Ovine Samples Research Articles

Terrestrial animal livers as a source of PCDD/Fs, PCBs and PBDEs in the diet

The livers of terrestrial animals are an important source of vitamins and minerals, but with the liver being the major filtering and detoxifying organ, it retains xenobiotic material, especially persistent organic pollutants (POPs). The aim of the study was human health risk assessment related to the presence of certain POPs, namely polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), polychlorinated biphenyls (PCBs), and polybrominated diphenyl ethers (PBDEs) in terrestrial animals' livers. HRGC-HRMS was used to determine analyte concentrations in 99 liver samples of chicken, pork, beef, and mutton livers obtained from Polish farms. The ovine livers were the most contaminated by ∑PCDD/F/dl-PCBs (0.06–2.21 pg WHO-TEQ g−1 wet weight), ∑ndl-PCBs (0.036–1.54 ng g−1 wet weight) and ∑PBDEs (0.21–2.27 ng g−1 wet weight). Samples were found of ovine and pork livers that exceeded the maximum concentration limits established for PCDD/Fs. The Kruskal–Wallis test showed that there were statistical differences between regions in the concentrations of the ∑PCDD/Fs and ∑PCDD/F-PCBs, but none in those of the ∑dl-PCBs, ∑ndl-PCBs or ∑PBDEs. According to Dunnett's test results, ovine liver samples from the Podlaskie province contained fewer ∑PCDD/F than those from the Dolnośląskie (p < 0.027) and Małopolskie (p < 0.0342) voivodeships. Taking into account PCDD/F contents and Tolerable Weekly Intake (2 pg WHO-TEQ/kg BW) (TWI2018), the lowest risk was associated with chicken liver consumption (TWI up to 11 % in the 95th percentile), pork (62 %) and bovine (92 %) livers should be eaten in limited amounts for risk avoidance, while mutton liver (400 %) should not be eaten at all. Additionally, there is no risk of ingestion of a harmful amount of PBDEs associated with consumption of the livers of terrestrial animals from Poland based on the margin of exposure values (>15).

Establishment of a method for detecting Trichinella spiralis in ovine muscle tissues using real-time fluorescence quantitative PCR

Trichinellosis is caused by Trichinella spiralis, a meat-borne zoonotic disease transmitted to humans through the consumption of infected undercooked or raw meat. Surveillance using safe and precise diagnostic tools to diagnose T. spiralis in sheep is needed to assess the incidence and probability of transmission from sheep to humans. In this study, we developed a real-time PCR assay to detect T. spiralis DNA in ovine muscle samples that can be used as an alternative surveillance tool to ensure food safety using newly designed primers. The assay is specific for the Scfld4 gene of Trichinella (T1) and enables the detection of larvae in ovine muscle tissue samples with high sensitivity and specificity. Trichuris ovis, Oesophagostomum dentatum, Haemonchus contortus, and Bunostomum trigonocephalum showed no nonspecific amplification. The assay could detect Trichinella DNA concentrations as low as 0.0026 ng/μL, equivalent to 0.0064 larvae, indicating a high sensitivity for T. spiralis detection. We used this real-time PCR to detect 73 ovine muscle samples from an ovine abattoir, and five samples tested positive via real-time PCR but negative via microscopy. This assay may provide a more specific and sensitive method for rapidly detecting Trichinella larvae in ovine muscle tissues.

Effect of Lyophilization Process on Nutritional Value of Meat By-Products

The meat industry generates large amounts of by-products, and their mass represents approximately one-third of the live weight of animals. Most by-products are disposed of and not used for processing, although they meet hygiene requirements and are suitable for human consumption or for the processing of food supplements. The aim of this study was to investigate the effect of lyophilization on the nutritional value and retention of functional ingredients in ovine and porcine by-products: liver, kidneys, hearts, and lungs. For this purpose, meat by-products of the third category were selected in X and Y slaughterhouses and divided into two parts: one part was freeze-dried at −80 °C for 72 h, and the other part was left raw. Fatty acid composition was determined by gas chromatography–mass spectrometry (GC-MS), and amino acid analysis was performed by AccQ Tag technology (Waters Corp., Milford, MA, USA) and HPLC. Our study shows that the lyophilization process did not significantly affect protein and fat content. The largest decrease in the amount of proteins was determined in samples of ovine kidneys, at 0.8%, while the difference in protein in ovine liver samples before and after lyophilization was 0.38%. The composition of essential amino acids did not change after lyophilization, except a decrease in Leu and Thr in porcine samples and Leu, Thr, and Met in ovine samples (p &lt; 0.05). The lyophilization process did not significantly affect the polyunsaturated fatty acid content, including the amounts of omega 3 and omega 6 fatty acids. The optimal ratio of omega 6 and 3 fatty acids was determined in samples of lyophilized ovine livers (2.65), and the largest ratio was found in samples of lyophilized porcine hearts (16.67). The study results show that, after lyophilization, meat by-products of the third category (according to Regulation (EC) No. 1069/2009, Categorization, Article 10), especially ovine liver, can be used as a source of amino acids and polyunsaturated fatty acids for functional food processing. The process of lyophilization is also appropriate for preserving meat by-products without losing the nutritional value and beneficial components.

