Search results for: food-borne pathogens

Authors: Haile Berihulay, Chenglong Luo

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Avian influenza, commonly known as bird flu, is a highly contagious viral disease primarily affecting poultry and wild birds, with significant implications for both animal health and public safety. The influenza virus (IV) is notorious for its ability to mutate and infect multiple species, including humans, leading to severe consequences. Avian influenza poses considerable pandemic risks due to the potential evolution of low pathogenic avian influenza (LPAI) into highly pathogenic avian influenza (HPAI), which can cause rapid outbreaks in domestic flocks. While AVI viruses typically do not replicate well in humans, strains such as H5N1 and H7N9 have crossed the species barrier, raising alarm over human infections. The recent documentation of human transmission of the H5N8 strain from birds underscores the ongoing threat posed by avian influenza. This review necessitates a thorough discussion about the genetic foundation of viral pathogens, identifying key candidate genes linked to disease resilience, and discussing powerful tools. This review can help researchers to comprehensively overview the disease severity and combat related to AIV, which causes significant economic impact and set effective control strategies to mitigate the risks associated with avian influenza outbreaks.

Keywords: Avian, candidate genes, chicken, molecular, pathogen, virus

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Authors: U.N. Emiri, E. Moroyei

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Mycoflora associated with the seed rot disease of Mucuna sloanei and their effects on nutrient and phytochemical composition of the seeds were investigated. The fungal pathogens implicated in the seed rot disease were Rhizopus stolonifer, Aspergillus flavus, Aspergillus niger, and Fusarium oxysporum. The fungal isolates were aseptically inoculated into healthy M. Sloanei seeds and incubated for 7 days at room temperature of 25 ± 30c. The results of the proximate and mineral analysis in mg/100g of fungal infected and non-infected (control) seeds that were carried out revealed that there was an increase in Moisture and Carbohydrate content of the fungal infected seeds relative to the non-infected seeds (control). However, there was a decrease in Ash, Fibre, Lipid, and Protein content of the fungal infected seeds relative to the non-infected (control). It was observed that moisture had increased from 10.50 ± 0.16 in the non-infected seeds to 17.60 ± 0.20 in the infected samples and Carbohydrate content had also increased from 49.6 ± 0.25 in the non-infected to 52.50 ± 0.29 in the infected seeds. The following parameters decreased in the infected than in the non-infected seeds. They include Ash 2.60 ± 0.12, Crude fibre 1.9 ± 0.08, Lipid 6.50 ± 0.16, and Protein content 18.50 ± 0.06. Similarly, Calcium 2.50 ± 0.12, Phosphorus 1.80 + 0.12 and Potassium 1.80 + 0.09 increased in the infected than in the non-infected seed, while iron 0.20 ± 0.05, Sodium 0.02 ± 0.01 and Magnesium 0.06 ± 0.02 decreased in the infected seeds. All phytochemical contents analyzed increased in the infected seeds viz Tannim 0.50 ± 0.12, Oxalate 1.60 ± 0.05, Hydrogen cyanide 1.82 ± 0.06, and Saponin 2.50+0.28. However, the nutrient compositions and Phytochemical between the infected and non-infected seeds are not significantly different (p > 0.05).

Keywords: Mycoflora, mucuna sloanei, seeds, phytochemical, nutrient composition

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Authors: Adela Diepoltova, Klara Konecna, Ondrej Jandourek, Petr Nachtigal

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A loss of control over antibiotic-resistant pathogens has become a global issue due to severe and often untreatable infections. This state is reflected in complicated treatment, health costs, and higher mortality. All these factors emphasize the urgent need for the discovery and development of new anti-infectives. One of the most common pathogens mentioned in the phenomenon of antibiotic resistance are bacteria of the genus Staphylococcus. These bacterial agents have developed several mechanisms against the effect of antibiotics. One of them is biofilm formation. In staphylococci, biofilms are associated with infections such as endocarditis, osteomyelitis, catheter-related bloodstream infections, etc. To author's best knowledge, no validated and standardized methodology evaluating candidate compound activity against staphylococcal biofilms exists. However, a variety of protocols for in vitro drug activity testing has been suggested, yet there are often fundamental differences. Based on our experience, a key methodological step that leads to credible results is to form a robust biofilm with appropriate attributes such as firm adherence to the substrate, a complex arrangement in layers, and the presence of extracellular polysaccharide matrix. At first, for the purpose of drug antibiofilm activity evaluation, the focus was put on various conditions (supplementation of cultivation media by human plasma/fetal bovine serum, shaking mode, the density of initial inoculum) that should lead to reproducible and robust in vitro staphylococcal biofilm formation in microtiter plate model. Three model staphylococcal reference strains were included in the study: Staphylococcus aureus (ATCC 29213), methicillin-resistant Staphylococcus aureus (ATCC 43300), and Staphylococcus epidermidis (ATCC 35983). The total biofilm biomass was quantified using the Christensen method with crystal violet, and results obtained from at least three independent experiments were statistically processed. Attention was also paid to the viability of the biofilm-forming staphylococcal cells and the presence of extracellular polysaccharide matrix. The conditions that led to robust biofilm biomass formation with attributes for biofilms mentioned above were then applied by introducing an alternative method analogous to the commercially available test system, the Calgary Biofilm Device. In this test system, biofilms are formed on pegs that are incorporated into the lid of the microtiter plate. This system provides several advantages (in situ detection and quantification of biofilm microbial cells that have retained their viability after drug exposure). Based on our preliminary studies, it was found that the attention to the peg surface and substrate on which the bacterial biofilms are formed should also be paid to. Therefore, further steps leading to the optimization were introduced. The surface of pegs was coated by human plasma, fetal bovine serum, and L-polylysine. Subsequently, the willingness of bacteria to adhere and form biofilm was monitored. In conclusion, suitable conditions were revealed, leading to the formation of reproducible, robust staphylococcal biofilms in vitro for the microtiter model and the system analogous to the Calgary biofilm device, as well. The robustness and typical slime texture could be detected visually. Likewise, an analysis by confocal laser scanning microscopy revealed a complex three-dimensional arrangement of biofilm forming organisms surrounded by an extracellular polysaccharide matrix.

