Bio-transformation of poultry litter and activated sewage sludge to produce biomixtures for the remediation of water polluted with pesticides.

The aim of this study was to formulate novel biomixtures with the ability to dissipate globally used pesticides. For this, an effective stabilization of two wastes, poultry litter and activated sewage sludge, was achieved through a combination of composting and vermicomposting, with the aid of the earthworm Eisenia fetida. Hence, two different mixtures were prepared combining the wastes with and without the addition of sewage sludge, and their physicochemical and microbiological characterization was examined during both processes. Earthworms reproduction was promoted by more than fourteen times the initial number of individuals introduced. This step made it possible to obtain substrates rich in organic matter, stable and non-pathogenic. The resulting vermicomposted substrates (V-C1 and V-C2) were used to produce two different biomixtures with wheat stubble (WS) and soil (S): SWSV-C1 and SWSV-C2, and they were tested for the remediation of a solution of five pesticides (2,4-D, cypermethrin, imidacloprid, acetochlor and dimethoate) in a 119-days assay. Comparisons were made with a WS-only biomixture (SWS) and a soil control. All biomixtures were more successful in dissipating the pesticides than soil; 2,4-D, dimethoate, and acetochlor degradation reached more than 99% in the three biomixtures after 28-56 days of assay. Biomixtures containing either vermicomposts acted faster than SWS, particularly for 2,4-D, dimethoate and cypermethrin. The total microbial activity was found to be higher in the two biomixtures containing vermicompost, which can be linked to their enhanced performance in the degradation of pesticides. Although the germination of Lactuca sativa proved that neither of the three spent biomixtures were phytotoxic at the end (germination index >60%), only SWSV-C1 and SWSV-C2 proved to be safe for the survival of E. fetida. This work confirms that vermicompost improves the success of biomixtures, not only in terms of pesticide removal, but also providing non-toxic spent biomixtures.

Escherichia coli ST117: exploring the zoonotic hypothesis.

Extraintestinal pathogenic Escherichia coli (ExPEC) can lead to severe infections, with additional risks of increasing antimicrobial resistance rates. Genotypic similarities between ExPEC and avian pathogenic E. coli (APEC) support a possible role for a poultry meat reservoir in human disease. Some genomic studies have been done on the ST117 lineage which contaminates poultry meat, carries multidrug resistance, can be found in the human intestinal microbiota, and causes human extraintestinal disease. This study analyzed the genomes of 61 E. coli from Brazilian poultry outbreaks focusing on ST117, to further define its possible zoonotic characteristics by genotypic and phylogenomic analyses, along with 1,699 worldwide ST117 isolates originating from human, animal, and environment sources. A predominance of ST117 was detected in the Brazilian isolates (n = 20/61) frequently carrying resistance to critical antibiotics (>86%) linked to IncFII, IncI1, or IncX4 replicons. High similarities were found between IncX4 from Brazilian outbreaks and those from E. coli recovered from imported Brazilian poultry meat and human clinical cases. The ST117 phylogeny showed non-specificity according to host and continent and an AMR index score indicated the highest resistance in Asia and South America, with the latter statistically more resistant and overrepresented with resistance to extended-spectrum beta-lactamases (ESBL). Most ST117 human isolates were predicted to have a poultry origin (93%, 138/148). In conclusion, poultry is a likely source for zoonotic ExPEC strains, particularly the ST117 lineage which can also serve as a reservoir for resistance determinants against critical antibiotics encoded on highly transmissible plasmids. IMPORTANCE: Certain extraintestinal pathogenic Escherichia coli (ExPEC) are particularly important as they affect humans and animals. Lineages, such as ST117, are predominant in poultry and frequent carriers of antibiotic resistance, presenting a risk to humans handling or ingesting poultry products. We analyzed ExPEC isolates causing outbreaks in Brazilian poultry, focusing on the ST117 as the most detected lineage. Genomic comparisons with international isolates from humans and animals were performed describing the potential zoonotic profile. The Brazilian ST117 isolates carried resistance determinants against critical antibiotics, mainly on plasmids, in some cases identical to those carried by international isolates. South American ST117 isolates from all sources generally exhibit more resistance, including to critical antibiotics, and worldwide, the vast majority of human isolates belonging to this lineage have a predicted poultry origin. As the world's largest poultry exporter, Brazil has an important role in developing strategies to prevent the dissemination of multidrug-resistant zoonotic ExPEC strains.

