Evaluation of single and comparative intradermal tuberculin tests for tuberculosis eradication in caprine flocks in Castilla y León (Spain)

Goats can act as reservoirs for tuberculosis (TB) infection. The main etiological agents of TB in goats are Mycobacterium caprae and Mycobacterium bovis and they infect also a wide range of domestic and wild animals and humans. Control programmes based mainly on the application of single and comparative intradermal tuberculin (SIT and SCIT respectively) tests are being implemented in certain regions of Spain with a high density of caprine flocks as Castilla y León, including goats with epidemiological relationship with cattle. The aim of this study was to evaluate the performance of the intradermal tests in naturally TB-infected caprine flocks from this region. The study was performed using data from 17,450 goats in 54 different flocks that were classified as TB-infected in the control programmes executed in 2010 and 2011. Data from 1237 goats from 7 dairy flocks depopulated after the first intradermal testing were used to estimate the sensitivity (Se) using bacteriology as the gold-standard. Overall Se of the SIT test using the severe interpretation was 43.9% (CI 95%, 40.4–47.4) and decreased to 38.8% (CI 95%, 35.5–42.3) using the standard interpretation. Overall Se of the SCIT test ranged between 21.3% (CI 95%, 17.6–25.4) and 7% (CI 95%, 4.9–9.8) depending of the interpretation criteria. A significant weak positive correlation was found between age and skin fold thickness (Spearman’s test p < 0.05). Results from this study yielded, in general, low Se values probably due the systematic detection and slaughter of reactors as a consequence of the eradication programme in previous years or the presence of factors that may interfere in the diagnosis. Therefore, these results suggest the necessity of including ancillary diagnostic tools and/or strict interpretation criteria to maximize detection of positive animals in infected settings.     

Body development, carcass, and meat quality of confined lambs fed increasing levels of whole rice meal

The objective of this trial was to evaluate the effect of whole rice meal (WRM) inclusion in the concentrate upon body development, carcass traits, and meat quality of lambs. Twenty-four castrated lambs with an average initial body weight of 17.90?±?2.72 kg were randomly blocked according to two genetic groups (Corriedale and Texel by Corriedale crossbreds). Three isocaloric (11.3 MJ/kg of metabolizable energy) and isonitrogenous (17 % crude protein) diets were offered to the animals for 74 days. Diets consisted of 40 % forage and 60 % concentrate diet, on a dry matter basis, with 0, 15, or 30 % of WRM inclusion into the concentrate. Body growth (after slaughter), carcass, and meat traits were evaluated on each animal. Results obtained indicated that genotype did not affect body growth, carcass, and meat traits except for yellowness. No significant interaction between diet and genotype were detected. Inclusion of up to 30 % WRM did not significantly (P?>?0.05) affect body growth, carcass, and meat traits, except for meat color. Meat luminosity progressively increased (36.32?+?0.055X) while redness (15.13???0.03X) decreased with the inclusion of WRM in the diet, but still remained within acceptable values. The study indicates that WRM may be included up to 30 % in the concentrate replacing corn without adverse effects upon body development, carcass traits, and meat quality of lambs.     

Evaluation in mice of Brucella ovis attenuated mutants for use as live vaccines against B. ovis infection

Brucella ovis causes ram contagious epididymitis, a disease for which a specific vaccine is lacking. Attenuated Brucella melitensis Rev 1, used as vaccine against ovine and caprine brucellosis caused by B. melitensis, is also considered the best vaccine available for the prophylaxis of B. ovis infection, but its use for this purpose has serious drawbacks. In this work, two previously characterized B. ovis attenuated mutants (Δomp25d and Δomp22) were evaluated in mice, in comparison with B. melitensis Rev 1, as vaccines against B. ovis. Similarities, but also significant differences, were found regarding the immune response induced by the three vaccines. Mice vaccinated with the B. ovis mutants developed anti-B. ovis antibodies in serum of the IgG₁, IgG_2a and IgG_2b subclasses and their levels were higher than those observed in Rev 1-vaccinated mice. After an antigen stimulus with B. ovis cells, splenocytes obtained from all vaccinated mice secreted similar levels of TNF-α and IL12(p40) and remarkably high amounts of IFN-γ, a crucial cytokine in protective immunity against other Brucella species. By contrast, IL-1α -an enhancer of T cell responses to antigen- was present at higher levels in mice vaccinated with the B. ovis mutants, while IL-10, an anti-inflammatory cytokine, was significantly more abundant in Rev 1-vaccinated mice. Additionally, the B. ovis mutants showed appropriate persistence, limited splenomegaly and protective efficacy against B. ovis similar to that observed with B. melitensis Rev 1. These characteristics encourage their evaluation in the natural host as homologous vaccines for the specific prophylaxis of B. ovis infection.     