Keywords: anti-biofilm drug activity screening, in vitro biofilm formation, microtiter plate model, the Calgary biofilm device, staphylococcal infections, substrate modification, surface coating

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Authors: Walaa Al-Maghraby

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Endophytes are organisms that colonize internal plant tissues without causing apparent harm to their host. Almost all groups of microorganisms have been found in endophytic association with plants may be fungi. They stimulate the production of secondary metabolites with a diverse range of biological activities. Leptadenia pyrotechnica is a more or less leafless, erect shrub with straight stems which is highly distributed in Saudi Arabia. Four endophytes fungi were isolated from Leptadenia pyrotechnica and identified using 18S ribosomal RNA sequences, which revealed four fungi genuses, namely Aspergillus terreus; Aspergillus welwitschiae; Aspergillus fumigatus and Aspergillus flavus. In this present study, four endophytic fungi from Leptadenia pyrotechnica were used for obtaining crude aqueous and ethyl acetate extracts for antimicrobial screening against 6 human pathogens, the antibacterial tests presented satisfactory results, where the pathogenic bacteria were inhibited by the four extracts tested, except for Escherichia coli that was inhibited by all extracts except ethyl acetate extract of Aspergillus terreus. Analysis of variance showed that the extract produced by endophyte Leptadenia pyrotechnica was the most effective against all bacteria, either gram-negative or positive. However, the extract was not efficient against pathogenic fungi. Therefore, this study indicates that endophytes from medicinal plant Leptadenia pyrotechnica could be potential sources of antibacterial substances.

Keywords: antimicrobial activity, Aspergillus sp, endophytes, Leptadenia pyrotechnica

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Authors: Maria Nejjari, Michel Cloutier, Guylaine Talbot, Martin Lanthier

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The fluorescence in situ hybridization (FISH) is a staining technique that allows the identification, detection and quantification of microorganisms without prior cultivation by means of epifluorescence and confocal laser scanning microscopy (CLSM). Oligonucleotide probes have been used to detect bacteria and archaea that colonize the cattle and swine digestive systems. These bacterial strains have been obtained from fecal samples issued from cattle manure and swine slurry. The collection of these samples has been done at 3 different pit’s levels A, B and C with same height. Two collection depth levels have been taken in consideration, one collection level just under the pit’s surface and the second one at the bottom of the pit. Cells were fixed and FISH was performed using oligonucleotides of 15 to 25 nucleotides of length associated with a fluorescent molecule Cy3 or Cy5. The double hybridization using Cy3 probe targeting bacteria (Cy3-EUB338-I) along with a Cy5 probe targeting Archaea (Gy5-ARCH915) gave a better signal. The CLSM images show that there are more bacteria than archaea in swine slurry. However, the choice of fluorescent probes is critical for getting the double hybridization and a unique signature for each microorganism. FISH technique is an easy way to detect pathogens like E. coli O157, Listeria, Salmonella that easily contaminate water streams, agricultural soils and, consequently, food products and endanger human health.

Keywords: archaea, bacteria, detection, FISH, fluorescence

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Authors: Osama M. Darwesh, Sahar H. Hassan, Abd El-Raheem R. El-Shanshoury, Shawky Z. Sabae

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Nanotechnology as multidisciplinary technology is growing rapidly with important applications in several sectors. Also, nanobiotechnology is known for the use of microorganisms for the synthesis of targeted nanoparticles. The present study deals with the green synthesis of silver nanoparticles using aquatic bacteria and the development of a biogenic nanocomposite for environmental applications. Twenty morphologically different colonies were isolated from the collected water samples from eight different locations at the Rosetta branch of the Nile Delta, Egypt. The obtained results illustrated that the most effective bacterial isolate (produced the higher amount of AgNPs after 24 h of incubation time) is isolate R3. Bacillus tequilensis was the strongest extracellular bio-manufactory of AgNPs. Biosynthesized nanoparticles had a spherical shape with a mean diameter of 2.74 to 28.4 nm. The antimicrobial activity of silver nanoparticles against many pathogenic microbes indicated that the produced AgNPs had high activity against all tested multi-antibiotic resistant pathogens. Also, the stabilized prepared AgNPs-SA nanocomposite has greater catalytic activity for the decolourization of some dyes like Methylene blue (MB) and Crystal violet. Such results represent a promising stage for producing eco-friendly, cost-effective, and easy-to-handle devices for the bioremediation of contaminated industrial wastewater.

Keywords: bioremediation, AgNPs, AgNPs-SA nanocomposite, Bacillus tequilensis, nanobiotechnology

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Authors: Waode Nurmayani, Nikmatul Riswanda

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Broilers are chickens which are kept with the most efficient time and hoped get a good body weight. All things are done, for example with the improvement of feed and use antibiotics. Feed cost is the most cost to be spent. Nearly 80% of the cost is spent just for buy feed. Earthworm (Lumbricus rubellus) is a good choice to reduce the cost of feed protein source. The Earthworm has a high crude protein content of about 48.5%-61.9%, rich with proline amino acid about 15% of the 62 amino acids. Not only about protein, this earthworm also has a role in disease prevention. Prevention of disease in livestock usual with use feed supplement. Earthworm (Lumbricus rubellus) is one of the natural materials used as feed. In addition, several types of earthworms that have been known to contain active substances about antibacterial pathogens namely Lumbricus rubellus. The earthworm could be used as an antibiotic because it contain the antibody of Lumbricine active substance. So that, this animal feed from Lumbricus rubellus could improve the performance of broilers. Bioactive of anti-bacterial is called Lumbricine able to inhibit the growth of pathogenic bacteria in the intestinal wall so that the population of pathogenic bacteria is reduced. The method of write in this scientific writing is divided into 3 techniques, namely data completion, data analysis, and thinking pan from various literature about earthworm (Lumbricus rubellus) as broiler feed. It is expected that innovation of feed material of earthworm (Lumbricus rubellus) could reduce the cost of protein feed and the use of chemical antibiotics.