MLVA and com1 genotyping of Coxiella burnetii in farmed ruminants in Great Britain

Coxiella burnetii, the causative agent of the zoonotic disease Q fever, has been shown to be endemic in Great Britain, but information on the prevailing genomic lineages or Genomic Groups (GGs) of Coxiella burnetii is limited. The aim of this study was to genotype C. burnetii isolates from infected farmed ruminants by Multiple Locus Variable Number Tandem Repeat Analysis (MLVA) and identify their associated Genomic Group. A total of 51 Coxiella-containing abortion samples from farmed ruminants (sheep, goats, and cattle), which were collected in Great Britain during 2013–2018, were included in the study, 34 of which returned a C. burnetii MLVA genotype. All bovine samples (n = 18), 5/7 of the ovine samples, and 3/9 of the caprine samples belonged to an MLVA cluster which we could link to the MST20 genotype of GG III, whereas 6/9 of the caprine samples and 2/7 of the ovine samples belonged to MLVA clusters which we could link to the MST33 or MST32 genotypes of GG II (7 vs 1 sample(s), respectively). We also noted that the Coxiella-specific com1 gene contained unique mutations that could genomotype isolates, i.e. assign them to a Genomic Group. In conclusion, both goats and sheep in Great Britain (from 2014 onward) were found to carry the same MLVA genotypes (MST33-like; GG II) that were linked to a human Q fever outbreak in the Netherlands. This knowledge in combination with the usage of genotyping/genomotyping methods should prove useful in future surveillance programs and in the management of outbreaks.

Preparation and evaluation of a lateral flow immunochromatographic nanogold diagnostic kit for brucellosis in sheep.

Brucellosis is a zoonotic disease with a worldwide distribution. It has a serious impact on the health of humans and animals, along with a negative impact on the economy. This study aimed to prepare and evaluate the diagnostic performance of a lateral flow immunochromatographic test (LFIT) nanogold diagnostic kit for detecting brucellosis in sheep. A rapidly developed LFIT, in which lipopolysaccharide conjugates with nanogold molecules, was placed on the conjugate pad. One hundred ovine serum samples were tested to detect Brucella antibodies (Ab) using the prepared lateral flow immunochromatography assay (LFA) kit and Rose Bengal test. The evaluation of specificity, sensitivity, and accuracy for LFIT and Rose Bengal plate test was conducted using the P04310-10 IDEXX brucellosis ovine/caprine Ab enzyme-linked immunosorbent assay (ELISA) test (gold standard). The lower amount of Brucella Ab in the ovine serum samples was detected and was 1.58 S/P ratio ELISA titer/100 μL using LFIT and with Rose Bengal to detect 1.86 S/P ratio ELISA. The results showed that the developed LFIT had high specificity with no cross-reactivity with other tested bacteria. The calculated sensitivity, specificity, and accuracy of LFIT and Rose Bengal test using the P04310-10 IDEXX brucellosis ovine/caprine Ab ELISA test (gold standard) were 74% and 89%, 81% and 59%, and 76.9% and 66%, respectively. The present results showed interesting results implying that the LFIA strip test could be used as a substantial diagnostic tool for field screening ovine Brucella as an essential step in the control of brucellosis. However, further studies for the validation of the present findings are necessary.

