Vagal reflex inhibition of motility in the abomasal body of sheep by antral and duodenal tension receptors

Vagally-mediated regulation of motility in the abomasal body by duodenal and abomasal antral motility was demonstrated in acutely prepared anaesthetized sheep. The enteric plexuses between the abomasal body, antrum and duodenum were interrupted by transection. Antral contractions were more effective than duodenal contractions at causing inhibition of the abomasal body, and antral isometric conditions were more effective than antral isotonic conditions. Inhibition of motility in the abomasal body was reduced by unilateral cervical vagotomy, was abolished by bilateral cervical vagotomy, and was reversibly inhibited by cervical vagus cold block. The demonstration of vagal pathways in abomaso-abomasal reflexes confirms a functional homology of the mechanisms in the ruminant and animals with simple forms of stomach.Abbreviations b-t balloon-tipped - EMG electromyography - NANC non-adrenergic, non-cholinergic - o-t open-tipped     

Scrapie resistance alleles are not associated with lower prolificity in Rasa Aragonesa sheep

Scrapie is a prion disease characterised by the accumulation of the pathological associated form of cellular prion protein (PrP(SC)) in the central nervous system. Susceptibility to scrapie is associated with polymorphism in the ovine prion protein (PrP) gene. The European Union has implemented scrapie control programs, relying on selective breeding for scrapie resistance; the use of ARR-carrier and the exclusion of VRQ-carrier were recommended. In this study, 4323 individuals from Rasa Aragonesa Sheep breed were genotyped for the PrP gene and the individual estimated breeding values (EBV) for prolificity were calculated. Most represented PrP alleles do not work against prolificity. Only a significant association between VRQ/VRQ genotype and a lower EBV was observed (p = 0.027, eta2 = 0.002). Therefore, avoiding reproduction of VRQ/VRQ individuals would not cause negative effect regarding prolificity.     

Dietary protein for young grazing sheep: Interactions with gastrointestinal parasitism

Effects of protein supplementation and of nematode control on production responses in young grazing sheep and on nematode population dynamics were assessed. Young Merino wether sheep (n = 270) were allocated to one of three supplementation (Su) treatments and one of three regimes of nematode control (drench, Dr) in a factorial design. Each of the nine treatments was replicated three times in a randomised complete block. Animals received no supplement (SuO) or were given supplements at a level equivalent to 200 g day⁻¹, fed three times a week. The supplements contained 25% lucerne meal with either 75% sunflower meal (Su1) or 75% formaldehyde-treated sunflower meal (Su2). Nematode control regimes were: not drenched unless survival of individual animals was threatened (DrO); drenched according to a strategic drenching programme (Dr1); treatment with a controlled-release albendazole capsule (Dr2). Infections with Haemonchus contortus were suppressed by use of closantel. Due to drought conditions and lack of pasture growth, lucerne hay was fed to all sheep from Week 14 onwards (350 g per head per day, fed twice a week). Supplementation reduced the need for ‘survival drenching’ in Dr0 sheep considerably. Live-weight gain was increased significantly by supplementation with Su1 or Su2 during the 36-week experiment. Undrenched animals given Su1 or Su2 tended to grow faster than unsupplemented animals in the Dr1 and Dr2 groups. Greasy wool production and fibre diameter were increased by supplementation and anthelmintic treatment. Faecal worm egg counts (FEC) in undrenched sheep were significantly lower for Su1 and Su2 sheep than for Su0 sheep. There were no effects of supplementation on FEC in Dr1 or Dr2 sheep. Worm burden was generally unaffected by supplementation, while the drenching treatments Dr1 and Dr2 reduced worm burden at Week 16 and to a lesser extent at Week 27. Trichostrongylus colubriformis was the predominant nematode species, with smaller numbers of Nematodirus spp., Trichostrongylus vitrinus, Trichostrongylus axei and Ostertagia circumcincta also present. It was concluded that supplementary feeding with protein meal substantially reduced production losses attributable to nematode infections in young grazing sheep. This appears to be due to an enhanced resilience of the host, rather than any major changes in development of protective immunity.     