Keywords: earthworm, broiler, protein, antibiotic

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Authors: Mark Osborn

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Clustered, regularly interspaced short palindromic repeat (CRISPR/Cas) proteins can be designed to bind specified DNA and RNA sequences and hold great promise for the accurate detection of nucleic acids for diagnostics. Commercially available reagents were integrated into a CRISPR/Cas9-based lateral flow assay that can detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sequences with single-base specificity. This approach requires minimal equipment and represents a simplified platform for field-based deployment. A rapid, multiplex fluorescence CRISPR/Cas9 nuclease cleavage assay capable of detecting and differentiating SARS-CoV-2, influenza A and B, and respiratory syncytial virus in a single reaction was also developed. These findings provide proof of principle for CRISPR/Cas9 point-of-care diagnosis that can detect specific SARS-CoV-2 strain(s). Further, Cas9 cleavage allows for a scalable fluorescent platform for identifying respiratory viral pathogens with overlapping symptomology. Collectively, this approach is a facile platform for diagnostics with broad application to user-defined sequence interrogation and detection.

Keywords: CRISPR/Cas9, lateral flow assay, SARS-Co-V2, single-nucleotide resolution

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Authors: Ankita Singh, Chandan Gorain, Amirul I. Mallick

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Successful adherence of the mucosal epithelial cells is the key early step for Campylobacter jejuni pathogenesis (C. jejuni). A set of Surface Exposed Colonization Proteins (SECPs) are among the major factors involved in host cell adherence and invasion of C. jejuni. Among them, constitutively expressed surface-exposed lipoprotein adhesin of C. jejuni, JlpA, interacts with intestinal heat shock protein 90 (hsp90α) and contributes in disease progression by triggering pro-inflammatory response via activation of NF-κB and p38 MAP kinase pathway. Together with its ability to express in the bacterial surface, higher sequence conservation and predicted predominance of several B cells epitopes, JlpA protein reserves its potential to become an effective vaccine candidate against wide range of Campylobacter sps including C. jejuni. Given that chickens are the primary sources for C. jejuni and persistent gut colonization remain as major cause for foodborne pathogenesis to humans, present study explicitly used chickens as model to test the immune-protective efficacy of JlpA protein. Taking into account that gastrointestinal tract is the focal site for C. jejuni colonization, to extrapolate the benefit of mucosal (intragastric) delivery of JlpA protein, a food grade Nisin inducible Lactic acid producing bacteria, Lactococcus lactis (L. lactis) was engineered to express recombinant JlpA protein (rJlpA) in the surface of the bacteria. Following evaluation of optimal surface expression and functionality of recombinant JlpA protein expressed by recombinant L. lactis (rL. lactis), the immune-protective role of intragastric administration of live rL. lactis was assessed in commercial broiler chickens. In addition to the significant elevation of antigen specific mucosal immune responses in the intestine of chickens that received three doses of rL. lactis, marked upregulation of Toll-like receptor 2 (TLR2) gene expression in association with mixed pro-inflammatory responses (both Th1 and Th17 type) was observed. Furthermore, intragastric delivery of rJlpA expressed by rL. lactis, but not the injectable form, resulted in a significant reduction in C. jejuni colonization in chickens suggesting that mucosal delivery of live rL. lactis expressing JlpA serves as a promising vaccine platform to induce strong immune-protective responses against C. jejuni in chickens.

Keywords: chickens, lipoprotein adhesion of Campylobacter jejuni, immuno-protection, Lactococcus lactis, mucosal delivery

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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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Authors: Tahra El-Obeid, Reham Mousa, Amal Alzahiri

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During the past few years the traditional market of Qatar has become an attraction to many customers who eat from the numerous women street vendors selling Qatari traditional dishes. To gain an understanding on the safety of these street vended foods, we designed the study to test microbiological quality of 14 different Qatari foods sold in Souk Wagif, the main traditional market in Qatar. This study was conducted to mainly identify presence or absence of microbial pathogens. A total of 56 samples were purchased from 10 different street vendors and the samples were collected randomly on different days. The samples were tested for microbial contaminants at Central Food Laboratories, Doha, Qatar. The qualitative study was conducted using Real Time-PCR to screen for; Salmonella spp., Listeria monocytogenes, Escherichia coli and E. coli 0157:H7. Out of the 56 samples, only two samples “Biryani” and “Khabess” contained E. coli. However, both samples tested negative for E. coli O157:H7. The microbial contamination of the Qatari traditional street vended foods was 3%. This result may be attributed to the food safety training requirement set by the regulatory authorities before issuing any license to food handlers in Qatar as well as the food inspection conducted by the food health inspectors on a regular basis.

