Chicken Feces Research Articles

Infectious phage particles packaging antibiotic resistance genes found in meat products and chicken feces

Bacteriophages can package part of their host’s genetic material, including antibiotic resistance genes (ARGs), contributing to a rapid dissemination of resistances among bacteria. Phage particles containing ARGs were evaluated in meat, pork, beef and chicken minced meat, and ham and mortadella, purchased in local retailer. Ten ARGs (blaTEM, blaCTX-M-1, blaCTX-M-9, blaOXA-48, blaVIM, qnrA, qnrS, mecA, armA and sul1) were analyzed by qPCR in the phage DNA fraction. The genes were quantified, before and after propagation experiments in Escherichia coli, to evaluate the ability of ARG-carrying phage particles to infect and propagate in a bacterial host. According to microbiological parameters, all samples were acceptable for consumption. ARGs were detected in most of the samples after particle propagation indicating that at least part of the isolated phage particles were infectious, being sul1the most abundant ARG in all the matrices followed by β-lactamase genes. ARGs were also found in the phage DNA fraction of thirty-seven archive chicken cecal samples, confirming chicken fecal microbiota as an important ARG reservoir and the plausible origin of the particles found in meat. Phages are vehicles for gene transmission in meat that should not be underestimated as a risk factor in the global crisis of antibiotic resistance.

A study of the relationship between human infection with avian influenza a (H5N6) and environmental avian influenza viruses in Fujian, China

BackgroundAvian influenza A (H5N6) virus poses a great threat to the human health since it is capable to cross the species barrier and infect humans. Although human infections are believed to largely originate from poultry contaminations, the transmissibility is unclear and only limited information was available on poultry environment contaminations, especially in Fujian Province.MethodsA total of 4901 environmental samples were collected and tested for Avian Influenza Virus (AIV) from six cities in Fujian Province through the Fujian Influenza Surveillance System from 2013 to 2017. Two patient-related samples were taken from Fujian’s first confirmed H5N6 human case and his backyard chicken feces in 2017. Chi-square test or Fisher’s exact probability test was used to compare the AIV and the viral subtype positive rates among samples from different Surveillance cities, surveillance sites, sample types, and seasons. Phylogenetic tree analysis and molecular analysis were conducted to track the viral transmission route of the human infection and to map out the evolutions of H5N6 in Fujian.ResultsThe overall positive rate of the H5 subtype AIVs was 4.24% (208/4903). There were distinctive differences (p < 0.05) in the positive rates in samples from different cities, sample sites, sample types and seasons. The viruses from the patient and his backyard chicken feces shared high homologies (99.9–100%) in all the eight gene segments. Phylogenetic trees also showed that these two H5N6 viruses were closely related to each other, and were classified into the same genetic clade 2.3.4.4 with another six H5N6 isolates from the environmental samples. The patient’s H5N6 virus carried genes from H6N6, H5N8 and H5N6 viruses originated from different areas. The R294K or N294S substitution was not detected in the neuraminidase (NA). The S31 N substitution in the matrix2 (M2) gene was detected but only in one strain from the environmental samples.ConclusionsThe H5 subtype of AIVs has started circulating in the poultry environments in Fujian Province. The patient’s viral strain originated from the chicken feces in his backyard. Genetic reassortment in H5N6 viruses in Fujian Province was indicated. The H5N6 viruses currently circulating in Fujian Province were still commonly sensitive to Oseltamivir and Zanamivir, but the resistance against Amantadine has emerged.