Interaction between nutrition and gastrointestinal parasitism in sheep

Effects of gastrointestinal nematode infection on metabolism and nutrient utilisation in sheep are reviewed. Infection induces protein deficiency by increasing the demand for amino acids in the alimentary tract while reducing supply through depression of appetite. Mechanisms through which improved protein nutrition could improve the performance of the host are then discussed. Opportunities for capitalising on such effects are limited by our rudimentary understanding of the cell-mediated immune response in gastrointestinal epithelial tissue. Both resistance of the animal to larval establishment and performance in the face of larval challenge can be enhanced by improved protein nutrition. However, enhanced immune responses may not necessarily be synonymous with improved productivity except at luxurious levels of protein intake, because of apparently competing demands for protein. Such levels of protein nutrition are difficult to achieve in pasture-based systems, because of the protein limiting role of the rumen. Work with protein-protecting tannins to overcome this limitation is discussed. The much more limited evidence for effect of mineral nutrition, particularly copper (Cu), molybdenum (Mo), cobalt (Co) and phosphorus (P), on outcome of larval challenge is also reviewed.     

Neural and chemical control of abomasal secretion in sheep.

Abomasal secretion has been collected from Grosskopftype pouches prepared in five adult Karrakul ewes. Feeding caused a marked increase in volume, acidity and pepsin activity, whereas these factors were reduced by starvation. Both insulin-induced hypoglycaemia and electrical stimulation of the vagal nerve supply to the pouch increased the volume, total acid and pepsin secretion. It is concluded that vagal activity is an important factor in the control of abomasal secretion in sheep.     

Interactions between nutrition and gastrointestinal parasitism in sheep

Effects of gastrointestinal nematode infection on metabolism and nutrient utilisation in sheep are reviewed. Infection induces protein deficiency by increasing the demand for amino acids in the alimentary tract while reducing supply through depression of appetite. Mechanisms through which improved protein nutrition could improve the performance of the host are then discussed. Opportunities for capitalising on such effects are limited by our rudimentary understanding of the cell-mediated immune response in gastrointestinal epithelial tissue. Both resistance of the animal to larval establishment and performance in the face of larval challenge can be enhanced by improved protein nutrition. However, enhanced immune responses may not necessarily be synonymous with improved productivity except at luxurious levels of protein intake, because of apparently competing demands for protein. Such levels of protein nutrition are difficult to achieve in pasture-based systems, because of the protein limiting role of the rumen. Work with proteinprotecting tannins to overcome this limitation is discussed. The much more limited evidence for effect of mineral nutrition, particularly copper (Cu), molybdenum (Mo), cobalt (Co) and phosphorus (P), on outcome of larval challenge is also reviewed.     

Serum and abomasal antibody response of sheep to infections with Haemonchus contortus

Serum and abomasal IgA, IgG and IgM antibody response against adult worm, L3 and egg antigens of Haemonchus contortus was monitored by the ELISA technique after one or two infections with this nematode. Following the first infection, antibody levels in serum did not change materially. After administration of a challenge dose of infective larvae, antibodies of the three immunoglobulin classes in infected animals rose slightly, but this rise appeared later than the fall in the faecal egg counts. In contrast, in abomasal mucosa, IgA anti-larval antibody levels, which did not increase materially after the primary infection, rose rapidly after a transient inhibition when sheep were challenged. A close temporal relationship was observed between the rise in local anti-worm IgA antibodies and the self-cure reaction, but antibody levels fell rapidly after worm diminution. The local antibody response was thus considered to be related to immunity of sheep to H. contortus.     