10 Utilizing1H NMR Spectroscopy to Quantify and Identify Biomarkers in Finishing Sheep

Abstract Backfat and carcass fatness is an important quality attribute in meat animals. A substantial economic gain in the livestock industry may result from the discovery of metabolites, which can improve methods for predicting desirable livestock traits before harvest. Complex phenotypes can be identified for genetic selection or precision management using metabolic and genetic biomarkers. An analysis of ovine samples collected before harvest from wethers with different degrees of back fat was conducted using serum samples obtained from 15 sheep (5 from each back fat class), 60 d, 30 d, and just before harvest. This study quantified 55 serum metabolites that play important roles in ovine metabolism by 1H nuclear magnetic resonance (NMR) spectroscopy. An RNAseq dataset was generated from each sheep's muscle and adipose tissue at harvest. A statistical analysis of the data was performed using MetaboAnalyst Version 5.0 software in R. The results of multiple analyses showed that three metabolites, betaine, choline, and dimethylamine, are associated with sheep's fat class at a significance of (P &amp;lt; 0.01). Integration with gene expression datasets is still underway due to the limitations of annotating sheep genes. In addition, these metabolites are novel and have never been shown to modulate fat class in sheep. To elucidate methods of predicting which livestock produce ideal carcasses, further research is needed to investigate how betaine and choline impact fatness in sheep.

Reconstruction of ovine axonal cytoarchitecture enables more accurate models of brain biomechanics

There is an increased need and focus to understand how local brain microstructure affects the transport of drug molecules directly administered to the brain tissue, for example in convection-enhanced delivery procedures. This study reports a systematic attempt to characterize the cytoarchitecture of commissural, long association and projection fibres, namely the corpus callosum, the fornix and the corona radiata, with the specific aim to map different regions of the tissue and provide essential information for the development of accurate models of brain biomechanics. Ovine samples are imaged using scanning electron microscopy combined with focused ion beam milling to generate 3D volume reconstructions of the tissue at subcellular spatial resolution. Focus is placed on the characteristic cytological feature of the white matter: the axons and their alignment in the tissue. For each tract, a 3D reconstruction of relatively large volumes, including a significant number of axons, is performed and outer axonal ellipticity, outer axonal cross-sectional area and their relative perimeter are measured. The study of well-resolved microstructural features provides useful insight into the fibrous organization of the tissue, whose micromechanical behaviour is that of a composite material presenting elliptical tortuous tubular axonal structures embedded in the extra-cellular matrix. Drug flow can be captured through microstructurally-based models using 3D volumes, either reconstructed directly from images or generated in silico using parameters extracted from the database of images, leading to a workflow to enable physically-accurate simulations of drug delivery to the targeted tissue.

Investigation of Bovine, Caprine and Ovine Brucellosis in North Bahri, Sudan

The study was conducted at Bahri North, Khartoum State, Sudan, during the year 2021. The study was aiming at detection of bovine, caprine and ovine brucellosis in North Bahri. A total of 1050 bovine, 2081 caprine and 870 ovine blood samples were collected. Sera were separated from blood samples and kept at -20o at the laboratory of College of Veterinary Medicine, University of Bahri. Sera were tested using Rose Bengal Plate Test (RBPT) and modified Rose Bengal Plate Test (mRBPT) for detection of Brucella antibodies. By using RBPT for testing the sera for brucellosis, the prevalence of bovine brucellosis was 0.95, caprine brucellosis 11.24% and ovine brucellosis 1.14%. By using mRBPT for testing the sera for brucellosis, the prevalence of bovine brucellosis was 1.42, caprine brucellosis 13.83% and ovine brucellosis 1.95%. mRBPT is more sensitive than RBPT in detection of Brucella antibodies. Investigation of more animals covering the whole state, using of other serological and immunological test and vaccination of animals are recommended.

Analytical Approaches in Official Food Safety Control: An LC-Orbitrap-HRMS Screening Method for the Multiresidue Determination of Antibiotics in Cow, Sheep, and Goat Milk.

