens, and the Vibrio cholerae is the most virulent pathogens, according to the hit-single models followed by the Ascaris lumbricoides and norovirus. The sensitivity analysis carried out in this work pointed out that in the whole QMRA, the most important input variable was the reduction of pathogens (Spearman rank value was 0.69) between harvest and consumption followed by water quality (Spearman rank value was 0.69). The decision-makers (Mozambique Government) must strengthen the prevention measures related to pathogens reduction in lettuce (i.e., washing) and engage in wastewater treatment engineering.

Keywords: annual infections risk, lettuce, non-parametric bootstrapping, quantitative microbial risk assessment tools

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15 Incidence and Etiology of Neonatal Calf Diarrhea in the Region of Blida, Algeria

Authors: A. Dadda, D. Khelef, K. Ait-Oudia, R. Kaidi

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Neonatal calf diarrhea is the most important disease of neonatal calves and results in the greatest economic losses due to disease in this age group in both dairy and beef calves. The objectives of the present study were to estimate the morbidity and the mortality of neonatal diarrhea in dairy calves also to determine aetiology and risk factors were caused diarrhea in dairy veal under 60 days old. A total of 324 claves, housed in 30 dairy breeding were followed during two velage season from January to Juan 2013. The total mortality was 5,9% and was significantly higher in calves had less than 15 days of age. The incidence rate of diarrhea was 31,5% and peaked in the first two weeks after velage. The main causes were breeding controls, defect of passive immunity, old of calf, production season, and nutrient of pregnant cattle, veal’s housing and infectious agents. ELISA test on 22 fecal samples revealed that the 31, 82% of dairy breeding were infected, by cryptosporidium parvum in 13, 6% of study population, E.Coli F5 in 9% and Rotavirus with rate of 4, 5%.

Keywords: diarrhoea, neonatal, mortality, aetiology, risk factors, incidence

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14 Parasitic Infection among Farmers Dealing with Treated Wastewater in Al-Zaitoun Area, Gaza City

Authors: Haneen Nabil Al-Sbaihi, Adnan Al-Hindi, Khalid Qahman

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Treated wastewater irrigation is associated with several benefits but can also lead to significant health risks. The main objective of this study is to investigate the parasitic infection (PI) among farmers dealing with treated wastewater (TWW) in Al-Zaitoun area- Gaza City. This study included two farmer groups: farmers who dealing with TWW (Mixed water users (MWUs)), and farmers who irrigate by using groundwater (GW) (Ground water users (GWUs)). Each participant was asked to provide stool samples on two phases. The two farmer groups were use GW in the 1st phase while the MWUs were use TWW in the 2nd phase which was after using TWW in irrigation for three months. Prevalence of PI was 30.9% and increased to be 47.3% in the 2nd phase. Negative association not statistically significant (OR= 0.659, CI 0.202-2.153)) was found in the 1st phase, while a positive association not statically significant was found between PI and TWWR in the 2nd phase (OR=1.37, CI 0.448-4.21). In this study six parasites species were identified among participants: Entamoeba ''histolytica/dispar and coil'', Cryptosporidium, Microsporidia, Giardia lamblia, Strongyloides stercoralis, and Ascaris lumbricoides.

Keywords: wastewater, groundwater, treated wastewater, parasitic infection, parasites

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13 Findings in Vascular Catheter Cultures at the Laboratory of Microbiology of General Hospital during One Year

Authors: P. Christodoulou, M. Gerasimou, S. Mantzoukis, N. Varsamis, G. Kolliopoulou, N. Zotos