Evidence for a parasite-mediated inhibition of abomasal acid secretion in sheep infected with Ostertagia leptospicularis

The acid secretory capacity of the abomasal mucosa was studied in sheep experimentally infected with Ostertagia leptospicularis. The acidity of the abomasal contents, permanently recorded by a pH probe located inside the abomasum, decreased markedly to mean levels between pH 5 and 6. Subcutaneous administration of histamine or carbachol successfully stimulated acid secretion (pH 3.4). The results indicate that the abomasal mucosa harboured a population of functional parietal cells which were also identified immunohistochemically (H(+)/K(+)-ATPase). Ultrastructural investigation before stimulation revealed that the majority of these cells was in a resting state. Despite high serum gastrin levels, the acid secretion was blocked either at the level of the parietal cell or the enterochromaffin-like cell by an unknown factor, possibly mediated by the parasites. This is the first report of a parietal cell dysfunction associated with a nematode infection in the abomasum. It is suggested that the parasites induce changes in their environment which favour their survival and/or increase their reproduction.     

Moxidectin and ivermectin metabolic stability in sheep ruminal and abomasal contents

The oral administration of macrocyclic lactones to sheep leads to poorer efficacy and shorter persistence of the antiparasitic activity compared to the subcutaneous treatment. Gastrointestinal biotransformation occurring after oral treatment to ruminant species has been considered as a possible cause of the differences observed between routes of administration. The current work was addressed to evaluate on a comparative basis the in vitro metabolism of moxidectin (MXD) and ivermectin (IVM) in sheep ruminal and abomasal contents. Both compounds were incubated under anaerobic conditions during 2, 6 and 24 h in ruminal and abomasal contents collected from untreated adult sheep. Drug concentrations were measured by high-performance liquid chromatography with fluorescence detection after sample clean up and solid phase extraction. Neither MXD nor IVM suffered metabolic conversion and/or chemical degradation after 24-h incubation in ruminal and abomasal contents collected from adult sheep. Unchanged MXD and IVM parent compounds represented between 95.5 and 100% of the total drug recovered in the ruminal and abomasal incubation mixtures compared with those measured in inactive control incubations. The partition of both molecules between the solid and fluid phases of both sheep digestive contents was assessed. MXD and IVM were extensively bound (>90%) to the solid material of both ruminal and abomasal contents collected from sheep fed on lucerne hay. The results reported here confirm the extensive degree of association to the solid digestive material and demonstrates a high chemical stability without evident metabolism and/or degradation for both MXD and IVM in ruminal and abomasal contents.     

Absence of hypobiosis in abomasal nematodes of sheep and goats in Egypt

A total of 96 abomasa from sheep and goats at the Zagazig abattoir, Sharkia Province, Egypt, were examined, during the four climatic seasons from April 1986 to March 1987, for the presence of adult and larval nematodes. Adults of Trichostrongylus axei were most numerous followed by adult Haemonchus contortus. Adult Ostertagia sp. and Parabronema skrjabini were less prevalent. There was no significant difference between sheep and goats in infection rates, but worm burdens of the nematodes other than Ostertagia sp. were significantly higher (P less than 0.05) in sheep than in goats. The animals were uniformly infected with these parasites throughout the year as there were no significant differences in the incidence or intensity of infections between the four seasons. The numbers of larvae recovered from the wall of the abomasa were small in relation to the total adult worm population throughout the year with a slight increase during the summer. Hypobiosis, therefore, seems not to be important in the life cycle of abomasal nematodes of sheep and goats in Sharkia Province, Egypt.     

Abomasal impaction associated with anorexia and mortality in lambs

Anorexia in forty-six (6.1%) out of 753 neonatal lambs was investigated. Thirty-five of these lambs (76.1%) died while eleven (23.9%) recovered after treatment. Coagulated rubber-like milk clots were found in fifteen (42.9%), phytobezoars in nine (25.7%) and trichophytobezoars and coarse straws in eleven (31.4%) of the dead lambs.     

Vagal afferent activities and respiratory reflexes during drug-induced bronchoconstriction in the guinea pig.

