Epidemiological pattern and risk factors associated with bovine viral-diarrhoea virus (BVDV) infection in a non-vaccinated dairy-cattle population from the Asturias region of Spain

A survey of bovine viral-diarrhoea virus (BVDV) infection was carried out in a non-vaccinated cattle population from the Asturias region of Spain in 1997 to assess seroprevalence and identify risk factors associated with infection. Twenty-eight herds were included; 529 cows were bled. Information regarding the herd and each animal sampled were recorded through a personal interview with the farmer. The true prevalence was estimated to be 21%. According to the antibody-age profiles and the herd-management characteristics, no persistently infected animals were suspected at that time within the herds sampled. Random-effects logistic regression found two major factors associated with seropositivity: age and cow origin. Results suggested that BVDV infection could be controlled in that area by livestock-trade control (without vaccines). In addition, an increasing risk of abortion was not observed when cows were seropositive to both BVDV and Neospora caninum infections.     

Buffalo (Bubalus bubalis) ES Cell–Like Cells are Capable of In Vitro Skeletal Myogenic Differentiation

When buffalo embryonic stem (ES) cell–like cells that expressed surface markers SSEA‐4, TRA‐1‐60, TRA‐1‐81, CD9 and CD90 and intracellular markers OCT4, SOX2 and FOXD3, as shown by immunofluorescence, and that expressed REX‐1 and NUCLEOSTEMIN as confirmed by RT‐PCR, were subjected to suspension culture in hanging drops in absence of LIF and buffalo foetal fibroblast feeder layer support, they differentiated to form three‐dimensional embryoid bodies (EBs). Of 231 EBs examined on Day 3 of suspension culture, 141 (61.3 ± 3.09%) were of compact type, whereas 90 (38.4 ± 3.12%) were of cystic type. The cells obtained from EBs were found to express NF‐68 and NESTIN (ectodermal lineage), BMP‐4 and α‐skeletal actin (mesodermal lineage), and α‐fetoprotein, GATA‐4 and HNF‐4 (endodermal lineage). When these EBs were cultured on gelatin‐coated dishes, they spontaneously differentiated to several cell types such as epithelial‐ and neuron‐like cells. When EBs were cultured in the presence of 1 or 2% DMSO or 10^?8 m or 10^?7 m retinoic acid for 25 days, ES cells could be directed to form muscle cell–like cells, the identity of which was confirmed by expression of α‐actinin by immunofluorescence and of MYF‐5, MYOD and MYOGENIN genes by RT‐PCR. MYOD was first detected on Day 10 in both treatment groups and on Day 15 in controls, whereas MYOGENIN was first detected on Day 10, Day 15 and Day 25 in the presence of retinoic acid, in the presence of DMSO and in controls, respectively. The present study demonstrates the ability of buffalo ES cell–like cells to undergo directed differentiation to cells of skeletal myogenic lineage.     

Potassium permanganate elicits a shift of the external fish microbiome and increases host susceptibility to columnaris disease

The external microbiome of fish is thought to benefit the host by hindering the invasion of opportunistic pathogens and/or stimulating the immune system. Disruption of those microbial communities could increase susceptibility to diseases. Traditional aquaculture practices include the use of potent surface-acting disinfectants such as potassium permanganate (PP, KMnO₄) to treat external infections. This study evaluated the effect of PP on the external microbiome of channel catfish and investigated if dysbiosis leads to an increase in disease susceptibility. Columnaris disease, caused by Flavobacterium columnare, was used as disease model. Four treatments were compared in the study: (I) negative control (not treated with PP nor challenged with F. columnare), (II) treated but not challenged, (III) not treated but challenged, and (IV) treated and challenged. Ribosomal intergenic spacer analysis (RISA) and pyrosequencing were used to analyze changes in the external microbiome during the experiment. Exposure to PP significantly disturbed the external microbiomes and increased catfish mortality following the experimental challenge. Analysis of similarities of RISA profiles showed statistically significant changes in the skin and gill microbiomes based on treatment and sampling time. Characterization of the microbiomes using 16S rRNA gene pyrosequencing confirmed the disruption of the skin microbiome by PP at different phylogenetic levels. Loss of diversity occurred during the study, even in the control group, but was more noticeable in fish subjected to PP than in those challenged with F. columnare. Fish treated with PP and challenged with the pathogen exhibited the least diverse microbiome at the end of the study.     