Morphologic and genomic characterization of a broad host range Salmonella enterica serovar Pullorum lytic phage vB_SPuM_SP116

For effective use of phages as antimicrobial agents for controlling multidrug resistant S. Pullorum, it is important to understand phage biology. A lytic S. Pullorum phage was isolated and characterized from chicken feces, and its whole genome was sequenced and analyzed. A new lytic phage-vB_SPuM_SP116 (in brief SP116)- isolated and characterized using S. Pullorum SPu-116 as its host belongs to Myoviridae A1 group. Phage SP116 had a lytic effect on 27 of 37 (72.9%) different serotypes of clinical Salmonella strains. It showed a high bactericidal activity in killing all pathogens in cultures containing 5 × 105 cfu/mL and achieved more than 6.58 and 5.97 log unit reductions in cultures containing 5 × 106 cfu/mL and 5 × 107 cfu/mL, respectively. The one-step growth curve showed that the burst size was up to 118 pfu/bacterial cell. Complete genome sequence analysis revealed a linear, double-stranded DNA genome of 87,510 bp with an average G + C content of 38.84%, including 128 predicted open reading frames (ORFs) and 22 tRNA genes. SP116 was classified as a Felix O1 virus based upon the general phage characterization and the genomic information. Regarding its high efficacy in preventing especially S. Pullorum infection and its lack of any bacterial virulence, antimicrobial resistance, and lysogenesis genes, it could be a potential alternative candidate for the treatment of S. Pullorum infections.

In Vitro Evaluation of Hen Eggshells Addition in Rice Husk on Litter Properties

Abstract. Ammonium (NH3) is released from uric acid of chicken feces. Good management practices are required to depress ammonium concentration in a litter. Several research findings showed that application of CaCO3 (calcium carbonate) can decrease ammonium concentration in a litter. Eggshell as a by-product of hennery (hen stable) and hatchery is mainly constructed from CaCO3. This research aimed to evaluate the addition of hen eggshell powder at different concentration in litter on water content, pH, microorganism, and NH3 concentration of litter. Rice husk as litter material, hen eggshell powder at a concentration of 0%, 5%, 10%, and 15% and hen manure were thoroughly mixed. After 24 hours, the concentration of water content, pH, microorganisms amount and NH3 in each treatment were measured. This research was completely randomized designed with 3 repetitions. Results showed the progressive addition of hen eggshell powder in litter until 15% increased pH and decreased water content, number of microorganisms and NH3 concentration. The addition of 15% hen eggshell powder in litter revealed best litter properties.Keywords: ammonium, hen eggshell, litter, pH, water content

Novel bifidobacteria strains isolated from nonconventional sources. Technological, antimicrobial and biological characterization for their use as probiotics.

To characterize four novel autochthonous bifidobacteria isolated from monkey faeces and a Bifidobacterium lactis strain isolated from chicken faeces by evaluating their technological and biological/functional potential to be used as probiotics. Different stressors, including food process parameters and storage, can affect their viability and functionality. The resistance to frozen storage, tolerance to lyophilization and viability during storage, thermal, acidic and simulated gastric resistance, surface hydrophobicity and antimicrobial activity against pathogens were studied. Bifidobacterium lactis Bb12 and INL1 were used as reference strains. The results obtained demonstrated that the new isolates presented strain-dependent behaviour. Good results were obtained for thermal resistance, frozen storage at -80°C and lyophilized powders maintained at 5°C. Cell viability during refrigerated storage was higher when the strains were resuspended in milk at pH 5·0 than at 4·5. The surface hydrophobicity ranged between 7 and 98% depending on the strain. The simulated gastric resistance was improved for the strains incorporated in cheese. Regarding antimicrobial activity, bifidobacteria isolated from monkey presented higher inhibitory capacity than the reference strains. This research provides a deeper insight into new strains of bifidobacteria isolated from primates and chicken that have not been previously characterized for their potential use in dairy products and confirm the most robust stress tolerance of B. lactis. The possibility of expanding the available bifidobacteria with the potential to be added to a probiotic food necessarily implies characterizing them from different points of view, especially when considering unknown species. For monkey isolates (which showed higher antimicrobial activity against pathogens), more in-depth knowledge is needed before applying strategies to improve their performance. On the contrary, the chicken isolate B. lactis P32/1 showed similar behaviour to the references B. lactis strains; therefore, it could be considered as a potential probiotic candidate.

