Identifikacija ptičjih vrsta lančanom reakcijom polimerazom i analizom slijeda mitohondrijskoga gena 12S rRNA

Authentic identification and differentiation of avian species is a vital step in conservative, taxonomic, forensic, legal and other ornithological interventions. The present investigation involved the application of molecular biological approach to identify and differentiate avian species i.e. two species of birds, namely black kite (Milvus migrans) and parakeet (Psittacula krameri). The DNA was isolated from blood samples of each species and a part of the mitochondrial 12S rRNA gene was amplified through polymerase chain reaction (PCR). The PCR products were sequenced and aligned using Basic Local Alignment Search Tool (BLAST) of the GenBank (NCBI). Based on the alignment and similarity/divergence, these avian species were accurately identified and differentiated.Autentična identifikacija i razlikovanje ptičjih vrsta od presudnoga su značenja u različitim konzervirajućim, taksonomskim, sudbenim, zakonskim i drugim ornitološkim aktivnostima. Ovo istraživanje bavi se molekularnobiološkim pristupom identifikaciji i razlikovanja dviju ptičjih vrsta: crvenkaste lunje (sokola) (Milvus migrans) i papige (Psittacula krameri). DNA je bila izdvojena iz uzoraka njihove krvi te je dio mitohondrijskoga 12S rRNA bio umnožen lančanom reakcijom polimerazom. Proizvodi PCR-a bili su sekvencirani i analizirani upotrebom Basic Local Alignment Search Tool (BLAST) genske banke GenBank (NCBI). Na osnovi sličnosti odnosno različitosti nalaza identificirane su te dvije pretraživane vrste

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Book chapter published in Meat Quality Analysis published by ElsevierDNA-based molecular techniques have emerged as the techniques of choice for species identification of meat and meat products in recent years due to their robustness, accuracy, and repeatability. Molecular techniques can be used for the species identification of both fresh and processed meat products and even for species identification of by-products derived from meat animals. DNA hybridization was one of the initial DNA-based molecular techniques used for meat species identification. Nevertheless, the widespread application of molecular techniques for the purpose of meat species identification was possible only after the advent of polymerase chain reaction (PCR); wherein million-fold in vitro amplification of target(s) was possible by repeated heating and cooling cycles. Consequently, several PCR-based methods, such as species-specific PCR, forensically informative nucleotide sequencing, restriction fragment length polymorphism, and DNA microarray, evolved as routine tests for the purpose of meat species identification in food, forensic, and referral laboratories worldwide. Quantification of adulteration is now possible using real-time PCR kinetic chemistry. Recently, techniques based on loop-mediated isothermal amplification principle were also developed for the purpose of species identification for field level or onsite speciation. In spite of these advancements, the need for a cost-effective, rapid, robust, and onsite test for the detection of varied animal species is still required. Therefore the scientific community must work to develop such platform tests of species identification that could even be integrated with recent ICT tools for enhanced field applications.Not Availabl

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Not AvailableMeat species identification is warranted to enforce product labelling, ensure traceability and for prevention of adulteration. Samples collected across meat supply chain requires to be submitted to a distant laboratory for authentication of the origin of species. Cryopreservation is an ideal method of sample preservation or shipping, however often it is not feasible under field conditions. Although several non-cryogenic tissue preservatives are in vogue, yet they have potential limitations for their field-level usage. India is the world’s largest exporter of buffalo meat but there are restrictions on slaughter of cattle and export of cattle meat (beef) is completely banned. Nevertheless, issues linked to the adulteration of buffalo meat with beef demands prompt authentication of the origin of species, hasten export and protect consumer sentiments. The present study deals with the preservation of buffalo meat samples in formalin and saturated salt solutions, extraction of DNA and polymerase chain reaction (PCR) to detect meat species. Universal and buffalo specific primers targeting mitochondrial DNA sequences were used during PCR of samples stored for two and six months at ambient temperatures in formalin and salt solution. Results showed successful target DNA amplification using PCR from meat samples stored in the salt solution for six months. Under the field conditions, preservation and transportation of meat samples in the saturated salt solution are recommended for buffalo meat authentication using DNA based PCR techniques.Not Availabl

