Sheep Serum Research Articles

Characterisation of molten globule-like state of sheep serum albumin at physiological pH

Sheep serum albumin (SSA) is a 583 amino acid residues long multidomain monomeric protein which is rich in cysteine and low in tryptophan content. The serum albumins (from human, bovine and sheep) play a vital role among all proteins investigated until now, as they are the most copious circulatory proteins. We have purified SSA from sheep kidneys by a simple and efficient two-step purification procedure. Further, we have studied urea-induced denaturation of SSA by monitoring changes in the difference absorption coefficient at 287nm (Δε287), intrinsic fluorescence emission intensity at 347nm (F347) and mean residue ellipticity at 222nm ([θ]222) at pH 7.4 and 25°C. The coincidence of denaturation curves of these optical properties suggests that urea-induced denaturation is a bi-phasic process (native (N) state↔intermediate (X) state↔denatured (D) state) with a stable intermediate populated around 4.2–4.7M urea. The intermediate (X) state was further characterized by the far-UV and near-UV CD, dynamic light scattering (DLS) and fluorescence using 1-anilinonaphthalene-8-sulfonic acid (ANS) binding method. All denaturation curves were analyzed for Gibbs free energy changes associated with the equilibria, N state↔X state and X state↔D state in the absence of urea.

HIGH SEROPREVALENCE OF BLUETONGUE IN SHEEP, GOATS AND CATTLE IN ASSIUT GOVERNORATE, EGYPT

A total of 552 serum samples of sheep, goats and cattle from different localities at Assiut governorate, Egypt were screened for qualitative analysis of the BTV antibodies using a commercial competitive ELISA (cELISA) kit. The results showed an overall percentage of BTV positive sheep, goats and cattle serum samples were 41.86%, 24% and 85.42% respectively. A highest percentage of seropositivity was found inArab El- Awamer farm (100%), followed by Abnoub( 63. 64%), Sedfa(11. 43%), veterinary teaching hospital (11.11%), Aboteeg (3.57%) and Bani zeed el akrad (0% ) respectively. From examined sera all over the four years from 2012 to 2015 indicates prevalence of BTV antibodies by ( 62. 69%) during the non hot months and (10%) during the hot months, Female ruminants were more susceptible to the infection (61.29%) than male( 21.67%). The association between sex and prevalence of BT disease among ruminants observed in table (4). The results showed that the prevalence of BT was higher in female (61.29%) than male (21.67%). In conclusion, this study confirms the presence of blue tongue infection in Assiut Governorate, and cELISA were found to be sensitive and effective for screening of BTV group specific antibodies. The disease should be notifying its presence in Egypt since it was free from this disease and more studies should be attempets to estimate the serotype of blue tongue virus

A study of the correlation between Coxiella burnetii seropositivity and abortions in sheep in Eastern and Southeastern Turkey

Q fever is a zoonotic disease that occurs worldwide and is caused by the obligate intracellular bacterium Coxiella burnetii. Infected animals are usually asymptomatic, but infection can cause abortion and stillbirth in ruminants. The main purpose of this study was to evaluate prevalance of Coxiella burnetii infection in aborted and nonaborted sheep serum samples in Eastern Anatolia region by using enzyme-linked immunosorbent assay (ELISA). The determine of prevalance in sheep flocks from four provinces (Elazig, Malatya, Tunceli, Bitlis) and tested for anti-C.burnetii antibody detection, by means of Chekit Q fever Elisa kit. 350 serum samples obtained from flocks belonging aborted sheep showed that a total of 56 (16%) were detected seropositivity, whereas 171 serum samples obtained from nonaborted sheep flocks in 13 of the 171 (7.60%) for C.burnetii in seropositivity were observed. Coxiellosis should be considered an important cause of sheep with abortion history and nonaborted in Elazig and neighboring provinces.

Cellular immune responses induced in vitro by Ehrlichia ruminantium secreted proteins and identification of vaccine candidate peptides.