Infectious bursal disease virus suppresses H9N2 avian influenza viral shedding in broiler chickens

ABSTRACT1. Infectious bursal disease virus (IBDV) causes immunosuppression in chickens, increasing their susceptibility to other infectious diseases and resulting in vaccination failure. Here, we investigated the immune-depressing effect of IBDV on H9N2 avian influenza viral infection in the broiler chickens.2. For this purpose, chickens were divided into four groups. In group A, chickens were inoculated with IBDV at 21 days of age and H9N2 avian influenza virus (AIV) 5 days later. Groups B and C only received AIV at 26 days of age and IBDV at 21 days, respectively. The control group (D) were inoculated with normal saline at the same times. Tissue samples from different organs were collected on the days 1, 3, 6, 9, and 12 after H9N2 infection.3. Macroscopic observation showed IBD lesions in groups A and C, including swollen bursa with the presence of gelatinous exudates, haemorrhages in the thigh muscle, edema, and nephritis.4. Reverse Transcription-PCR was used to study H9N2 AIV dissemination, and qRT-PCR to determine viral genome copy number in different organs. A considerable titre of AIV was found in the trachea, lungs, cecal tonsils, spleens, and feces of infected chickens. The genome copy number of the virus in the trachea and lungs of group A was significantly higher than that in group B on the first day after inoculation. But in the other days post inoculation, RT-PCR did not detect the AIV genome in group A. Although there might have been some immunosuppression in group A, IBDV could interfere with AIV replication in the chickens of this group.5. In conclusion, we propose that pre-exposure to IBDV at 3 weeks of age reduces the replication and shedding of H9N2 in broiler chicken.

Media Access is Associated with Knowledge of Optimal Water, Sanitation and Hygiene Practices in Tanzania.

The importance of water, sanitation and hygiene (WASH) behaviors in low- and middle-income countries in preventing childhood illness is well established. Tanzania is known to have high rates of chronic malnutrition and childhood stunting—both of which have been linked to poor WASH practices. Interviews were conducted with 5000 primary caregivers of children aged 0–23 months. Four composite WASH knowledge variables were created to assess the relationship between WASH knowledge and access to different forms of media, such as television, radio, and mobile phones. WASH knowledge variables measure knowledge of when to wash hands, the need for soap when washing hands, when to wash a baby’s hands, and how eating soil or chicken feces can affect a baby’s health. Logistic and linear regression analyses were conducted to measure the association between media access and WASH knowledge. Having watched television was positively associated with higher WASH knowledge indicators (all p < 0.05). Higher WASH knowledge was positively associated with more frequent handwashing after cleaning a baby’s bottom (all p < 0.0001). The quantity of media access also had a positive linear effect on handwashing; more media items owned was associated with increases in handwashing. Study findings indicate media access is associated with WASH knowledge among caregivers in resource-poor settings.

SYNTHESIS OF LIQUID ORGANIC FERTILIZER BASED ON CHICKEN MANURE USING BIOSCA AND FUNGUS BIOACTIVATOR Trichoderma harzianum

Production of liquid organic fertilizer from chicken feces with addition bioaktivator Biosca and Trichoderma harzianum fungus aims to determine the influence of time against pH and macro elements (N, P, and K), analyze the influence and determine the bioactivator composition against pH, macro element content, growth and production of tomato plants. The method of making this liquid organic fertilizer is chicken feces dissolved using water in a composter, then composted with addition of bioactivator which is consisting B1 (40 mL Biosca), B2 (40 mL T. harzianum), B3 (10 mL Biosca + 30 mL T. harzianum), B4 (20 mL Biosca + 20 mL T. harzianum) and B5 (30 mL Biosca + 10 mL T. harzianum). Composting is done by time variations of 7 days (M1), 14 days (M2) and 21 days (M3). The result of research shows that the process of making organic liquid fertilizer with time variation of composting and effective bioactivator composition in increasing macro element level and good pH is B5 with 21 days of composting time. The influence of bioactivator composition against pH and macro element indicator is showed by value of N: 5,33%, P: 2,41%, K: 1,34% at pH: 7,1. While the influence against tomato plants on the observation at the 10th week is for the growth of plant height: 197,7 cm, average of plant diameter: 16,27 mm, average of leaf production: 41 sheets, and for the average of fruit production: 32 pieces, average of fruit weight: 32,4 g.

