Selected Aspects of Advanced Porcine Reproductive Technology

In vitro fertilization (IVF) of in vitro matured (IVM) oocytes in pigs has become the most popular method of studying gametogenesis and embryogenesis in this species. Furthermore, because of recent advances in in vitro culture (IVC) of IVM–IVF embryos, in vitro production (IVP) of embryos now enables us to generate viable embryos as successfully as for in vivo -derived embryos and with less cost and in less time. These technologies contribute not only to developments in reproductive physiology and agriculture but also to the conservation of porcine genetic resources and the production of cloned or genetically modified pigs. However, in IVP, there still remains the problem of abnormal ploidy, which is caused by performing procedures under non-physiological conditions. In recent years, unique technologies such as intracytoplasmic sperm injection (ICSI) or xenografting of gonadal tissue into immunodeficient experimental animals have been developed to help conserve gamete resources. These technologies combined with IVP are expected to be useful for the conservation of gametes from important genetic resources. Here, we discuss the developmental ability and normality of porcine IVP embryos and also the utilization of ICSI and xenografting in advancing biotechnology in pigs.     

Feeding Acacia etbaica and Dichrostachys cinerea fruits to smallholder goats in northern Ethiopia improves their performance during the dry season

Acacia etbaica Schweinf. and Dichrostachys cinerea (L.) Wight et Arn. are among the common woody browse plants that naturally grow in many arid and semi-arid rangelands in sub Saharan Africa. An experiment was conducted to compare the voluntary dry matter intake, body weight gain, nitrogen balance, carcass composition and sensory attributes of goats supplemented with different levels of A. etbaica and D. cinerea fruits. Average initial body weight of the goats was 20.4 kg (± 0.8SE) and received one of the following fruit supplementation regimes for 120 days (on body weight basis, n = 4 goats per group): (1) Control, no supplement; (2) 0.5% of A. etbaica; (3) 0.5% of D. cinerea; (4) 1.0% of A. etbaica; (5) 1.0% of D. cinerea; (6) 1.5% of A. etbaica; (7) 1.5% of D. cinerea. All groups were allowed to graze/browse during the day. Compared to A. etbaica, D. cinerea fruits contained higher amounts of crude protein (182 vs 135 g/kg DM), metabolizable energy (11 vs 8.4 MJ/kg DM), and in vitro dry matter digestibility coefficient (0.7 vs 0.6). The concentrations of acid detergent fiber (ADF), sulphuric acid solubulized lignin, and tannin (soluble and condensed) were lower (P < 0.05) in D. cinerea than in A. etbaica fruits. A. etbaica fruits, however, contained higher amounts of Na, K, Fe, and Zn concentrations than D. cinerea fruits. Dry matter intake was markedly higher in supplemented groups than in the control group without supplement. Similarly, body weight gain in the control group was negative and lower (P < 0.05) than any of the supplemented groups. During the course of the experimental period (120 days) the group placed in the 1.5% D. cinerea fruit diet gained 2.6 kg whereas the control group lost 2.0 kg. Thus, the difference between these two groups was 4.6 kg, which suggests about 22.5% increment of the goats' initial body weight. N retention was negative for the control group and varied little among the remaining treatment groups. Dressing percent increased with increased level of either type of fruit supplement, the highest (51.8%) being achieved at the 1.5% D. cinerea level. There was no difference in bone tissue across treatment groups. D. cinerea fruit intake also resulted in juicy/watery flavor and tender carcass. D. cinerea fruits appeared to have no adverse effects when included at the highest level (1.5%) and could be collected and stored as dry season supplement to smallholder goats. This study shows that poor farmers can increase goat performance substantially through supplement feeding with wild fruits. Areas in Ethiopia originally set aside for provision of ecological services and biodiversity protection can also be used for wild fruit production and feeding of small stock. The effect of increasing D. cinerea fruits above the 1.5% body weight level should be a focus of future investigation. Further work on deactivation mechanism of tannins is required for A. etbaica fruits included at levels higher than 1.0%.     

Physiological and immunological responses to weaning and transport in the young pig: modulation by administration of porcine somatotropin

To examine the effects of exogenous porcine (p) ST on measures of stress and immune function in weaned pigs with or without transport, pigs (20 +/- 1 d of age) received daily injections of pST (0.5 mg/kg; n = 16) or saline (n = 16) for 5 d. On d 5, a blood sample was collected immediately before injection. At 4 h postinjection, pigs were weighed, sampled for blood, injected with di-nitrophenyl-conjugated keyhole limpet hemocyanin, and weaned. One half of the pigs in each group were transported for 3 h before placement in the nursery. Pigs were weighed, and blood was collected on 1, 7, and 14 d postweaning. Statistical significance was set at P < 0.05. Serum IGF-I concentrations were increased by pST and decreased by weaning, but not affected by transport. The free cortisol index was elevated in all pigs 1 d postweaning, although less in transported versus nontransported pigs. By 7 d postweaning, the free cortisol index returned to prewean values. Serum concentrations of immunoglobulin (Ig) G increased in all pigs by 14 d postweaning, but were not affected by pST or transport. Serum IgM concentrations were elevated at 7 and 14 d postweaning. Before weaning and again 1 d postweaning, pigs treated with pST had greater concentrations of IgM than did control animals. Circulating neutrophils increased in pST-treated pigs 4 h after the final pST injection. Improved immune function in weaned pigs by pST may lead to greater health and growth in a commercial setting.     