Secreted proteins are reported to induce cell-mediated immunity characterised by the production of interferon-gamma (IFN)-γ. In this study three open reading frames (ORFs) (Erum8060, Erum7760, Erum5000) encoding secreted proteins were selected from the Ehrlichia ruminantium (Welgevonden) genome sequence using bioinformatics tools to determine whether they induce a cellular immune response in vitro with mononuclear cells from needle and tick infected animals. The whole recombinant protein of the three ORFs as well as four adjacent fragments of the Erum5000 protein (Erum5000A, Erum5000B, Erum5000C, Erum5000D) were successfully expressed in a bacterial expression system which was confirmed by immunoblots using anti-His antibodies and sheep sera. These recombinant proteins were assayed with immune sheep and cattle peripheral blood mononuclear cells (PBMCs), spleen and lymph node (LN) cells to determine whether they induce recall cellular immune responses in vitro. Significant proliferative responses and IFN-γ production were evident for all recombinant proteins, especially Erum5000A, in both ruminant species tested. Thus overlapping peptides spanning Erum5000A were synthesised and peptides that induce proliferation of memory CD4+ and CD8+ T cells and production of IFN-γ were identified. These results illustrate that a Th1 type immune response was elicited and these recombinant proteins and peptides may therefore be promising candidates for development of a heartwater vaccine.

A novel technique for in vitro maturation of sheep oocytes in a liquid marble microbioreactor.

The aim of this work was to develop a microbioreactor using liquid marble (LM) as a novel system for oocyte in vitro maturation (IVM) in small volumes. Cumulus-oocyte complexes (COCs) obtained from slaughterhouse sheep ovaries were in vitro matured in a LM system prepared by placing a drop (30μl containing 10 COCs) suspended in TCM 199 supplemented with 10% (v/v) oestrus sheep serum (OSS) and 0.1IU FSH and LH onto a polytetrafluoroethylene (PTFE) particle bed (LM group). As a control group (CTRL group), COCs were in vitro matured in standard volume and conditions (600μl of IVM medium in a four-well dish). After 24-h culture at 38.5°C in 5% CO2 in air, COCs were released from LM and the following parameters were evaluated: (a) percentage of MII oocytes, (b) oocyte developmental competence following in vitro fertilization (IVF) or parthenogenetic activation (PA) and embryo culture for 8days in synthetic oviductal fluid (SOF) medium at 38.5°C in 5% O2, 5% CO2, and 90% N2. The results indicated similar percentage of MII oocytes in LM and CTRL groups (88.0 vs. 92.0%). No differences were observed in blastocyst rate after IVF (LM 47.5% vs. CTRL 50.2%, P=0.637) or PA (LM 44.4% vs. CTRL 48.3%, P=0.426). The results indicate that LM microbioreactor is a viable technique that provides a suitable microenvironment to induce oocyte in vitro maturation.

Motorized actuation system to perform droplet operations on printed plastic sheets.

We developed an open microfluidic system to dispense and manipulate discrete droplets on planar plastic sheets. Here, a superhydrophobic material is spray-coated on commercially-available plastic sheets followed by the printing of hydrophilic symbols using an inkjet printer. The patterned plastic sheets are taped to a two-axis tilting platform, powered by stepper motors, that provides mechanical agitation for droplet transport. We demonstrate the following droplet operations: transport of droplets of different sizes, parallel transport of multiple droplets, merging and mixing of multiple droplets, dispensing of smaller droplets from a large droplet or a fluid reservoir, and one-directional transport of droplets. As a proof-of-concept, a colorimetric assay is implemented to measure the glucose concentration in sheep serum. Compared to silicon-based digital microfluidic devices, we believe that the presented system is appealing for various biological experiments because of the ease of altering design layouts of hydrophilic symbols, relatively faster turnaround time in printing plastic sheets, larger area to accommodate more tests, and lower operational costs by using off-the-shelf products.