FlaA-SVR Based Genetic Diversity of Multiresistant Campylobacter jejuni Isolated From Chickens and Humans.

Campylobacter jejuni is one of the most common causes of human foodborne bacterial infections worldwide. The objective of this study was to assess the molecular diversity, using flaA sequencing, of 602 C. jejuni isolated from chicken food chain, i.e., chicken feces (n = 151), chicken carcasses (n = 150), chicken meat (n = 150), and from humans (n = 151) and to determine antimicrobial multiresistant profiles of the isolates as well as to analyze the relationship of the isolate genotypes with their antimicrobial resistance profiles and source of isolation. Multidrug resistant patterns were identified in 110 (18.3%) C. jejuni isolates recovered from all sources and most isolates were resistant to ciprofloxacin (CIP), nalidixic acid (NAL), streptomycin (STR), and tetracycline (TET) (92; 15.3%) or ciprofloxacin, streptomycin, and tetracycline (13; 2.2%). Only a few isolates were multiresistant to ciprofloxacin, nalidixic acid, tetracycline, and erythromycin (3; 0.5%) or ciprofloxacin, nalidixic acid, streptomycin, tetracycline, and erythromycin (2; 0.3%). A total of 79 flaA-SVR subtypes were identified, including 40 (50.6%) unique to the isolates’ origins, with the most common sequence types 16, 54, 36, 34, and 287 which covered 56 (9.3%), 50 (8.3%), 48 (8.0%), 35 (5.8%), and 32 (5.3%) of C. jejuni isolates, respectively. It was found that 13 isolates had the novel flaA-SVR subtypes which were not present in the pubMLST database. These isolates were recovered from chicken feces (6 isolates), carcasses (2 isolates), meat (one isolate) and from humans (4 isolates). Multiresistant C. jejuni were classified into 26 different sequence subtypes. Among the most numerous multidrug resistant profile CIP+NAL+STR+TET 21 different flaA-SVR subtypes, with total of 92 isolates, were identified. Most of them were classified to 287 (18; 19.6% isolates), 100 (13; 14.1%), 34 (9; 9.8%), 208 (8; 8.7%), and 781 (8; 8.7%) molecular variants. Isolates resistant to CIP, STR and TET (13 isolates) were mainly from chicken feces (12 isolates) and classified into 5 flaA-SVR sequence types, with the most common 36 (8 isolates). The obtained results show a broad molecular diversity of multiresistant C. jejuni isolates and suggest chickens as a possible source of human Campylobacter infections in Poland.

The role of the American cockroach (Periplaneta americana) as transport host of Eimeria tenella to chickens.

The role of the American cockroach, Periplaneta americana as transport host for Eimeria tenella was evaluated. Twenty-four cockroaches were orally fed with sporulated oocysts of E. tenella. Their feces and digestive tract were examined for oocysts by sugar centrifugal flotation technique and PCR. Infectivity of the oocysts recovered from the digestive tract of infected cockroaches as well as from their feces was evaluated by orally inoculating them into Boris Brown chickens. E. tenella oocysts were found in the digestive tract and feces of infected cockroaches up to day 4 after ingestion of oocysts. Furthermore, oocysts that were recovered from the digestive tract and feces of cockroaches remained infective for 4 and 3days after ingestion of oocysts, respectively. Presence of oocysts in the feces of chicken that had been inoculated with either digestive tract or feces of P. americana demonstrated the infectivity of E. tenella oocysts from digestive tract or feces, suggesting that P. americana may play a role in the transmission of E. tenella among chicken and between chicken flocks.