Effect of training location and time period on racehorse performance in New Zealand. 2. Multivariable analysis

AIM: To investigate training location (horses trained in Matamata vs those trained at all other venues in New Zealand), and time period (1996–1997 and 1998–1999), while controlling for other horse- and race- or trial-related factors, as a means of assessing the possible impact of construction of a new training surface at the Matamata Racing Club on indirect measures of racehorse performance (number of starts, and failure to race within 6 months of any start).

METHODS: Multivariable logistic regression and poisson analysis were used to analyse data derived using a retrospective cohort approach. Multivariable logistic regression was also used to analyse a case-control study. All data were derived from New Zealand Thoroughbred Racing (NZTR), records of race and trial results for racehorses trained in Matamata and other venues in New Zealand, covering two 19-month time periods (1996–1997 and 1998–1999). Outcome variables included whether a horse started again in the 6 months following any start that occurred in the first 13 months of either time period, and a count of the total starts for every horse.

RESULTS: Factors associated with increased risk of a start being followed by a 6-month no-race period included training location other than Matamata in comparison to horses trained in Matamata in the 1996–1997 time period, increasing age, 1998–1999 over 1996–1997, starting in a trial rather than a race, placing fourth or worse in a start, softer track conditions, summer vs autumn, increasing cumulative exercise intensity in the 60 days prior to a start, and increasing race distance. Factors associated with an increase in the total number of starts included horses trained at Matamata in 1996–1997 compared with other time period-location combinations, younger age of horses at the time of a start, longer race distance, and an increasing proportion of starts in stakes races.

CONCLUSIONS: Official race and trial results data provided a valuable resource for epidemiological studies of factors influencing racehorse performance. Results of analyses performed here provided little evidence of any adverse impact of a new training surface at the Matamata Racing Club on indirect measures of racehorse performance.     

Forage Quality of Plant Species Consumed by Capybaras (Hydrochoerus hydrochaeris) in the Paraná River Delta,Argentina

The capybara (Hydrochoerus hydrochaeris) is a selective herbivore that may show a preference for certain plant species. Such a preference is likely to be explained by the nutritional benefits hypothesis, stating that the nutritional value of species selected by capybaras is higher than that of avoided ones. Therefore, the objectives of this work are to evaluate the nutritional value of the plant species eaten by capybaras in the Lower Delta of the Paraná River, and to test the nutritional benefits hypothesis by analyzing forage quality. Samples of consumed plant species were collected from habitats located in the study area. We also collected plant species that are very abundant in the field but absent in the capybara’s diet, some of which are consumed by other native herbivores such as coypus (Myocastor coypus). Their nutritional quality was analyzed by estimating the following variables: water content, ash percentage, percentage of organic matter, acid detergent fiber, nitrogen percentage, crude protein, and caloric energy. The protein to fiber ratio was calculated and the water content was determined. The species consumed and not consumed by capybaras had a similar nutritional composition for all the variables analyzed (P  <  0.05). A similar result was observed when comparing within consumed species, except for the caloric energy content, which was significantly higher in species consumed in greater proportion than availability (P  <  0.05). Water content was at its minimum in species consumed in greater proportion than availability, and at its maximum in unconsumed species. No significant differences were found between consumed Poaceae and the rest of the consumed plant species for any of the analyzed variables. Diet selection by capybaras in the studied area may only be partially related to nutritional quality, and there would be other factors involved in foraging, such as physiological mechanisms of the animal or chemical and structural characteristics of food.     

Cholinergic reactivity of the bovine fetal ruminal wall in vitro

Strips of rumen wall from bovine fetuses were incubated in an organ bath with acetylcholine only (0.16 to 5.12 g/ml) or in the presence of neostigmine (0.20 g/ml) or atropine (0.05g/ml). The highest reactivity was observed in the period of 4.0–5.9 months of fetal age. This reactivity could be associated with the starting point of rumen papillary development.     