Effects of Rapid Cooling Prior to Freezing on the Quality of Canine Cryopreserved Spermatozoa

The aim of this study was to evaluate the effects of rapid cooling prior to freezing on frozen-thawed canine sperm quality. In experiment 1, centrifuged ejaculates from 6 dogs were pooled, split into 4 aliquots and cryopreserved by the Uppsala procedure using different cooling rates (control, cooling speed 18 C/90 min and average cooling rate 0.2 C/min; rapid, cooling speed 18 C/8 min and average cooling rate 2.25 C/min) in combination with 2 glycerol addition protocols (fractionated or unfractionated). In experiment 2, centrifuged ejaculates from 4 dogs were processed individually using the same cooling rates described in experiment 1 in combination with an unfractionated glycerol addition protocol. Each of the experiments was replicated 5 times. Sperm quality was evaluated after 30 and 150 min of post-thawing incubation at 38 C. Total motility (TM), progressive motility (PM) and quality of movement parameters were assessed using a computerized system, and sperm viability (spermatozoa with intact plasma and acrosome membranes) was assessed using flow cytometry (H-42/PI/FITC-PNA). Values for TM, PM, viable spermatozoa and the quality of movement parameters after thawing were not significantly affected by the cooling rate. The interaction between the cooling rate and the added glycerol protocol was not significant. There were significant differences among the males (P<0.01) in the sperm quality parameters evaluated after thawing. The interaction between the males and the cooling rate was not significant. In conclusion, canine spermatozoa can be cryopreserved using the Uppsala method at an average cooling rate of 2.25 C/min prior to freezing together with addition of fractionated or unfractionated glycerol.     

Inter-specific competition and management modify the morphology, nutrient content and resorption in Scots pine needles

Influence of tree-to-tree competition on nutrient resorption is still not well understood. To contribute filling this gap, we assessed the effects of thinning (0, 20 and 30 % extraction of basal area) and canopy type (beech–pine vs. pine subplots) on needle dry weight, needle length, nutrient content and nutrient resorption (N, P and K) in Scots pine (Pinus sylvestris) needles of different cohorts, 8–9 years after thinning. Thinning and canopy type often concurrently affected needle morphology (e.g. lighter and shorter needles in 30 % thinning, heavier and longer needles in pine canopy on the first year of study) and nutrient content (e.g. decrease in N, P and K in 30 %; N, P and K higher in pine canopy on the first year of study). However, effects of thinning appeared only in older cohorts for N and P but were found in old and new cohorts for needle dry weight, needle length and K, indicating that some thinning effects remained after 8–9 years. Canopy effects on morphology and nutrient content were more frequent in recent cohorts, in relation to an increase in beech cover over time. While no clear effects of thinning on the nutrient resorption were observed, higher values were observed in the pine than in the mixed canopy, which could be related to a higher Scots pine stem growth in those patches. The observed differences between treatments will likely increase as the stand develops, probably leading to beech trees being dominant, and as future thinnings are carried out.     

Effect of a size-based selection program on blood metabolites and immune response of Litopenaeus vannamei juveniles fed different dietary carbohydrate levels

Blood metabolites in wild and seventh-generation cultivated shrimp were measured to determine how size-based selection could alter the nutritional and immunological conditions of Litopenaeus vannamei. Wild L. vannamei juveniles and a sample of seventh-generation cultured shrimp were acclimated under identical conditions. During 55 days, shrimp were fed a high (HCHO: 44%) or a low (LCHO: 3%) carbohydrate diet for 55 days. Wild shrimp showed a direct relation between dietary CHO and lactate, protein and hemocyte levels indicating that dietary CHO was used for protein synthesis via transamination pathways. In seventh-generation cultured shrimp these parameters were inversely proportional to dietary CHO level, indicating the capacity to synthesize protein from dietary CHO was repressed in cultured shrimp. Farmed shrimp showed a limited capacity to respond to LCHO diets demonstrating high protein dependence in their metabolism and immune response. These results demonstrate that during size-based breeding programs other metabolic process than CHO catabolism can be selected. The incapacity of shrimp to use dietary CHO could limit protein reduction of diets and limit the efforts of the shrimp industry to be ecologically and environmentally profitable.     