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Abstract— Purpose: The Intensive Care Unit (ICU) environment is conducive to the growth of microorganisms. A variety of microorganisms gain access to the intravascular area and are transported throughout the circulatory system. Therefore, examination of the catheters used in ICU patients is of paramount importance. Material and Method: The culture medium is a catheter tip, which is enriched with Tryptic soy broth (TSB). After one day of incubation, the broth is passaged in the following selective media: Blood, Mac conkey No. 2, chocolate, Mueller Hinton, Chapman, and Saboureaud agar. The above selective media is incubated for 2 days. After this period, if any number of microbial colonies is detected, gram staining is performed and then the microorganisms are identified by biochemical techniques in the automated Microscan (Siemens) system followed by a sensitivity test in the same system using the minimum inhibitory concentration (MIC) technique. The sensitivity test is verified by a Kirby Bauer test. Results: In 2017, the Microbiology Laboratory received 84 catheters from the ICU. 42 were found positive. Of these, S. epidermidis was identified at 8, A. baumannii in 10, K. pneumoniae in 6, P. aeruginosa in 6, P. mirabilis in 3, S. simulans in 1, S. haemolyticus in 4, S. aureus in 3 and S. hominis in 1. Conclusions: The results show that the placement and maintenance of the catheters in ICU patients are relatively successful, despite the unfavorable environment of the unit.

Keywords: culture, intensive care unit, microorganisms, vascular catheters

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12 Survey on Prevalence of Endo and Ecto-Parasites of Rattus rattus in Mazandaran Province, North of Iran

Authors: Fatemeh Rezaei, Afsaneh Amouei, Iman Bakouei, Mahdi Sharif, Shahabeddin Sarvi, Mohammad Taghi Rahimi, Ahmad Daryani

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Background: Rodents act as reservoir host and important potential source for many zoonotic pathogens which pose a public health risk to humans. Therefore, it is necessary to investigate the prevalence of gastrointestinal and ectoparasites among rodents. Materials and Methods: 118 Rattus rattus were captured using snap live traps. Each rat was combed with a fine tooth comb to dislodge ectoparasite and studied. Various samples were collected from feces, examined wet smear, formalin-ether method and stained with modified acid-fast staining and trichrome. Result: The overall prevalence of gastrointestinal parasites of examined rats was 75.4%. Cryptosporidium 30.5%, was the most prevalent protozoan which was followed by Giardia 20.3% and Entamoeba muris 13.5%, Trichomonas muris 10.1% and Spironucleus muris 3.3%. The prevalence of helminth egg was as following Syphacia obvelata 24.5%, Hymenolepis diminuta 10.1% and Trichuris muris 9.3%. 86.4% rodents were found to be infested with ectoparasites including mite 35.6%, flea 28.4%, and lice 42.7%. A significant statistical difference was observed between prevalence and gender of infected individuals. Conclusions: The prevalence of gastrointestinal and ectoparasites of collected rats in studied area is remarkably high. In addition, Rattus rattus can be considered as potential risk for human health.

Keywords: prevalence, rodent, intestinal parasites, ecto-parasites, zoonose

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11 The Microflora Assessment of the Urethra Area of Children with Newly Diagnosed Type 1 Diabetes

Authors: Ewa Rusak, Sebastian Seget, Aleksandra Mroskowiak, Mirosław Partyka, Ewa Samulska, Julia Strózik, Anna Wilk, Przemysława Jarosz-Chobot

