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PubMed1989

The effects of immunization of sheep with Trichostrongylus colubriformis larvae on worm burdens acquired during grazing.

The effects of immunization of sheep with Trichostrongylus colubriformis larvae on worm burdens acquired during grazing. International Journal for Parasitology 19: 177-181. Romney sheep, reared helminth-free in pens to 5 months of age, were immunized against Trichostrongylus colubriformis by giving two doses of 200,000 T. colubriformis infective larvae at 15 day intervals to assess protection from natural challenge during grazing. Five immunized sheep and five unimmunized sheep were grazed on infested pasture for 4 weeks, and were then returned to the pens for 4 weeks before slaughter. Worm burdens, gastrointestinal histology and mucus antiparasite activity were examined at slaughter. Faecal egg counts and haematological examinations were carried out at regular intervals throughout the trial. Significant protection (P less than 0.05) was afforded immunized sheep against adult T. colubriformis (87%), T. axei (67%), Nematodirus spathiger (91%) and Ostertagia spp. (42%). Greater numbers of immature Nematodirus spp. and Ostertagia spp. were present in immunized sheep Overall, a significant (P less than 0.05) 42% reduction in total nematode burdens was afforded by immunization of the sheep with T. colubriformis larvae. Immunized sheep had significantly (P less than 0.05) more globule leukocytes, mast cells and eosinophils in gastrointestinal tissue and significantly (P less than 0.05) higher levels of mucous antiparasite activity than unimmunized sheep. Haematological observations showed some sheep had transient eosinophilia during immunization or grazing. Both immunized and unimmunized sheep showed depressed (P less than 0.05) total leukocyte counts during grazing which returned to pre-grazing levels within 1 week of return of the sheep to the pens. Overall, haematological parameters reflected parasite challenge and were unrelated to worm burdens acquired.

Animals↗
PubMed1995

Proliferating cell nuclear antigen expression in sheep infected with bovine leukemia virus.

Proliferating cell nuclear antigen (PCNA) or cyclin (C), a major nuclear protein, has been shown to be associated with human leukemia and malignancies. PCNA protein was quantitated in this study, in lymphocytes from bovine leukemia virus (BLV) infected and non-infected sheep, using two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and silver staining. The PCNA mean levels in lymphocytes of BLV-infected sheep (27 months post-infection) was significantly (P < 0.001) higher than in the lymphocytes of the non-infected sheep. The mean of PCNA levels in lymphocytes of sheep, 21 days after BLV infection, showed a two-fold increase compared with the non-infected sheep. Phytohemagglutinin (PHA) (3 days) treatment of lymphocytes from the non-infected and from the BLV-infected sheep resulted in a significant (P < 0.01) increase in the mean of PCNA levels only in the non-infected sheep. The mean lymphocyte counts of the BLV-infected sheep were not significantly different from the mean counts of the non-infected sheep at the time of lymphocyte protein analysis. Thus, these findings showed, similar to human leukemia and malignancies, that high levels of PCNA were found in lymphocytes from BLV-infected sheep compared with those from the non-infected sheep, and this was independent of high cell count. Our results also suggest that PCNA protein may play a role in the process of lymphoid transformation as a result of BLV infection of sheep.

Amino Acid Sequence↗
PubMed1995

The effect of propylthiouracyl-induced low thyroid function on secretion response and action of insulin in sheep.