Epidemiology of Ovine Pasteurellosis in Lume District, East Shewa Zone of Oromiya Region, Ethiopia

Open Access Journal of Veterinary Science & Research ISSN: 2474 - 92 22 Epidemiology of Ovine Pasteurellosis in Lume District, East Shewa Zone of Oromiya Region, Ethiopia J Vet Sci Res Epidemiology of Ovine Pasteurellosis in Lume District, East Shewa Zone of Oromiya Region, Ethiopia Sadia H 1 , Abunna F 2 and Jarso D 2 * 1 Oromia Livestock and Fishery Resource Development Bureau, Addis Ababa, Ethiopia 2 Department of Veterinary Medicine and Agriculture, Addis Ababa University, Ethiopia *Corresponding author: Jarso D, Department of Veterinary Medicine and Agriculture, Addis Ababa University, Ethiopia, E - mail: djmojo74.ethio@mail.com Abstract The study of ovine pasteurellosis was conducted in Lume districts, East Shoa Zone of Oromia region, Ethiopia to determine the prevalence of M annheimia haemolytica , Pasteurella trehalosi and Pasteurella multocida from nasal swabs (384), abattoir specimens (145 ), and the serotype diversity among the species from sheep sera (150). A total of 115 isolates of M. haemolytica , P. treha losi and P. multocida were isolated from nasal swabs of apparently health and clinically sick sheep and from pneumonic lungs. The M. haemolytica, P. trehalosi and Pasteurella multocida were isolated from the nasal swabs (11.2%), (7.6%) and (2.1) respective ly, whereas M. haemolytica and P. trehalosi were isolated from pneumonic lungs (11.7%) , ( 10.3 ) respectively. However, Pasteurella multocida was the lowest among species isolated (2.1%). The overall isolation rate of M. haemolytica, P. trehalosi and Pasteurella multocida was 15.7%, 11.5% and 2.9%, respectively. From 145 lung samples collected and cultured, Pasteurella was isolated successfully in 35 (24.1%) sheep. Out of 35 sheep lung lesion sample, the percentage recovery rate of M. haemolytica 17(11 .7%), P. trehalose 15(10.3%) and P. multocida 3(2.1) % respectively. On the basis of these results, M. haemolytica and P.trehalose were the most common cause of pasteurellosis in sheep at the study area. A total of 150 sheep sera were examined for serotype specific antibodies using indirect haemagglutination test for M. haemolytica, P. trehalose and P. multocida serotypes. Variation in prevalence among the different serotypes was observed (P<0.001). The IHA test revealed that serotype A1, A2, A7, T3, T10, a nd T15 were the dominant serotypes with 23.3%, 42.6%, 32, 51.3, 29.3 and 30% positive by IHA whereas serotypes P. multocida biotype A and T4 were the least positive with 14.6% and16% respectively. Generally, both bacterial and serological results of this s tudy revealed that the causal agents of pasteurellosis are prevalent in the area, and serotypes A1, A2, A7, T3, T10 and T15 were dominant over the other serotypes

First report of seroprevalence of Toxoplasma gondii infection in sheep in Pomerania, northern Poland.

Toxoplasmosis is parasitic disease which has economic relevance for both veterinary and human medicine. In sheep, toxoplasmosis is a major cause of abortion and can thus cause reproductive problems. The current study aimed to determine the occurrence of anti-Toxoplasma gondii IgG antibodies in sheep from 13 districts of northern Poland and thereby obtain actual data about T. gondii seroprevalence in this population of animals. Blood samples from 1,646 animals from 99 herds were collected, and an in-house enzyme-linked immunosorbent assay (ELISA) based on native Toxoplasma lysate antigen (TLA) was used for serological testing. The diagnostic sensitivity of diagnostic test used in this study was 98.6%, and specificity 94.9% for the group of 113 sheep sera (74 seropositive and 39 seronegative) previously characterized by using an commercial agglutination test. Antibodies against T. gondii were found in 921 (55.9%) of all tested animals. The percentage of infected sheep was the highest (67.6%) for older animals (>6 years), whereas for younger ones it was significantly lower (50.1% - 57.2% for 1-5-year-old animals, respectively). Furthermore, a higher percentage of seropositive animals was noted among males (63%) than females (55.5%). The results also showed that the size of the herd is not a factor which may affect the seroprevalence of T. gondii infection in the examined population of sheep. The results of this study indicate that T. gondii infection in sheep from region of northern Poland is relatively high, and consumption of ovine meat and milk can be regarded as a significant source of infection for humans.

