Safety and efficacy of an experimental reovirus vaccine for in ovo administration

A commercial reovirus vaccine alone or experimental reovirus vaccine plus antibody complex were inoculated into 18-day-old specific pathogen free (SPF) broiler embryos at 0.1 of the recommended chick dose. The following groups were used: group 1A was not vaccinated or challenged; group 1B was not vaccinated, but was challenged with virulent reovirus; group 2 received the vaccine complexed with 1/4 dilution of antiserum; group 3 received the vaccine with 1/8 dilution of antiserum; group 4 received the vaccine with 1/16 dilution of antiserum, and group 5 received vaccine alone. At 1, 3, 6, 9, and 12 days of age, serum was collected and antibody against avian reovirus was analyzed by enzyme-linked immunosorbent assay (ELISA). At the same times, spleens were collected and vaccine virus detected by inoculating chicken embryo fibroblasts (CEFs) and examining for cytopathic effect. At 15 days of age, chickens in groups 2-5 were challenged with reovirus. At 22 days of age, birds were euthanatized and weighed. Efficacy of the vaccines was based on safety, percent protection, and antibody response. In ovo vaccination with the commercial or experimental vaccines did not adversely affect hatchability of SPF chickens. The vaccine complexed with antibody resulted in significantly less posthatch mortality (3.7%) when compared to mortality of chickens that received vaccine alone (17%). Both vaccine virus recovery and antibody response were delayed at least 3 days in birds receiving the experimental vaccines. In evo administration of reovirus antibody complex vaccines provided at least 70% protection. The experimental reovirus-antibody complex vaccines were safe and efficacious when given in ovo to SPF broiler embryos.     

Beta2-agonists and cAMP inhibit protein degradation in isolated chick (Gallus domesticus) skeletal muscle

1. The role of beta2-agonist and of cAMP in chick skeletal muscle proteolytic pathways and protein synthesis was investigated using an in vitro preparation that maintains tissue glycogen stores and metabolic activity for several hours. 2. In extensor digitorum longus (EDL) muscle total proteolysis decreased by 15 to 20% in the presence of equimolar concentrations of epinephrine, clenbuterol, a selective hbetaagonist, or dibutyryl-cAMP. Rates of protein synthesis were not altered by clenbuterol or dibutyryl-cAMP. 3. The decrease in the rate of total protein degradation induced by 10(-5)M clenbuterol was paralleled by a 44% reduction in Ca2+-dependent proteolysis, which was prevented by 10(-5)M ICI 118.551, a selective fbeta2antagonist. 4. No change was observed in the activity of the lysosomal, ATP-dependent, and ATP-independent proteolytic systems. Ca2+-dependent proteolytic activity was also reduced by 58% in the presence of 10(-4)M dibutyryl-cAMP or isobutylmethylxanthine. 5. The data suggest that catecholamines exert an inhibitory control of Ca2+-dependent proteolysis in chick skeletal muscle, probably mediated by fbeta2adrenoceptors, with the participation of a cAMP-dependent pathway.     

Effects of bacterial direct-fed microbials and yeast on site and extent of digestion,blood chemistry,and subclinical ruminal acidosis in feedlot cattle

