Ovine Origin Research Articles

Evaluation of In Vitro Antihypertensive and Anti-Inflammatory Properties of Dairy By-Products

Sweet whey (SW) and yogurt acid whey (YAW) are dairy by-products of the cheese-making process and Greek-style yogurt production, respectively. Both of them are considered pollutants with huge volumes of SW and YAW produced due to the growing demand for dairy products worldwide. Moreover, whey-derived peptides, resulting from fermentation as well as from further hydrolysis during digestion, have been associated with various biological activities. In the present study, the angiotensin-converting enzyme (ACE)-inhibitory activity of 48 SW samples and 33 YAW samples from bovine, ovine, caprine, and ovine/caprine milk obtained were evaluated. Additionally, the SW and YAW digestates and two of their fractions (smaller than 10 kDa, SW-D-P10 and YAW-D-P10, and smaller than 3 kDa, SW-D-P3 and YAW-D-P3), which were obtained after in vitro digestion and subsequent ultrafiltration, were also subjected to evaluation. Our data indicated that the D-P10 and D-P3 fractions exhibited higher ACE-inhibitory activity compared to the corresponding values before digestion. The ACE-inhibitory capacity after in vitro digestion was higher for the ovine SW samples compared to their bovine and caprine counterparts. The effect of the D-P3 fraction on the inhibition of nitric oxide (NO) production and the expression of a selected panel of immune-response-related genes in LPS-stimulated RAW 264.7 macrophages was also evaluated. Fractions from both dairy by-products inhibited NO production in LPS-stimulated RAW 264.7 cells. Especially, ovine SW-D-P3 showed a strong NO inhibitory activity and suppressed inducible nitric oxide synthase (Nos2) mRNA levels. However, YAW-D-P3 could not trigger neither the gene expression of inflammatory macrophage mediators Nos2 and cyclooxygenase-2 (Ptgs2) nor tumor necrosis factor-α (Tnf) and interleukin 6 (Il6) in LPS-stimulated murine macrophages regardless of animal origin. These findings suggest that in vitro digestion could enhance the production of ACE-inhibitory peptides in both dairy by-products, while SW from ovine origin displays higher potential as an anti-inflammatory agent, effectively preventing excessive NO production.

Retrospective study on the occurrence of Salmonella serotypes in veterinary specimens of Atlantic Canada (2012-2021).

This study aimed to summarize the frequency and the antimicrobial susceptibility profiles of the Salmonella serotypes identified from the specimens of companion animals, livestock, avian, wildlife and exotic species within Atlantic Canada. The retrospective electronic laboratory data of microbiological analyses of a selected subset of samples from 03 January 2012 to 29 December 2021 submitted from various animal species were retrieved. The frequency of Salmonella serotypes identified, and their antimicrobial susceptibility results obtained using the disk diffusion or broth method were analysed. The test results were interpreted according to the Clinical and Laboratory Standards Institute standard. The Salmonella serotypes were identified by slide agglutination (Kauffman-White-Le-Minor Scheme) and/or the Whole Genome Sequencing for the Salmonella in silico Serovar Typing Resource-based identification. Of the cases included in this study, 4.6% (n=154) had at least one Salmonella isolate, corresponding to 55 different serovars. Salmonella isolation was highest from exotic animal species (n=40, 1.20%), followed by porcine (n=26, 0.78%), and canine (n=23, 0.69%). Salmonella subsp. enterica serovar Typhimurium was predominant among exotic mammals, porcine and caprine samples, whereas S. Enteritidis was mostly identified in bovine and canine samples. S. Typhimurium of porcine origin was frequently resistant (>70.0%) to ampicillin. In contrast, S. Typhimurium isolates from porcine and caprine samples were susceptible (>70.0%) to florfenicol. S. Oranienburg from equine samples was susceptible to chloramphenicol, but frequently resistant (>90.0%) to azithromycin. In avian samples, S. Copenhagen was susceptible (>90.0%) to florfenicol, whereas Muenchen was frequently resistant (>90.0%) to florfenicol. S. subsp. diarizonae serovar IIIb:61:k:1,5 of ovine origin was resistant (50.0% isolates) to sulfadimethoxine. No significant changes were observed in the antibiotic resistance profiles across the study years. This report provides data for surveillance studies, distribution of Salmonella serotypes and their antimicrobial resistance among veterinary specimens of Atlantic Canada.