The effect of propylthiouracyl (PTU)-induced low thyroid function on insulin responsiveness to glucose and glucose responsiveness to insulin in sheep was studied by performing hyperglycemic and euglycemic clamp experiments. All sheep were maintained at a level of 125% daily metabolizable energy intake and were housed in an environmental room that was maintained at 20 degrees C with a 16-hr lighting period. In the first study, eight female Suffolk sheep were divided equally into two groups and were subjected to oral PTU treatments of 4 mg/kg body weight (BW) per day for 7 d (low PTU) and 8 mg/kg BW per day for 14 d (high PTU). A hyperglycemic clamp experiment was conducted in each group on both control and PTU treatment periods. Plasma concentrations of triiodothyronine and thyroxine decreased (P < 0.05) in high PTU-treated sheep compared with those of low PTU-treated and control sheep. Both PTU treatments did not significantly influence basal insulin and glucose levels. Results of the hyperglycemic clamp experiment indicated that the mean plasma insulin increment and the ratio of mean plasma insulin increment to glucose infusion rate were significantly higher (P < 0.01) in high PTU-treated sheep than in low PTU-treated and control sheep. In the second study, the PTU treatment (8 mg/kg BW per day) was applied for 17 d in four male Suffolk sheep. A euglycemic clamp experiment with two insulin infusion rates (1.0 and 10.0 mU/kg per minutes) for two sequential periods of 2 hr each and thyroid hormone responses to intrajugular injection of thyrotropin-releasing hormone (1 microgram/kg BW) were performed in each sheep on both control and PTU treatment periods. In the euglycemic clamp experiment, the glucose infusion rate and the ratio of glucose infusion rate to mean plasma insulin increment were significantly reduced (P < 0.05) during the PTU treatment period for 10.0 mU/kg BW per minute of insulin infusion rate. The response areas of plasma thyroxine and triiodothyronine to thyrotropin-releasing hormone injection were blunted (P < 0.01) in PTU-treated sheep compared with those of control sheep. The high PTU treatment induced low thyroid function, enhanced insulin secretion response, and impaired insulin action in sheep.

Animals↗
PubMed2001

Paratuberculosis in sheep: its possible role in the epidemiology of paratuberculosis in cattle.

A total of 50 sheep originating from 15 Dutch farms with a known paratuberculosis infection in their cattle herd, but with no history of paratuberculosis infection in their sheep flock, were examined for infection with Mycobacterium avium subsp. paratuberculosis (Map). The sheep had been grazing on the same pastures as the cattle or on pastures fertilised with manure from these cows. The sheep were screened for paratuberculosis by serum biochemistry, serology and intradermal skin tests. At necropsy they were examined macroscopically, microscopically and bacteriologically for paratuberculosis. From 10 sheep, originating from eight flocks, Map could be isolated from various tissues but not from the intestinal contents, after an incubation period of 2.5-4 months. Six of these culture-positive sheep had no macroscopic signs of paratuberculosis at necropsy. Seven sheep were Map culture negative but showed macroscopic and microscopic lesions consistent with a paratuberculosis infection. Results of serology and skin tests did not correlate with the results of bacteriological culture. Serum concentrations of calcium, albumin and total protein of the infected, suspected and negative sheep were not different. These results indicate that a substantial number of the sheep examined were infected with Map. Even though this bacterium was not isolated from their faeces, the possibility that these sheep could have been shedding Map with their faeces below detection level or at a later stage of the disease cannot be eliminated. Map infected sheep should, therefore, be considered as a possible factor in the epidemiology of with Map infected cattle herds in The Netherlands. At necropsy bacteriological culture of Map should be performed on a routine basis to improve the diagnosis of paratuberculosis in sheep.

Animals↗
PubMed2000

Transmission of ovine herpesvirus 2 among adult sheep.

Previous studies from this laboratory have defined the pattern of acquisition of ovine herpesvirus 2 (OHV-2) in lambs under natural flock conditions. This study examined the question of whether OHV-2 could be transmitted between adult sheep. Two potential routes of transmission were examined: (1) direct inoculation of either viable leukocytes or whole blood from OHV-2 positive sheep, and (2) horizontal transmission through natural contact with OHV-2 positive sheep. Two groups of OHV-2 negative adult sheep were inoculated with material from infected sheep, one with 5x10(8) viable peripheral blood leukocytes (PBL), and the other with 100 ml of whole peripheral blood. No PCR signals were detected in any of the three sheep inoculated with the PBL during the 20 weeks following inoculation. In the group of five sheep inoculated with whole blood, two became PCR-positive at 7 and 8 weeks post-inoculation, respectively, and the remaining three sheep maintained their negative status until termination of the experiment at 20 weeks post-inoculation. In two experiments conducted in different flocks, a total of 20 adult sheep were used to examine horizontal transmission by contact; all animals became PCR-positive within 12 months of mixing the uninfected and infected animals. The results of these experiments support two conclusions. First, the susceptibility to OHV-2 is not limited to young lambs; adult sheep remain fully susceptible. Second, the fact that whole blood, but not PBL, from infected sheep was able to transmit the infection to only two of five inoculated sheep suggests that the infection in peripheral blood cells may be largely non-productive.

