The role of maternal triglycerides in the supply of lipids to the ovine fetus.

Maternal and fetal blood samples were obtained for up to 20 min after the injection of an emulsion of 14C-labelled glycerol tripalmitate into the circulation of pregnant ewes. No evidence was obtained for the direct transfer of intact triglycerides across the placenta. Results obtained suggest that there was a very rapid uptake of the triglyceride from the maternal circulation followed by hydrolysis and release of free fatty acids back into the maternal circulation.     

Serology of Campylobacter fetus fetus strains from four outbreaks of ovine abortion

As a result of a Ministry of Agriculture & Fisheries survey on ovine abortions, 76 isolates of Campylobacterfetus fetus were obtained. These isolates were from four farms in the southern Hawkes Bay, with an abortion incidence of 2.8% to 9.1%. Antisera to eight different strains of C. fetus fetus were made in rabbits. Strains were then examined using whole cell tube agglutination tests and sensitised Staphylococcal Protein A slide agglutination tests. Heat labile antigens were examined by absorbing antisera with heat-treated bacteria. Two broad serogroups were found, but within-group variation was demonstrated by cross-absorbing antisera. The isolates from one farm were all of a single broad serogroup. Both serogroups were found on the remaining three farms. Evidence for the presence of two major serogroups was also obtained by immobilisation tests and antigen analysis by gel diffusion.     

Histological development of the small intestinal mucosa in the ovine fetus.

Histological changes in the small intestine of 85 ovine fetuses were studied, with particular reference to the mucosa. In 24-day-old fetuses the endoderm of the intestine consisted of a single layer of epithelium. By the 26th day the epithelium was flattened and stratified. The first evidence of rudimentary villus formation was noted in 39-day-old fetuses. Intestinal glands were first observed at 56 days. Other mucosal structures were present by 101 days. At birth the intestinal mucosa was fully mature.     

Effect of passive transfer of maternal immune components on infection with ovine herpesvirus 2 in lambs

OBJECTIVE: To define the role of passively tranferred immunity in protection against early infection with ovine herpesvirus 2 (OvHV-2) in lambs. ANIMALS: 15 adult sheep and 34 lambs. PROCEDURES: 2 groups of animals were used, including 15 lambs born to OvHV-2-free ewes and 19 lambs born to OvHV-2-positive ewes. After nursing colostrum, all lambs and their dams were introduced into a flock positive for OvHV-2. Blood was obtained from the lambs every 2 weeks and examined by PCR assay and competitive inhibition ELISA. RESULTS: None of the animals had positive results by PCR analysis for samples obtained approximately 2 weeks after introduction into the flock. In the group of lambs from OvHV-2-infected ewes, 5 of 19 had positive results at 1 month of age and 17 of 19 by 5 months of age. In the group of offspring from OvHV-2-negative ewes, only 1 of 15 had positive results at 1 month of age, and the number reached 12 of 15 by 5 months of age. All lambs in both groups had positive results by 6 months. An active antibody response to the virus was detected in animals within 3 weeks after viral DNA became detectable in the blood. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis suggests that passively transferred immunity does not play an important role in the delay of infection with OvHV-2 in lambs. Age also does not seem to influence susceptibility. The rate of infection in young lambs may simply be a reflection of the intensity of viral exposure in their environment.     

Sequential sterile autolysis in the ovine fetus: macroscopic changes.

Sterile intrauterine autolysis was experimentally produced in 28 ovine fetuses in the last 3rd of gestation by umbilical artery ligation. The fetuses were retrieved by cesarean section after periods of sterile intrauterine autolysis ranging from 0.5 to 168 hours. The following autolytic changes, frequently reported as significant lesions in infectious abortions in cattle and sheep, were observed: subcutaneous blood-tinged gelatinous edema, blood-tinged fluid in the serous cavities, renal cortical softening, uniform reddish brown tissues, hepatic friability and mottling, and cloudy yellow to cloudy red abomasal contents. The appearance of these and other autolytic changes corresponded to sterile sequential autolytic changes reported in the rabbit fetus and used to determine duration of retention of stillborn infants and are of comparative biomedical significance. The sequential autolytic changes may be used as an index to duration of retention after death of a ruminant fetus and provide a basis for reappraising various infective abortifacients common to ruminants by allowing separation of specific lesions from strictly autolytic changes.     