Simultaneous analysis of antimicrobial residues and contaminants in poultry droppings by HPLC-MS/MS: a tool for environmental and food safety monitoring.

Animal waste is a potential pollution hazard as it can harbour contaminants, such as antimicrobial residues, mycotoxins, and pesticides, becoming a risk to the public, animal, and environmental health. To assess this risk, 15 experimental broiler chickens orally received contaminants to evaluate excretion levels. An analytical method was previously developed to detect 18 substances in poultry droppings using high-performance liquid chromatography coupled to a tandem mass spectrometer (HPLC-MS/MS). Contaminants including tetracycline, 4-epi-tetracycline, oxytetracycline, 4-epi-oxytetracycline, chlortetracycline, 4-epi-chlortetracycline, tylosin, erythromycin, enrofloxacin, ciprofloxacin, flumequine, florfenicol, sulfachloropyridazine, sulfadiazine, 2,4-dichlorophenoxyacetic acid, zearalenone, alpha- and beta-zearalenol, were extracted with EDTA-McIlvain and acetonitrile. This method showed a p-value < 0.05, RSD < 25%, and R2 > 0.95 in the calibration curves linearity for all analytes. The limit of quantification, selectivity, decision limit for confirmation, matrix effect, precision, and recovery parameters were validated according to European Union document 2021/808/EC, technical report CEN/TR 16059, SANTE/11813/2017 and according to the Veterinary International Conference on Harmonization: VICH GL2 and GL49. This method confirmed the detection of most analytes 12-36 h post-administration and simultaneously detected and quantified mixed contaminants. Thereby, poultry droppings are a potential matrix for spreading contaminants in animal production before slaughter and their control will minimize environmental impacts and mitigate antimicrobial resistance.

Infection of South American coatis (Nasua nasua) with highly pathogenic avian influenza H5N1 virus displaying mammalian adaptive mutations.

Deadly outbreaks among poultry, wild birds, and carnivorous mammals by the highly pathogenic H5N1 virus of the clade 2.3.4.4b have been reported in South America. The increasing virus incidence in various mammal species poses a severe zoonotic and pandemic threat. In Uruguay, the clade 2.3.4.4b viruses were first detected in February 2023, affecting wild birds and backyard poultry. Three months after the first reported case in Uruguay, the disease affected a population of 23 coatis (Nasua) in an ecological park. Most animals became infected, likely directly or indirectly from wild birds in the park, and experienced sudden death. Five animals from the colony survived, and four of them developed antibodies. The genomes of the H5N1 strains infecting coatis belonged to the B3.2 genotype of the clade 2.3.4.4b. Genomes from coatis were closely associated with those infecting backyard poultry, but transmission likely occurred through wild birds. Notable, two genomes have a 627K substitution in the RNA polymerase PB2 subunit, a hallmark amino acid linked to mammalian adaptation. Our findings support the ability of the avian influenza virus of the 2.3.4.4b clade to infect and transmit among terrestrial mammals with high pathogenicity and undergo rapid adaptive changes. It also highlights the coatis' ability to develop immunity and naturally clear the infection.

Risk for Waterborne Transmission and Environmental Persistence of Avian Influenza Virus in a Wildlife/Domestic Interface in Mexico.

Aquatic habitats provide a bridge for influenza transmission among wild and domestic species. However, water sources pose highly variable physicochemical and ecological characteristics that affect avian influenza virus (AIV) stability. Therefore, the risk of survival or transmissibility of AIV in the environment is quite variable and has been understudied. In this study, we determine the risk of waterborne transmission and environmental persistence of AIV in a wild/domestic bird interface in the Central Mexico plateau (North America) during the winter season using a multi-criteria decision analysis (MCDA). A total of 13 eco-epidemiological factors were selected from public-access databases to develop the risk assessment. The MCDA showed that the Atarasquillo wetland presents a higher persistence risk in January. Likewise, most of the backyard poultry farms at this wild-domestic interface present a high persistence risk (50%). Our results suggest that drinking water may represent a more enabling environment for AIV persistence in contrast with wastewater. Moreover, almost all backyard poultry farms evidence a moderate or high risk of waterborne transmission especially farms close to water bodies. The wildlife/domestic bird interface on the Atarasquillo wetland holds eco-epidemiological factors such as the presence of farms in flood-prone areas, the poultry access to outdoor water, and the use of drinking-water troughs among multiple animal species that may enhance waterborne transmission of AIV. These findings highlight the relevance of understanding the influence of multiple factors on AIV ecology for early intervention and long-term control strategies.