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Introduction: Various infections can affect children suffering from Type 1 Diabetes (T1D) because of dysfunctions of the immune system. The urinary tract and urethra of these children can be easily infected areas because of glycosuria. Aim: The microflora assessment of the urethra area of children with newly diagnosed T1D. Methods: The materials of the study were swabs taken prospectively from the urethral area of 63 children at the time of diagnosis of T1D (37 boys), then the results were correlated to the clinical parameters. In the statistical analysis, there were T student, Chi square, and U Mann-Whitney tests used. Results: The mean age was 9.4 years (6 months-17.4 years). The mean HbA1c value was 12.1% (5,6% - 20.1%). The mean value of glycosuria was 4463.2 mg/dl (0 - 9770 mg/dl). Ketoacidosis was diagnosed in 29 children (49%). The following microbial species were isolated in the collected materials: Staphylococcus epidermidis in 18 children (28.6%), Enterococcus faecalis in 17 children (27%), Candida albicans in 15 children (23.8%), coagulase-negative staphylococciin 11 children (17.5%), group B Streptococcus beta-hemolysis in 10 children (15.9%), S. aureus, E. coli, S. anginosus, C. glucuronolyticum, and A. urinae in 7 children each (11.1%), group B Streptococcus beta-hemolysis and S. hominis in 6 children each (9.5%), L. gasseri in 5 children (7.5%), C. dubliniensis in 4 children (6.3) and other, isolated cases. 2 of diagnosed patients were cultured negatively (3.2%). There were statistical correlations between the type of colonisation and patients’ sex and HbA1C value. Conclusions: It is extremely important to examine the urethral area at the time of diagnosis of T1D in order to detect inflammation and to undertake the appropriate and effective intervention.

Keywords: diabetology, skin disorders, microbiology, microflora

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10 Retrospective Study of Positive Blood Cultures Carried out in the Microbiology Department of General Hospital of Ioannina in 2017

Authors: M. Gerasimou, S. Mantzoukis, P. Christodoulou, N. Varsamis, G. Kolliopoulou, N. Zotos

Abstract:

Purpose: Microbial infection of the blood is a serious condition where bacteria invade the bloodstream and cause systemic disease. In such cases, blood cultures are performed. Blood cultures are a key diagnostic test for intensive care unit (ICU) patients. Material and method: The BacT/Alert system, which measures the production of carbon dioxide with metabolic organisms, is used. The positive result in the BacT/Alert system is followed by culture in the following selective media: Blood, Mac Conkey No 2, Chocolate, Mueller Hinton, Chapman and Sabaureaud agar. Gram staining method was used to differentiate bacterial species. The microorganisms were identified by biochemical techniques in the automated Microscan (Siemens) system and followed by a sensitivity test on the same system using the minimum inhibitory concentration MIC technique. The sensitivity test is verified by a Kirby Bauer-based test. Results: In 2017 the Laboratory of Microbiology received 3347 blood cultures. Of these, 170 came from the ICU. 116 found positive. Of these S. epidermidis was identified in 42, A. baumannii in 27, K. pneumoniae in 12 (4 of these KPC ‘Klebsiella pneumoniae carbapenemase’), S. hominis in 8, E. faecium in 7, E. faecalis in 5, P. aeruginosa in 3, C. albicans in 3, S. capitis in 2, K. oxytoca in 2, P. mirabilis in 2, E. coli in 1, S. intermidius in 1 and S. lugdunensis in 1. Conclusions: The study of epidemiological data and microbial resistance phenotypes is essential for the choice of therapeutic regimen for the early treatment and limitation of multivalent strains, while it is a crucial factor to solve diagnostic problems.

Keywords: blood culture, bloodstream, infection, intensive care unit

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9 Intestinal Parasites Detected by Fecal Examination in Cats in the Konya Province, Turkey