Keywords: microbiological quality, street vended food, traditional dishes, Qatar

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Authors: Anusha Bhardwaj, Shekhar Shinde

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Nitric oxide (NO•) has been documented in research papers as one of the most versatile player in the therapeutics. It is identified as a biological multifunctional messenger molecule which is synthesized by the action of nitric oxide synthase (NOS) enzyme from L-arginine. The protective and the toxic effect in conjunction form the complete picture of the biological function of nitric oxide in humans. The dual nature is because of various factors such as concentration of NO, the isoform of NOS involved, type of cells in which it is synthesized, reaction partners like proteins, reactive oxygen intermediates, prosthetic groups, thiols etc., availability of the substrate L-arginine, intracellular environment in which NO is produced and generation of guanosine 3, 5’- cyclic monophosphate (cGMP). Activation of NOS through infection or trauma leads to one or more systemic effects including enhanced immune activity against invading pathogens, vaso/bronchodilatation in the cardiovascular and respiratory systems and altered neurotransmission which can be protective or toxic. Hence, NO affects the balance between healthy signaling and neurodegeneration in the brain. In lungs, it has beneficial effects on the function of airways as a bronchodilator and acts as the neurotransmitter of bronchodilator nerves. Whereas, on the other hand, NO may have deleterious effects by amplifying the asthmatic inflammatory response and also act as a vasodilator in the airways by increasing plasma exudation. But NOS Inhibitors and NO donors hamper the signalling pathway and hence a therapeutic application of NO is compromised.

Keywords: nitric oxide, multifunctional, dual nature, therapeutic applications

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Authors: Mohammad Saifuddin, Shahjada Selim

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Patients with diabetes mellitus (DM) are prone to develop infection, especially urinary tract infection (UTI) in comparison with non-diabetics. Due to the emergence of multidrug resistant (MDR) uropathogenic strains, the choice of antimicrobial agent is sometimes difficult. This study is designed to reveal the distribution of uropathogens in Diabetic patients and corresponding sensitivity patterns and to correlate the microbiological results with various clinical parameters. A nine-month retrospective review of 100 urine culture reports of Diabetic patients from January 2015 to September 2015 from semiurbanmultispeciality hospital of Feni, Bangladesh were analyzed. Only Diabetic patients were included in this study who were clinically diagnosed as UTI patients with a corresponding urine culture showing a bacterial count of ˃105cfu/ml.Out of 100 patients with UTI, 39 (39%) were male, and 61 (61%) were female. Organisms grown in urine culture were Escherichia coli (64) followed by Klebsiella (11), Proteus (7), Staph Aureus (4), Pseudomonas (4), Acinetobacter (3), Sreptococcus(3), Enterococcus (2 ) and one each of Enterobacter and Fungi. Overall sensitivity pattern in decreasing order of various commonly used antibiotics was Meropenem (89%), Nitrofurantoin (86%), Amikacin (81%), Ceftriaxone (68%), Cefuroxime (61%), Cefixime (39%), Quinolones (28%), Amoxicillin (16%). The significance of the study lies in the determination of common pathogens in diabetic patients with UTI and the resistance pattern of antibiotics so that physicians and pharmacists get the proper information rationalizing the rational use of antibiotics.

Keywords: Bangladesh, Diabetes Mellitus, E. coli, urinary tract infection

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Authors: Rajni Kant Panik, Deependra Singh, Manju Singh

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Despite significant advances in the treatment of severe burn injury, infection and sepsis persist as frequent causes of morbidity and mortality for burn victims due to extensive compromise of the skin and contiguous tissue that serve as a protective barrier against microbial invasion. In the setting of a burn wound infection, Staphylococcus aureus is the most commonly isolated pathogens from bloodstream infections in burn care hospitals. We aimed to develop a biocompatible system of Poly vinyl alcohol (PVA)-sodium alginate hydrogel carrying multiple drugs- catalase and mupirocin in controlled manner for effective and complete burn wound healing. PLGA nanoparticles of Catalase and mupirocin were prepared by homogenization method and optimized system was incorporated in PVA-sodium alginate slurry. PVA-sodium alginate hydrogels were prepared by freeze thaw method. The prepared dispersion was casted into films to prepare multiphase hydrogel system and characterized by in vitro and in vivo studies. The study clearly showed the beneficial effect of antioxidant enzyme and antibiotic in the treatment of infected burn wound, as evidenced by the reduced incidence of wound infection and the shortening of healing time.

Keywords: burn wound, catalase, mupirocin, wound healing

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Authors: Ricardo Munguia-Perez, Nayeli Remigio-Alvarado, M.Miriam Hernandez-Arroyo, Elsa Castañeda-Roldan

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Fungi have emerged as important pathogens causing morbidity and mortality mainly in immunosuppressed, malnourished and elderly patients. It has detected an increase in resistance to azoles primarily to fluconazol. The fungal infections have become a problem of public health for the resistance to antifungal agents, they have developed new antifungals with broad-spectrum. The aim of this study was determine the antifungal susceptibility of yeasts isolated from clinical samples (respiratory secretions, exudates, wounds, blood cultures, urine cultures) obtained from inpatients and outpatients of a tertiary hospital from State of Puebla. The antifungal susceptibility of the yeast from several clinical samples were determined by the CLS M44-A disk diffusion methods. 149 samples of yeast were analyzed. All species were 100% susceptible to nystatin and amphotericin B. Candida albicans showed resistance of 95.5 % to fluconazole, 50.7 % to 5-flurocytosine and 55.2 % intermediate susceptibility to ketoconazole. Candida glabrata 81.3 % was susceptibility to ketoconazole and 75 % to fluconazole, for the case of 5-flurocytosine the 56.3 % was susceptible. Candida krusei 100 % was susceptible to ketoconazole, 50 % to fluconazole and 37.5 % to 5-flurocytosine. The internal medicine have greater diversity of yeast, the samples have susceptibility of 64.7% to ketoconazole, 47.1 % to fluconazole and 27.5 % to 5-flurocytosine. Hospitalized patients are more resistant to fluconazole and nystatin, but in the case of outpatients presents resistance to ketoconazole.