Vagal afferent activities and respiratory reflexes during drug-induced bronchoconstriction were studied in 31 anesthetized, spontaneously breathing or artificially ventilated guinea pigs. Histamine (5, 10, 20 micrograms/kg), ACh (10, 20, 40 micrograms/kg) and endothelin-1 (2 micrograms/kg) were intravenously injected to the animals in order to induce the bronchoconstriction. In spontaneously breathing and vagi intact animals, a considerable respiratory change characterized by rapid-shallow breathing was elicited by histamine. Such respiratory change was abolished by bilateral vagotomy, indicating that the vagal pathway fairly participated in the respiratory change during bronchoconstriction. Indeed, recordings of electrical activities of single vagal afferent nerve fibers from pulmonary stretch and irritant receptors elucidated that the bronchoconstriction by the three drugs markedly influenced these receptor activities. The response of stretch receptors to bronchoconstriction was grouped into four types: two of those types showed a marked increase in their activities and the other two a decrease or no change. Such uneven response was assumed to be derived from heterogenous contraction and aeration among the intrapulmonary small airway. On the other hand, irritant receptors were invariably stimulated by increased transmural pressure during bronchoconstriction. Administration of isoproterenol (20 micrograms/kg) which inhibited the smooth muscle contraction abolished stimulatory effect of the drugs to irritant receptors, suggesting that the effect was due to indirect action through the muscle contraction rather than their direct action to the nerve endings.     

Effect of parasitism with Ostertagia circumcincta on pharmacokinetics of fenbendazole in sheep

Plasma and abomasal fluid concentrations of fenbendazole and its two major metabolites in sheep experimentally infected with Ostertagia circumcincta were compared with those in the same sheep when non-parasitised. Bio-availability of the drug was reduced in the parasitised state. There was also a reduction in the proportion of drug present in the form of metabolites in parasitised as compared with non-parasitised animals.     

Laboratory findings associated with abomasal ulcers/tympany in range calves

The etiology of abomasal ulcers/tympany was investigated in 48 animals from 36 ranches in Wyoming and Nebraska. Results indicate that subclinical trace mineral deficiencies of copper and/or selenium exist in the range cattle in west central Nebraska and Wyoming. Etiological agents most frequently incriminated by bacteriologic cultures and/or histopathic examination were Clostridium perfringens and Campylobacter species. Histopathologic evaluation of abomasums revealed 31 of 38 cases contained abundant gram-positive bacteria associated with the damaged abomasal mucosa. Campylobacter-like organisms were demonstrated in 9 of 38 cases using the modified Dieterle stain. Clostridium perfringens was isolated in 14 of 38 cases, and Campylobacter jejuni was recovered from 5 of 38 cases.     

A method for studying interbreeding of nematodes of the subfamily Ostertagiinae in sheep with abomasal fistulae.

Interbreeding in two nematode species, Teladorsagia circumcincta and Ostertagia leptospicularis, was studied in lambs using small populations. Offspring from a single mated female were investigated for T. circumcincta. The following protocol was found to be satisfactory for small populations: virgin females and adult males were obtained from donor lambs infected artificially and transferred into the abomasa of fistulated sheep or lambs being treated with a long-acting corticosteroid. Infective larvae obtained from cultures of faeces of fistulated sheep were given to a recipient lamb and when this animal was slaughtered it was possible to examine the adult nematodes resulting from the interbreeding. Interbreeding between one male and one female was not successful while there was limited recovery of infective larvae from the implantation of a single mated female.     

Effect of cimetidine on abomasal pH and Haemonchus and Ostertagia species in sheep

Cimetidine, a histamine H2-receptor blocking agent which reduces gastric acid flow in simple stomach animals was administered directly into the abomasum of sheep at 100 mg kg-1 twice daily for five consecutive days and again on the eighth day. The drug caused a significant increase in the pH values of the abomasal contents and was highly effective in reducing the adult population of Haemonchus contortus (100 per cent) and Ostertagia circumcincta (70 per cent) in the abomasum of sheep. The possibility of using cimetidine as a method of treating abomasal nematode parasitism is discussed.     