Animals↗
PubMed1991

Dermal and serological response against Pseudomonas aeruginosa in sheep bred for resistance and susceptibility to fleece-rot.

Genetically select lines of Merino sheep have been bred at Trangie (NSW Agriculture and Fisheries) for resistance (R) or susceptibility (S) to fleece-rot and flystrike. It is believed that fleece characters are primarily responsible for the R or S phenotype. When transferred to the wetter coastal environment of Sydney, R and S sheep with no more than 6 weeks wool cover, continued to show significant differences in the incidence and severity of fleece-rot dermatitis. To test the hypothesis that these sheep might also exhibit differences in their local skin reactions and immune responsiveness, 3 intradermal injections of killed Pseudomonas aeruginosa were administered at monthly intervals. After primary intradermal challenge, R sheep had a higher incidence of skin induration and a stronger inflammatory response (increased induration diameter) than S sheep. Compared to S sheep, R sheep also developed higher levels of circulating antibodies against whole cell antigen and both inner and outer membrane proteins of P. aeruginosa. These responses were maintained in R sheep with each consecutive challenge while S sheep showed a decline in their immune responsiveness. Differences in antibody response against outer membrane proteins were also detected when antigenically naive sheep from each genetic line were sensitised by epicutaneous challenge with P. aeruginosa under experimental wetting conditions. Intradermal challenge of these animals 6 months later with outer membrane proteins, revealed a late maximum (72 h) in the development of induration diameters for R sheep while S animals showed maximal induration diameters by 24 h. However, there was no significant difference in induration response between 24 h and 72 h within each group of sheep.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2006

The impact of malnutrition and other causes of losses of adult sheep in 12 flocks during drought.

OBJECTIVE: To establish the range and cost of losses in Merino flocks in southern New South Wales during drought conditions by determining the cause of death, morbidity or wasting in adult sheep. DESIGN AND POPULATION: Pathological studies were performed on 392 dead or moribund adult sheep from 12 Ovine Johne's disease (OJD)-infected flocks during 2002 and a further 58 sheep culled due to wasting from one of these flocks in 2003. Flocks ranged between 3,500 and 20,000 sheep. METHOD: The most likely cause of death, morbidity or wasting was determined following consideration of the environment in which the animal was found, clinical and gross pathological findings, plus histopathology of tissues collected during necropsy. RESULTS: A most likely cause of death, morbidity or wasting was determined for 362 sheep in 2002 and 58 sheep in 2003. OJD contributed to the death of 250 sheep in 2002, and wasting of 48 sheep in 2003. Of the sheep that died or were euthanased due to other causes, malnutrition was a contributing factor in the death of 70 sheep (63%) in 2002 and 2 sheep (20%) in 2003. Losses were not evenly distributed across flocks, with 57% of mortalities caused by malnutrition in 2002 occurring in one flock. Malnutrition accounted for 18% of the annual cost of all deaths among adult sheep in 2002 with an average cost of $16,882 per farm. Losses not attributed to malnutrition included a range of infectious and non-infectious disorders. These included balanoposthitis, clostridial enterotoxaemia, cutaneous myiasis, endoparasitism, enteritis, intestinal adenocarcinoma, misadventure, peritonitis, periparturient death of ewes, photosensitisation, pneumonia, post-shearing stress and squamous cell carcinoma of the perineum. CONCLUSION: Almost one third of mortalities in OJD-infected flocks during drought were unrelated to OJD and could be reduced by improving nutritional and disease management practices. The importance of close supervision of the flock is highlighted so that early management intervention can be instituted, including the culling of cases of welfare concern.