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Not AvailableSubclinical mastitis (SCM) is one of the major economic diseases of dairy cows. In the present cross-sectional study, dairy cows (n=205) of small holding dairy farms were screened using California mastitis test (CMT) and prevalence of 12.19% was recorded. Holstein Friesian and Jersey crossbred cows showed highest CMT score of 2 followed by 3, 1 and T; linear scores of somatic cell counts (SCC) of 3.14±0.08, 4.72±0.25, 6.17±0.12, 7.99±0.11 and 9.84±0.45 were observed for CMT scores N, T, 1, 2 and 3, respectively. Risk factors associated with SCM were studied in 25 randomly selected cows. Staphylococcus species were detected in 54% of quarters based on PCR amplification of 16s rRNA gene; thermonuclease (nuc gene) PCR categorized Staphylococcus species as S. aureus (SA, 37%) and non-aureus Staphylococci (NAS, 26%). Significant association between CMT score 2 and Staphylococcus species was established. Staphylococcus species, SA and NAS, were detected in 30.7, 20.5 and 17.9% of SCM negative as well as 68.85, 47.54 and 31.14% SCM positive quarters, respectively. Significant association between SCM and Staphylococcus species and Staphylococcus aureus was observed. Study indicated strong association between SCM, Staphylococcus aureus and certain risk factors among cows reared under small holding dairy farms of the tropical region.Not Availabl

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Not AvailableFood-producing animals act as reservoirs of non-typhoidal Salmonella (NTS) serovars with potential food safety and public health implications. The present cross-sectional study aimed at determining the prevalence of Salmonella serotypes in non-diarrhoeic pigs and characterizing the isolates using molecular tools. Salmonella isolates (n = 22) recovered from faecal samples of 194 randomly selected pigs were characterized for virulence and antimicrobial resistance and subtyped using XbaI-PFGE. The prevalence of Salmonella in apparently healthy non-diarrhoeic pigs was 11.3% (95%CI, 4.3–19.5%), with S. Weltevreden (81.8%) and S. Enteritidis (18.2%) being the serotypes detected. Salmonella isolates harboured virulence genes such as invA (100%), stn (100%), spvR/spvC (86.3%) and fimA (22.7%). Phenotypically, isolates showed sensitivity to chloramphenicol, levofloxacin and ciprofloxacin and resistance to tetracycline and ampicillin (100%), streptomycin (86.4%), amoxicillin-clavulanate (63.6%), cefotaxime (22.7%) and ceftriaxone (9.1%). Notably, 18.2% isolates were multidrug-resistant (≥ 3 antimicrobial class) with multiple antimicrobial resistance (MAR) index of 0.56–0.67 (18.2%), 0.44 (45.5%), 0.33 (31.8%) and 0.22 (4.5%). Genotypically, isolates carried various antibiotic resistance genes: ESBL (blaTEM and blaOXA), aminoglycoside (strA, strB and aadA1), sulphonamide (sul1, sul2 and dfrA1), tetracycline (tetA and tetB) and plasmid AmpC beta-lactamase (ACC, FOX, MOX, DHA, CIT and EBC). The present investigation emphasizes the epidemiological significance of PFGE typing in the detection of emerging strains of highly virulent and multidrug-resistant S. Weltevreden and S. Enteritidis in non-diarrhoeic pigs that pose serious public health implications in the pork supply chain environment. More extensive longitudinal study is warranted to provide epidemiological links between environmental reservoirs and animal and human infections in piggery settings.Not Availabl