Preliminary snapshot reveals a relationship between multidrug-resistance and biofilm production among enterobacteriaceae isolated from fecal samples of farm-raised poultry in ceará, Brazil.

Antimicrobial resistance and biofilm formation by microbial pathogens pose a significant challenge to poultry production systems due to the persistent risk of dissemination and compromise of bird health and productivity. In this context, the study aimed to investigate the occurrence of different multiresistance phenotypes and the biofilm-forming ability of Enterobacteriaceae isolated from broiler chicken excreta in poultry production units in Ceará, Brazil. Samples were collected from three distinct broiler breeding facilities and subjected to isolation, identification, antibiotic susceptibility testing, phenotypic screening for ß-lactamases enzymes, and biofilm formation evaluation. Seventy-one strains were identified, being Escherichia coli (37 %) and Proteus mirabilis (32 %), followed by Klebsiella pneumoniae (11 %), Providencia stuartii (9 %), Klebsiella aerogenes (6 %), Alcaligenes faecalis (4 %), and Salmonella sp. (1 %). A significant proportion (87 %) of multiresistant strains were detected. For the phenotypic evaluation of ß-lactamases production, strains with resistance to second and third-generation cephalosporins and carbapenems were tested. About 4 of 6 and 10 of 26 were positive for inducible chromosomal AmpC ß-lactamase and extended-spectrum ß-lactamase (ESBL), respectively. Regarding biofilm formation, it was observed that all MDR strains were capable of forming biofilm. In this sense the potential of these MDR bacteria to develop biofilms becomes a significant concern, representing a real threat to both human and animal health, as biofilms offer stability, antimicrobial protection, and facilitate genetic transfer.

Agronomic performance and technological quality of sugarcane submitted to different poultry litter dosages.

Sugarcane is a central crop for sugar and ethanol production. Investing in sustainable practices can enhance productivity, technological quality, mitigate impacts, and contribute to a cleaner energy future. Among the factors that help increase the productivity of sugarcane, the physical, chemical and biological parameters of the soil are amongst the most important. The use of poultry litter has been an important alternative for soil improvement, as it acts as a soil conditioner. Therefore, this work aimed to verify the best doses of poultry litter for the vegetative, reproductive and technological components of sugarcane. The experiment was carried out at Usina Denusa Destilaria Nova União S/A in the municipality of Jandaia, GO. The experimental design used was a complete randomized block design with four replications: 5 × 4, totaling 20 experimental units. The evaluated factor consisted of four doses of poultry litter plus the control (0 (control), 2, 4, 6 and 8 t ha-1). In this study, were evaluated the number of tillers, lower stem diameter, average stem diameter, upper stem diameter, plant height, stem weight and productivity. The technological variables of total recoverable sugar, recoverable sugar, Brix, fiber, purity and percentage of oligosaccharides were also evaluated. It was observed, within the conditions of this experiment, that the insertion of poultry litter did not interfere significantly in most biometric, productive and technological variables of the sugarcane. But it can also be inferred that there was a statistical trend toward better results when the sugarcane was cultivated with 4 t ha-1 of poultry litter.

Enhancing biodegradation of vegetable oil-contaminated soil with soybean texturized waste, spent mushroom substrate, and stabilized poultry litter in microcosm systems.