Animals↗
PubMed2002

Active surveillance for scrapie by third eyelid biopsy and genetic susceptibility testing of flocks of sheep in Wyoming.

Control of scrapie, an ovine transmissible spongiform encephalopathy or prion disorder, has been hampered by the lack of conventional antemortem diagnostic tests. Currently, scrapie is diagnosed by postmortem examination of the brain and lymphoid tissues for PrP(Sc), the protein marker for this group of disorders. For live, asymptomatic sheep, diagnosis using tonsil or third-eyelid lymphoid tissue biopsy and PrP(Sc) assay has been described. To evaluate the feasibility and efficacy of third-eyelid testing for identification of infected flocks and individual infected sheep, 690 sheep from 22 flocks were sampled by third-eyelid lymphoid tissue biopsy and immunohistochemistry. Sheep were further evaluated for relative genetic susceptibility and potential contact exposure to scrapie. Third-eyelid testing yielded suitable samples for 80% of the sheep tested, with a mean of 18.1 lymphoid follicles (germinal centers) per histologic section. Three hundred eleven of the sheep were sampled through passive surveillance programs, in which only sheep with potential contact with an infected sheep at a lambing event were tested, regardless of their scrapie susceptibility genotype. In addition, 141 genetically susceptible sheep with no record of contact with an infected animal at a lambing event were sampled through a targeted active surveillance program. Ten PrP(Sc)-positive sheep were identified through the passive surveillance program, and an additional three PrP(Sc)-positive sheep, including two from flocks with no history of scrapie, were identified through the active surveillance program. All PrP(Sc)-positive sheep had the highly susceptible PrP genotype. Third-eyelid testing is a useful adjunct to flock monitoring programs, slaughter surveillance, and mandatory disease reporting in a comprehensive scrapie eradication and research program.

Animals↗
PubMed1996

Natural transmission of orf virus from clinically normal ewes to orf-naive sheep.

The apparent natural transmission of orf virus from clinically normal ewes to susceptible sheep was observed during a border disease vaccine experiment. The 14 susceptible sheep were persistently infected with border disease virus and had been reared indoors in isolation from other sheep since birth. Their ages ranged from two to four years and they were housed in two groups; group 1 consisted of four sheep persistently infected with the Moredun strain of border disease virus and group 2 consisted of 10 sheep persistently infected with the Oban strain of the virus. On day 0, six sheep were removed from group 2 and rehoused. To the remaining four sheep in each group were added eight four- to six-year-old pregnant conventionally reared ewes at 48 days gestation. Fourteen days later the four sheep in group 1 were moved to another pen housing eight similar five-year-old pregnant ewes at 48 days' gestation, and the four sheep from group 2 were rehoused with their original stallmates. Twenty-one days later lip lesions typical of orf were first observed on the sheep from both groups and the disease spread to all the sheep persistently infected with border disease virus over the next four weeks. Virological and serological evidence demonstrated that the source of infection for the sheep was almost certainly the conventionally reared ewes, on which no lesions resembling orf were observed at any time during the study.

Animals↗
PubMed1990

Granulocyte depletion attenuates sustained pulmonary hypertension and increased pulmonary vasoreactivity caused by continuous air embolization in sheep.