Prevalence of colistin resistance and mcr-1/mcr-2 genes in extended-spectrum β-lactamase/AmpC-producing Escherichia coli isolated from chickens in Canada, Senegal and Vietnam.

This study investigated the prevalence of Escherichia coli (E. coli) colistin resistance and mcr-1 and mcr-2 genes among extended-spectrum β-lactamase (ESBL)/AmpC-producing E. coli isolates recovered from chicken feces in Canada (Quebec), Senegal and Vietnam, and evaluated the susceptibility pattern of the colistin-resistant E. coli isolates to other clinically relevant antimicrobials. A total of 327 potential ESBL/AmpC-producing E. coli isolates from chicken farms in Canada (Quebec), Senegal and Vietnam were analysed for colistin susceptibility by broth microdilution method and for the presence of mcr (1-2) genes by PCR. The pmrA and pmrB genes of colistin-resistant E. coli isolates, in the absence of mcr (1-2) genes, were sequenced. Antimicrobial resistance phenotypes of colistin-resistant E. coli isolates were determined by disk diffusion. None of the 108 potential ESBL/AmpC-producing E. coli isolates from seven farms in Canada were colistin-resistant or possessed mcr-1 or mcr-2 gene. A low prevalence of 2.2% of colistin resistance was observed in 93 Senegalese isolates from the 15 sampled farms, although neither mcr-1 nor mcr-2 gene was found. A prevalence of 8.7% of colistin resistance was observed among 126 Vietnamese isolates from two of the four sampled farms. The mcr-1 gene was detected in 85% of the 13 phenotypically colistin-resistant isolates. Moreover, all colistin-resistant isolates presented a multidrug-resistant phenotype. The co-existence of the mcr-1 and ESBL/AmpC genes and the very high level of multiple drug resistance in all colistin-resistant E. coli isolates obtained from sampled chicken farms in Vietnam is a major concern.

Poultry husbandry, water, sanitation, and hygiene practices, and child anthropometry in rural Burkina Faso.

Poultry production in low income countries provides households with nutrient‐rich meat and egg products, as well as cash income. However, traditional production systems present potential health and nutrition risks because poultry scavenging around household compounds may increase children's exposure to livestock‐related pathogens. Data from a cross‐sectional survey were analysed to examine associations between poultry, water, sanitation, and hygiene practices, and anthropometric indicators in children (6–59 months; n = 3,230) in Burkina Faso. Multilevel regression was used to account for the hierarchical nature of the data. The prevalence of stunting and wasting in children 6–24 months was 19% and 17%, respectively, compared with a prevalence of 26% and 6%, respectively, in children 25–60 months. Over 90% of households owned poultry, and chicken faeces were visible in 70% of compounds. Caregivers reported that 3% of children consumed eggs during a 24‐hr recall. The presence of poultry faeces was associated with poultry flock size, poultry‐husbandry and household hygiene practices. Having an improved water source and a child visibly clean was associated with higher height‐for‐age z scores (HAZ). The presence of chicken faeces was associated with lower weight‐for‐height z scores, and no associations were found with HAZ. Low levels of poultry flock size and poultry consumption in Burkina Faso suggest there is scope to expand production and improve diets in children, including increasing chicken and egg consumption. However, to minimize potential child health risks associated with expanding informal poultry production, research is required to understand the mechanisms through which cohabitation with poultry adversely affects child health and design interventions to minimize these risks.