Andexanet Alfa Neutralizes the Anticoagulant Effects of Unfractionated Heparin of Bovine, Ovine and Porcine Origin Almost as Protamine Sulfate.

Andexanet alfa (AA) - zhzo, recombinant coagulation factor Xa, is an approved antidote for oral Xa inhibitors (apixaban and rivaroxaban). Unfractionated heparin (UFH) is commonly used for therapeutic, interventional, and surgical indications. Protamine sulfate (PrSO4) is frequently used to neutralize UFH. This study aimed to investigate the comparative neutralization profiles of AA and PrSO4 for heparins of bovine, ovine, and porcine origin. The neutralization effect of PrSO4 at 25 µg/ml and AA at 100 µg/ml was studied on an approximate surgical/interventional concentration of heparin by supplementing whole blood with each of the heparins at 25 µg/ml. For the clotting profile (activated partial thromboplastin time: aPTT), amidolytic (anti-Xa and anti-IIa), and thrombin generation assay each of the heparin were supplemented from -10-0.62 µg/ml. In the whole blood ACT studies, all three heparins produced strong anti-coagulant effects (400-450 seconds) compared to saline (130-150 seconds). Both AA and PrSO4 almost fully neutralized the anti-coagulant effects of heparins (140-160 seconds). Both antidotes completely reversed the anticoagulant effects of all three heparins in the aPTT and thrombin generation assay. However, PrSO4 was more effective in neutralizing the anti-Xa, and anti-IIa effects than AA, which only partially neutralized these effects. Andexanet alfa at 100 µg/ml effectively neutralizes the therapeutic and surgical/interventional concentrations of heparins in in-vitro settings. While differences in the anti-Xa, and anti-IIa effects between heparins were noted, anti-coagulant effect of these agents in the aPTT assay were comparable. A similar neutralization profile was observed in the ACT and thrombin generation assays by both agents.

Effect of Milk Origin and Seasonality of Yogurt Acid Whey on Antioxidant Activity before and after In Vitro Gastrointestinal Digestion.

Yogurt acid whey (YAW) is a by-product of Greek strained yogurt production. The disposal of YAW constitutes an environmental problem, and given the increasing demand of Greek yogurt worldwide, its handling is a challenge. However, whey-derived peptides, resulting from microbial fermentation as well as those resulting from further hydrolysis during the digestion process, have been linked to enhanced biological activities. In this study, the antioxidant capacity of 33 samples of YAW obtained from Greek dairy companies of bovine, ovine or caprine origin was investigated using both cell-free and cell-based assays. The YAW samples, their in vitro digestion products (YAW-Ds) and a fraction of the digests (less than 3 kDa; YAW-D-P3) were assessed using four biochemical assays, namely ORAC, ABTS, FRAP and P-FRAP. Our data revealed a higher antioxidant capacity for digested samples compared with undigested samples, with all four methods. ORAC values after in vitro digestion were higher for the ovine samples compared to their bovine (YAW-D and YAW-D-P3) and caprine (YAW-D-P3) counterparts. Furthermore, the YAW-D-P3 fraction derived from samples collected in the summer months exhibited higher ORAC values when compared to the respective fraction from the winter months' samples. The cellular antioxidant activity of ovine YAW-D-P3 was improved in H2O2-treated HT29 cells compared to the control H2O2-treated cells. However, YAW-D-P3 could not trigger either the pathways involving the transcription factors NF-κB or NFE2L2 or the gene expression of SOD1, CAT and HMOX1 in LPS-challenged THP-1-derived macrophages. These results suggest that YAW, and particularly YAW from ovine origin, could be used as a natural source for its antioxidant potential in human and animal nutrition.

First report on genotypic estimation of MRSA load in udder of nomadic sheep flocks affected with subclinical mastitis in Pakistan