A PROPOSED RADIOGRAPHIC CLASSIFICATION SCHEME FOR CONGENITAL THORACIC VERTEBRAL MALFORMATIONS IN BRACHYCEPHALIC “SCREW‐TAILED” DOG BREEDS

Congenital vertebral malformations are common in brachycephalic “screw‐tailed” dog breeds such as French bulldogs, English bulldogs, Boston terriers, and pugs. The aim of this retrospective study was to determine whether a radiographic classification scheme developed for use in humans would be feasible for use in these dog breeds. Inclusion criteria were hospital admission between September 2009 and April 2013, neurologic examination findings available, diagnostic quality lateral and ventro‐dorsal digital radiographs of the thoracic vertebral column, and at least one congenital vertebral malformation. Radiographs were retrieved and interpreted by two observers who were unaware of neurologic status. Vertebral malformations were classified based on a classification scheme modified from a previous human study and a consensus of both observers. Twenty‐eight dogs met inclusion criteria (12 with neurologic deficits, 16 with no neurologic deficits). Congenital vertebral malformations affected 85/362 (23.5%) of thoracic vertebrae. Vertebral body formation defects were the most common (butterfly vertebrae 6.6%, ventral wedge‐shaped vertebrae 5.5%, dorsal hemivertebrae 0.8%, and dorso‐lateral hemivertebrae 0.5%). No lateral hemivertebrae or lateral wedge‐shaped vertebrae were identified. The T7 vertebra was the most commonly affected (11/28 dogs), followed by T8 (8/28 dogs) and T12 (8/28 dogs). The number and type of vertebral malformations differed between groups (P = 0.01). Based on MRI, dorsal, and dorso‐lateral hemivertebrae were the cause of spinal cord compression in 5/12 (41.6%) of dogs with neurologic deficits. Findings indicated that a modified human radiographic classification system of vertebral malformations is feasible for use in future studies of brachycephalic “screw‐tailed” dogs.     

Genetic improvement of farmed tilapias: Biochemical characterization of strain differences in Nile tilapia

Four African wild strains (Egypt, Ghana, Senegal and Kenya) and four established Asian farmed strains of Nile tilapia, Oreochromis niloticus (popularly known in the Philippines as Taiwan, Thailand, Singapore and Israel) were analysed electrophoretically at 30 protein loci to estimate genetic differences among the strains. All strains shared alleles at 14 monomorphic and 16 variable loci. Among the African strains, characteristic allele frequency differences were observed at AAT-1 ^* 46 for Ghana and Senegal, ADH ^* 83 for Kenya, ADH ^* 120 for Senegal, G3PDH-2 ^* 300 for Egypt, IDDH ^* 67 for Senegal, sMDH-1 ^* 120 for Kenya and SOD ^* 150 for Senegal. Genetic distance values among the strains revealed a clustering of the farmed strains with Egypt and Ghana O. niloticus, a slight separation of the Senegal strain and a larger separation of the Kenya strain. This profile may reflect the origins of the few founder populations of this species previously introduced to Asia. It also confirms the wider genetic divergence of the Kenya strain (O. niloticus vulcani) from the others studied here, which are all O. n. niloticus. Observed heterozygosities of the strains ranged from 0.026 to 0.071, with the African wild strains the lower values (mean Ho = 0.036) and the farmed strains the higher ones (mean Ho = 0.056). The implications of these results to the ongoing tilapia genetic improvement programme in the Philippines are discussed.     

Common bottlenose dolphin (Tursiops truncatus) interaction with fish farms in the Gulf of Ambracia,western Greece

1. Marine aquaculture, and its fast-growing development, has the potential for wider environmental, ecosystem, and biodiversity impacts. This study assesses the impact of fish farming on a bottlenose dolphin (Tursiops truncatus) population within the Gulf of Ambracia (western Greece) between 2007 and 2018. Two different study areas were defined in order to investigate differences on occurrence, abundance, behaviour, and seasonal fluctuations between them: a ‘control area’, in the north-western side of the Gulf; and a ‘fish farm area’, in the south west, where the highest density of fish farm cages is found.
2. A total of 169 daily surveys were dedicated to the control area and 74 days were dedicated to the fish farm area, yielding 104 and 37 sightings, respectively. Both the probability of detecting dolphins (U = 6,763.000, P = 0.002) and the group sizes (U = 578.000, P < 0.001) were smaller around fish farms.
3. Seasonality analyses were restricted to 2007–2008, as this was the period with year-round effort. Results revealed that dolphins were more frequently seen around fish farms in the winter (Kruskal–Wallis test, P = 0.036).
4. From the 40 identified individuals that were re-sighted at least 10 times during the study period, 21 used the fish farm area less than expected according to sampling effort, and 10 of them were never observed in that area (binomial test, P < 0.05).
5. The results revealed an impact of fish farms on the distribution of bottlenose dolphins in the Gulf of Ambracia. This information should be taken into consideration when defining ecosystem-based management measures within the management plan, which is currently in preparation for this Natura 2000 site.