Industrial activities contribute to environmental pollution, particularly through unregulated effluent discharges, causing adverse effects on ecosystems. Vegetable oils, as insoluble substances, exacerbate this pollution, forming impermeable films and affecting the oxygen transfer, leading to serious habitat disruption. Organic wastes, such as soybean texturized waste, spent mushroom substrate, and stabilized poultry litter, were assessed for their efficacy in enhancing the degradation of vegetable oil in contaminated soil. For this purpose, contaminated soil was amended with each of the wastes (10% w/w) using microcosm systems, which were monitored physico-chemically, microbiologically and toxicologically. Results indicate that the wastes promoted significant oil degradation, achieving 83.1, 90.7, and 86.2% removal for soybean texturized waste, spent mushroom substrate, and stabilized poultry litter, respectively, within a 90-day period. Additionally, they positively influenced soil microbial activity, as evidenced by increased levels of culturable microorganisms and hydrolytic microbial activity. While bioassays indicated no phytotoxicity in most cases, soybean texturized waste exhibited inhibitory effects on seed germination and root elongation of Lactuca sativa. This study significantly enhances our comprehension of remediation techniques for sites tainted with vegetable oils, highlighting the critical role of organic waste as eco-friendly agents in soil restoration. Emphasizing the practical implications of these findings is imperative to underscore the relevance and urgency of addressing vegetable oil contamination in soil. Moving forward, tailored strategies considering both contaminant characteristics and soil ecosystem traits are vital for ensuring effective and sustainable soil remediation.

Phylodynamics of avian influenza A(H5N1) viruses from outbreaks in Brazil.

Our study identified strains of the A/H5N1 virus in analyzed samples of subsistence poultry, wild birds, and mammals, belonging to clade 2.3.4.4b, genotype B3.2, with very high genetic similarity to strains from Chile, Uruguay, and Argentina. This suggests a migratory route for wild birds across the Pacific, explaining the phylogenetic relatedness. The Brazilian samples displayed similarity to strains that had already been previously detected in South America. Phylogeographic analysis suggests transmission of US viruses from Europe and Asia, co-circulating with other lineages in the American continent. As mutations can influence virulence and host specificity, genomic surveillance is essential to detect those changes, especially in critical regions, such as hot spots in the HA, NA, and PB2 sequences. Mutations in the PB2 gene (D701N and Q591K) associated with adaptation and transmission in mammals were detected suggesting a potential zoonotic risk. Nonetheless, resistance to neuraminidase inhibitors (NAIs) was not identified, however, continued surveillance is crucial to detect potential resistance. Our study also mapped the spread of the virus in the Southern hemisphere, identifying possible entry routes and highlighting the importance of surveillance to prevent outbreaks and protect both human and animal populations.

Mentha spp. essential oils: toxicity to Alphitobius diaperinus, activity against poultry pathogenic bacteria, and Beauveria bassiana compatibility.

The botanical insecticide market is growing because of limitations placed on the use of certain synthetic chemical insecticides. In this sense, the lesser mealworm Alphitobius diaperius (Coleoptera: Tenebrionidae) is the main poultry pest. The insect causes weight loss and damage to the digestive system of poultry, and it is a vector and reservoir of pathogens. Consequently, this study explored the following hypotheses: (i) essential oils (EOs) derived from Mentha spp. are toxic to A. diaperius; (ii) these EOs are compatible with Beauveria bassiana, the natural enemy of the poultry pest, that parasite A. diaperinus; (iii) these EOs also exhibit activity against bacteria that are pathogenic to poultry. In topical applications and ingestion tests, EOs from Mentha arvensis, Mentha spicata, and Mentha piperita were toxic to A. diaperinus. Chromatographic analyses revealed that menthol is the predominant compound in M. arvensis and M. piperita, whereas carvone is the major compound in M. spicata. Both (-)- and (+)-menthol, along with (-)- and (+)-carvone, underwent testing with A. diaperinus. Nevertheless, their activity was not as potent as those of the EOs, suggesting a possible synergistic and/or additive effect. The EOs did not have any adverse effects on the conidial germination, vegetative growth, or conidia production per colony of the entomopathogenic fungus B. bassiana. Consequently, these EOs are compatible with this natural enemy. The EO extracted from M. spicata exhibited significant toxicity against Staphylococcus aureus (ATCC 25923), whereas the remaining EOs displayed moderate toxicity against this bacterium. The EOs derived from Mentha spp., as assessed in this study, hold promise for the development of botanical insecticides tailored for the control of A. diaperinus. These insecticides are selective in favor of the natural enemy B. bassiana and can also serve as effective sanitizers, thanks to their antibacterial properties.