Chronic pulmonary hypertension can develop in diseases associated with acute or repeated inflammation in the lungs, e.g., adult respiratory distress syndrome, chronic bronchitis. Inflammation has also been associated with some animal models of chronic pulmonary hypertension. We have previously shown that 12 days of continuous air embolization into sheep results in the functional and structural changes of chronic pulmonary hypertension. To determine whether granulocytes contribute to these changes, five sheep were granulocyte-depleted with hydroxyurea immediately before and during air embolization (AIR-PMN) and were compared with sheep receiving air embolization (AIR only). Air embolization was discontinued briefly every 4 days for monitoring of pulmonary vascular pressures and assessment of pulmonary vasoreactivity to a bolus injection of PGH2-A. After 12 days of air embolization, the lungs were removed for structural studies. AIR-PMN sheep did not develop the sustained increase in pulmonary artery pressure seen in the AIR sheep (Day 12, AIR-PMN = 20 +/- 3 cm H2O; AIR = 29 +/- 2; mean +/- SE). Similarly, the increased pulmonary pressor response to PGH2-A seen in AIR sheep was not found in the AIR-PMN group. Structural studies of the barium-injected lungs of AIR-PMN sheep revealed a twofold increase in medial thickness of normally muscular arteries and a significant increase in the percent of muscular intraacinar arteries (similar to findings in lungs from AIR sheep). The number of barium-filled arteries was increased in AIR-PMN sheep when compared with that in AIR sheep, but the number was still less than in the control sheep. We conclude that granulocytes may contribute to the functional changes of chronic pulmonary hypertension after continuous air embolization in sheep, but they do not play a role in structural changes involving pulmonary arterial smooth muscle cells and their precursors. The present data also suggest that the reduction in peripheral arterial filling is the structural alteration that contributes most to the sustained rise in pulmonary artery pressure. The data further suggest that pulmonary hypertension after air embolization may have a vasoconstrictive component that is granulocyte-dependent.

Animals↗
PubMed2004

Experimental transmission of ovine herpesvirus-2 in sheep.

Transmission of ovine herpesvirus-2 (OvHV-2) in sheep via natural contact and nasal secretions was examined. OvHV-2-free lambs were produced by separating newborn lambs from their mothers within 5 days of birth and raising them in an isolation facility. Transmission experiments via natural contact were conducted by keeping OvHV-2-free lambs with OvHV-2-infected sheep of different ages. Six of the infected ewes in this experiment were pregnant and gave birth during the experimental period. OvHV-2 was not transmitted from the adult sheep, though viral DNA was consistently detected in their peripheral blood leukocytes (PBL). On the other hand, OvHV-2 was transmitted from recently infected lambs to sheep at 10 or 12 weeks after the onset of contact. In addition, we attempted the experimental transmission of OvHV-2 via nasal secretions, by transferring nasal washings from infected sheep to the nostrils of uninfected sheep. Sheep receiving the nasal washings from infected adult sheep maintained their negative status for 15 months, whereas sheep receiving nasal washings from recently infected lambs acquired OvHV-2 by 8 months. The results of these experiments support that OvHV-2 is more easily transmitted to negative sheep by recently infected lambs than by adult sheep. Further, it is supposed that the nasal cavity is a portal for entry and shedding of infectious OvHV-2 in sheep.

Animals↗
PubMed2001

Influence of sheep breed and application site on the efficacy of a flumethrin pour-on formulation against ticks.

The objectives of this study were to determine the influence of application site and sheep breed on the efficacy of a flumethrin (1% m/v) solution for the control of 'bont'-legged (Hyalomma spp.) and red-legged ticks (Rhipicephalus evertsi evertsi). This study was conducted from November 1996 to February 1997 on 3 farms in the southwestern Free State Province. Two trials were conducted on Dorper sheep and 2 on Merino sheep. For each specific application 30 sheep were selected and allocated to 3 groups of 10 animals each using randomisation through minimisation, with pre-treatment total tick count as only criterion. Groups consisted of an untreated control group, a group treated with 3 ml of a flumethrin (1% m/v) solution applied only to the anogenital region, and a group treated at a dose rate of 1 mf flumethrin (1% m/v)/5 kg host body mass. The total dose volume for animals in the last group was divided into 3 equal parts and applied to the brisket/axillae, groin and anogenital regions respectively. Animals grazed under extensive farming conditions and were infested by ticks that occurred naturally in the environment. Ticks were counted and removed weekly over a 6-week period. In all 4 trials, Rhipicephalus e. evertsi was the dominant tick species, followed, in 3 of the trials, by Hyalomma spp. Efficacy (%) of control against ticks for Dorper sheep, treated only on the anogenital region, was variable, ranging between 29.5 and 97%. In Merino sheep the efficacy values ranged between 23.1 and 90 %. The site-spcific (anogenital region) efficacy of control against ticks infesting Merino sheep was in general 100% or almost 100%. In Dorper sheep the efficacy values were >80 % for 3-5 weeks. The efficacy (%) of control against ticks for sheep treated on the brisket/axillae, groin and anogenital regions was always higher compared to sheep treated only on the anogenital region. In Dorper sheep, efficacy of control was >80 % for up to 4 weeks and in Merino sheep >80 % for 6 weeks.