Evaluation of a bovine pregnancy-associated glycoprotein enzyme-linked immunosorbent assay kit for serological diagnosis of pregnancy in sheep

ABSTRACT: Pregnancy diagnosis is an important tool for farm management. Ultrasonography is the main technique used for pregnancy diagnosis in ewes. As an alternative, radioimmunoassay (RIA) allows accurate and early detection of pregnancy-associated glycoproteins (PAGs) in sheep blood. However, radioactive-based techniques, as RIA, have been increasingly inadvisable due to environmental risk. Homology between ovine and bovine PAGs is high, and ELISA kits used for PAGs detection in cattle are safer than RIA. Thus, this study aimed to evaluate the feasibility of PAGs detection for pregnancy diagnosis in sheep serum samples using an ELISA kit produced for cattle. The sensitivity and specificity of the ELISA kit were 93.5% and 98.9%, respectively, whereas positive and negative predictive values were 99.0% and 93.1%, respectively, in comparison to ultrasonography diagnostic (control). PAGs reached consistently detectable concentrations in ovine serum around 33 days after mating. Accuracy of the ELISA test was 96.1% from 33 days of pregnancy until lambing. After parturition, PAGs were still detectable seven days post-lambing. However, from 21 days post-parturition, PAGs from the previous pregnancy were no longer detected in serum samples. In conclusion, the bovine ELISA kit can accurately detect pregnancy in sheep 33 days following mating, while PAGs levels from the previous gestation are no longer detected from 21 days post-partum. The evaluated ELISA test is a reliable tool for pregnancy diagnosis in sheep at random stages and as a complementary exam at early gestation.

Cross antigenicity of immunodominant polypeptides of somatic antigen of Oesophagostomum columbianum with other helminths by western blotting.

Aim:Oesophagostomum columbianum in small ruminants in India is found as mixed infection commonly in sheep and goat. Haemonchus contortus, an abomasal nematode is found as concurrent infection with it. Eggs of Haemonchus and O. columbianum cannot be easily distinguished. Diagnosis of O. columbianum may only be possible if a non-cross antigenic polypeptide was available for immunodiagnosis.Materials and Methods:Somatic antigen (SoAg) of O. columbianum was fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunodominant polypeptides were identified by western blotting with homologous hyperimmune serum (HIS) and experimental sera of sheep or goat infected with other helminths.Results:SoAg of O. columbianum was immunoaffinity purified. Sharp polypeptide bands of 130, 72 and 68 KDa were observed along with several faint bands of lower molecular weight. Western blot of purified SoAg of O. columbianum with homologous HIS showed reaction with all the protein bands of 17, 28, 30, 32, 35, 38, 50, 68, 100, 130, 150, and 170 kDa. For identification of non-cross antigenic polypeptide, immunoaffinity purified SoAg of O. columbianum was reacted to heterologous HIS against H. contortus, Paramphistomum epiclitum, and Fasciola gigantica in western blotting utilizing completely dry method (i-blot). Among high molecular weight polypeptides 100 and 150 kDa were non-cross antigenic and among low molecular weight except 50 kDa polypeptide, 17, 30, 32, 35, and 38 kDa of O. columbianum were not cross antigenic with other helminths.Conclusions:Hence, polypeptides of 17, 30, 32, 35 and 38 kDa as well as 100 and 150 kDa polypeptides of O. columbianum may be exploited for immunodiagnosis of the infection in sheep and goat with extensive studies on cross antigenicity.