Metagenomic analysis of isolation methods of a targeted microbe, Campylobacter jejuni, from chicken feces with high microbial contamination

BackgroundOriginating from poultry, particularly chickens, Campylobacter jejuni is the leading foodborne pathogen worldwide and a major cause of campylobacteriosis. Isolating C. jejuni is difficult due to its specific growth requirements, the presence of viable but non-culturable bacteria, and because it is often masked by competing flora. Currently, there is no optimized method for isolating C. jejuni from chicken feces. Here, we evaluated the method for isolating C. jejuni from chicken feces using culture-independent sequence-based metagenomics and culture-dependent tools. Further, we assessed changes in microbial communities during microbe isolation to determine how the process can be improved.ResultsFourteen different variations of C. jejuni isolation procedures were applied to all 35 chicken fecal samples. These variations included using different enrichment broths (without enrichment or enrichment in Bolton or Preston broth), different ratios of sample-to-enrichment broth (1:101, 1:102, and 1:103), and different selective agars (modified charcoal-cefoperazone-deoxycholate agar (mCCDA) or Preston agar). Enrichment during isolation of C. jejuni was evaluated on the basis of microbial diversity and taxonomic composition using metagenomics tools. The effect of selective media was evaluated using a combination of metagenomics and culture-dependent tools. Microbial diversity significantly decreased during the enrichment process, regardless of the type of enrichment broth, with the most significant decrease observed at a feces-to-broth ratio of 1:103. Particularly, in 103-Preston broth, the relative abundance of Campylobacter increased, while extended-spectrum beta-lactamase-producing Escherichia coli, which interfere with Campylobacter isolation, decreased. Metagenomics results were validated by quantitative PCR and culture-dependent analysis. Additionally, selective media affected the isolation results, although microbes with high relative abundance during enrichment were also frequently isolated using culture-dependent methods. Significantly more C. jejuni was isolated from mCCDA than from Preston agar enriched in 103 Preston broth.ConclusionsEnrichment in Preston broth at a ratio of 1:103 followed by spreading onto mCCDA was the most effective method for isolating C. jejuni. This is the first study to apply metagenomics to evaluate a method for isolating a targeted microbe, C. jejuni, from chicken feces, a source with high microbial contamination. Thus, metagenomics can be applied to improve methods for isolating bacteria that are difficult to separate.

Identification and characterization of Diutina rugosa SD-17 for potential use as a probiotic

In this study, a new strain of D. rugosa isolated from chicken feces was identified and characterized. Phylogenetic analyses using the 5.8 internal transcribed spacer and 18S rDNA sequencing showed that this strain belongs to the species D. rugosa, and was thus named D. rugosa SD-17. Its general biological features were studied and compared with its closest related strain, D. rugosa ATCC 10571. The results showed that D. rugosa SD-17 formed a depression in the middle of the colony; furthermore, the cell wall was significantly thicker (P = 0.004) and the production of β-glucan was significantly higher (P < 0.001) compared to D. rugosa ATCC 10571. Intestinal environmental tolerance tests showed that D. rugosa SD-17 could thrive at the host intestinal temperature; approximately 45% of D. rugosa SD-17 survived at pH = 2 for 4 h and 0.3% of bile salts had no effect on growth. Colonization and adhesion tests showed that elimination of D. rugosa SD-17 was achieved after 20 h of oral administration, and the adhesion rate to Caco-2 cells reached 61.5% (P < 0.001). The results of this study indicate that D. rugosa SD-17 could be considered an appropriate probiotic candidate and may have potential use as a novel probiotic in animals.