The presence of unauthorized substances, such as residues of veterinary medicines or chemical contaminants, in food can represent a possible health concern. For this reason, a complete legislative framework has been established in the European Union (EU), which defines the maximum limits allowed in food and carries out surveillance programs to control the presence of these substances. Official food control laboratories, in order to ensure a high level of consumer protection, must respond to the challenge of improving and harmonizing the performance of the analytical methods used for the analysis of residues of authorized, unauthorized, or prohibited pharmacologically active substances. Laboratories must also consider the state of the art of the analytical methodologies and the performance requirements of current legislation. The aim of this work was to develop a multiresidue method for the determination of antibiotics in milk, compliant with the criteria and procedures established by Commission Implementing Regulation (EU) 2021/808. The method uses an LC-Orbitrap-HRMS for the determination of 57 molecules of antibiotic and active antibacterial substances belonging to different chemical classes (beta-lactams, tetracyclines, sulfonamides, quinolones, pleuromutilins, macrolides, and lincosamides) in bovine, ovine, and goat milk samples. It provides a simple and quick sample pretreatment and a subsequent identification phase of analytes, at concentrations equal to or lower than the maximum residual limit (MRL), in compliance with Commission Regulation (EU) 2010/37. The validation parameters: selectivity, stability, applicability, and detection capability (ccβ), are in agreement with the requirements of Commission Implementing Regulation (EU) 2021/808 and demonstrated the effectiveness of the method in detecting veterinary drug residues at the target screening concentration (at the MRL level or below), with a false positive rate of less than 5%. This method represents an effective solution for detecting antibiotics in milk, which can be successfully applied in routine analyses for official food control plans.

Detection of Anaplasma phagocytophilum in Ovine Serum Samples—A Retrospective Study

Tick-borne fever in sheep is caused by Anaplasma phagocytophilum and this obligate intracellular bacterium multiplies mostly in neutrophil granulocytes of its host. In Europe, the pathogen is mainly transmitted by the tick species Ixodes ricinus. Data on the dissemination of A. phagocytophilum in the German sheep population are scarce. Hence, this pilot study aimed to investigate the presence of A. phagocytophilum in ovine serum samples retrospectively. The use of sera is beneficial because the specimens are available in large numbers and usually stored over a long period of time. Ten sheep flocks located in five federal states of Germany (Baden-Württemberg, Bavaria, Lower Saxony, North Rhine-Westphalia, Schleswig-Holstein) were included in the investigations, due to the high intra-flock seroprevalence (&gt;63%) of antibodies against the Anaplasma species. In total, 357 serum samples from seropositive sheep were analysed by 16S rRNA real-time PCR. DNA of A. phagocytophilum was detected in two ewes from the same farm in the northern German federal state of Schleswig-Holstein and represented an overall detection rate of 0.6%. Serum samples can be used to determine A. phagocytophilum, but sensitivity might be reduced in comparison to whole blood samples. Moreover, the sampling time may influence the detection of A. phagocytophilum in sheep, due to the seasonal pattern of tick activity in Germany. Our results support the findings of previous studies and demonstrate that A. phagocytophilum is also present in sheep from northern Germany (Schleswig-Holstein).

An ex-vivo model for the biomechanical assessment of cement discoplasty.

Percutaneous Cement Discoplasty (PCD) is a surgical technique developed to relieve pain in patients with advanced degenerative disc disease characterized by a vacuum phenomenon. It has been hypothesized that injecting bone cement into the disc improves the overall stability of the spinal segment. However, there is limited knowledge on the biomechanics of the spine postoperatively and a lack of models to assess the effect of PCD ex-vivo. This study aimed to develop a biomechanical model to study PCD in a repeatable and clinically relevant manner. Eleven ovine functional spinal units were dissected and tested under compression in three conditions: healthy, injured and treated. Injury was induced by a papain buffer and the treatment was conducted using PMMA cement. Each sample was scanned with micro-computed tomography (CT) and segmented for the three conditions. Similar cement volumes (in %) were injected in the ovine samples compared to volumes measured on clinical PCD CT images. Anterior and posterior disc heights decreased on average by 22.5% and 23.9% after injury. After treatment, the anterior and posterior disc height was restored on average to 98.5% and 83.6%, respectively, of their original healthy height. Compression testing showed a similar stiffness behavior between samples in the same group. A decrease of 51.5% in segment stiffness was found after injury, as expected. The following PCD treatment was found to result in a restoration of stiffness—showing only a difference of 5% in comparison to the uninjured state. The developed ex-vivo model gave an adequate representation of the clinical vacuum phenomena in terms of volume, and a repeatable mechanical response between samples. Discoplasty treatment was found to give a restoration in stiffness after injury. The data presented confirm the effectiveness of the PCD procedure in terms of restoration of axial stiffness in the spinal segment. The model can be used in the future to test more complex loading scenarios, novel materials, and different surgical techniques.

Circulating miRNAs in maternal plasma as potential biomarkers of early pregnancy in sheep.