TGIF1 and SF1 polymorphisms are associated with litter size in Small Tail Han sheep

TGF-β induced factor homeobox 1 (TGIF1) and splicing factor 1 (SF1) are important for mammalian reproduction; however, the effects of these genes on litter size in sheep remain unexplored. In this study, we genotyped 768 ewes from seven sheep breeds at two loci: g.37871539C>T, a synonymous mutation of TGIF1; and g.42314637T>C, a 3′UTR variant of SF1. Our analysis of polymorphism revealed only two genotypes at locus g.37871539C>T in TGIF1, with most sheep populations being moderately polymorphic (0.25 < PIC < 0.5) at this site. In contrast, most breeds exhibited low polymorphism (PIC ≤0.25) at the SF1 locus g.42314637T>C. The association analysis revealed that a synonymous mutation at g.37871539C>T in TGIF1 was highly associated with litter size in Small Tail Han sheep, in which it causes a significant decrease in litter size. Conversely, while the SF1 3′UTR variant g.42314637T>C was also highly associated with litter size in sheep, it causes a significant increase in the number of litter size. Combined, these data provide valuable information regarding candidate genetic markers for sheep breeding programs.     

Cellular profiles in the abomasal mucosa and lymph node during primary infection with Haemonchus contortus in sheep

Cellular changes in the abomasal tissue and draining abomasal lymph nodes were examined after primary infection of lambs with Haemonchus contortus for 3, 5 or 27-36 days.Infection with H. contortus larvae resulted in a rapid and selective increase in the percentage of CD4(+) T-cells in the abomasal lymph node at 3 days post-infection (PI). By 5 days PI, the lymph node weight had increased two-fold; however, the percentage of lymphocyte populations in the abomasal lymph node resembled that seen in uninfected sheep. Lymph node weights remained at increased levels in the adult nematode infected sheep and down-regulation of B-cell surface markers (sIg and MHC Class II) was apparent in this group. Significant increases in the percentage of CD4(+) T-cells co-expressing MHC Class II, but not CD25, were observed in the larval infected groups except in adult nematode infected sheep. Increased numbers of eosinophils, CD4(+), gamma delta(+) T-cells and B-cells were found in the abomasal tissue by 5 days PI, but no further increases in these cell populations were observed in the adult nematode infected group. In contrast, the level of both lamina propria and intraepithelial mast cells observed in the abomasal mucosa was highest in the sheep carrying an adult nematode burden. These findings indicate that sheep are able to generate an early immune response to infection with H. contortus larvae, characterised by the activation of CD4 T-cells and B-cells in the draining lymph nodes and recruitment of eosinophils, CD4(+) and gamma delta-TCR,WC1(+) T-cells and B-cells in larval infected tissues. However, these changes do not seem to be maintained during infection with the adult parasite where increases in mast cell numbers dominate the local response, indicating that different parasite stages may induce distinct and possibly counteractive immune responses.     

Extenders containing dimethylformamide associated or not with glycerol are ineffective for ovine sperm cryopreservation

The study aimed at testing the effectiveness of dimethylformamide, alone or combined with glycerol, as cryoprotectant for freezing ram semen. Ejaculates from nine rams were cryopreserved in Tris-based extenders, containing 5% of glycerol, association of dimethylformamide with glycerol, in four proportions achieving 5% of cryoprotectors in the media and pure dimethylformamide (2, 3, 4 and 5%) in replacement to glycerol. The samples were diluted to 100 × 10(6) sptz/ml and stored in 0.25-ml straws in liquid nitrogen. After thawing (37 °C for 30 s), motility was preserved better by the extender containing 5% of glycerol (p < 0.05). The extenders containing pure dimethylformamide, or more than 2% in combination with glycerol, provided sperm motilities close to zero. Plasma and acrosomal membrane integrity were preserved better (p < 0.05) in the extender containing 5% glycerol. It can be concluded that dimethylformamide, alone or combined with glycerol, has no beneficial effects on ovine semen cryopreservation.