Comparison of Two Doses of the GnRH Antagonist, Acyline, for Pregnancy Termination in Bitches

Gonadotropin-releasing hormone (GnRH) antagonists are particularly useful when a rapid inhibitory effect on the gonadal axis is required. The aim of this study was to test the efficacy and clinical safety of a low and high dose of the third generation GnRH antagonist, acyline, on pregnancy termination in female dogs. The effect of the antagonist on the progesterone (P₄) serum concentration was also described. Twenty-one mid-pregnant bitches were randomly assigned to a single subcutaneous (SC) dose of a placebo (PLACE; n = 7), a low (ACY-L; 110 μg/kg; n = 6) or high (ACY-H; 330 μg/kg; n = 8) dose of acyline. The animals were followed up for 15 days. All ACY treated but no placebo-treated animals terminated their pregnancy by abortion (p < 0.01). The ACY-L and ACY-H groups interrupted their pregnancy 7 ± 1.9 and 6.4 ± 1.3   days after treatment, respectively (p = 0.7). A significant interaction between treatment and day was found (p < 0.01) for P₄ serum concentrations when PLACE was compared with both ACY groups. No difference was found for this hormone between both ACY groups (p > 0.05) where P₄ diminished throughout the study. The decreasing rate varied among animals and was closely related to the time of abortion when P₄ reached basal concentrations. In PLACE animals, gestation progressed normally and P₄ did not change throughout the study (p > 0.05). None of the bitches presented side effects. It was concluded that acyline safely terminated mid-pregnancy by permanently decreasing P₄ serum concentrations.     

Effect of beta‐aminoproprionitrile on eggshell formation

1. Eggshells are bioceramic‐biopolymer composites made by a cell‐mediated deposition of an extracellular matrix which drives the organisation of the inorganic phase. Ultrastructurally, eggshells are composed of shell membranes, mammillary knobs, palisade, and cuticle. Shell membranes are two nets of type X collagen‐containing fibrils. On to these membranes, the mammillary knobs, that is, the crystal nucleation sites, are deposited. Type X collagen is highly cross‐linked and insoluble.

2. In order to evaluate the role of type X collagen cross‐linking on eggshell formation, hens were injected with different doses of β‐aminoproprionitrile, which specifically interferes with cross‐link formation.

3. Changes in egg size and shape were observed. Scanning electron micrographs analysis of these eggs demonstrated marked changes in crystal growth and shell membrane structure and arrangement. A dot‐blot analysis, using a monoclonal antibody against chicken type X collagen, shows a dose‐dependent increase in shell membrane collagen extractability.

4. It is concluded that the formation of β‐aminoproprionitrile‐sensitive cross‐links among the type X collagen molecules of the shell membranes play an essential role in normal eggshell formation.     

Assessment of DNA Damage during In Vitro Development of Buffalo (Bubalus bubalis) Embryos: Effect of Cysteamine

Comet assay was used in the present study to examine DNA damage to buffalo oocytes and embryos during in vitro culture. Embryos were produced in vitro from oocytes obtained from slaughterhouse ovaries in presence of cysteamine (IVM and IVC media supplemented with 50 and 100 μm , respectively) or in its absence (controls). Compared to controls, cysteamine supplementation increased (p < 0.01) cleavage rate and proportion of oocytes that developed to 8‐ to 16‐cell stage. The incidence of DNA damage was lower (p < 0.01) in cysteamine group than that in controls at 8‐ to 16‐ (19.3 ± 4.24 vs 72.0 ± 5.22%) but not in 2‐cell stage embryos (11.7 ± 5.63 vs 20.8 ± 5.49%) or in mature oocytes (5.3 ± 3.43 vs 10.3 ± 4.73%). The tail length, which indicates magnitude of DNA damage, was shorter (p < 0.01) in cysteamine group than in controls in mature oocytes (25.5 ± 0.5 vs 36.0 ± 0.71 pixels) and 8‐ to 16‐cell stage (49.2 ± 1.64 vs 152.7 ± 1.28 pixels) but not in 2‐cell stage embryos (36.3 ± 1.54 vs 36.4 ± 0.75 pixels). Also, exposure of oocytes/embryos to UV radiation or H₂O₂ caused extensive DNA damage. In conclusion, these results suggest that oocytes/embryos suffer from DNA damage during progress of in vitro culture, which can be partly ameliorated by cysteamine supplementation of culture media.