MicroRNA (miRNA) plays an important role in the control of gene expression and is implied in many biological functions, including embryo implantation and development. The aim was to assess plasma miRNA profiles during the peri-implantation and ascertain potential candidate miRNA markers for early pregnancy diagnosis in ovine plasma. The plasma samples were obtained from a total of 24 ewes on days 12 (pre-implantation; P12, n = 4), 16 (implantation; P16, n = 4) and 22 (post-implantation; P22, n = 4) after mating, and on their corresponding days of 12 (Pre-C; C12, n = 4), 16 (Imp-C; C16, n = 4) and 22 (Post-C; C22, n = 4) of the estrous cycle. The miRNA profiles in plasma were assessed by microarray technology. We detected the presence of 60 ovine-specific miRNAs in plasma samples. Of these miRNAs, 22 demonstrated a differential expression pattern, especially between the estrous cycle and early pregnancy, and targeted 521 genes. Two miRNAs (oar-miR-218a and oar-miR-1185-3p) were confirmed using RT-qPCR in the ovine plasma samples. Protein-protein interaction (PPI) network of target genes established six functional modules, of which modules 1 and 3 were enriched in the common GO terms, such as inflammatory response, defense response, and regulation of immune response. In contrast, module 2 was enriched in the developmental process involved in reproduction, embryo development, embryonic morphogenesis, and regulation of the developmental process. The results indicate that miRNAs profiles of plasma seemed to be modulated during the peri-implantation stage of pregnancy in ewes. Circulating miRNAs could be promising candidates for diagnosis in early ovine pregnancy.

Ribotyping and assessment of toxigenic genes of clostridium difficile strains isolated from raw meat

Background: Clostridium difficile is an anaerobic, spore forming, rod shaped bacterium that can produce toxins and may transmit through some types of food samples. The present survey was done to assess the prevalence, toxigenic gene profile and ribotyping of C. difficile strains isolated from raw bovine and ovine meat samples. Methods: Two hundred and fifty raw meat samples were collected from Isfahan, Iran and analyzed for presence of C. difficile using the microbial culture. Genomic DNA was extracted from C. difficile isolates. Extracted DNA samples were subjected to PCR for determination of tcdA, tcdB, and cdtA toxigenic genes. Ribotyping was done using the 16S and 23S rRNA genes amplification in a separated PCR assay. Results: Fifteen out of 250 (6%) raw meat samples were contaminated with C. difficile. Prevalence of C. difficile strains amongst the raw bovine and ovine meat samples were 8% and 4%, respectively. The most commonly detected toxigenic genes were tcdA (33.33%) and cdtA (20%). Distribution of tcdB was 13.33%. Amongst the combined toxigenic genes, tcdA+cdtA had the highest distribution (13.33%).

On the microstructurally driven heterogeneous response of brain white matter to drug infusion pressure

Delivering therapeutic agents into the brain via convection-enhanced delivery (CED), a mechanically controlled infusion method, provides an efficient approach to bypass the blood–brain barrier and deliver drugs directly to the targeted focus in the brain. Mathematical methods based on Darcy’s law have been widely adopted to predict drug distribution in the brain to improve the accuracy and reduce the side effects of this technique. However, most of the current studies assume that the hydraulic permeability and porosity of brain tissue are homogeneous and constant during the infusion process, which is less accurate due to the deformability of the axonal structures and the extracellular matrix in brain white matter. To solve this problem, a multiscale model was established in this study, which takes into account the pressure-driven deformation of brain microstructure to quantify the change of local permeability and porosity. The simulation results were corroborated using experiments measuring hydraulic permeability in ovine brain samples. Results show that both hydraulic pressure and drug concentration in the brain would be significantly underestimated by classical Darcy’s law, thus highlighting the great importance of the present multiscale model in providing a better understanding of how drugs transport inside the brain and how brain tissue responds to the infusion pressure. This new method can assist the development of both new drugs for brain diseases and preoperative evaluation techniques for CED surgery, thus helping to improve the efficiency and precision of treatments for brain diseases.