Unveiling the traits of antibiotic resistance and virulence in Escherichia coli obtained from poultry waste.

Antibiotic resistance and virulence factors in avian pathogenic Escherichia coli (APEC) have become significant concerns, contributing to adverse environmental effects. The extensive use of antibiotics in poultry farming has resulted in the emergence of antibiotic-resistant APEC strains. This study prioritizes the molecular screening of APEC to uncover their antibiotic resistance and virulence attributes, with specific attention to their environmental impact. To address the imperative of understanding APEC pathogenesis, our study analyzed 50 poultry waste samples including 10 poultry litter, 15 fecal matter, 15 wastewater, and 10 anatomical waste samples. For the presence of virulence genes, 35 Escherichia coli isolates were subjected to molecular characterization. Amongst these, 27 were APEC strains demonstrating the presence of at least four virulence genes each. Notably, virulence genes such as fimH, ompA, ybjX, waaL, cvaC, hlyF, iss, ompT, and iroN were observed among all the E. coli isolates. Furthermore, eleven of the APEC strains exhibited resistance to tetracycline, ampicillin, sulphonamides, and fluoroquinolones.These findings highlight the role of APEC as a potential source of environmental pollution serving as a reservoir for virulence and resistance genes. Understanding the dynamics of antibiotic resistance and virulence in APEC is essential due to its potential threat to broiler chickens and the broader population through the food chain, intensifying concerns related to environmental pollution. Recognizing the ecological impact of APEC is essential for developing effective strategies to mitigate environmental pollution and safeguard the health of ecosystems and human populations.

Mobile Robot + IoT: Project of Sustainable Technology for Sanitizing Broiler Poultry Litter.

The traditional aviary decontamination process involves farmers applying pesticides to the aviary's ground. These agricultural defenses are easily dispersed in the air, making the farmers susceptible to chronic diseases related to recurrent exposure. Industry 5.0 raises new pillars of research and innovation in transitioning to more sustainable, human-centric, and resilient companies. Based on these concepts, this paper presents a new aviary decontamination process that uses IoT and a robotic platform coupled with ozonizer (O3) and ultraviolet light (UVL). These clean technologies can successfully decontaminate poultry farms against pathogenic microorganisms, insects, and mites. Also, they can degrade toxic compounds used to control living organisms. This new decontamination process uses physicochemical information from the poultry litter through sensors installed in the environment, which allows accurate and safe disinfection. Different experimental tests were conducted to construct the system. First, tests related to measuring soil moisture, temperature, and pH were carried out, establishing the range of use and the confidence interval of the measurements. The robot's navigation uses a back-and-forth motion that parallels the aviary's longest side because it reduces the number of turns, reducing energy consumption. This task becomes more accessible because of the aviaries' standardized geometry. Furthermore, the prototype was tested in a real aviary to confirm the innovation, safety, and effectiveness of the proposal. Tests have shown that the UV + ozone combination is sufficient to disinfect this environment.

Molecular characterization of antibiotic resistant Salmonella enterica across the poultry production chain in Costa Rica: A cross-sectional study.

Antibiotic resistant Salmonella enterica are on the increase, worldwide. Given the scarcity of data, this study aimed to investigate its occurrence, virulence, and antibiotic resistance in Costa Rica's food chain. In total, 65 chicken meat- and 171 chicken caecal samples were collected and examined for Salmonella. High frequencies of Salmonella were found in chicken meat (58.5 %, n/N = 38/65) and poultry farms (38.0 %, n/N = 65/171). The majority of Salmonella from chicken meat (89.5 %, n/N = 34/38) and caecum samples (93.6 %, n/N = 59/63) exhibited multidrug resistance (MDR). Serovar Infantis was the most prevalent (94 %, n/N = 67/71), followed by serovars Anatum and Kentucky (3 %, n/N = 2/71). A pESI-like plasmid (92 %, n/N = 65/71) containing virulence and resistance markers was found in S. Infantis. Given the high prevalence of MDR Salmonella, this study emphasizes the need to enhance surveillance systems for foodborne pathogens and antimicrobial resistance in Costa Rica's food production chain.