Specific detection of peste des petits ruminants virus antibodies in sheep and goat sera by the luciferase immunoprecipitation system

Peste des petits ruminants (PPR) is a contagious and often fatal transboundary animal disease affecting mostly sheep, goats and wild small ruminants. This disease is endemic in most of Africa, the Middle, Near East, and large parts of Asia. The causal agent is peste des petits ruminants virus (PPRV), which belongs to the genus Morbillivirus in the family Paramyxoviridae. This genus also includes measles virus (MV), canine distemper virus (CDV) and rinderpest virus (RPV). All are closely related viruses with serological cross reactivity.In this study, we have developed a Luciferase Immunoprecipitation System (LIPS) for the rapid detection of antibodies against PPRV in serum samples and for specific differentiation from antibodies against RPV.PPR and rinderpest (RP) serum samples were assayed by PPR-LIPS and two commercially available PPR cELISA tests. The PPR-LIPS showed high sensitivity and specificity for the samples tested and showed no cross reactivity with RPV unlike the commercial PPR cELISA tests which did cross react with RPV. Based on the results shown in this study, PPR-LIPS is presented as a good candidate for the specific serosurveillance of PPR.

A new immunoreactive recombinant protein designated as rBoSA2 from Babesia ovis: Its molecular characterization, subcellular localization and antibody recognition by infected sheep

Ovine babesiosis, caused by the intra-erythrocytic protozoan parasite Babesia ovis, is an infectious and economically important tick-borne disease of sheep. Diagnostic testing is an essential tool used for the control of the disease. In order to identify and characterize the immunoreactive proteins which are useful in serological diagnosis of the disease, a complementary DNA (cDNA) expression library was constructed from B. ovis merozoite mRNA. A cDNA clone designated as BoSA2 was identified by immunoscreening of a cDNA library using immune sheep serum. The sequence of the BoSA2 cDNA had a partial open reading frame of 1156 nucleotides encoding a polypeptide of 384 amino acid residues. Theoretical molecular mass for the mature protein was 43.5kDa. The sequence of the BoSA2 was inserted into the expression vector pGEX-4T-1 and then expressed in Escherichia coli DH5α cells as a glutathione S-transferase (GST)-tagged fusion protein. This recombinant fusion protein (rBoSA2) was purified by GST-affinity chromatography. Immunoreactivity of the rBoSA2 was evaluated by indirect enzyme-linked immunosorbent assay (ELISA) using the sera from the animals naturally and experimentally infected with B. ovis. ELISA results demonstrated that this antigen was useful for the diagnosis of ovine babesiosis. The localization of the BoSA2 protein was shown in and on the parasite and in the cytoplasm of the infected erythrocyte by confocal laser microscope. To our knowledge, rBoSA2 is the second immunoreactive recombinant protein of B. ovis until the present.

Serological evidence for the presence of influenza D virus in small ruminants

Influenza D virus (FLUDV) was isolated from diseased pigs with respiratory disease symptoms in 2011, and since then the new virus has also been spread to cattle. Little is known about the susceptibility of other agricultural animals and poultry to FLUDV. This study was designed to determine if other farm animals such as goats, sheep, chickens, and turkey are possible hosts to this newly emerging influenza virus. 648 goat and sheep serum samples and 250 chicken and turkey serum samples were collected from 141 small ruminant and 25 poultry farms from different geographical locations in the United States and Canada. Serum samples were examined using the hemagglutination inhibition (HI) assay and the sheep and goat samples were further analyzed using the serum neutralization assay. Results of this study showed FLUDV antibodies were detected in 13.5% (17/126) of the sampled sheep farms, and 5.2% (29/557) of tested sheep serum samples were positive for FLUDV antibodies. For the goat results, the FLUDV antibodies were detected in 13.3% (2/15) of the sampled farms, and 8.8% (8/91) of the tested goat serum samples were positive for FLUDV antibodies. Furthermore, all tested poultry serum samples were negative for FLUDV antibodies. Our data demonstrated that sheep and goat are susceptible to FLUDV virus and multiple states in U.S. have this virus infection already in these two species. This new finding highlights a need for future surveillance of FLUDV virus in small ruminants toward better understanding both the origin and natural reservoir of this new virus.