Comparative profiling of agr locus, virulence, and biofilm-production genes of human and ovine non-aureus staphylococci

BackgroundIn a collaboration between animal and human health care professionals, we assessed the genetic characteristics shared by non-aureus staphylococci (NAS) infecting humans and dairy ewes to investigate their relatedness in a region concentrating half of the total National sheep stock. We examined by PCR 125 ovine and 70 human NAS for biofilm production, pyrogenic toxins, adhesins, autolysins genes, and accessory gene regulator (agr) locus. The microtiter plate assay (MPA) was used for the phenotypic screening of biofilm production. Ovine NAS included S. epidermidis, S. chromogenes, S. haemolyticus, S. simulans, S. caprae, S. warneri, S. saprophyticus, S. intermedius, and S. muscae. Human NAS included S. haemolyticus, S. epidermidis, S. hominis, S. lugdunensis, S. capitis, S. warneri, S. xylosus, S. pasteuri, and S. saprophyticus subsp. bovis.ResultsPhenotypically, 41 (32.8%) ovine and 24 (34.3%) human isolates were characterized as biofilm producers. Of the ovine isolates, 12 were classified as biofilm-producing while the remaining 29 as weak biofilm-producing. All 24 human isolates were considered weak biofilm-producing. Few S. epidermidis isolates harbored the icaA/D genes coding for the polysaccharide intercellular adhesin (PIA), while the bhp, aap, and embp genes coding biofilm accumulation proteins were present in both non-producing and biofilm-producing isolates. Fifty-nine sheep NAS (all S. epidermidis, 1 S. chromogenes, and 1 S. haemolyticus) and 27 human NAS (all S. epidermidis and 1 S. warneri) were positive for the agr locus: agr-3se (57.8%) followed by agr-1se (36.8%) predominated in sheep, while agr-1se (65.4%), followed by agr-2se (34.6%) predominated in humans.Concerning virulence genes, 40, 39.2, 47.2%, 52.8, 80 and 43.2% of the sheep isolates carried atlE, aae, sdrF, sdrG, eno and epbS respectively, against 37.1, 42.8, 32.8, 60, 100 and 100% of human isolates. Enterotoxins and tsst were not detected.ConclusionsConsiderable variation in biofilm formation ability was observed among NAS isolates from ovine and human samples. S. epidermidis was the best biofilm producer with the highest prevalence of adhesin-encoding genes.

Evaluation of Heller's reaction and the urine dipstick method alone or in combination to detect proteinuria in sheep

Urine dipstick and Heller's reaction are easy first-line screening tests for the detection of proteinuria; however, the performance of these methods in alkaline ovine urine is largely unknown. The aim of this study was to evaluate the diagnostic performance of Heller's reaction alone or in combination with dipstick for the detection of proteinuria in sheep, using the urine protein to creatinine ratio (UP/C) with two cut-off values as the reference method. Ninety-eight urine samples were collected from sheep using the transient apnea method. Heller's reaction, the dipstick method, and the UP/C ratio were used to assess proteinuria. The results were statistically analyzed twice, based on two different UP/C cut-off values of 0.2 and 0.5. Cohen's kappa value was used to determine the agreement between the UP/C ratios and Heller's reaction, the dipstick method, or the combination of methods. Sensitivity, specificity, and positive and negative likelihood ratios were calculated. ROC curves were also generated, and the areas under the curve (AUC) were evaluated to determine the optimal threshold for the numerical values of the two methods. Heller's reaction is more specific (96.67% and 96.00% when the cut-off value is 0.2 and 0.5, respectively) than the dipstick method, while the dipstick method was more sensitive (91.18% and 91.30%, when the cut-off value was 0.2 and 0.5, respectively) than Heller's reaction for the detection of proteinuria. Both tests were accurate when any grade >0 was considered positive. Proteinuria can almost be excluded in ovine urine samples with negative Heller's reaction and dipstick test.

Successful restoration of archived ovine formalin fixed paraffin-embedded tissue DNA and single nucleotide polymorphism analysis

Archived formalin fixed paraffin-embedded (FFPE) tissues are powerful tools in medicine, capable of harboring diagnostic and genetic answers to challenging clinical questions. Successful utilization of DNA derived from FFPE samples is dependent upon repairing DNA damage generated from the fixation process. Methods to repair FFPE DNA have been successful in human medicine for a variety of research and clinical applications, yet remain underutilized in veterinary medicine. Despite the available technology, our study is the first to evaluate the repair of FFPE derived DNA from veterinary species for single-nucleotide polymorphism (SNP) analysis using the Illumina OvineSNP50 BeadChip and Illumina FFPE QC and DNA Restore kit. To accomplish this, 48 ovine FFPE samples were run using the Illumina OvineSNP50 BeadChip with and without restoration. Compared to pre-restore data, we found increased sample call rates, SNP call frequency, and assay metrics for all samples post-restoration. Further, we utilized four sheep with available parallel fresh DNA and FFPE DNA to compare assay metrics and genotype calls between the two starting sample types. Although fresh samples generated increased call rates, we found 99% concordance in allele calls between restored FFPE and fresh DNA for all four samples. Our results indicate successful restoration and genotyping of ovine FFPE samples using this technology, with potential for utilization in other veterinary species.