Molecular characterization of the meq oncogene of Marek's disease virus in vaccinated Brazilian poultry farms reveals selective pressure on prevalent strains.

Marek's disease virus (MDV) has become an increasingly virulent pathogen in the poultry industry despite vaccination efforts to control it. Brazil has experienced a significant rise of Marek's disease (MD) outbreaks in recent years. Our study aimed to analyze the complete meq gene sequences to understand the molecular epidemiological basis of MD outbreaks in Brazilian vaccinated layer farms. We detected a high incidence rate of visceral MD (67.74%) and multiple circulating MDV strains. The most prevalent and geographically widespread genotype presented several clinical and molecular characteristics of a highly virulent strain and evolving under positive selective pressure. Phylogenetic and phylogeographic analysis revealed a closer relationship with strains from the USA and Japan. This study sheds light on the circulation of MDV strains capable of infecting vaccinated birds. We emphasize the urgency of adopting preventive measures to manage MDV outbreaks threatening the poultry farming industry.

Evaluating antibiotic occurrence, degradation, and environmental risks in poultry litter within Argentina's agricultural hub.

This study explores the relationship between poultry farming's antibiotic administration practices and residual antibiotic levels in the litter before its application onto agricultural soils. Twenty-three antibiotics were performed across 19 Argentinean farms representing diverse antibiotic management practices. Analysis revealed up to 20 antibiotics from eight chemical classes in poultry litter samples, with tylosin, enrofloxacin, and salinomycin being the most relevant drugs. Farms with restricted antibiotic use in feed exhibited lower residual concentrations. A self-heating treatment was tested to reduce litter antibiotic levels. Although a 60 % reduction of antibiotics was found after treatment, prevalent compounds persisted at residual levels. Regulatory measures and comprehensive litter treatments pre-application are crucial to mitigate environmental risks. This is the first study that provides insight on the occurrence of >20 drugs in real poultry production scenarios from Latin America and demonstrates how relatively simple treatments can be readily applied to decrease the associated environmental risks.

Low temperatures do not impair the bacterial plasmid conjugation on poultry meat.

Conjugation plays an important role in the dissemination of antimicrobial resistance genes. Besides, this process is influenced by many biotic and abiotic factors, especially temperature. This study aimed to investigate the effect of different conditions of temperature and storage (time and recipient) of poultry meat, intended for the final consumer, affect the plasmid transfer between pathogenic (harboring the IncB/O-plasmid) and non-pathogenic Escherichia coli organisms. The determination of minimal inhibitory concentrations (MIC) of ampicillin, cephalexin, cefotaxime, and ceftazidime was performed before and after the conjugation assay. It was possible to recover transconjugants in the poultry meat at all the treatments, also these bacteria showed a significant increase of the MIC for all antimicrobials tested. Our results show that a non-pathogenic E. coli can acquire an IncB/O-plasmid through a conjugation process in poultry meat, even stored at low temperatures. Once acquired, the resistance genes endanger public health especially when it is about critically and highly important antimicrobials to human medicine.

Salmonella enterica serovars linked with poultry in India: antibiotic resistance profiles and carriage of virulence genes.

Salmonella is an important poultry pathogen with zoonotic potential. Being a foodborne pathogen, Salmonella-contaminated poultry products can act as the major source of infection in humans. In India, limited studies have addressed the diversity of Salmonella strains of poultry origin. This study represented 26 strains belonging to Salmonella serovars Typhimurium, Infantis, Virchow, Kentucky, and Agona. The strains were tested for resistance to 14 different antimicrobial agents using the Kirby-Bauer disk-diffusion assay. The presence of the invA, hilA, agfA, lpfA, sopE, and spvC virulence genes was assessed by polymerase chain reaction (PCR), and the genetic diversity was assessed by Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR). The highest resistance to tetracycline (n = 17; 65.38%) followed by nalidixic acid (n = 16; 61.53%) was detected among the strains. Among the strains (n = 17) phenotypically resistant to tetracycline, 94% (n = 16) were also positive for the tetA gene. Based on the presence of virulence genes, the strains were characterized into three virulence profiles (PI, P2, and P3). Among the investigated virulence genes, invA, hilA, agfA, and lpfA were present in all strains. The sopE gene was mostly associated with serovars Virchow (n = 3; 100%) and Typhimurium (n = 8; 80%), whereas spvC gene was exclusive for two Typhimurium strains that lacked sopE gene. ERIC-PCR profiling indicated clusters correlating their serovar, geographical, and farm origins. These results demonstrate that Salmonella isolates with a wide genetic range, antibiotic resistance, and virulence characteristics can colonize poultry. The presence of such strains is crucial for both food safety and public health.