Administration, Topical↗
PubMed1996

Experimental contact of bighorn sheep (Ovis canadensis) with horses and cattle, and comparison of neutrophil sensitivity to Pasteurella haemolytica cytotoxins.

Peripheral blood neutrophils from horses, cattle, and Rocky Mountain bighorn sheep (Ovis canadensis canadensis) were evaluated for susceptibility to cytotoxin-dependent lysis of different biotypes and serotypes of Pasteurella haemolytica of domestic sheep, cattle, bighorn sheep, or mountain goat (Oreamnos americana) origin utilizing a cytotoxicity assay which measures the degree of bacteria cytotoxin-killing of neutrophils. All isolates of P. haemolytica (biotypes A and T) were noncytotoxic to horse neutrophils. Thirteen of 18 R haemolytica biotype A isolates were cytotoxic (> 50% neutrophil death in vitro) to bighorn sheep neutrophils, and four of 10 P. haemolytica biotype A isolates were cytotoxic to neutrophils of cattle; P. haemolytica biotype T (= Pasteurella trehelosi) isolates were noncytotoxic to neutrophils of bighorn sheep and cattle. When six bighorn sheep were pastured with three horses, only P. haemolytica biotype T isolates were recovered from the bighorn sheep throughout the study; Pasteurella spp. organisms were not isolated from the three horses. At initiation of a study where five bighorn sheep were pastured with three cattle, P. haemolytica biotype A, serotype 1, 2 was isolated from all three cattle, and only P. haemolytica biotype T isolates were recovered from the bighorn sheep. One bighorn sheep died in each of the horse and cattle copasturing experiments. Pasteurella haemolytica was not isolated from the bighorn sheep which died in the horse copasturing experiment. A noncytotoxic P. haemolytica biotype A, serotype 2 was isolated at necropsy from the bighorn which died in the cattle contact experiment. Based on these experiments, we believe bighorn sheep and horse association would not be detrimental to bighorns due to P. haemolytica induced pneumonia.

Animals↗
PubMed1991

Prevalence of pigment gallstones in sheep.

In a survey of 666 sheep at a slaughterhouse, gallstones (concretions with a diameter greater than or equal to 1 mm) were found in the gallbladder of 50 sheep (7.5%), sludge (concretions with a diameter less than 1 mm) was found in 9 sheep (1.4%), and sludge plus gallstones were found in 7 sheep (1.1%). Gallstones and sludge were associated, and were more frequent in lambs and females, compared with adults and males. Qualitative analysis of the stones revealed all to be pigment (bilirubin) stones. There was a statistically significant increase of biliary bilirubin (total and indirect quota) only in sheep with gallstones plus sludge, compared with control sheep without sludge or gallstones. Concentrations of bilirubin, cholesterol, phospholipids, total and single bile aids, and total and ionized calcium were similar in the bile of sheep with gallstones, sludge, or both and control sheep. Bacteriologic analysis of the bile in 10 sheep with gallstones and 10 controls revealed bacteria in 50% of the first group and in 75% of the second group (Escherichia coli in all sheep and Salmonella spp also in 1 sheep with gallstones). These findings confirm our earlier findings of a high prevalence of black pigment gallstones in sheep. On that basis, we suggest that gallstones are associated with high total bilirubin concentration in the bile, and deconjugating bacteria are common in the biliary tract of these animals.

Animals↗
PubMed1996

Evaluation of an agar gel immunodiffusion test kit for detection of antibodies to Mycobacterium paratuberculosis in sheep.