A quantitative interspecies comparison of the respiratory mucociliary clearance mechanism

Collectively coordinated ciliary activity propels the airway mucus, which lines the luminal surface of the vertebrate respiratory system, in cranial direction. Our contemporary understanding on how the quantitative characteristics of the metachronal wave field determines the resulting mucociliary transport is still limited, partly due to the sparse availability of quantitative observational data. We employed high-speed video reflection microscopy to image and quantitatively characterize the metachronal wave field as well as the mucociliary transport in excised bovine, porcine, ovine, lapine, turkey and ostrich samples. Image processing techniques were used to determine the ciliary beating frequency (CBF), the velocity and wavelength of the metachronal wave and the mucociliary transport velocity. The transport direction was found to strongly correlate with the mean wave propagation direction in all six species. The CBF yielded similar values (10–15 Hz) for all six species. Birds were found to exhibit higher transport speeds (130–260 upmum/s) than mammals (20–80 upmum/s). While the average transport direction significantly deviates from the tracheal long axis in mammals, no significant deviation was found in birds. The metachronal waves were found to propagate at about 4–8 times the speed of mucociliary transport in mammals, whereas in birds they propagate at about the transport speed. The mucociliary transport in birds is fast and roughly follows the TLA, whereas the transport is slower and proceeds along a left-handed spiral in mammals. The longer wavelengths and the lower ratio between the metachronal wave speed and the mucociliary transport speed provide evidence that the mucociliary clearance mechanism operates differently in birds than in mammals.

Sex Determination of Unknown Ovine Samples Gathered from Slaughters

Accurate and specific methods for sex determination in farm animals play a vital role in animal resources. Therefore, this study was conducted to differentiate between males and females for unknown sheep samples collected from slaughters based on the amelogenin (AMELX/ AMELY) gene by using the polymerase chain reaction (PCR) technique. A total of 200 blood samples of Awassi sheep, collected from slaughterhouses, with an average age ranging between 3-6 years. Blood samples from the sheep were used to isolate the DNA. Specific PCR primers were used to amplify one fragment (262 bp) from the X- chromosome in ewes, and two fragments (262 and 202 bp) from the X and Y- chromosome in rams, respectively. The specificity of the primers was evaluated by using amelogenin amplicons for samples of known sex. The results revealed a single band for ewes (262 bp) and two bands for the rams (202 and 262 bp) after electrophoresis. In conclusion, this assay is proved to be a precise, inexpensive, and favorable method in sex determination, especially when unknown samples are collected from the slaughterhouse, and the samples are mixed so that it is difficult for the researcher to determine the sex of the samples. This method can be employed in other vertebrates and sexing of offspring in animal breeding because the amelogenin gene is conserved among mammals.

An Investigation of Contagious Agalactia Disease of Sheep and Goats in Isparta and Afyonkarahisar in Turkey

Background: Contagious agalactia causes significant economic losses. The aim of this study is to investigate the presence of contagious agalactia disease in cities of Isparta and Afyonkarahisar, Turkey. Methods: The study includes 45.500 animals in 220 ovine enterprises and samples were taken from those suspected of contagious agalactia disease. 202 animals in the 21 ovine enterprises comprised of 139 goats, 56 sheep, 3 kid goats, 2 goats, 2 lambs in total suspected of the disease were sampled. A total of 289 samples were collected, including 91 milk samples, 28 nasal swabs, 101 eye swabs, 8 joint fluids and 61 ear swabs. The isolates obtained after incubation were identified with polymerase chain reaction by using specific primers to assess film and spot formation, glucose fermentation, growth inhibition tests. Result: Three Mycoplasma spp. isolates obtained from 28 nasal swabs turned out to be negative for M. agalactiae after PCR analysis. Colony morphology, biochemical tests and growth inhibition tests revealed that one agent was M. arginine and the two factors were identified as M. ovipneumoniae with centerless colony morphology. The obtained results were confirmed with the polymerase chain reaction. None of the four factors causing contagious agalactia were isolated and identified.