Development of a fast and sensitive RT-qPCR assay based on SYBR® green for diagnostic and quantification of Avian Nephritis Virus (ANV) in chickens affected with enteric disease.

BACKGROUND: Enteric viruses are among the most prominent etiological agents of Runting-Stunting Syndrome (RSS). The Avian Nephritis Virus (ANV) is an astrovirus associated with enteric diseases in poultry, whose early diagnosis is essential for maintaining a good poultry breeding environment. ANV is an RNA virus that rapidly mutates, except for some conserved regions such as ORF1b. Therefore, the approach of a diagnostic method based on fast-RT-qPCR using SYBR® Green that focuses on the amplification of a fragment of ORF1b is presented as a feasible alternative for the diagnosis of this viral agent. In this study, the proposed assay showed a standard curve with an efficiency of 103.8% and a LoD and LoQ of 1 gene viral copies. The assay was specific to amplify the ORF 1b gene, and no amplification was shown from other viral genomes or in the negative controls. 200 enteric (feces) samples from chickens (broilers) and laying hens with signs of RSS from Ecuadorian poultry flocks were examined to validate the proposed method. RESULTS: Using our method, 164 positive results were obtained out of the total number of samples run, while the presence of viral RNA was detected in samples collected from one day to 44 weeks old in both avian lines. CONCLUSIONS: Our study presents a novel, rapid, robust, and sensitive molecular assay capable of detecting and quantifying even low copy numbers of the ANV in commercial birds, therefore introducing a handy tool in the early diagnosis of ANV in enteric disease outbreaks in poultry.

Identification and bioinformatic analysis of invA gene of Salmonella in free range chicken / Identificação e análise bioinformática do gene invA de Salmonella em frango caipira

Salmonella is a serious cause of the health issues in human and animal worldwide. Salmonella has been isolated from different biological samples and it considers as the key role in induction of inflammation of gastrointestinal tract which in turn cause diarrhoea in different species. To further understand the involvement of Salmonella in contaminating and infecting fresh eggs and meat of free-range chicken. This study aimed to establish the microbiological and molecular detections of Salmonella in the cloaca of the free-range chicken and to identify predicted biological functions using Kyoto Encyclopedia of Gene and Genomic (KEGG) pathways and protein-protein interaction. Cloacal swabs were collected from free range chicken raised in the local farm in Duhok city. The isolates were cultured and biochemical test performed using XLD and TSI, respectively. Molecular detection and functional annotation of invA gene was carried out using Conventional PCR and bioinformatics approaches. The present study found that Salmonella was detected in 36 out of 86 samples using microbiological methods. To confirm these findings, invA gene was utilised and 9 out of 36 Salmonella isolates have shown a positive signal of invA by agarose gel. In addition, bioinformatic analysis revealed that invA gene was mainly associated with bacterial secretion processes as well as their KEGG terms and Protein-Protein Interaction were involved in bacterial invasion and secretion pathways. These findings suggested that invA gene plays important role in regulating colonization and invasion processes of Salmonella within the gut host in the free range chicken.