Authors: Nermin Isik, Ozlem Derinbay Ekici

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The cat is one of the potential hosts for parasitic zoonoses, such as Toxocara cati, Ancylostoma braziliense, A. tubaeforme, Uncinaria stenocephala, Cryptosporidium sp, Giardia sp. This study was performed to determine the prevalence and intensity of intestinal parasites in household cats in Konya, Turkey. In this study, a total of 100 stool samples with different ages and sex were used as a material. They were examined for infections with endoparasites by the use of native, Fulleborn flotation and Benedek sedimentation methods in University of Selcuk, Faculty of Veterinary Medicine, Department of Parasitology. The overall prevalence of intestinal parasites was 15%. A total of 6 parasite species was recorded: Giardia sp (6%), Toxocara cati (4%), Isospora sp (3%), Joyeuxiella pasqualei, Taenia sp (1%), Trichuris sp (1%). The most common intestinal parasites in cats were Giardia sp (6%) and Toxocara cati (4%). Younger cats up to 1 year of age were more frequently infected with endoparasites than animals over 1 year of age (p < 0.05). Giardia sp and Toxocara cati were detected significantly more often in younger than 1 year of age (p < 0.05). In fecal examination, Toxocara cati, Ancylostoma sp. Joyeuxiella pasqualei, Dipylidium caninum, Trichuris sp were found in cats in Turkey. In this study, based on microscopic and macroscopic fecal examinations, Giardia sp (6%), Toxocara cati (4%), Isospora sp (3%), Joyeuxiella pasqualei (%2), Taenia sp (1%), Trichuris sp (1%) was detected in cats. In conclusion, zoonotic intestinal parasites in household cats such as Giardia sp and Toxocara cati should be considered more seriously and it is necessary to take precautions against these infections. Cats should be routinely checked by faecal examination for endoparasite infections.

Keywords: cat, intestinal parasites, faecal, Turkey

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8 The Epidemiological Study on Prevalence of Giardia lamblia among Children in Esfahan City of Iran

Authors: Shahla Rostamirad

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Purpose: Giardiasis is a widespread infection in humans caused by Giardia lamblia. The prevalence of this parasite among children in Isfahan of Iran is unknown. This study intended to estimate Giardia lamblia infection prevalence and identify possible associated risk factors in a healthy pediatric population living in the Isfahan, a metropolitan city of Iran. Methods: Between September 2010 and March 2012, 1448 stool sample from children with clinical manifestation that refer to clinical lab in Isfahan city for stool examination were collected and analyzed. About 1218 samples were positive for parasitic disease. All of samples were examined and diagnosed by direct examination and formalin-ether concentration of stools. Results: A total of 1218 positive cases were analyzed in this study. The findings showed that 92.5% of patients were infected by protozoa and 7.5 percent with helminth infection. The highest and lowest rate of infection belongs to Giardia lamblia and Entamoeba histolytica with 75% and 1.1%, respectively. Other infection cases were included of Blastocystys hominis 9.9%, E. coli 6.5%, H. nana 1.3%, Enterobious vermicolaris 4% and Ascaris lumbricoides 2.2% percent. The population studied revealed a gender distribution of 53.2% male and 46.8% female. Age distribution was 57.3% between 0-5 years and 42.7% between 6-15 years.The prevalence was higher among children aged 0-5 years (57.8%), than among older children (42.2%). Conclusion: The prevalence of protozoan parasite, especially Giardiasis, in children residing in the region of Isfahan is high. Several risk factors were associated with this prevalence and highlight the importance of parents' education and sanitation conditions in the children's well being. The association between Giardia lamblia and H. pylori seems an important issue deserving further investigation in order to promote prevention or treatment strategies. Other risk factor include presence of Helicobacter pylori infection, living in houses with own drainage system and reported household, pet contact, especially with cat and dog.

Keywords: Giardia duodenalis, prevalence, risk factors, children, Isfahan, Iran

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7 Species Distribution and Incidence of Inducible Clindamycin Resistance in Coagulase-Negative Staphylococci Isolated from Blood Cultures of Patients with True Bacteremia in Turkey