Keywords: antifungal, susceptibility, yeast, clinical samples

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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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Authors: Usha Kiran, M. Z. Abdin

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Osmotin is an abundant cationic multifunctional protein discovered in cells of tobacco (Nicotiana tabacum L. var Wisconsin 38) adapted to an environment of low osmotic potential. It provides plants protection from pathogens, hence placed in the PRP family of proteins. The osmotin induced proline accumulation has been reported in plants including transgenic tomato and strawberry conferring tolerance against both biotic and abiotic stresses. The exact mechanism of induction of proline by osmotin is however, not known till date. These observations have led us to hypothesize that osmotin induced proline accumulation could be due to its involvement as transcription factor and/or cell signal pathway modulator in proline biosynthesis. The present investigation was therefore, undertaken to analyze the osmotin protein as transcription factor /cell signalling modulator using bioinformatics tools. The results of available online DNA binding motif search programs revealed that osmotin does not contain DNA-binding motifs. The alignment results of osmotin protein with the protein sequence from DATF showed the homology in the range of 0-20%, suggesting that it might not contain a DNA binding motif. Further to find unique DNA-binding domain, the superimposition of osmotin 3D structure on modeled Arabidopsis transcription factors using Chimera also suggested absence of the same. We, however, found evidence implicating osmotin in cell signaling. With these results, we concluded that osmotin is not a transcription factor but regulating proline biosynthesis and accumulation through cell signaling during abiotic stresses.

Keywords: osmotin, cell signaling modulator, bioinformatic tools, protein

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Authors: Young-Ji Kim, Jin-Hyeong Park, Hong-Seok Kim, Jung-Whan Chon, Kwang-Yeop Kim, Dong-Hyeon Kim, Il-Byeong Kang, Da-Na Jeong, Jin-Hyeok Yim, Ho-Seok Jang, Kwang-Young Song, Kun-Ho Seo

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Stenotrophomonas maltophilia is one of the emerging opportunistic pathogens, and also known to have extensive drug resistance intrinsically including carbepenems which is last resort for most serious infections. One possible way for S. maltophilia to infect human is via wastewater treatment plant (WWTP). In the period between October 2016 and February 2017, effluent samples of WWTP from 3 different pig farms were collected once a month and screened for isolation of S. maltophilia. Total 16 strains of S. maltophilia were isolated and, the antibiotic susceptibility phenotypes were determined by Vitek 2 system for 16 antibiotics, ampicillin (AMP), amoxicillin/clavulanic acid (AMC), piperacillin/tazobactam (TZP), cefazolin (CZ), cefoxitin (FOX), cefotaxime (CTX), ceftazidime (CAZ), cefepime (FEP), aztreonam (AZT), ertapenem (ETP), imipenem (IMP), amikacin (AK), gentamicin (GN), ciprofloxacin (CIP), tigecycline (TGC) and trimethoprim/sulfamethoxazole (SXT). All isolates showed high resistance to AMP (100%), CZ (100%), FOX (100%), CTX (100%), CAZ (100%), FEP (94%), AZT (100%), ETP (100%), IMP (100%), AK (100%), GN (100%) whereas were susceptible to CIP (0%), TGC (0%), SXT (6%). All strains harbored at least one of the antibiotic resistance determinant such as spgM, rmlA, and rpfF. Some isolates had similar MLST (multilocus sequence typing) types with clinical isolates, suggesting WWTP could have potential role in the transmission of S. maltophilia to aquatic environment and, possibly, to humans.

Keywords: Stenotrophomonas maltophilia, Carbapenem resistance, waste water treatment plant, pig farm

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Authors: V. Baweja, K. K. Gupta, V. Dubey, C. Keshavam

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Antimicrobial activity of six indigenous plants such as Tulsi Ocimum sanctum, Neem Azadirachta indica, Aloe vera, Turmeric Curcuma longa, Lantana Lantana camara, and Clove Syzygium aromaticum was assessed against the gut microbiota of the dengue fever mosquito Aedes aegypti, keeping in view that the presence of midgut bacteria may affect the ability of the vector to transmit pathogens. Eleven different types of bacterial clones were isolated from the midgut of lab-reared fourth instar larvae of Aedes aegypti and were grown on LB agar medium at an optimum temperature of 25 ºC. Identification of these bacteria was done on the basis of their colony characteristic such as colony size, shape, opacity, elevation, consistency, and growth. Light microscopic studies of the gut microbiota revealed dominance of Gram-negative cocci over gram positive cocci and bacilli and Gram-negative bacilli. Identification of species was done by chemical characterization of the colonies. Crude extracts of all test plants were screened for their antimicrobial activities against gut microbiota by disc diffusion assay. The zone of exclusion seen after 24 hr of incubation in different assays revealed the most potent antibacterial activities in neem followed by clove and turmeric. Lantana and Aloe vera were least effective.

Keywords: plant extract, aedes, dengue, antimicrobial activity

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Authors: P. Praveen Kumar, P. Vimal Prabhu, Renu John

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In biology most microscopy specimens, in particular living cells are transparent. In cell imaging, it is hard to create an image of a cell which is transparent with a very small refractive index change with respect to the surrounding media. Various techniques like addition of staining and contrast agents, markers have been applied in the past for creating contrast. Many of the staining agents or markers are not applicable to live cell imaging as they are toxic. In this paper, we report theoretical and experimental results from quantitative phase imaging of yeast cells with a commercial bright field microscope. We reconstruct the phase of cells non-interferometrically based on the transport of intensity equations (TIE). This technique estimates the axial derivative from positive through-focus intensity measurements. This technique allows phase imaging using a regular microscope with white light illumination. We demonstrate nano-metric depth sensitivity in imaging live yeast cells using this technique. Experimental results will be shown in the paper demonstrating the capability of the technique in 3-D volume estimation of living cells. This real-time imaging technique would be highly promising in real-time digital pathology applications, screening of pathogens and staging of diseases like malaria as it does not need any pre-processing of samples.