Estrous sheep serum enables in vitro capacitation of ram spermatozoa while preventing caspase activation

Estrous sheep serum (ESS) is considered the most efficient agent for in vitro capacitation of ram spermatozoa. We have explored the relationship between caspase activation and capacitation in ram. Semen samples from 17 rams were cryopreserved. In vivo fertility was evaluated after intrauterine artificial insemination. Samples were submitted to four treatments: control, ESS (10%), caspase inhibitor (Z-VAD-FMK), and estrous ewe serum plus caspase inhibitor (I + E). Sperm samples were incubated for 30 minutes at 38.5 °C and 5% CO2 and analyzed with flow cytometry for mitochondrial membrane potential (MitoTracker deep red), sperm viability and apoptosis-like changes (YO-PRO-1/propidium iodide), acrosomal status (peanut agglutinin–fluorescein isothiocyanate), membrane fluidity (merocyanine 540), and caspase activity (Vybrant FAM kits for polycaspases, caspase-8, and caspases 3–7). Estrous sheep serum induced changes compatible with capacitation, doubling the proportion of viable spermatozoa with increased merocyanine 540 and increasing YO-PRO-1+ and acrosome-reacted spermatozoa (P < 0.05). Incubation increased the proportion of spermatozoa with activated caspases (P < 0.05), which was abolished by the treatments. We detected a simultaneous decrease in the proportion of the viable and caspase− spermatozoa after the incubation, which was prevented by the presence of estrous ewe serum (P < 0.05). The analysis of caspases 3/7 and 8 resulted in less marked differences. Fertility was positively related to viability and inactivated caspases and negatively to viable-capacitated spermatozoa and active caspases. In vitro induction of capacitation in thawed ram spermatozoa by using ESS suggests a downregulation in apoptotic pathways. However, males with the lowest fertility showed parameters similar to high-fertility males, suggesting that other factors were involved apart from capacitation and/or caspase activation.

An Innovative Pseudotypes-Based Enzyme-Linked Lectin Assay for the Measurement of Functional Anti-Neuraminidase Antibodies.

Antibodies (Ab) to neuraminidase (NA) play a role in limiting influenza infection and might help reduce the disease impact. The most widely used serological assay to measure functional anti-NA immune responses is the Enzyme-Linked Lectin Assay (ELLA) which relies on hemagglutinin (HA) mismatched virus reassortants, or detergent treated viruses as the NA source to overcome interference associated with steric hindrance of anti-HA Ab present in sera. The difficulty in producing and handling these reagents, which are not easily adapted for screening large numbers of samples, limits the routine analysis of functional anti-NA Ab in clinical trials. In this study, we produced influenza lentiviral pseudoparticles (PPs) containing only the NA antigen (NA-PPs) with a simple two-plasmid co-transfection system. NA-PPs were characterized and tested as an innovative source of NA in the NA inhibition (NI) assay. Both swine A/California/07/2009 (H1N1) and avian A/turkey/Turkey/01/2005 (H5N1) N1s within NA-PPs retained their sialidase activity and were specifically inhibited by homologous and N1 subtype-specific, heterologous sheep sera. Moreover, A/California/07/2009 N1-PPs were a better source of NA compared to whole live and detergent treated H1N1 viruses in ELLA, likely due to lack of interference by anti-HA Ab, and absence of possible structural modifications caused by treatment with detergent. This innovative assay is safer and applicable to all NAs. Taken together, these results highlight the potential of NA-PPs-based NI assays to be developed as sensitive, flexible, easy to handle and scalable serological tests for routine NA immune response analysis.