Prevalence of dental disorders in 483 horses and ponies

Prevalence of dental disorders in 483 horses and ponies Five equine veterinarians working at three equine referral hospitals examined the oral cavity of 483 horses and ponies prior to surgery, to gain information about the prevalence of common dental disorders in the horse. A thorough examination, using hands/fingers, a full-mouth speculum, a dental mirror and a good light source, is essential to detect such disorders. Many horses examined needed dental attention even though there were no problems with mastication or riding. The most detected abnormalities were sharp enamel points on the buccal side of the upper arcade and the lingual side of the lower arcade, and hooks at 106/206 and 311/411. Regular dental care would improve the condition and welfare of horses.     

Quantifying the aging response and nutrient composition for muscles of the beef round

The objective of this study was to determine the optimal postmortem aging period and nutrient composition for Beef Value Cuts of the round. Forty USDA Select and 40 Premium USDA Choice beef carcasses were selected from a commercial beef packing plant in Colorado over a 12-wk period. The bottom and inside rounds were collected from both sides of each carcass for further fabrication into the following muscles: adductor, gastrocnemius, gracilis, pectineus, and superficial digital flexor. Each pair of muscles was cut into 7 steaks and randomly assigned to 1 of the following aging periods: 2, 4, 6, 10, 14, 21, and 28 d, and placed in refrigerated storage (2°C, never frozen). Upon completion of the designated aging period, steaks were removed from storage, cooked to a peak internal temperature of 72°C, and evaluated using Warner-Bratzler shear force (WBSF). A 2-way interaction was detected (P < 0.05) between individual muscle and postmortem aging period. The WBSF of all muscles except the superficial digital flexor decreased with increased time of postmortem aging. Quality grade did not affect (P > 0.05) WBSF values for the adductor, gastrocnemius, pectineus, and superficial digital flexor muscles. Exponential decay models were used to predict the change in WBSF from 2 to 28 d postmortem (aging response). The adductor, gastrocnemius, Select gracilis, Premium Choice gracilis, and pectineus required 21, 14, 23, 23, and 25 d, respectively, to complete the majority of the aging response. To determine the nutrient composition of the adductor, gastrocnemius, gracilis, pectineus, semimembranosus, and superficial digital flexor, bottom and inside rounds were collected from 10 USDA Select and 10 Premium USDA Choice carcasses and fabricated into the respective muscles, cut into 2.54-cm cubes, frozen (-20°C), and then homogenized. The adductor, gracilis, pectineus, semimembranosus, and superficial digital flexor were analyzed for DM, moisture, CP, and ash percentages. All muscles were evaluated for total lipid, fatty acid, and cholesterol composition. When quality grades were combined, all muscles fell into the extra lean or lean categories specified by USDA guidelines. Results of this study illustrate the potential for Beef Value Cuts of the round to be sold in food service operations and retail stores with marketing emphasis being placed on the exceptional leanness and acceptable tenderness of these cuts.     

Molecular phylogenetic analysis of Onchocerca lupi and its Wolbachia endosymbiont

The morphology of Onchocerca lupi, responsible for canine ocular onchocercosis, is unique within the genus. Earlier analyses of the 5S ribosomal RNA gene spacer region sequence of the parasite and the 16S ribosomal RNA gene sequence of its Wolbachia endosymbiotic bacteria (Rickettsiales) supported the morphological and biological arguments that O. lupi is a distinct species. However, the exact phylogenetic position of O. lupi and its endosymbiont could not be unambiguously determined. Herein we report analyses based on the mitochondrial cytochrome oxidase I (COI) gene of the filarial species and the Wolbachia surface protein (wsp) and the bacterial cell-cycle ftsZ genes of their wolbachiae. Our results indicate that O. lupi separated from other Onchocerca spp. early in evolution. This is in line with the previous morphological analysis demonstrating that O. lupi is an atypical Onchocerca species showing both primitive and evolved characters. The phylogenetic trees generated for the COI sequences of filariae and the wsp and ftsZ sequences of their wolbachiae were congruent with each other, which supports the hypothesis that nematodes and their Wolbachia endobacteria share a long co-evolutionary history.     