Mastitis is one of the highly devastating issues responsible for production and economic losses in all dairy animals including sheep. This study was designed to investigate subclinical mastitis (SCM) associated with S. aureus in lactating nomadic ewes, along with the associated risk factors analysis. Furthermore, molecular characterization and antibiogram profiling of local methicillin-resistant S. aureus (MRSA) isolates of ovine origin were also performed. A total of 384 milk samples (n = 384) were collected from 13 nomadic sheep flocks using a convenient sampling technique. SCM was evaluated using a Surf Field Mastitis test and the S. aureus was isolated using standard microbiological techniques. Kirby-Bauer disc diffusion assay was used for phenotypic identification of MRSA while the mecA gene was tested through PCR. Study results revealed that SCM was prevalent at 34.37% while S. aureus association was recorded at 39.39%. MRSA prevalence was 36.53% and 21.15% using phenotypic and genotypic tests, respectively. The mecA gene sequences of study isolates showed maximum resemblance with already reported sequences from Pakistan, China, and Myanmar. MRSA isolates showed maximum resistance towards penicillin, ceftriaxone sodium, and trimethoprim + sulphamethoxazole while gentamicin, ciprofloxacin, and tylosin showed maximum efficacy. Risk factors analysis revealed that various flock management, housing, and host-related factors positively influenced the incidence of S. aureus-associated SCM. This study is the first report on the prevalence of S. aureus and MRSA associated with SCM in lactating ewes in Pakistan. This study will help to devise effective treatment and control strategies for S. aureus-associated SCM.

PB0551 A Comparison of USP Potency Adjusted and Mass Adjusted Heparins of Bovine, Ovine, and Porcine Origin in Various Assays

PB0551 A Comparison of USP Potency Adjusted and Mass Adjusted Heparins of Bovine, Ovine, and Porcine Origin in Various Assays

Expanding the rumen Prevotella collection: The description of Prevotella communis, sp. nov. of ovine origin

27 strains representing eight new Prevotella species were isolated from rumen of a single sheep in eight weeks interval. One of the putative species encompassing the highest number of isolated strains which also exhibited some genetic variability in preliminary data, was then selected for description of a novel species. We examined six strains in genomic and phenotypic detail, two of which may actually be the same strain isolated nearly three weeks apart. Other strains formed clearly diverged intraspecies lineages as evidenced by core genome phylogeny and phenotypic differences. Strains of the proposed new Prevotella species are strictly saccharolytic as is usual for rumen Prevotella, and use plant cell-wall xylans and pectins for growth. However, the range of cell-wall polysaccharides utilised for growth is rather limited compared to rumen generalists such as Prevotella bryantii or Prevotella ruminicola and this extends also to the inability to utilise starch, which is unexpected for the members of the genus Prevotella. Based on the data obtained, we propose Prevotella communis sp. nov. to accommodate strain E1-9T as well as other strains with the similar properties. The proposed species is widespread: two other strains were previously isolated from sheep in Japan and is also common in metagenomic data of cattle and sheep rumen samples from Scotland and New Zealand. It was also found in a collection of metagenome-assembled genomes originating from cattle in Scotland. Thus, it is a ubiquitous bacterium of domesticated ruminants specialising in degradation of a somewhat restricted set of plant cell wall components.

Synthetic antibodies block receptor binding and current-inhibiting effects of α-cobratoxin from Naja kaouthia.

Each year, thousands of people fall victim to envenomings caused by cobras. These incidents often result in death due to paralysis caused by α-neurotoxins from the three-finger toxin (3FTx) family, which are abundant in elapid venoms. Due to their small size, 3FTxs are among the snake toxins that are most poorly neutralized by current antivenoms, which are based on polyclonal antibodies of equine or ovine origin. While antivenoms have saved countless lives since their development in the late 18th century, an opportunity now exists to improve snakebite envenoming therapy via the application of new biotechnological methods, particularly by developing monoclonal antibodies against poorly neutralized α-neurotoxins. Here, we describe the use of phage-displayed synthetic antibody libraries and the development and characterization of six synthetic antibodies built on a human IgG framework and developed against α-cobratoxin - the most abundant long-chain α-neurotoxin from Naja kaouthia venom. The synthetic antibodies exhibited sub-nanomolar affinities to α-cobratoxin and neutralized the curare-mimetic effect of the toxin in vitro. These results demonstrate that phage display technology based on synthetic repertoires can be used to rapidly develop human antibodies with drug-grade potencies as inhibitors of venom toxins.

Molecular Survey on Kobuviruses in Domestic and Wild Ungulates From Northwestern Italian Alps.