Authors: Fatma Koksal Cakirlar, Murat Gunaydin, Nevri̇ye Gonullu, Nuri Kiraz

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During the last few decades, the increasing prevalence of methicillin resistant-CoNS isolates has become a common problem worldwide. Macrolide-lincosamide-streptogramin B (MLSB) antibiotics are effectively used for the treatment of CoNS infections. However, resistance to MLSB antibiotics is prevalent among staphylococci. The aim of this study is to determine species distribution and the incidence of inducible clindamycin resistance in CoNS isolates caused nosocomial bacteremia in our hospital. Between January 2014 and October 2015, a total of 484 coagulase-negative CoNS isolates were isolated from blood samples of patients with true bacteremia who were hospitalized in intensive care units and in other departments of Istanbul University Cerrahpasa Medical Hospital. Blood cultures were analyzed with the BACTEC 9120 system (Becton Dickinson, USA). The identification and antimicrobial resistance of isolates were determined by Phoenix automated system (BD Diagnostic Systems, Sparks, MD). Inducible clindamycin resistance was detected using D-test. The species distribution was as follows: Staphylococcus epidermidis 211 (43%), S. hominis 154 (32%), S. haemolyticus 69 (14%), S. capitis 28 (6%), S. saprophyticus 11 (2%), S. warnerii 7 (1%), S. schleiferi 5 (1%) and S. lugdunensis 1 (0.2%). Resistance to methicillin was detected in 74.6% of CoNS isolates. Methicillin resistance was highest in S.hemoliticus isolates (89%). Resistance rates of CoNS strains to the antibacterial agents, respectively, were as follows: ampicillin 77%, gentamicin 20%, erythromycin 71%, clindamycin 22%, trimethoprim-sulfamethoxazole 45%, ciprofloxacin 52%, tetracycline 34%, rifampicin 20%, daptomycin 0.2% and linezolid 0.2%. None of the strains were resistant to vancomycin and teicoplanin. Fifteen (3%) CoNS isolates were D-test positive, inducible MLSB resistance type (iMLSB-phenotype), 94 (19%) were constitutively resistant (cMLSB -phenotype), and 237 (46,76%) isolates were found D-test negative, indicating truly clindamycin-susceptible MS phenotype (M-phenotype resistance). The incidence of iMLSB-phenotypes was higher in S. epidermidis isolates (4,7%) compared to other CoNS isolates.

Keywords: bacteremia, inducible MLSB resistance phenotype, methicillin-resistant, staphylococci

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6 Inactivation of Root-Knot Nematode Eggs Meloidogyne enterolobii in Irrigation Water Treated with Ozone

Authors: I. A. Landa-Fernandez, I. Monje-Ramirez, M. T. Orta-Ledesma

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Every year plant-parasitic nematodes diminish the yield of high-value crops worldwide causing important economic losses. Currently, Meloidogyne enterolobii has increased its importance due to its high aggressiveness, increasing geographical distribution and host range. Root-knot nematodes inhabit the rhizosphere soil around plant roots. However, they can come into contact with irrigation water. Thus, plant-parasitic nematodes can be transported by water, as eggs or juveniles. Due to their high resistance, common water disinfection methods are not effective for inactivating these parasites. Ozone is the most effective disinfectant for microbial inactivation. The objective of this study is to demonstrate that ozone treatment is an alternative method control in irrigation water of the root-knot nematode M. enterolobii. It has been shown that ozonation is an effective treatment for the inactivation of protozoan cysts and oocysts (Giardia and Cryptosporidium) and for other species of the genus Meloidogyne (M. incognita), but not for the enterolobii specie. In this study, the strain of M. enterolobii was isolated from tomatoes roots. For the tests, eggs were used and were inoculated in water with similar characteristics of irrigation water. Subsequently, the disinfection process was carried out in an ozonation unit. The performance of the treatments was evaluated through the egg's viability by assessing its structure by optical microscopy. As a result of exposure to ozone, the viability of the nematode eggs was reduced practically in its entirety; with dissolved ozone levels in water close to the standard concentration (equal to 0.4 mgO₃/L), but with high contact times (greater than 4 min): 0.2 mgO₃/L for 15 minutes or 0.55 mgO₃/L for 10 minutes. Additionally, the effect of temperature, alkalinity and organic matter of the water was evaluated. Ozonation is effective and a promising alternative for the inactivation of nematodes in irrigation water, which could contribute to diminish the agricultural losses caused by these organisms.