Pathogenic potential and the role of clones and plasmids in beta-lactamase-producing E. coli from chicken faeces in Vietnam

BackgroundAntimicrobial resistance (AMR) in food-producing animals is a global public health issue. This study investigated AMR and virulence profiles of E. coli isolated from healthy chickens in Vietnam. E. coli were isolated from fecal samples collected in five chicken farms located in the provinces of Hoa Binh, Thai Nguyen and Bac Giang in the North of Vietnam. These isolates were examined by disk diffusion for their AMR, PCR for virulence and AMR genes, pulsed-field gel electrophoresis for relatedness between blaCMY-2/blaCTX-M–positive isolates, electroporation for transferability of blaCMY-2 or blaCTX-M genes and sequencing for mutations responsible for ciprofloxacin resistance.ResultsUp to 99% of indicator isolates were multidrug resistant. Resistance to third-generation cephalosporins (3GC) was encoded by both blaCTX-M and blaCMY-2 genes; blaCTX-M genes being of genotypes blaCTX-M-1, − 14, − 15, − 17, − 57 and − 87, whereas ciprofloxacin resistance was due to mutations in the gyrA and parC genes. Some isolates originating from farms located in different provinces of Vietnam were found to be closely related, suggesting they may have been disseminated from a same source of contamination. Plasmids may also have played a role in the diffusion of 3GC-resistance as the blaCMY-2 gene was located on plasmids A/C and I1, and the blaCTX-M gene variants were carried by I1, FIB, R and HI1. Plasmids carrying the blaCMY-2/blaCTX-M genes also co-transferred resistance to other antimicrobials. In addition, isolates potentially capable of infecting humans, of which some produced blaCMY-2/blaCTX-M, were identified in this study.ConclusionsBoth clones and plasmids could be involved in the dissemination of 3GC-resistant E. coli within and between chicken farms in Vietnam. These results demonstrate the necessity to monitor AMR and control antimicrobial use in poultry in Vietnam.

Metagenomic characterization of antibiotic resistance genes in Antarctic soils

Antibiotic resistance genes (ARGs) are considered environmental pollutants. Comprehensive characterization of the ARGs in pristine environments is essential towards understanding the evolution of antibiotic resistance. Here, we analyzed ARGs in soil samples collected from relatively pristine Antarctica using metagenomic approaches. We identified 79 subtypes related to 12 antibiotic classes in Antarctic soils, in which ARGs related to multidrug and polypeptide were dominant. The characteristics of ARGs in Antarctic soils were significantly different from those in active sludge, chicken feces and swine feces, in terms of composition, abundance and potential transferability. ARG subtypes (e.g., bacA, ceoB, dfrE, mdtB, amrB, and acrB) were more abundant than others in Antarctic soils. Approximately 60% of the ARGs conferred antibiotic resistance via an efflux mechanism, and a low fraction of ARGs (∼16%) might be present on plasmids. Culturable bacterial consortiums isolated from Antarctic soils were consistently susceptible to most of the tested antibiotics frequently used in clinical therapies. The amrB and ceoB carried by culturable species did not express the resistance to aminoglycoside and fluoroquinolone at the levels of clinical concern. Our results suggest that the wide use of antibiotics may have contributed to developing higher antibiotic resistance and mobility.

Initial Characterization of Alkaline Protease from Pseudomonas sp Isolated from Chicken Feces (POSTER)

Abstract Proteases are ubiquitous enzymes and it can be found in all living organisms. Extracellular proteases have high economic value, where the enzymes can be applied to various industrial sectors. In order to find an alternative source of protease, a search for microbe producing extracellular protease has been done using a selective medium. An extracellular producing bacterium that is identified as Pseudomonas sp has been isolated from chicken feces, that is Gallus gallus bankiva, collected from residential areas at Lowokwaru, Malang, Indonesia. The analysis shows that the bacterium produces extracellular protease with optimum activity after incubation of 66 hours at 35oC. Initial characterization of the extracellular protease shows that the extracellular protease produced by Pseudomonas sp has an optimum activity at pH 12 and an optimum temperature at 39oC. At these conditions, the crude extract enzyme has a specific activity of 0,522 U/mg. The high pH resistance allows its application to variety of industrial processes that require high pH operation. Therefore further studies need to be done to get better understanding of enzyme properties. Keywords: Alkaline protease, Chicken feces, Gallus gallus bankiva, Pseudomonas sp