Two studies were conducted to determine whether a bacterial direct-fed microbial (DFM) alone or with yeast could minimize the risk of acidosis and improve feed utilization in feedlot cattle receiving high-concentrate diets. Eight ruminally cannulated steers, previously adapted to a high-concentrate diet, were used in crossover designs to study the effects of DFM on feed intake, ruminal pH, ruminal fermentation, blood characteristics, site and extent of digestion, and microbial protein synthesis. Steers were provided ad libitum access to a diet containing steam-rolled barley, barley silage, and a protein-mineral supplement (87, 8, and 5% on a DM basis, respectively). In Exp. 1, treatments were control vs. the lactic-acid producing bacterium Enterococcus faecium EF212 (EF; 6 x 10(9) cfu/d). In Exp. 2, treatments were control vs EF (6 x 10(9) cfu/d) and yeast (Saccharomyces cerevisiae; 6 x 10(9) cfu/d). Supplementing feedlot cattle diets with EF in Exp. 1 increased (P < 0.05) propionate and (P < 0.05) decreased butyrate concentrations, decreased the nadir of ruminal pH (P < 0.05), enhanced the flow of feed N (P < 0.10) to the duodenum but reduced that of microbial N (P < 0.10), reduced (P < 0.10) intestinal digestion of NDF, and increased (P < 0.10) fecal coliform numbers. Other than the increase in propionate concentrations that signify an increase in energy precursors for growth, the other metabolic changes were generally considered to be undesirable. In Exp. 2, providing EF together with yeast abolished most of these undesirable effects. Combining EF with yeast increased the DM digestion of corn grain incubated in sacco, but there were no effects on altering the site or extent of nutrient digestion. The diets used in this study were highly fermentable, and the incidence of subclinical ruminal acidosis, defined as steers with ruminal pH below 5.5 for prolonged periods of time, was high. Supplementing the diet with EF, with or without yeast, had limited effects on reducing ruminal acidosis. It seems that cattle adapted to high-grain diets are able to maintain relatively high feed intake and high fiber digestion despite low ruminal pH. The Enterococcus faecium bacterium and yeast used in this study were of limited value for feedlot cattle already adapted to high-grain diets.     

Supplemental norgestomet,progesterone, or melengestrol acetate increases pregnancy rates in suckled beef cows after timed inseminations

In Exp. 1, 187 lactating beef cows were treated with injections of GnRH 7 d before and 48 h after prostaglandin F2alpha (PGF2alpha; Cosynch) or with Cosynch plus a 7-d treatment with an intravaginal progesterone (P4)-releasing insert (CIDR-B; Cosynch + CIDR). In Exp. 2, 183 lactating beef cows were treated with the Cosynch protocol or with Cosynch plus a 7-d treatment with norgestomet (Cosynch + NORG). In Exp. 1 and 2, blood samples for later P4 analyses were collected on d -17, -7 (first GnRH injection), 0 (PGF2alpha injection), and at timed artificial insemination (TAI; 48 h after PGF2alpha). In Exp. 3, 609 lactating beef cows were treated with the Cosynch + CIDR protocol or were fed 0.5 mg of melengestrol acetate (MGA) per day for 14 d before initiating the Cosynch protocol 12 d after the 14th d of MGA feeding (MGA + Cosynch). Blood samples were collected as in Exp. 1 and 2, plus additional samples on d -33 and -19 before PGF2alpha. In Exp. 4, 360 lactating beef cows were treated with a Cosynch + CIDR protocol, with TAI occurring at either 48 or 60 h after PGF2alpha, while receiving either GnRH or saline to form four treatments. Blood samples were collected as in Exp. 1 and 2. In Exp. 1, addition of P4 reduced the ability of the first GnRH injection to induce ovulation in anestrous cows with low P4 before PGF2alpha but improved (P = 0.06) pregnancy rates (61 vs 66%). In Exp. 2, the addition of NORG mimicked P4 by likewise increasing (P < 0.01) pregnancy rates (31 vs 51%) beyond those after Cosynch. In Exp. 3, the Cosynch + CIDR protocol increased (P < 0.001) pregnancy rates from 46 to 55% compared to the MGA + Cosynch protocol. In Exp. 4, administration of GnRH at TAI improved (P < 0.05) pregnancy outcomes (50 vs 42%), whereas timing of TAI had limited effects. We conclude that a progestin treatment concurrent with the Cosynch protocol improved pregnancy outcomes in all experiments, but pretreatment of cows with MGA was not as effective as the CIDR insert or NORG implants in this Cosynch-TAI model. Most of the improvement in pregnancy rates was associated with the increase in pregnancy rates of anestrous cows, regardless of whether ovulation was successfully induced in response to GnRH 7 d before PGF2alpha. Injection of GnRH at TAI following the Cosynch + CIDR protocol increased pregnancy rates in cycling cows with high P4 before the PGF2alpha injection and in anestrous cows with low P4 before PGF2alpha injection.     