Keywords: inactivation process, irrigation water treatment, ozonation, plant-parasite nematodes

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5 Seasonal Prevalence of Gastrointestinal Parasites and Their Association with Trace Element Contents in Sera of Sheep, Grazing Forages and Soils of Sialkot District, Punjab, Pakistan

Authors: Hafiz M. Rizwan, Muhammad S. Sajid, Zafar Iqbal, Muhammad Saqib

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Gastro-intestinal (GI) helminths infection in sheep causes a substantial loss in terms of productivity and constitutes serious economic losses in the world. Different types of forages are rich in trace element contents and may act as a natural resource to improve the trace element deficiencies leading to immunity boost-up in general and against gastrointestinal parasitic infections in particular. In the present study, the level of trace elements (Cu, Co, Mn, Zn) determined in sera of different breeds of sheep, available feedstuffs, respective soil samples and their association with GI helminths in Sialkot district, Punjab, Pakistan. Almost similar prevalence of GI helminths was recorded (32.81%) during spring 2015 and (32.55%) during autumn 2014. The parasitic species identified from the microscopically scanned faecal samples of district Sialkot were Fasciola (F.) hepatica, F. gigantica, Haemonchus contortus, Eimeria crandallis, Gongylonema pulchrum, Oesophagostomum sp., Trichuris ovis, Strongyles sp., Cryptosporidium sp. and Trichostrongylus sp. Among variables like age, sex, and breed, only sex was found significant in district Sialkot. A significant (P < 0.05) variation in the concentration of Zn, Cu, Mn, and Co was recorded in collected forages species. Soils of grazing field showed insignificant (P > 0.05) variation among soils of different tehsils of Sialkot district. Statistically, sera of sheep showed no variation (P > 0.05) during autumn 2014, While, variation (P < 0.05) among different tehsils of Sialkot district during spring 2015 except Co. During autumn 2014 the mean concentration of Cu, Zn, and Co in sera was inversely proportional to the mean EPG of sheep while during spring 2015 only Zn was inversely proportional to the mean EPG of sheep. The trace element-rich forages preferably Zn were effective ones against helminths infection. The trace element-rich forages will be recommended for their utilization as an alternate to improve the trace element deficiencies in sheep which ultimately boost up the immunity against gastrointestinal parasitic infections.

Keywords: coprological examination, gastro-intestinal parasites, prevalence, sheep, trace elements

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4 Assessment of Current and Future Opportunities of Chemical and Biological Surveillance of Wastewater for Human Health

Authors: Adam Gushgari

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The SARS-CoV-2 pandemic has catalyzed the rapid adoption of wastewater-based epidemiology (WBE) methodologies both domestically and internationally. To support the rapid scale-up of pandemic-response wastewater surveillance systems, multiple federal agencies (i.e. US CDC), non-government organizations (i.e. Water Environment Federation), and private charities (i.e. Bill and Melinda Gates Foundation) have funded over $220 million USD supporting development and expanding equitable access of surveillance methods. Funds were primarily distributed directly to municipalities under the CARES Act (90.6%), followed by academic projects (7.6%), and initiatives developed by private companies (1.8%). In addition to federal funding for wastewater monitoring primarily conducted at wastewater treatment plants, state/local governments and private companies have leveraged wastewater sampling to obtain health and lifestyle data on student, prison inmate, and employee populations. We explore the viable paths for expansion of the WBE m1ethodology across a variety of analytical methods; the development of WBE-specific samplers and real-time wastewater sensors; and their application to various governments and private sector industries. Considerable investment in, and public acceptance of WBE suggests the methodology will be applied to other future notifiable diseases and health risks. Early research suggests that WBE methods can be applied to a host of additional “biological insults” including communicable diseases and pathogens, such as influenza, Cryptosporidium, Giardia, mycotoxin exposure, hepatitis, dengue, West Nile, Zika, and yellow fever. Interest in chemical insults is also likely, providing community health and lifestyle data on narcotics consumption, use of pharmaceutical and personal care products (PPCP), PFAS and hazardous chemical exposure, and microplastic exposure. Successful application of WBE to monitor analytes correlated with carcinogen exposure, community stress prevalence, and dietary indicators has also been shown. Additionally, technology developments of in situ wastewater sensors, WBE-specific wastewater samplers, and integration of artificial intelligence will drastically change the landscape of WBE through the development of “smart sewer” networks. The rapid expansion of the WBE field is creating significant business opportunities for professionals across the scientific, engineering, and technology industries ultimately focused on community health improvement.