OBJECTIVE: To determine whether a commercially available agar gel immunodiffusion test approved for detecting antibodies to Mycobacterium paratuberculosis in cattle could be used for sheep. DESIGN: Experimental trial. SAMPLE POPULATION: Serum samples from 27 sheep confirmed to have paratuberculosis by means of acid-fast staining of smears of ileal mucosa, histologic examination of tissues, or bacteriologic culture; 7 sheep with clinical signs of paratuberculosis; and 55 sheep from 5 uninfected flocks. PROCEDURE: Serum samples were tested concurrently with the commercially available test and with a previously validated agar gel immunodiffusion test. Multiple samples collected from 13 infected sheep over a period of 6 years were also tested so that each test's ability to detect onset of seropositivity could be compared. RESULTS: For both tests, results for samples from all 55 uninfected sheep were negative, results for samples from 32 of the 34 sheep with paratuberculosis were positive, and results for the remaining 2 sheep with paratuberculosis were negative. Results of both tests were in agreement for 50 of 54 samples obtained from 13 infected sheep over time. The 4 samples for which results of the 2 tests disagreed were the fourth, eighth, and ninth of 10 samples from 1 sheep and the first of 6 samples from a second sheep. For all 4 samples, the commercially available assay yielded a weak-positive result, but the previously described test yielded a negative result. CLINICAL IMPLICATIONS: The commercially available agar gel immunodiffusion test approved for use in cattle may be useful in the differential diagnosis of paratuberculosis in sheep.

Animals↗
PubMed2025

Genomic insights into the population history of fat-tailed sheep and identification of two mutations that contribute to fat tail adipogenesis.

INTRODUCTION: Since their domestication, domestic sheep (Ovis aries) have been culturally and economically significant farming animals worldwide. Fat-tailed sheep serve as a unique genetic resource for understanding adipogenesis and adaptive evolution in livestock. OBJECTIVES: Several genomic analyses have been conducted on various sheep breeds to elucidate the genome and regulation mechanism of the fat tail trait, prior genomic studies have failed to reconcile conflicting evidence about the genetic basis of tail morphology, particularly regarding the roles of PDGFD and BMP2. METHODS: Here, we conducted whole-genome resequencing of 283 sheep, encompassing 66 domestic breeds and 5 wild ovine species, to investigate the domestication history and selection signatures of fat-tailed sheep. Additionally, we performed transcriptome sequencing on adipose tissue to identify differentially expressed genes and cellular assays to validate these results. RESULTS: Demographic analysis revealed that domestic sheep descended from Asiatic mouflon and fat-tailed sheep began to diverge from thin-tailed sheep approximately 4.4-7.5 thousand years ago in East Asia. Chinese indigenous sheep were classified into Mongolian, Kazakh, Tibetan, and Yunnan populations. The Yunnan population may have experienced more recent genetic introgression from wild species, rather than an independent domestication event. Moreover, many potential regions associated with the fat-tailed phenotype (DDI1, PDGFD, and BMP2) were identified by selective sweep and genome-wide association analyses. Additionally, a fine-scale analysis of fat-tailed and thin-tailed sheep revealed two novel mutations: a G/A missense variant of PDGFD (Chr15: 3900312) and a C/T missense variant of BMP2 (Chr13: 48462350), both of which were significantly associated with tail adiposity. Functional validation demonstrated that mutant A-PDGFD significantly activated PFGFD expression and reduced fat deposition compared to wildtype. The C-BMP2 mutant activated BMP2 expression and promoted preadipocyte fat deposition. CONCLUSION: Our study provides the first evidence that these genes jointly regulate fat tail development through complementary mechanisms: PDGFD promotes adipose expansion, whereas BMP2 modulates energy partitioning. These findings offer new insights into the evolutionary history of fat-tailed sheep and identify potential targets for precision breeding in small ruminants.

Animals↗
PubMed1992

Developmental regulation of insulin-like growth factor binding protein production: studies in fetal, postnatal, and pregnant sheep.