Since the first identification in 1989 in humans, kobuviruses (KoVs) have been identified from a wide range of animal species including carnivores, rodents, birds, ungulates, rabbits, and bats. Several studies have described the identification of genetically related KoVs in the fecal virome of domestic and wild animals suggesting a mutual exchange of viruses. By screening a total of 231 fecal samples from wild and domestic ungulates, KoVs RNA was detected in wild boars (3.2%; 2/63), chamois (4.6%; 2/43), and goats (2.6%; 2/77). On phylogenetic analysis of the partial RdRp sequence, the wild boar strains clustered within the species Aichivirus C whilst the strains identified in domestic and wild ruminants grouped into the species Aichivirus B. The complete VP1 gene was obtained for chamois and goat KoVs. Interestingly, upon phylogenetic analysis the strains grouped together with a KoV of ovine origin within a distinct genetic type (B3) of the species Aichivirus B.

Diversity and characterization of temperate bacteriophages induced in Pasteurella multocida from different host species

BackgroundBacteriophages play important roles in the evolution of bacteria and in the emergence of new pathogenic strains by mediating the horizontal transfer of virulence genes. Pasteurella multocida is responsible for different disease syndromes in a wide range of domesticated animal species. However, very little is known about the influence of bacteriophages on disease pathogenesis in this species.ResultsTemperate bacteriophage diversity was assessed in 47 P. multocida isolates of avian (9), bovine (8), ovine (10) and porcine (20) origin. Induction of phage particles with mitomycin C identified a diverse range of morphological types representing both Siphoviridae and Myoviridae family-types in 29 isolates. Phage of both morphological types were identified in three isolates indicating that a single bacterial host may harbour multiple prophages. DNA was isolated from bacteriophages recovered from 18 P. multocida isolates and its characterization by restriction endonuclease (RE) analysis identified 10 different RE types. Phage of identical RE types were identified in certain closely-related strains but phage having different RE types were present in other closely-related isolates suggesting possible recent acquisition. The host range of the induced phage particles was explored using plaque assay but only 11 (38%) phage lysates produced signs of infection in a panel of indicator strains comprising all 47 isolates. Notably, the majority (9/11) of phage lysates which caused infection originated from two groups of phylogenetically unrelated ovine and porcine strains that uniquely possessed the toxA gene.ConclusionsPasteurella multocida possesses a wide range of Siphoviridae- and Myoviridae-type bacteriophages which likely play key roles in the evolution and virulence of this pathogen.

Occurrence and characterization of ciprofloxacin‐resistantEscherichia colifrom bovine and ovine bulk tank milk samples in Turkey

AbstractThe objective of this study was to examine the occurrence and characteristics of ciprofloxacin‐resistant (CipR)Escherichia coliisolates on bulk tank milk (BTM) samples (bovine and ovine origins) in Turkey. A total of 91 BTM samples (41.7%, 95% confidence intervals 35.2–48.6%) out of 218 were found to be positive for CipRE. coliisolates (MIC values of ≥4 μg/ml). Analysis of PFGE fingerprint profile forE. coliisolates resulted in the 55 different pulsotypes based on >85% homology. All isolates were resistant to enrofloxacin and nalidixic acid and the resistance rates in bovine and ovine origin isolates were 94.9 and 78.1% for norfloxacin (p < .05) and 27.1 and 34.4% for levofloxacin, respectively. Additionally, resistance to non‐quinolone antibiotics was commonly observed against tetracycline (resistance rates in bovine and ovine = 91.5 and 87.5%, respectively), trimethoprim‐sulfamethoxazole (83.1 and 93.8%, respectively), gentamycin (15.3 and 40.6%, respectively,p < .05) and chloramphenicol (23.7 and 65.6%, respectively,p < .05). TheqnrS1gene (3.1 and 6.8%, respectively) was the most prevalent PMQR genes in isolates from ovine and bovine origins, followed byaac (6′)‐Ib‐cr(0 and 5.1%, respectively) andqnrB19(0 and 1.7%, respectively). The other resistance genes includingtetA,tetB,strA/B,aPozhA1,aadA,aadB,blaCTX‐M, andblaTEMwere also identified in various frequencies. The most frequently observed virulence trait wasfimH. The low‐level presence of PMQR genes and as well as some virulence traits is an important finding, yet the results of this study are worrisome because quinolone antibiotics are still the drugs of choice for severe infections in humans.