A salmonela é uma causa séria de problemas de saúde em humanos e animais em todo o mundo. Salmonella tem sido isolada de diferentes amostras biológicas e é considerada como o principal papel na indução da inflamação do trato gastrointestinal que por sua vez causa diarreia em diferentes espécies. Compreender melhor o envolvimento da Salmonella na contaminação e infecção de ovos frescos e carne de frango caipira. Este estudo teve como objetivo estabelecer as detecções microbiológicas e moleculares de Salmonella na cloaca da galinha caipira e identificar as funções biológicas previstas usando as vias da Enciclopédia de Gene e Genômica de Kyoto (KEGG) e interação proteína-proteína. Suabes cloacais foram coletados de galinhas criadas ao ar livre em fazenda local na cidade de Duhok. Os isolados foram cultivados e o teste bioquímico realizado com XLD e TSI, respectivamente. A detecção molecular e anotação funcional do gene invA foi realizada usando PCR convencional e abordagens de bioinformática. O presente estudo descobriu que Salmonella foi detectada em 36 das 86 amostras usando métodos microbiológicos. Para confirmar esses achados, o gene invA foi utilizado e 9 dos 36 isolados de Salmonella mostraram um sinal positivo de invA pelo gel de agarose. Além disso, a análise bioinformática revelou que o gene invA estava associado principalmente a processos de secreção bacteriana, assim como seus termos KEGG e interação proteína-proteína estavam envolvidos na invasão bacteriana e nas vias de secreção. Esses achados sugeriram que o gene invA desempenha um papel importante na regulação dos processos de colonização e invasão de Salmonella dentro do hospedeiro intestinal na galinha caipira.

Duty cycle and high-frequency effects on welfare and meat quality of broilers chicken: compliance with European animal stunning regulation / Efeito do duty cycle e altas frequências no bem estar animal e qualidade de carne de frango: atendimento do regulamento europeu de insensibilização animal

European market regulates that poultry electronarcosis stunning in abattoirs must be performed with a minimal required current (mA per animal) and correlative frequency (Hz), to promote animal welfare and meat quality. In this way, Brazilian abattoirs must adjust the stunning parameters so that they can meet the requirements of that market. This study evaluated the effect of stunning parameters using frequency and duty cycle variables. For this, nine treatments were performed, whose results showed that the analyzed frequencies (700, 1100, and 1500 Hz) had a higher incidence of indicators of animal welfare (AW). Thus, stunning had low efficiency in AW terms. However, the 25% and 40% duty cycles had the best stunning efficiency. There was no significant difference (P > 0.05) for pH 24 h, bruises, and drip loss. Color and pH 2 h were significantly affected (P < 0.05). The shear force was higher in birds subjected to higher frequencies and interaction between 1500 Hz and 15% and 25% duty cycles. Water-holding capacity was lower at 1100 Hz and in the 25% duty cycle interactions. The treatments affected the evaluated indicators, except for the 40% duty cycle, which had a positive influence. Therefore, the duty cycle applicability must be elucidated due to its direct influence on stunning efficiency.

O mercado europeu regulamenta que a insensibilização de aves por eletronarcose nos abatedouros deva acontecer com uma corrente mínima necessária (mA por animal) e frequência correlata (Hz), com corrente e frequência alta. O intuito é promover o bem estar animal e qualidade de carne. Desta forma, os abatedouros brasileiros devem ajustar os parâmetros de insensibilização para que possam atender este mercado. O objetivo do trabalho foi avaliar o efeito dos parâmetros de insensibilização usando as variáveis frequência e duty cycle. Foram realizados nove ensaios. Os resultados demostraram que as frequências avaliadas (700 Hz, 1100 Hz e 1500 Hz) apresentaram maior incidência nos indicadores de Bem Estar Animal (BEA). Assim, houve baixa eficiência na insensibilização das aves para este fator. Entretanto, o duty cycle 25% e 40% promoveu a melhor eficiência na insensibilização das aves. Não foi observada diferença significativa (P > 0,05) para pH 24h, hematomas e a perda por gotejamento. A cor e o pH 2h foram afetados significativamente (P < 0,05) nos ensaios avaliados. A força de cisalhamento foi maior nas frequências mais altas e na interação da frequência de 1500 Hz com o duty cycle de 15% e 25% A capacidade de retenção de água foi menor na frequência de 1100 Hz, e na interação duty cycle 25%. Os ensaios estudados apresentaram comprometimento dos indicadores avaliados, exceto a variável duty cycle a 40%, que demonstrou influência positiva nos indicadores avaliados. Portanto, a aplicabilidade do duty cycle deve ser elucidada devido a sua influência direta na eficiência da insensibilização.