Poor maternal nutrition inhibits muscle development in ovine offspring

Background:Maternal over and restricted nutrition has negative consequences on the muscle of offspring by reducing muscle fiber number and altering regulators of muscle growth.To determine if over and restricted maternal nutrition affected muscle growth and gene and protein expression in offspring,36 pregnant ewes were fed 60%,100%or 140%of National Research Council requirements from d 31 ± 1.3 of gestation until parturition.Lambs from control-fed(CON),restricted-fed(RES) or over-fed(OVER) ewes were necropsied within 1 d of birth(n = 18) or maintained on a control diet for 3 mo(n = 15).Semitendinosus muscle was collected for immunohistochemistry,and protein and gene expression analysis.Results:Compared with CON,muscle fiber cross-sectional area(CSA) increased in RES(58%) and OVER(47%)lambs at 1 d of age(P 0.01);however at 3 mo,CSA decreased 15%and 17%compared with CON,respectively(P 0.01).Compared with CON,muscle lipid content was increased in OVER(212.4%) and RES(92.5%) at d 1(P 0.0001).Muscle lipid content was increased 36.1%in OVER and decreased 23.6%in RES compared with CON at 3mo(P 0.0001).At d 1,myostatin mRNA abundance in whole muscle tended to be greater in OVER(P = 0.07) than CON.Follistatin mRNA abundance increased in OVER(P = 0.04) and tended to increase in RES(P = 0.06) compared with CON at d 1.However,there was no difference in myostatin or follistatin protein expression(P 0.3).Phosphorylated Akt(ser473) was increased in RES at 3 mo compared with CON(P = 0.006).Conclusions:In conclusion,maternal over and restricted nutrient intake alters muscle lipid content and growth of offspring,possibly through altered gene and protein expression.     

Poor maternal nutrition inhibits muscle development in ovine offspring

Background

Maternal over and restricted nutrition has negative consequences on the muscle of offspring by reducing muscle fiber number and altering regulators of muscle growth. To determine if over and restricted maternal nutrition affected muscle growth and gene and protein expression in offspring, 36 pregnant ewes were fed 60%, 100% or 140% of National Research Council requirements from d 31 ± 1.3 of gestation until parturition. Lambs from control-fed (CON), restricted-fed (RES) or over-fed (OVER) ewes were necropsied within 1 d of birth (n = 18) or maintained on a control diet for 3 mo (n = 15). Semitendinosus muscle was collected for immunohistochemistry, and protein and gene expression analysis.

Results

Compared with CON, muscle fiber cross-sectional area (CSA) increased in RES (58%) and OVER (47%) lambs at 1 d of age (P < 0.01); however at 3 mo, CSA decreased 15% and 17% compared with CON, respectively (P < 0.01). Compared with CON, muscle lipid content was increased in OVER (212.4%) and RES (92.5%) at d 1 (P < 0.0001). Muscle lipid content was increased 36.1% in OVER and decreased 23.6% in RES compared with CON at 3 mo (P < 0.0001). At d 1, myostatin mRNA abundance in whole muscle tended to be greater in OVER (P = 0.07) than CON. Follistatin mRNA abundance increased in OVER (P = 0.04) and tended to increase in RES (P = 0.06) compared with CON at d 1. However, there was no difference in myostatin or follistatin protein expression (P > 0.3). Phosphorylated Akt (ser473) was increased in RES at 3 mo compared with CON (P = 0.006).

Conclusions

In conclusion, maternal over and restricted nutrient intake alters muscle lipid content and growth of offspring, possibly through altered gene and protein expression.     

Immunohistochemical identification of Campylobacter fetus in natural cases of bovine and ovine abortions

An immunohistochemistry (IHC) procedure for the detection of Campylobacter fetus antigens using an avidin-biotin complex technique was performed on formalin fixed bovine and ovine fetal tissues from 26 natural cases of Campylobacter spp. abortion (four ovine and 22 bovine). The species of Campylobacter isolated included C. fetus ssp. venerealis from 13 bovine fetuses, C. fetus ssp. fetus from two ovine and one bovine fetus, Campylobacter jejuni from seven bovine fetuses, Campylobacter lari from two ovine fetuses and an unspeciated Campylobacter species in one bovine fetus. Histologic lesions identified in the aborted fetuses included placentitis, serositis, pneumonia, gastroenteritis, hepatitis and encephalitis. Campylobacter fetus antigens were identified by IHC in 13 of 13 bovine fetuses from which C. fetus ssp. venerealis was isolated and in two of two ovine fetuses from which C. fetus ssp. fetus was isolated. The IHC stains were negative in tissues from seven bovine fetuses from which C. jejuni was isolated, one bovine fetus infected with C. fetus ssp. fetus, one bovine fetus infected with the unspeciated Campylobacter and two ovine fetuses infected with C. lari. In positive cases, the IHC stain most frequently identified bacteria in the lung and gastrointestinal tract. The C. fetus IHC procedure performed on formalin fixed tissues is a practical tool for the diagnosis of natural cases of ovine and bovine abortion caused by C. fetus.     