Studies on the Epidemiology of Tropical Theileriosis (Theileria annulata Infection) in Cattle in Central Anatolia,Turkey

An epidemiological survey for Theileria annulata infection was conducted in 12 selected villages around Ankara in Central Anatolia, Turkey, during the period April 1990 to January 1993. During the survey, 198 cattle of 30 local breeds, 84 Holstein-Friesian×local breeds and 84 Holstein-Friesian breed were examined for antibodies to T. annulata and the presence of the vector ticks. Four species of Hyalomma ticks were identified: Hyalomma anatolicum anatolicum, Hyalomma anatolicum excavtum, Hyalomma detritum and Hyalomma marginatum marginatum. Salivary gland staining indicated that infected adult ticks of all four species were present and, therefore, were implicated in the transmission of tropical theileriosis in the field. Generally, the Hyalomma infestation rate was low, with the heaviest infestations occurring on the older animals. Young adults and calves had very low infestation rates. Most ticks seen on cattle were adults, very few nymphs were found. The blood smear and serological examination of the 198 cattle conducted in March, before the start of the first disease season, showed that the prevalence of piroplasmosis was 11.1% (22 out of 198) and the seroprevalence of T. annulata was 10.6% (21 out of 198). Forty-three animals were then excluded from the study because they were seropositive and/or harboured piroplasms. Ninety-two seronegative animals showed piroplasmosis (92 out of 155) and 34 seronegative animals became seropositive for T. annulata (34 out of 155) during the three disease seasons. One animal became clinically ill with tropical theileriosis and required treatment. The incidence of cattle showing piroplasmosis and disease in the total study sample was 50.7% and 0.5% per disease season, respectively. The seroconversion rate of new infection with T. annulata in the total study was 14.3% per animal season. The number of cattle showing piroplasmosis was much greater than the number of seropositive cattle, which may indicate the presence of another species of Theileria. The two different management systems encountered in the study were considered to have influenced the tick infestation levels.     

The effects of low doses of Senna occidentalis seeds on broiler chickens

The effects of 0.5%, 0.3% and 0.1% w/w concentrations of Senna occidentalis (So) seed mixed with commercial ration were studied in 18 groups of 32 broiler chicks each, from 1 day to 49 days of age. Three groups were fed one of the rations throughout their lives (TL). Three other groups were fed one of the rations from the 1st to the 28th day of life (starter phase, SP), and the final 3 groups were fed one of the rations from the 29th to 49th day (finisher phase, FP). Each experimental group was matched by a control group fed the same diet over the same period but without the inclusion of So. All the animals were killed at 49 days of age, and blood was collected from 10 birds in each group for biochemical studies (ALT, AST, GGT, LDH, UA). A complete necropsy was performed on 3 birds from each group. No significant differences in the biochemical parameters in the serum were found between the control and experimental chicks, but animals treated with 0.5% So in groups FP and TL, gained less weight and chicks that received 0.3% So or 0.5% So in the ration throughout life (TL) had a larger feed conversion ratio. Besides this, degenerative changes were found in the striated skeletal muscle in the chest, in the myocardium and in the liver in the animals that received the higher concentrations of So seeds.     

Characterization of Cryptosporidium spp.--recent developments and future needs

Cryptosporidia, widely distributed protozoan parasites of vertebrates have recently attracted increasing interest due to several serious waterborne outbreaks, the life-threatening nature of infection in immunocompromised patients, and the realization of economic losses caused by these pathogens in livestock. Genetic polymorphism within Cryptosporidium spp. is being detected at a continuously growing rate, owing to the widespread use of modern molecular techniques. The aim of this paper is to review the current status of taxonomy, genotyping, molecular phylogeny, and characterization of cryptosporidia, and to highlight the need for polyphasic typing, i.e. an integrated approach comprising standardized morphologic, biologic, and molecular methods for describing Cryptosporidium species and isolates, and for establishing "virtual" reference strains.     