To assess the roles of developmental factors in the regulation of sheep IGFBP production at the cellular level, we characterized and compared the IGFBPs released by fetal, postnatal, and maternal sheep skin fibroblasts in culture with those in fetal, postnatal, and maternal sheep plasma. Sheep fibroblasts produced seven IGFBPs: a 36.5-41 kDa protein induced in vitro by IGF-I, likely representing oIGFBP-3; a 28.5 kDa protein that reacted with antisera to human IGFBP-2, likely representing oIGFBP-2; 25 and 27 kDa proteins induced in fetal fibroblasts by IGF-I; a 22 kDa protein that was inhibited by IGF-I, likely representing oIGFBP-4; and 21 and 23 kDa proteins labelled only by IGF-II, suggesting their similarities to IGFBP-6. The developmental pattern of IGFBP production by sheep fibroblasts in culture was similar in several respects to that observed in sheep plasma. For example, relative amounts of the 21, 22, and 28.5 kDa IGFBPs exceeded that of the 36.5-41 kDa protein in early fetal fibroblast conditioned media and in fetal plasma, while the relative concentrations of the 36.5-41 kDa protein increased markedly during the perinatal period. Sheep plasma differed, however, in two major respects from fibroblast conditioned media: First, fetal, and to a far lesser extent maternal, plasma contained a 200 kDa IGF-II-selective BP, likely to be the circulating form of the IGF-II receptor; and second, plasma, unlike conditioned media, contained a 26 kDa IGFBP, likely to be oIGFBP-1. The results of our studies suggest that the production and release of IGFBPs by isolated sheep fibroblasts is regulated by developmental factors operative under in vitro culture conditions. The differences in the relative levels of IGFBPs in conditioned media from fetal, postnatal, and maternal sheep fibroblasts resemble in several respects the differences in the relative concentrations of the various IGFBPs in fetal, postnatal, and maternal sheep plasma. Thus, sheep fibroblasts provide a useful though imperfect model system by which to examine the nutritional and hormonal regulation of sheep IGFBP production at various developmental stages.

Animals↗
PubMed1995

Thirteen loci physically assigned to sheep chromosome 2 by cell hybrid analysis and in situ hybridization.

Sheep x hamster cell hybrids containing sheep metacentric Chromosome (Chr) 2 were produced by fusing blood leukocytes from normal sheep with hamster auxotrophic Ade F-minus mutants. Cell clones that were isocitrate dehydrogenase 1 (IDH1) positive were cytogenetically characterized, confirming that they contained sheep Chr 2. The following loci were newly assigned by Southern hybridization to sheep Chr 2: lipoprotein lipase (LPL), glycoprotein-4-beta galactosyltransferase 2 (GGTB2), neurofilament light polypeptide (68 kDa; NEFL), surfactant-associated protein 2 (SFTP2), lymphocyte-specific protein tyrosine kinase (LCK), and nebulin (NEB). These new assignments and the in situ localization of gelsolin (GSN) to sheep Chr 2pter-p24 are consistent with the predicted homology of cattle Chr 8 (U18) with sheep Chr 2p, and of cattle Chr 2 (U17) with sheep 2q. In addition, the assignment by cell hybrid analysis of loci previously mapped to Chr 2 in sheep, viz., cholinergic receptor, nicotinic, delta polypeptide (CHRND), collagen type III alpha 1 (COL3A1), fibronectin 1 (FN1), isocitrate dehydrogenase (IDH1), and villin 1 (VIL1), confirmed the localization of sheep syntenic group U11 to this chromosome. By nutritional selection and complementation of the hamster auxotrophic Ade F mutation, the multifunctional enzyme locus phosphoribosylaminoimidazolecarboxamide formyltransferase (AICAR transformylase)/IMP cyclohydrolase (inosinicase) (provisionally given the symbol PRACFT) has also been newly assigned to sheep Chr 2. This report significantly extends the number of loci physically mapped to sheep Chr 2 and confirms its close homology with cattle Chrs 2 and 8.

Animals↗