Serogroups of Campylobacter fetus and Campylobacter jejuni isolated in cases of ovine abortion

Of 38 aborted ovine fetuses from 23 sheep flocks 29 C. fetus subsp. fetus and 22 C. jejuni were isolated and examined biochemically and serologically for heat-stable antigens. Serologic examinations were carried out by passive haemagglutination test. In case of C. fetus subsp. fetus strains alkaline antigen extraction was used. Antisera to two serogroups of C. fetus and to Penner serotype reference strains 1 to 60 were produced in rabbits. Abortion was caused in 18 (78.3%) flocks by C. fetus subsp. fetus and in 5 (21.7%) flocks by C. jejuni. Six C. fetus subsp. fetus strains grew well at both 43 and 25 degrees C. With one exception all C. fetus subsp. fetus were resistant, whereas all 29 C. fetus subsp. venerealis strains were sensitive to 30 micrograms/ml cefoxitin and cefamandole. These two cephalosporins can be used to differentiate the two subspecies of C. fetus. Passive haemagglutination test using alkaline antigen extraction is a proper method for the examination of heat-stable antigens of both C. fetus subspecies. Out of 24 C. fetus subsp. fetus strains 13 belonged to serogroup A(01), and 11 to serogroup B(02). C. jejuni strains examined belonged to Penner serogroup 1 (6 strains), to serogroup 5 (4 strains) and to serogroup 8 (4 strains).     

Comparison of surface antigens of some Campylobacter fetus subsp. fetus strains of ovine origin by polyacrylamide gel electrophoresis and immunoblotting.

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting were used to identify and to compare the surface antigens of eight C. fetus subsp. fetus strains. Seven strains (one of serogroup A and six of serogroup B) were isolated from aborted ovine fetuses, while one strain (serogroup A) originated from an aborted calf fetus. Saline extracts at 56 degrees C and 100 degrees C were used as antigens. Antisera were produced in rabbits. In saline extracts (56 degrees C) of the strains at least 19 fractions were identified by SDS-PAGE, with molecular masses ranging from approx. 4,800 to 205,000. The major bands appeared at 205,000, 66,000, 31,500, 25,000, 21,000 and 17,500. Despite the fact that the strains were cultured from 4 different sheep flocks and belonged to serogroup A or B, the SDS-PAGE profiles of the strains were very similar. When boiled (100 degrees C) extracts were used, a band migrating at 32,500 in sheep strains and a band at 97,500 in the calf isolate were missing. Most of the bands obtained by SDS-PAGE could be identified also by the immunoblot procedure. A or B type specificity of the ovine isolates was due to an LPS fraction, migrating at approx. 21,000, while the other LPS fractions appearing under this region although reacted with antisera did not influence the type specificity. Using alkaline extracts (pH 12) in SDS-PAGE, LPS fractions gave more pronounced profiles. In two of our C. fetus subsp. fetus isolates, plasmids with a molecular mass of 31,500 were identified.     

Ontogeny of immunity and leukocytes in the ovine fetus and elevation of immunoglobulins related to congenital infection

Certain humoral and cellular aspects of the immune response were observed during gestation in the ovine fetus. Emergence and development of leukocytes in peripheral blood were observed. Morphologically mature lymphocytes were consistently present at 32 days of gestation, but mature neutrophils were not neutrophils were not consistently present until gestational day 123. Monocytes were first observed at 63 days and eosinophils at 112 days of gestational age. Numbers of each class of leukocyte increased from the time of their appearance until just prior to birth when they declined. Production of immunoglobulin G (IgG) and immunoglobulin M (IgM) was monitored from the time of their appearance at 56 and 77 days, respectively, until birth. In contrast to consistently low base-line values of IgG and IgM (below 0.22 mg/ml and 0.21 mg/ml, respectively) found in clinically normal, unstimulated fetuses, the immunoglobulin values in experimentally infected animals increased with the age of the fetus. This finding can be utilized in diagnosis of several congenital infections and anomalies.     

Diagnostic studies of the fetus, placenta and maternal blood from 265 bovine abortions

During 1981, the fetus, placenta and maternal serum were received from each of 265 bovine abortions. These specimens were examined using histopathological, histochemical, bacteriological, mycological, endocrinological, immunological, serological and virological techniques. The cause of abortion was identified in 98 (37%) cases. Of these diagnosed abortions 27 (28%) were due to infection with fungi, 17 (17%) to Salmonella spp, 11 (11%) to Campylobacter fetus and 10 (10%) to Corynebacterium pyogenes infection. Of the remaining 33 (34%) diagnosed abortions 8 were due to Leptospira sp, 5 due to protozoan (probably Sarcocystis) encephalitis and 20 due to miscellaneous bacterial infections (including Brucella abortus) and fetal deformities. Bovine virus diarrhoea virus was isolated in 5 (2%) abortions, however the significance of the isolations was not known. Infectious bovine rhinotracheitis virus and chlamydia were not identified as causal agents in any abortion. In 18 (7%) abortions no definite etiologic agent was identified despite pathological findings suggestive of infection. Fetal heart blood serology and immunoglobulin values were not reliable indicators of infectious abortion. Of the remaining 149 (56%) abortions, 17% had high maternal serum cortisol levels and twin fetuses occurred in 9%.