Differentiated ovine tracheal epithelial cells support the colonisation of pathogenic and non-pathogenic strains of Mannheimia haemolytica

Mannheimia haemolytica is the primary bacterial species associated with respiratory disease of ruminants. A lack of cost-effective, reproducible models for the study of M. haemolytica pathogenesis has hampered efforts to better understand the molecular interactions governing disease progression. We employed a highly optimised ovine tracheal epithelial cell model to assess the colonisation of various pathogenic and non-pathogenic M. haemolytica isolates of bovine and ovine origin. Comparison of single representative pathogenic and non-pathogenic ovine isolates over ten time-points by enumeration of tissue-associated bacteria, histology, immunofluorescence microscopy and scanning electron microscopy revealed temporal differences in adhesion, proliferation, bacterial cell physiology and host cell responses. Comparison of eight isolates of bovine and ovine origin at three key time-points (2 h, 48 h and 72 h), revealed that colonisation was not strictly pathogen or serotype specific, with isolates of serotype A1, A2, A6 and A12 being capable of colonising the cell layer regardless of host species or disease status of the host. A trend towards increased proliferative capacity by pathogenic ovine isolates was observed. These results indicate that the host-specific nature of M. haemolytica infection may result at least partially from the colonisation-related processes of adhesion, invasion and proliferation at the epithelial interface.

Studies on Tissue Factor Pathway Inhibitor Antigen Release by Bovine, Ovine and Porcine Heparins Following Intravenous Administration to Non-Human Primates.

Unfractionated heparin (UFH) is a sulfated glycosaminoglycan that consists of repeating disaccharides, containing iduronic acid (or glucuronic acid) and glucosamine, exhibiting variable degrees of sulfation. UFHs release tissue factor pathway inhibitor (TFPI) which inhibits the extrinsic pathway of coagulation by inactivating factor Xa and the factor VIIa/TF complex. Most heparins used clinically are derived from porcine intestinal mucosa however, heparins can also be derived from tissues of bovine and ovine origin. Currently there are some concerns about the shortage of the porcine heparins as they are widely used in the manufacturing of the low molecular weight heparins (LMWHs). Moreover, due to cultural and religious reasons in some countries, alternative sources of heparins are needed. Bovine mucosal heparins (BMH) are currently being developed for re-introduction to the US market for both medical and surgical indications. Compared to porcine mucosal heparin (PMH), BMH exhibits a somewhat weaker anti-coagulant activity. In this study, we determined the TFPI antigen level following administration of various dosages of UFHs from different origins. These studies demonstrated that IV administration of equigravemetric dosages of PMH and ovine mucosal heparin (OMH) to non-human primates resulted in comparable TFPI antigen release from endothelial cells. In addition, the levels of TFPI were significantly higher than TFPI antigen levels observed after BMH administration. Potency adjusted dosing resulted in comparable TFPI release profiles for all 3 heparins. Therefore, such dosing may provide uniform levels of anticoagulation for the parenteral indications for UFHs. These observations warrant further clinical validation in specific indications.

1D and 2D-HSQC NMR: Two Methods to Distinguish and Characterize Heparin From Different Animal and Tissue Sources.

The US Food and Drug Administration has encouraged the reintroduction of bovine heparin drug product to the US market to mitigate the risks of heparin shortages and potential adulteration or contamination of the primary source which is porcine heparin. Here, a 1D-NMR method was applied to compare heparin sodium of bovine intestinal origin with that of bovine lung, porcine, or ovine intestinal origin. The results showed that a simple 1D test using NMR signal intensity ratios among diagnostic signals of the proton spectra uniquely identified the origin of heparin and concomitantly could be used to assure the correct sample labeling. However, a limitation of the use of only mono-dimensional spectra is that these spectra may not provide sufficiently detailed information on the composition of heparin batches to adequately determine the quality of this complex product. As an alternative, a higher resolution quantitative 2D-HSQC method was used to calculate the percentage of mono- and disaccharides, distinguish the origin of heparin and, simultaneously, assess the heparin composition. The 2D-HSQC method is proposed to provide sufficient information to evaluate the quality of industrial production process used to make the drug substance. Together, the 1D and 2D data produced by these measurements can be used to assure the identity and purity of this widely used drug.