Keywords: axial derivative, non-interferometric imaging, quantitative phase imaging, transport of intensity equation

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Authors: James Sinclair, Katy Soapi, Brad Carte

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The marine environment has continuously demonstrated to be a rich source of secondary metabolites and bioactive compounds that can address the many pharmaceutical problems facing mankind. The emergence of multidrug resistant pathogens has caused scientists to explore contemporary ways of combating these super bugs. A red-pigmented bacterial strain isolated from a marine alga collected in Fiji was identified to be Pseudoalteromonas rubra from 16s rRNA sequencing. This bacterial strain was cultured using a yeast-peptone media and incubated for five days. The ethyl acetate extract of this bacterium was subjected to chromatographic separation techniques such as vacuum liquid chromatography, flash chromatography, size exclusion chromatography and high-pressure liquid chromatography to yield the pure compound and a number of semi-pure fractions. The crude extract and subsequent purified fractions were analyzed by ultraviolet/visible spectroscopy and mass spectroscopy and was found to contain the compounds ivermectin, stenothricin, cyclo-L-pro-L-val, prodigiosin, mycophenolic acid, phenazine-1-carboxylic acid, eplerenone, staurosporine and pseudoalteromone A. The structure of the pure compound, pseudoalteromone A, was elucidated using NMR 1H, 13C, 1H-1H COSY, HSQC and HMBC spectroscopic data.

Keywords: Pseudoalteromonas rubra, Pseudoalteromone A, secondary metabolites, structure elucidation

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Authors: Carmen J. Savelli, Romy Conzade

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Background: Access to sufficient amounts of safe and nutritious food is a basic human necessity, required to sustain life and promote good health. Food safety and food security are therefore inextricably linked, yet the importance of food safety in this relationship is often overlooked. Methodologies: A literature review and desk study were conducted to examine existing frameworks for discussing food security, especially from an international perspective, to determine the entry points for enhancing considerations for food safety in national and international policies. Major Findings: Food security is commonly understood as the state when all people at all times have physical, social and economic access to sufficient, safe and nutritious food to meet their dietary needs and food preferences for an active and healthy life. Conceptually, food security is built upon four pillars including food availability, access, utilization and stability. Within this framework, the safety of food is often wrongly assumed as a given. However, in places where food supplies are insufficient, coping mechanisms for food insecurity are primarily focused on access to food without considerations for ensuring safety. Under such conditions, hygiene and nutrition are often ignored as people shift to less nutritious diets and consume more potentially unsafe foods, in which chemical, microbiological, zoonotic and other hazards can pose serious, acute and chronic health risks. While food supplies might be safe and nutritious, if consumed in quantities insufficient to support normal growth, health and activity, the result is hunger and famine. Recent estimates indicate that at least 842 million people, or roughly one in eight, still suffer from chronic hunger. Even if people eat enough food that is safe, they will become malnourished if the food does not provide the proper amounts of micronutrients and/or macronutrients to meet daily nutritional requirements, resulting in under- or over-nutrition. Two billion people suffer from one or more micronutrient deficiencies and over half a billion adults are obese. Access to sufficient amounts of nutritious food is not enough. If food is unsafe, whether arising from poor quality supplies or inadequate treatment and preparation, it increases the risk of foodborne infections such as diarrhoea. 70% of diarrhoea episodes occurring annually in children under five are due to biologically contaminated food. Conclusions: An integrated approach is needed where food safety and nutrition are systematically introduced into mainstream food system policies and interventions worldwide in order to achieve health and development goals. A new framework, “Food for Health” is proposed to guide policy development and requires all three aspects of food security to be addressed in balance: sufficiency, nutrition and safety.

Keywords: food safety, food security, nutrition, policy

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Authors: David Guillermo Piedrahita Marquez, Hector Suarez Mahecha, Jairo Humberto Lopez

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It was necessary to establish which formulation is suitable for the preservation of aquaculture products, that why edible films were made. These were to a characterization in order to meet their morphology physicochemical and mechanical properties, optical. Six Formulations of chitosan and propolis ethanolic extract encapsulated were developed because of their activity against pathogens and due to their properties, which allows the creation waterproof polymer networks against gasses, vapor, and physical damage. In the six Formulations, the concentration of comparison material (1% w/v, 2% pv) and the bioactive concentrations (0.5% w/v, 1% w/v, 1.5% pv) were changed and the results obtained were compared with statistical and multivariate analysis methods. It was observed that the matrices showed a mayor impermeability and thickness control samples and the samples reported in the literature. Also, these films showed a notorious uniformity of the films and a bigger resistance to the physical damage compared with other edible films made of other biopolymers. However the action of some compounds had a negative effect on the mechanical properties and changed drastically the optical properties, the bioactive has an effect on Polymer Matrix and it was determined that the films with 2% w / v of chitosan and 1.5% w/v encapsulated, exhibited the best properties and suffered to a lesser extent the negative impact of immiscible substances.