The effectiveness of bokashi against growth of mustard Brassica juncea L., Brassica rapa L. Pokcay And Maize Zea mays L

This research aims to find out the effectiveness of Bokashi on growing plant. The Research used quantitative methods approach to true experimental Random Design, with complete (RAL) factorial 4 X 3 pattern, that is a factor of growing media and plant type. The plant-ing medium consists of 100% bokashi, 50% bokashi, and two media controls i.e. control by the media and the media ground chicken feces. Kind of plant used are mustard (Bassica juncea L.), pokcay (Brassica rapa L.) and maize (Zea mays L.). In this study there was 12 experimental which each experiment planted at 30 polybags, so there was 360 polybags. The area of the research is in the village of Kebon Agung, Jember Regency. Plants are observated over the three over the three weeks after planting. In the first week, the second week and the third week after planting. The observations focused on the ability of seeds to grow once planted that is growing or not growing. The data were analyzed with ANOVA and Duncan uses of SPSS v20. The results showed that the most effective media was 50% bokashi on Zea mays L.

Phylogrouping and antimicrobial resistance analysis of extraintestinal pathogenic Escherichia coli isolated from poultry species

This study was undertaken to determine the prevalence and antimicrobial resistance pattern of extraintestinal pathogenic Escherichia coli (ExPEC) in the feces of healthy poultry and retail chicken. All 146 E. coli recovered from 351 samples were screened by PCR for detection of ExPEC strains. Nineteen (13.01%) isolates were confirmed as ExPEC. Distribution of ExPEC strains was revealed as follows: broiler (25%), layer (15.87%), and raw chicken (3.12%). Turkey, duck, and water samples were negative for ExPEC strains. ExPEC strains belonged to phylogenetic groups B2 (52.63%), A (36.84%), and D (10.53%). Twenty-three (15.75%) isolates were ESBL-positive, including four ExPEC strains. ESBL-positive E. coli were isolated from all the samples except turkey. A high degree of resistance to commonly used antimicrobials, namely nalidixic acid (95.89%), tetracycline (95.89%), trimethoprim (89.04%), colistin (82.88%), and ciprofloxacin (54.11%), including ß-lactam antimicrobials ampicillin (84.93%) and amoxicillin/clavulanic acid (81.51%), was expressed by the isolates. Out of fifteen randomly selected ESBL-positive E. coli isolates, ß-lactam genes, namely blaTEM, blaCTX-M, and blaOXA, were detected in three, six, and one, respectively. Poultry and raw chicken harbor multidrug-resistant, ESBL-type E. coli as well as ExPEC and these strains may be transmitted to humans via the food chain.

Evaluation of Novel Probiotic Bacillus Strains Based on Enzyme Production and Protective Activity Against Salmonellosis

Probiotic strains of Bacillus Species are used in industrial poultry production because of their ability to produce enzymes enhancing the absorption of food, and their ability to reduce the risk of Salmonella spp. infection. The aim of this study was to isolate native potential probiotic Bacillus spp. with the ability to produce enzymes and attachment to intestinal epithelial cells in order to prevent Salmonella typhimurium infection. First, 25 samples of chicken feces were collected from 7 industrial poultry in Golestan province located in north of Iran. Bacillus species from samples were isolated on nutrient agar. These strains were evaluated for the ability of producing amyalse and phytase and their probiotic characteristics such as bile salt, acid and antibiotic resistance, the ability to attach to intestinal epithelial cells and inhibit Salmonella typhimurium invasion. Then selected isolates were identified based on 16S rDNA. Results showed that from 86 isolated, 4 Bacillus strains‌ had desirable characteristics such as the ability to produce phytase and amylase and having suitable probiotics features. We identified K03, K02, and K20 isolates as Bacillus tequilensis and K20 as Bacillus subtilis. Bacillus tequilensis K03 showed the highest attachment ability to intestinal epithelium cells and could inhibited Salmonella typhimurium attachment.