Use of human intravenous immunoglobulin in dogs with primary immune mediated hemolytic anemia

Immune mediated hemolytic anemia (IMHA) in dogs is a severe disease with a high mortality rate. As human immunoglobulin (HIG) was reported to be beneficial for the treatment of IMHA in dogs we examined the influence of HIG on the course of the disease in our dogs with IMHA. Of 22 dogs with primary IMHA 9 dogs received in addition to routine immunosuppressive therapy HIG at a dose of 0.19 to 0.68 g/kg (median 0.35 g/kg), 13 dogs did not receive HIG (-HIG group). Both groups were similar in terms of age, weight, the presence of autoagglutination, spherocytosis, positive Coombs' test, icterus and pigmenturia. The lowest hematocrit measured during the disease was significantly lower in the +HIG group compared to the -HIG group and dogs in the +HIG group received significantly more transfusions than those of the -HIG group. This is an indication for more severe disease signs of the +HIG group dogs. Although mortality during hospitalization and the time from hospital admission to release or death was not significantly different between the two groups, we interpret this similar course of the IMHA despite more severe signs of the +HIG group dogs as a potential positive effect of the HIG therapy.     

Antigen-specific IgG(T) responses in natural and experimental cyathostominae infection in horses

Equine clinical larval cyathostominosis is caused by simultaneous mass emergence of previously inhibited larvae from the mucosa of the colon. Clinical signs include diarrhoea, colic, weight loss and malaise, and in up to 50% of cases, the disease results in death. Cyathostominae spend a large part of their life cycle as larval stages in the intestinal mucosa. Definitive diagnosis is difficult due to the lack of diagnostic methods for pre-patent infection. In the present study, the enzyme-linked immunosorbent assay (ELISA) was used to investigate isotype responses to larval cyathostominae somatic antigen. Measurement of anti-larval IgG(T) responses appeared to have the most immunodiagnostic potential. An increase in IgG(T) response was detected to crude larval antigen by 5 weeks post-infection (PI) in individual infected ponies. Subsequently, IgG(T) responses to larval and adult somatic extracts were examined by Western blotting using sera from experimentally-infected horses and helminth-naive animals (n=6). Two antigen complexes, designated A and B, in larval somatic antigen were recognised specifically by the infected animals by 7 weeks PI. Sera taken from 23 endemically-infected animals, whose cyathostominae burdens had been enumerated, were also used to identify putative diagnostic antigens. Eighteen horses had positive mucosal worm burdens (range 723-3,595,725) and all but two of these animals had serum IgG(T) antibody specific to either complex. Moreover, IgG(T) responses specific to antigen complexes A and B were absent in all five parasite negative horses that were tested. Serum IgG(T) responses to either of the two complexes were identified in five clinical cases tested. IgG(T) responses to adult antigen somatic extracts were more heterogeneous, with no clear pattern between experimentally-infected ponies and helminth-free controls. The results indicate that increases in serum IgG(T) to mucosal larvae occur in the pre-patent period and that two antigenic complexes within somatic preparations of these stages have immunodiagnostic potential.     

Effects of feeding ergovaline on lamb performance in a heat stress environment

Lambs exposed to a heat-stressed environment (33 degrees C, 50% relative humidity) were used in three experiments to determine whether ergovaline (EV) is the primary toxin involved in fescue toxicosis. The first study evaluated the effects of feeding diets containing increasing levels of endophyte-infected tall fescue seed (E+) and decreasing levels of endophyte-free tall fescue seed (E-). The second and third study evaluated the response to a diet that contained synthetic EV added to an E- diet and the response to a diet containing endophyte-infected ryegrass seed (R+) with an elevated concentration of EV. In Exp. 1, lambs were fed diets of: 1) 10% E- and 0% E+, 2) 5% E- and 5% E+, or 3) 0% E- and 10% E+. Increasing the percentage of E+ in the diet resulted in a linear decrease (P < 0.01) in feed intake (as-fed basis), skin temperature, thermocirculation index (TCI), and serum prolactin. Body weight gain also decreased (P < 0.06). Respiratory rate and core body temperature were not affected by the 5 or 10% E+ diets. In Exp. 2, lambs were fed diets that contained: 1) 10% E-, 2) 10% E- with synthetic EV added at a level equivalent to the 10% E+ diet, or 3) 10% E+. Feed intake (as-fed basis), body weight gain, and skin temperature did not differ for lambs fed the E- and EV diets. The EV diet elicited a decrease (P < 0.05) in TCI and prolactin compared with the E- diet. The TCI for lambs fed EV did not differ (P > 0.10) from the E+ lambs; however, serum prolactin was lower (P < 0.05) for lambs on the E+ diet than for those fed EV. Core body temperature was not affected (P > 0.10) by feeding EV or E+ fescue seed in Exp. 2. In Exp. 3, lambs were fed diets that contained: 1) 10% E-, 2) 3.24% R+ and 6.76% E-, which added an equivalent amount of EV to E+ diets but reduced concentrations of other ergot alkaloids, or 3) 10% E+. Lambs fed the E+ diet and maintained at 33 degrees C had lowered feed intake (as-fed basis), skin temperature, and TCI compared with lambs fed the E- or R+ diets (P < 0.05). Lambs fed the E+ diet had increased rectal temperatures and lowered serum prolactin compared with lambs on the R+ diet (P < 0.05). Lambs on the R+ diet had a greater rectal temperature and lower serum prolactin than lambs on the E- diet (P < 0.05). These results suggest that EV is a fescue toxin; however, other alkaloids might work synergistically with EV, causing the full expression of fescue toxicosis.