Keywords: chitosan, edible films, ethanolic extract of propolis, mechanical properties, optical properties, physical characterization, scanning electron microscopy (SEM)

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Authors: Sehrish Jalal, Choon-Mee Kim, Dong-Min Kim

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Background: Many pathogens causing hemorrhagic fevers of medical and veterinary importance have been identified and isolated from rodents in the Republic of Korea (ROK). Objective: We investigated the prevalence of emerging viruses causing hemorrhagic fevers, such as hemorrhagic fever with renal syndrome (HFRS), severe fever with thrombocytopenia syndrome (SFTS) and flaviviruses, from wild rodents. Methods: Striped field mice, Apodemus agrarius, (n=39) were captured during 2014-2015 in the south-west of ROK. Using molecular methods, lung samples were evaluated for SFTS virus, HFRS virus and flavivirus, and seropositivity was evaluated in the blood. Results: A high positive rate of Hantavirus (46.2%) was detected in A.agrarius lungs by reverse transcription-nested polymerase chain reaction (RT-N-PCR). The monthly prevalence of HFRS virus was 16.7% in October, 86.7% in November and 25% in August of the following year (p < 0.001). Moreover, 17.9% of blood samples were serologically positive for Hantavirus antibodies. The most prevalent strain in A. agrarius was Hantaan virus. All samples were positive for neither SFTS nor flavivirus. Conclusion: Hantan virus was detected in 86.7% of A. agrarius in November (autumn), and thus, virus shedding from A. agrarius can increase the risk of humans contracting HFRS. These findings may help to predict and prevent disease outbreaks in ROK.

Keywords: hemorrhagic fever virus, molecular diagnostic technique, rodents, Korea

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Authors: Ananthakrishna L. R., Ramesh D., Kumar Wodeyar, Kotresh A. M., Gururaj P. M.

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Malnad Gidda is the indigenous recognized cattle breed of Shivamogga District of Karnataka state, India is known for its disease resistance to many of the infectious diseases. There are 25 LRR (Leucine Rich Repeats) identified in bovine (Bos indicus) TLR9. The amino acid sequence of LRR is deduced to nucleotide sequence in BLASTx bioinformatic online tools. LRR2 to LRR10 are involved in pathogen recognition and binding in human TLR9 which showed a higher degree of nucleotide variations with respect to disease resistance to various pathogens. Hence, primers were designed to amplify the flanking sequences of LRR2 to LRR10, to discover the nucleotide variations if any, in Malnad Gidda breed of Cattle which is associated with disease resistance. The DNA isolated from peripheral blood mononuclear cells of ten Malnad Gidda cattle. A desired and specific amplification product of 0.8 kb was obtained at an annealing temperature of 56.6ᵒC. All the PCR products were sequenced on both sides by gene-specific primers. The sequences were compared with TLR9 sequence of cross breed cattle obtained from NCBI data bank. The sequence analysis between Malnad Gidda and crossbreed cattle revealed no nucleotide variations in the region LRR2 to LRR9 which shows the conserved in pathogen binding domain (LRR) of TLR9.

Keywords: leucine rich repeats, Malnad Gidda, cross breed, TLR9

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Authors: Miguel García-Ferrús, Ana González, María A. Ferrús

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The most demanded organic foods worldwide are those that are consumed fresh, such as fruits and vegetables. However, there is a knowledge gap about some aspects of organic food microbiological quality and safety. Organic fruits and vegetables are more exposed to pathogenic microorganisms due to surface contact with natural fertilizers such as animal manure, wastes and vermicompost used during farming. It has been suggested that some emergent pathogens, such as Helicobacter pylori or Arcobacter spp., could reach humans through the consumption of raw or minimally processed vegetables. Therefore, the objective of this work was to study the contamination of organic fresh green leafy vegetables by Arcobacter spp. and Helicobacter pylori. For this purpose, a total of 24 vegetable samples, 13 lettuce and 11 spinach were acquired from 10 different ecological supermarkets and greengroceries and analyzed by culture and PCR. Arcobacter spp. was detected in 5 samples (20%) by PCR, 4 spinach and one lettuce. One spinach sample was found to be also positive by culture. For H. pylori, the H. pylori VacA gene-specific band was detected in 12 vegetable samples (50%), 10 lettuces and 2 spinach. Isolation in the selective medium did not yield any positive result, possibly because of low contamination levels together with the presence of the organism in its viable but non-culturable form. Results showed significant levels of H. pylori and Arcobacter contamination in organic vegetables that are generally consumed raw, which seems to confirm that these foods can act as transmission vehicles to humans.

Keywords: Arcobacter sp., Helicobacter pylori, Organic Vegetables, Polymerase Chain Reaction (PCR)

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Authors: Beata Lachtara, Renata Szewczyk, Katarzyna Bielinska, Kinga Wieczorek, Jacek Osek

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Listeria monocytogenes is an important human and animal pathogen that causes foodborne outbreaks. The bacteria may be present in different types of food: cheese, raw vegetables, sliced meat products and vacuum-packed sausages, poultry, meat, fish. The most common method, which has been used for the investigation of genetic diversity of L. monocytogenes, is PFGE. This technique is reliable and reproducible and established as gold standard for typing of L. monocytogenes. The aim of the study was characterization by molecular serotyping and PFGE analysis of L. monocytogenes strains isolated from fresh fish and fish products in Poland. A total of 301 samples, including fresh fish (n = 129) and fish products (n = 172) were, collected between January 2014 and March 2016. The bacteria were detected using the ISO 11290-1 standard method. Molecular serotyping was performed with PCR. The isolates were tested with the PFGE method according to the protocol developed by the European Union Reference Laboratory for L. monocytogenes with some modifications. Based on the PFGE profiles, two dendrograms were generated for strains digested separately with two restriction enzymes: AscI and ApaI. Analysis of the fingerprint profiles was performed using Bionumerics software version 6.6 (Applied Maths, Belgium). The 95% of similarity was applied to differentiate the PFGE pulsotypes. The study revealed that 57 of 301 (18.9%) samples were positive for L. monocytogenes. The bacteria were identified in 29 (50.9%) ready-to-eat fish products and in 28 (49.1%) fresh fish. It was found that 40 (70.2%) strains were of serotype 1/2a, 14 (24.6%) 1/2b, two (4.3%) 4b and one (1.8%) 1/2c. Serotypes 1/2a, 1/2b, and 4b were presented with the same frequency in both categories of food, whereas serotype 1/2c was detected only in fresh fish. The PFGE analysis with AscI demonstrated 43 different pulsotypes; among them 33 (76.7%) were represented by only one strain. The remaining 10 profiles contained more than one isolate. Among them 8 pulsotypes comprised of two L. monocytogenes isolates, one profile of three isolates and one restriction type of 5 strains. In case of ApaI typing, the PFGE analysis showed 27 different pulsotypes including 17 (63.0%) types represented by only one strain. Ten (37.0%) clusters contained more than one strain among which four profiles covered two strains; three had three isolates, one with five strains, one with eight strains and one with ten isolates. It was observed that the isolates assigned to the same PFGE type were usually of the same serotype (1/2a or 1/2b). The majority of the clusters had strains of both sources (fresh fish and fish products) isolated at different time. Most of the strains grouped in one cluster of the AscI restriction was assigned to the same groups in ApaI investigation. In conclusion, PFGE used in the study showed a high genetic diversity among L. monocytogenes. The strains were grouped into varied clonal clusters, which may suggest different sources of contamination. The results demonstrated that 1/2a serotype was the most common among isolates from fresh fish and fish products in Poland.