Selenium speciation in paired serum and cerebrospinal fluid samples of sheep

This study was performed to characterise selenium (Se) and Se species in cerebrospinal fluid (CSF) of sheep and its relation to the respective Se concentrations in serum. Paired samples from 10 adult sheep were used for the study. Five sheep were fed a diet with a marginal Se concentration of <0.05mg Se/kg diet dry weight (dw, Se–), and five animals were fed the same diet supplemented with sodium selenite revealing a concentration of 0.2mg Se/kg diet dw (Se+). The feeding strategy was conducted for two years; The results on metabolic effects were published previously. At the end of the feeding period, paired samples of serum and CSF were collected and analysed using ion exchange chromatography inductively coupled plasma–dynamic reaction cell–mass spectrometry (IEC-ICP-DRC-MS) technique for total Se concentration and concentrations of Se species. Albumin concentrations were analysed additionally.The feeding strategy caused significant differences (p<0.01) in serum Se concentrations with 33.1±5.11μg Se/l in the Se– group and 96.5±18.3μg Se/l in the Se+ group, respectively. The corresponding total Se concentrations in CSF were 4.38±1.02μg Se/l and 6.13±1.64μg Se/l in the Se– and the Se+ group, respectively, missing statistical significance (p=0.077). IEC-ICP-DRC-MS technique was able to differentiate the Se species selenoprotein P-bound Se (SePP), selenomethionine, glutathione peroxidase-bound Se (Se-GPx), selenocystine, thioredoxin reductase-bound Se, ovine serum albumin-bound Se (Se-OSA), SeIV and SeVI in ovine serum and CSF. Quantitatively, SePP is the main selenoprotein in ovine serum followed by Se-GPx. The CSF/blood ratio of albumin (QAlbumin) reflected a physiological function of the blood-CSF barrier in all sheep. QSe-species were higher than QAlbumin both feeding groups, supporting the hypothesis of local production of Se species in the brain. Significant positive regression lines for CSF vs. serum were found for albumin and Se-OSA only, suggesting a role of albumin to convey Se across the blood-CSF barrier. The ovine model, together with the IEC-ICP-DRC-MS technique to characterise the Se species, might be a worthwhile model for further studies as repeated sample collection as well as modification of the nutritional status is feasible and effective.

Serologic detection of antibodies against Fasciola hepatica in sheep in the middle Black Sea region of Turkey.

The aim of the present study was to estimate the prevalence of Fasciola hepatica infection in sheep in the Black Sea region of Turkey. Samples from 213 sheep were collected randomly in Samsun, Tokat, and Sinop from September 2005 to January 2007 and tested by indirect enzyme-linked immunosorbent assay (ELISA) and Western blot analysis using F. hepatica excretory-secretory (E/S) antigens. The distribution of ELISA-positive samples for F. hepatica infections out of a total of 213 sheep serum samples was 23/71 (32.4%), 15/59 (25.4%), and 29/83 (34.9%) in Samsun, Sinop, and Tokat, respectively. The immunodominant proteins were determined by Western blot analysis using molecular weight markers of 14kDa, 20kDa, 24kDa, 27kDa, 33kDa, 45kDa, and 66kDa and extracted from sera of sheep that were positive for Fasciola spp. eggs and also hyperimmune sera from rabbits immunized with E/S antigens. The ELISA-positive results were confirmed by Western blot analysis. As a result, seroprevalence of F. hepatica infection was found in 31.4% of sheep from the Karayaka breed in the Middle Black sea region of Turkey.

Effects of different roughage sources and feeding levels on adipogenesis of ovine adipocytes.