Journal of Food Science and Technology

Not AvailableIdentification of meat species origin using reliable techniques is a critical requirement for ensuring label compliance, protection of consumer preference and prevention of fraudulence in the meat trade. Although a plethora of protein and DNA based meat species identification techniques are in vogue, need for rapid test suitable for under-resourced laboratories catering point-of-care (PoC) services was construed. Present study deals with development of rapid sheep (Ovis aries) meat identification technique using DNA extraction by alkaline lysis (AL) and loop-mediated isothermal amplification (LAMP) technique. The AL?LAMP specifically amplifies sheep-specific signal of mitochondrial D loop region under an isothermal temperature of 60 ?C with an analytical sensitivity of 0.5 ng sheep DNA. The test was highly specific to sheep and performed well even in the presence of DNA of closely related meat animal species such as goat, cattle, buffalo and chicken. The novel primers designed for the AL?LAMP successfully detected sheep meat in raw and cooked meat samples heated up to 121 ?C for 30 min. Sheep-specific AL?LAMP assay could detect 0.1% mutton-in-beef adulteration. Novel AL?LAMP assay being simple, rapid and reliable for sheep meat authentication in just 120 min; hence, it could be conveniently used by terminal laboratories engaged in rendering on-site or PoC services

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Not AvailableWe report a novel, rapid, economical and species-specific DNA-based assay for the authentication of pork. The technique specifically amplified porcine mitochondrial D loop region by combining Alkaline Lysis (AL) method of DNA extraction and Loop Mediated Isothermal Amplification (LAMP). Visual detection of the reaction was accomplished by color development in the reaction with the addition of SYBR Green I dye. Dependable amplification was possible in thermally processed meat samples heated up to 121 °C for 30 min. The assay was able to detect pork in beef up to the level of 0.1% admixture and limit of detection of DNA was at 0.5 ng/μL. Cross amplification of related species like cattle, buffalo, sheep, goat and chicken was excluded by incorporating their DNA in the reaction assay. The novel approach (AL-LAMP technique) was found to be robust and handy, suitable even for resource compromised laboratories engaged in the food analysisNot Availabl

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Not AvailableIdentification of meat species origin using reliable techniques is a critical requirement for ensuring label compliance, protection of consumer preference and prevention of fraudulence in the meat trade. Although a plethora of protein and DNA based meat species identification techniques are in vogue, need for rapid test suitable for under-resourced laboratories catering point-of-care (PoC) services was construed. Present study deals with development of rapid sheep (Ovis aries) meat identification technique using DNA extraction by alkaline lysis (AL) and loop-mediated isothermal amplification (LAMP) technique. The AL–LAMP specifically amplifies sheep-specific signal of mitochondrial D loop region under an isothermal temperature of 60 °C with an analytical sensitivity of 0.5 ng sheep DNA. The test was highly specific to sheep and performed well even in the presence of DNA of closely related meat animal species such as goat, cattle, buffalo and chicken. The novel primers designed for the AL–LAMP successfully detected sheep meat in raw and cooked meat samples heated up to 121 °C for 30 min. Sheep-specific AL–LAMP assay could detect 0.1% mutton-in-beef adulteration. Novel AL–LAMP assay being simple, rapid and reliable for sheep meat authentication in just 120 min; hence, it could be conveniently used by terminal laboratories engaged in rendering on-site or PoC services.Not Availabl

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Not AvailableNon-typhoidal Salmonella serotypes continue to be major food safety and public health threats worldwide. In the era of emerging antimicrobial resistance, it is imperative to search for alternative strategies of infection control. Present study deals with passive protection of young chicks using polyclonal egg yolk antibodies (IgY). Laying hens were actively immunized with heat inactivated Salmonella serotypes Salmonella enterica subsp. enterica serotypes S. Enteritidis (SE) and S. Typhimurium (ST) isolated from the poultry. Serotype specific polyclonal IgY isolated from the hen egg yolk were evaluated for in vitro neutralization potential. Chicks challenged with pathogenic SE and ST were evaluated for passive protection that were co-administered with polyclonal immune IgY. Heat killed antigens (SE/ST) induced protective immune response in the immunized laying hens; and purified immune IgY inhibited the logarithmic phase of Salmonella growth. Administration of egg yolk at 5% (w/w) in the feed containing specific anti-SE or anti-ST polyclonal IgY to Salmonella-challenged chicks resulted in 37.48 and 38.54% reduction in the caecal Salmonella counts. As antimicrobial resistance is emerging as a growing public health problem, findings of this study indicate benefits of prophylactic feeding of egg yolk containing polyclonal IgY for the passive protection of chicks and minimize caecal Salmonella carriage.Not Availabl