Morphological relationship between intercalated duct and pancreatic islet in streptozotocin and/or camostat mesilate administrations in the chicken

Present electron microscopical and immunocytochemistrical studies elucidated some morphological relationship between intercalated duct (ICD) and pancreatic islet cells in the chicken in streptozotocin (STZ) and/or camostat mesilate (CM) administrations. Twenty-one chickens were set into four experimental groups: (1) control group, (2) STZ administration group, (3) CM administration group, and (4) STZ + CM administration group. Cytoplasms of ICD cells stained more strongly with eosin in STZ administration group than other groups, and electron-dense materials and intercalated processes between ICD and islet cells were also increasing in time dependence in STZ administration. Number of pancreatic islet in STZ + CM co-administration was about 3.1 times larger than other groups. Many small sized cells were detected at surrounding area of ICD and they incorporated 5-bromo-2'-deoxyuridine better than other experimental groups. Present morphological data suggested that ICD cells might support some tolerances of pancreatic endocrine cells against toxic substances and also involve in regeneration of new pancreatic islet cells in STZ + CM co-administration.     

The effects of potassium diformate and its molecular constituents on the apparent ileal and fecal digestibility and retention of nutrients in growing-finishing pigs

Five 43-kg barrows [(Dutch Landrace x Yorkshire) x Yorkshire] were fitted with steered ileocecal valve cannulas to compare the effects of K-diformate (KDF), a specifically conjugated salt vs its molecular constituents, namely, formic acid and K-formate, as acidifiers in lysine-deficient diets on the apparent ileal (ID) and fecal digestibility, retention of nutrients, and manure production. The animals were randomly assigned to five dietary treatments according to a 5 x 5 Latin square design as follows: 1) control-no acidifier; 2) 1% KDF (= 0.65% K-formate + 0.35% formic acid, or 0.7% [HCOO-] + 0.3% [K+]); 3) 0.65% K-formate (= 0.35% [HCOO-] + 0.3% [K+]); 4) 0.35% formic acid (= 0.35% [HCOO-]); and 5) 1.3% K-formate (= 0.7% [HCOO-] + 0.6% [K+]). Diets were formulated with barley, wheat, soybean meal, and canola meal as major ingredients, and provided all nutrients at adequate levels, except for lysine (24% less than estimated requirement). Feeding level was equal to 2.5 x maintenance requirement (MR) for ME (MR = 418 kJ ME x BW(-0.75)), and daily rations were given in two portions after mixing with water in a ratio of 1:2.5. Chromic oxide was used as an indigestible marker. No clinical health problems due to the dietary treatments were observed. Irrespective of the additive, there were no differences (P < or = 0.10) in the ID of DM, OM, CP, or essential amino acids compared to the control, except for phenylalanine (P < or = 0.05). Among nonessential AA, only the ID of tyrosine tended (P = 0.092) to increase (up to 3.9 percentage units). The fecal digestibility of ash and K were greater (P < or = 0.001) in pigs fed supplemental K, irrespective of its source. The greater intake and fecal digestibility of K corresponded with greater (P < or = 0.05) losses of K in urine. Body retention of N, Ca, total P, and K was similar (P > or = 0.10) among treatments. As estimated from a separate nonorthogonal analysis, supplemental K improved (P < or = 0.05) body N by 3.7 percentage units compared to the control. The results of this study do not provide a clear explanation for the improved growth performance reported previously with KDF and its molecular constituents, and further research on their in vivo mode of action will require methodological refinement, especially with regard to the efficiency of AA utilization.