The objective of the present study was to conduct an adipogenic evaluation of different roughage sources and feeding levels during ruminant adipocyte differentiation in vitro. Six wether sheep were divided into a timothy hay feeding group (TFG, n = 3) and an Italian ryegrass straw feeding group (IFG, n = 3). The sheep were fed high-roughage (HR), medium roughage (MR) and low-roughage (LR) diets in a one-way layout design each over a 6-day period. Sheep serum samples collected on the last day of each dietary treatment were added to an adipogenic induction medium for differentiation of preadipocytes derived from sheep subcutaneous adipose tissue. The cytoplasmic lipid accumulations in the TFG serum-treated preadipocytes were significantly higher than those of the IFG-serum treated preadipocytes on day 12. Messenger RNA expression of CCAAT/enhancer-binding protein (C/EBP)-α, C/EBP-β, C/EBP-δ, fatty-acid-binding protein (aP2) and stearoyl-coenzyme A desaturase (SCD) were regulated by each serum treatment. This study shows that different roughage source diets and roughage-to-concentrate ratio diets can regulate adipocyte differentiation via ruminant blood composition.

First Report of Coxiella burnetii Seroprevalence in Tibetan Sheep in China.

Query (Q) fever is a vector-borne zoonosis caused by the obligate intracellular pathogen Coxiella burnetii. Animals, including dogs, cats, cattle, and sheep, can be infected by C. burnetii. However, little information is available about C. burnetii infection in Tibetan sheep in China. Antibodies against C. burnetii in Tibetan sheep serums, which were collected from Maqu County, Tianzhu County and Nyingchi Prefecture, China, between 2011 and 2013, were examined by enzyme-linked immunosorbent assay (ELISA). Overall, 14.39% (304/2112, 95% confidence interval [CI] 12.90-15.89) of the examined Tibetan sheep were positive for C. burnetii infection. The C. burnetii seroprevalence in male and female Tibetan sheep was 12.94% (95% CI 10.26-15.61) and 14.98% (95% CI 13.18-16.78), respectively. The seroprevalence varied from 13.19% (95% CI 10.08-16.30) to 14.96% (95% CI 12.31-17.62) in Tibetan sheep in different areas. In addition, the seroprevalence ranged from 13.16% (95% CI 9.83-16.50) to 15.10% (95% CI 13.15-17.04) in different age groups. The results of the present survey indicate that C. burnetii infection is highly prevalent in Tibetan sheep in China. This is the first report of C. burnetii infection in Tibetan sheep in China, extending the host range for C. burnetii. Further studies are warranted to understand better the epidemiological situation of C. burnetii transmission in these areas and elsewhere.

Diagnosis of Toxoplasmosis in Sheep Using Serological (Elisa) and Molecular Technique in Duhok Governorate-Kurdistan Region

Toxoplasmosis is an important zoonotic diseases in human and animals. The disease caused by the protozoan Toxoplasma gondii. It is an economically important disease of livestock, especially sheep and goats. The present work aimed to diagnose toxoplasmosis in sheep using two methods, serological (ELISA) and molecular tools (PCR) and comparing the serological data with the molecular results to determine the sensitivity and the specifity of the molecular tools. The study was carried out at Duhok Research Center, University of Duhok. Ninety sex whole blood samples were collected from in aborted ewes in Duhok governorate during the period September 2013- September 2014.The samples were collected from different localities including Aqra, Dohuk district, Shikhan and Zakho The serological tests showed that22(22.91 %) of the samples were positive from examined sheep serum by ELISA, while 23(23.93%) by using PCR assay. PCR was performed on all DNA of sheep blood samples to amplify B1 gene as a target sequence. Good correlation between the results of PCR and ELISA were detected, there’s no statistically significant difference, It can be concluded that ELISA combined with the PCR technique is a recommended tool for accurate diagnosis of Toxoplasmosis but PCR is more specific for detection of T. gondii with sensitivity of 95.45 % and specificity of 97.29%.When taking the ELISA as a reference test. The results of PCR assay showed it’s important in the diagnosis of the carrier infected cases more than ELISA techniques.