Keywords: Listeria monocytogenes, molecular characteristic, PFGE, serotyping

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Authors: Mudassar Iqbal, Riaz Hussain, Khalid Mehmood, Farah Ali, Fazal Mahmood, Abdul Ghaffar

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Corynebacterium pseudotuberculosis is an important cause of caseous lymphadenitis (CL), a complex, chronic devastating and destructive disease of small ruminants. In present study, postmortem examination of Chinkara deer (n=25) was conducted in year 2014. Pus samples suggestive of CL were collected from the superficial lymph nodes, liver, spleen and lungs during necropsy and subjected to standard microbiological procedures for isolation and molecular analysis of bacterial pathogens. Pus samples collected from carcasses (25) presenting clinical lesions of C. pseudotuberculosis infection was identified in 19 (76%) carcasses on the basis of culture characteristics. The frequency of C. pseudotuberculosis bacterium was higher in older animals as compared to young animals. Grossly, multiple tubercles of variable size having caseous material were observed in liver, lungs, spleen and lymph nodes. Histopathologically, tissue sections from all the visceral organs were extensively plugged with abscess. In present study specific prolineiminopeptidase (PIP) gene of the C. pseudotuberculosis was amplified by the Polymerase chain reaction technique (PCR) in 17(25) cases. The efficient and reliable molecular analysis along with necropsy findings in present study can be used as valuable approach for diagnosis of caseous lymphadenitis in small ruminants.

Keywords: Chinkara deer, Corynebacterium pseudotuberculosis, Caseous lymphadenitis, PCR

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Authors: Anikó Kaluzsa

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The article's author aims to introduce and analyze those microbiological safety hazards which indicate the presence of secondary contamination in the water supply system. Since drinking water belongs to primary foods and is the basic condition of life, special attention should be paid on its quality. There are such indicators among the microbiological features can be found in water, which are clear evidence of the presence of water contamination, and based on this there is no need to perform other diagnostics, because they prove properly the contamination of the given water supply section. Laboratory analysis can help - both technologically and temporally – to identify contamination, but it does matter how long takes the removal and if the disinfection process takes place in time. The identification of the factors that often occur in the same places or the chance of their occurrence is greater than the average, facilitates our work. The pathogen microbiological risk assessment by the help of several features determines the most likely occurring microbiological features in the Carpathian basin. From among all the microbiological indicators, that are recommended targets for routine inspection by the World Health Organization, there is a paramount importance of the appearance of Escherichia coli in the water network, as its presence indicates the potential ubietiy of enteric pathogens or other contaminants in the water network. In addition, the author presents the steps of microbiological risk assessment analyzing those pathogenic micro-organisms registered to be the most critical.

Keywords: drinking water, E. coli, microbiological indicators, risk assessment, water safety plan

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Authors: Jae Young Song, In-Ohk Ouh, Seyeon Park, Byeong Sul Kang, Soo Dong Cho, In-Soo Cho

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Canine parvovirus (CPV) is a major pathogen of diarrhea disease in dogs. CPV type 2 has three of antigenic variants such as 2a, 2b, and 2c. CPV constructs a small non-enveloped, icosahedral capsid that contains single-stranded DNA. It has capsids that two largely overlapping virion proteins (VP), VP1 (82 kDa), and VP2 (65 kDa). Baculoviruses are insect pathogens that regulate insect populations in nature and are being successfully used to control insect pests. The proteins produced in the baculovirus-expression system are used for instance for functional studies, vaccine preparations, or diagnostics. The vaccines produced by baculovirus-expression system showed elicitation of antibodies. The recombinant baculovirus infected SF9 cells showed broken shape. The recombinant VP2 proteins from cell pellet or supernatant were confirmed by western blotting. The result showed that the recombinant VP2 protein bands were appeared at 65 kDa molecular weight in both cell pellet and supernatant of infected SF9 cell. These results indicated that the recombinant baculovirus infected SF9 cell express the recombinant VP2 protein successfully. In addition, the expressed recombinant VP2 protein is secreted from cell to supernatant. The baculovirus expression system can be used to produce the VP2 protein of CPV 2c. In addition, the secretion property of the expression of VP2 protein may decrease the cost of production, because it can be skipped the cell breaking step. The produced VP2 protein could be used for vaccine and the agent of diagnostic tests. This study provides the foundation of the production of CPV 2c vaccine and the diagnostic agent.

Keywords: baculovirus, canine parvovirus 2c, dog, Korea

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