A Factor Xa Inhibitor Antidote (andexanet alfa) is Capable of Neutralizing the Anticoagulant Effects of Unfractionated Heparin of Bovine, Ovine and Porcine Origin in a Comparable Manner as Protamine Sulfate

IntroductionAndexanet alfa (zhzo, coagulation factor Xa recombinant) is approved for such factor Xa inhibitors as apixaban and rivaroxaban. Unfractionated heparin (UFH) is commonly used for therapeutic, interventional and surgical indications. Protamine sulfate is commonly used to neutralize UFH. The study investigated the comparative neutralization profiles of andexanet alfa and protamine sulfate for heparins of bovine, ovine or porcine origin.Materials and MethodPowdered form of pharmaceutical grade heparins of bovine (140 U/mg), ovine (200 U/mg) and porcine (190 U/mg) origin were obtained from commercial vendors. Andexanet alfa was obtained from the hospital pharmacy (Skokie Hospital, Skokie, Illinois) and reconstituted at 10 mg/ml according to the manufacturer's instructions. Powdered protamine sulfate (USP grade) was reconstituted at 1.0 mg/ml in saline. To investigate the neutralization profile of these two antidotes for various heparins, each of the bovine, ovine and porcine heparins was supplemented to plasma over a concentration range of 0–10 ug/ml. The effect of protamine sulfate at 10 ug/ml was studied in the activated partial thromboplastin time (aPTT), anti‐Xa and anti‐IIa assays. Similar studies were carried out with andexanet alfa at a concentration of 100 ug/ml. The neutralization effect of protamine sulfate and andexanet alfa was also studied on an approximate surgical/interventional concentration of heparin by supplementing freshly drawn whole blood from healthy individuals (n=5–10) with each of the heparins at a concentration of 25 ug/ml. The neutralization effect of andexanet alfa was studied at 100 ug/ml and protamine sulfate was studied at 25 ug/ml.ResultsBoth andexanet alfa (100 ug/ml) and protamine sulfate (10 ug/ml) completely neutralized the anticoagulant effects of all three heparins in the aPTT assay. However, protamine sulfate was more effective in neutralizing the amidolytic anti‐Xa and anti‐IIa effects in comparison to andexanet alfa which only partially neutralized these effects. In the whole blood clotting ACT studies, all three heparins produced a strong anti‐coagulant effects (400–450 seconds) in comparison to saline (130–150 seconds). Both andexanet alfa (100 ug/ml) and protamine sulfate (25 ug/ml) almost completely neutralized the anti‐coagulant effects of heparins (140–160 seconds).ConclusionThese results clearly suggest that andexanet alfa at 100 ug/ml is effective in neutralizing both the therapeutic and surgical/interventional concentrations of heparins in the in‐vitro settings. While differences in the anti‐Xa and anti‐IIa effects between heparins are noted, the anti‐coagulant effect of these agents in the aPTT assay were comparable and similar neutralization profile was observed in the ACT assays by both agents. The pharmacologic implication of these observations require further investigations.Support or Funding InformationN/AThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

In vitro analysis of genetically distinct Chlamydia pecorum isolates reveals key growth differences in mammalian epithelial and immune cells

Chlamydia (C.) pecorum is an obligate intracellular bacterium that infects and causes disease in a broad range of animal hosts. Molecular studies have revealed that this pathogen is genetically diverse with certain isolates linked to different disease outcomes. Limited in vitro or in vivo data exist to support these observations, further hampering efforts to improve our understanding of C. pecorum pathogenesis. In this study, we evaluated whether genetically distinct C. pecorum isolates (IPA, E58, 1710S, W73, JP-1-751) display different in vitro growth phenotypes in different mammalian epithelial and immune cells. In McCoy cells, shorter lag phases were observed for W73 and JP-1-751 isolates. Significantly smaller inclusions were observed for the naturally plasmid-free E58 isolate. C. pecorum isolates of bovine (E58) and ovine origin (IPA, W73, JP-1-751) grew faster in bovine cells compared to a porcine isolate (1710S). C. pecorum isolates could infect but appear not able to complete their developmental cycle in bovine peripheral neutrophil granulocytes. All isolates, except 1710S, could multiply in bovine monocyte-derived macrophages. These results reveal potentially important phenotypic differences that will help to understand the pathogenesis of C. pecorum in vivo and to identify C. pecorum virulence factors.

Simultaneous Detection of Carnosine and Anserine by UHPLC-MS/MS and Its Application on Biomarker Analysis for Differentiation of Meat and Bone Meal.