Keywords: wastewater surveillance, wastewater-based epidemiology, smart cities, public health, pandemic management, substance abuse

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3 Bacterial Diversity in Vaginal Microbiota in Patients with Different Levels of Cervical Lesions Related to Human Papillomavirus Infection

Authors: Michelle S. Pereira, Analice C. Azevedo, Julliane D. Medeiros, Ana Claudia S. Martins, Didier S. Castellano-Filho, Claudio G. Diniz, Vania L. Silva

Abstract:

Vaginal microbiota is a complex ecosystem, composed by aerobic and anaerobic bacteria, living in a dynamic equilibrium. Lactobacillus spp. are predominant in vaginal ecosystem, and factors such as immunity and hormonal variations may lead to disruptions, resulting in proliferation of opportunistic pathogens. Bacterial vaginosis (BV) is a polymicrobial syndrome, caused by an increasing of anaerobic bacteria replacing Lactobacillus spp. Microorganisms such as Gardnerella vaginalis, Mycoplasma hominis, Mobiluncus spp., and Atopobium vaginae can be found in BV, which may also be associated to other infections such as by Human Papillomavirus (HPV). HPV is highly prevalent in sexually active women, and is considered a risk factor for development of cervical cancer. As long as few data is available on vaginal microbiota of women with HPV-associated cervical lesions, our objectives were to evaluate the diversity in vaginal ecosystem in these women. To all patients, clinical and socio-demographic data were collected after gynecological examination. This study was approved by the Ethics Committee from Federal University of Juiz de Fora, Minas Gerais, Brazil. Vaginal secretion and cervical scraping were collected. Gram-stained smears were evaluated to establish Nugent score for BV determination. Viral and bacterial DNA obtained was used as template for HPV genotyping (PCR) and bacterial fingerprint (REP-PCR). In total 31 patients were included (mean age 35 and 93.6% sexually active). The Nugent score showed that 38.7% were BV. From the medical records, Pap smear tests showed that 32.3% had low grade squamous epithelial lesion (LSIL), 29% had high grade squamous epithelial lesion (HSIL), 25.8% had atypical squamous cells of undetermined significance (ASC-US) and 12.9% with atypical squamous cells that would not exclude high-grade lesion (ASC-H). All participants were HPV+. HPV-16 was the most frequent (87.1%), followed by HPV-18 (61.3%). HPV-31, HPV-52 and HPV-58 were also detected. Coinfection HPV-16/HPV-18 was observed in 75%. In the 18-30 age group, HPV-16 was detected in 40%, and HPV-16/HPV-18 coinfection in 35%. HPV-16 was associated to 30% of ASC-H and 20% of HSIL patients. BV was observed in 50% of HPV-16+ participants and in 45% of HPV-16/HPV-18+. Fingerprints of bacterial communities showed clusters with low similarity suggesting high heterogeneity in vaginal microbiota within the sampled group. Overall, the data is worrisome once cervical-cancer highly risk-associated HPV-types were identified. The high microbial diversity observed may be related to the different levels of cellular lesions, and different physiological conditions of the participants (age, social behavior, education). Further prospective studies are needed to better address correlations and BV and microbial imbalance in vaginal ecosystems which would be related to the different cellular lesions in women with HPV infections. Supported by FAPEMIG, CNPq, CAPES, PPGCBIO/UFJF.

Keywords: human papillomavirus, bacterial vaginosis, bacterial diversity, cervical cancer

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