A novel ultra-high performance liquid chromatography (UHPLC) procedure, coupled with tandem mass spectrometry (MS/MS), was established for the analysis of anserine (ANS) and carnosine (CAR) in meat and bone meal (MBM) (bovine, ovine, porcine, and poultry origins). The pretreatment strategies were optimized for four types of MBM samples prior to UHPLC-MS/MS analysis. This method allowed determining CAR and ANS in short analysis time (18 min per sample). The limits of detection (LODs) and limits of quantification (LOQs) of two analytes in four types of MBM samples were in the ranges of 0.41–3.07 ng/g and 0.83–5.71 ng/g, respectively. The recovery rates spiked with low, intermediate, and high levels of two analytes in four types of MBM samples were 48.53–98.93%, 60.12–98.94%, and 67.90–98.92%, respectively. Acceptable inter-day reproducibility (RSD < 12.63%) supported the application of this proposed method for determining CAR and ANS in MBM samples. Overall, this rapid, effective, and robust method was successfully applied for quantitative detection of CAR and ANS in MBM samples. Furthermore, The CAR/ANS ratio was found to be in the decreasing order: porcine > bovine > ovine > poultry MBM. This proposed methodology was novelly applied to identify the biomarker (CAR/ANS ratio) for species-specific identification of MBM.

Species discrimination of terrestrial processed animal proteins by laser-induced breakdown spectroscopy (LIBS) based on elemental characteristics

Description of the subject. This research presents a novel approach for species discrimination of terrestrial processed animal proteins (PAP) using laser-induced breakdown spectroscopy (LIBS) as an alternative technique for determining feed safety. Objectives. To explore the correlation between LIBS results and elemental composition. To assess the potential of LIBS spectroscopic methods for species discrimination of terrestrial PAP. Method. A total of 42 reliable terrestrial PAP samples of ovine, bovine, porcine, and poultry origin were analyzed and LIBS spectra were innovatively collected from the separated bone fraction. Partial least squares-discriminant analysis (PLS-DA) models were performed on the dataset. Results. Significant differences were found in the element compositions (carbon, nitrogen, hydrogen, calcium, sodium, potassium, magnesium, zinc, strontium and barium, p &lt; 0.05) of PAP from different sources, which indicated a high potential for species discrimination. High-marked LIBS spectral characteristics (21 variables) related to six elements (calcium, sodium, magnesium, zinc, strontium and barium) for different species of terrestrial PAP samples were found out. PAP from the four tested species could be well classified with high sensitivity and specificity (&gt; 0.90), while the ruminant and non-ruminant PAP could be thoroughly discriminated. For the discriminant analysis of ruminant-derived components in terrestrial PAPs, the detection of 10%-40% adulteration can be achieved and the correct rate is about 95%. Conclusions. This study demonstrates that LIBS can mostly exhibit the elemental composition of different terrestrial PAP, and can be a promising method for discriminating the different animal species or animal groups (ruminant/non-ruminant) involved in terrestrial PAPs.

Interspecies comparative epidemiology of Livestock Associated Staphylococcus aureus among bovine and ovine origins - A public health perspective

Interspecies comparative epidemiology of Livestock Associated Staphylococcus aureus among bovine and ovine origins - A public health perspective

Phenotypic and genotypic characteristics of Trueperella pyogenes isolated from ruminants.

Trueperella pyogenes is an opportunistic pathogen that causes suppurative infections in animals including humans. Data on phenotypic and genotypic properties of T. pyogenes isolated from ruminants, particularly goats and sheep, are lacking. We characterized, by phenotypic and genotypic means, T. pyogenes of caprine and ovine origin, and established their phylogenetic relationship with isolates from other ruminants. T. pyogenes isolates ( n = 50) from diagnostic specimens of bovine ( n = 25), caprine ( n = 19), and ovine ( n = 6) origin were analyzed. Overall, variable biochemical activities were observed among the T. pyogenes isolates. The fimbriae-encoding gene, fimE, and neuraminidase-encoding gene, nanH, were, respectively, more frequently detected in the large ( p = 0.0006) and small ( p = 0.0001) ruminant isolates. Moreover, genotype V ( plo/ nanH/ nanP/ fimA/ fimC) was only detected in the caprine and ovine isolates, whereas genotype IX ( plo/ nanP/ fimA/ fimC/ fimE) was solely present in the isolates of bovine origin ( p = 0.0223). The 16S rRNA gene sequences of all T. pyogenes isolates were clustered with the reference T. pyogenes strain ATCC 19411 and displayed a high degree of identity to each other. Our results highlight phenotypic and genotypic diversity among ruminant isolates of T. pyogenes and reinforce the importance of characterization of more clinical isolates to better understand the pathogenesis of this bacterium in different animal species.