Isolation and cryopreservation of functionally competent equine leucocytes

Sufficient numbers of functionally competent polymorphonuclear neutrophil granulocytes (PMN) seem to be of major importance during the course of equine endometritis. In this study, we wanted to establish a method for cryopreservation of functionally competent neutrophils for an intended local endometritis therapy in mares. The separation of leucocytes by hypotonic lysis of whole blood from clinically healthy mares was superior to the separation by dextrose sedimentation. After suspension of the cells in the cryoprotective solution [equine plasma with 5% (v/v) dimethyl sulphoxide (DMSO)], the leucocytes were frozen in liquid nitrogen. A temperature gradient with low cooling velocity (1 degree C/min between 4 and -70 degrees C) resulted in highest numbers of viable cells after thawing. Thawed PMN had a high phagocytic capacity for opsonized streptococci. Their ability to generate reactive oxygen species (ROS) after stimulation with a phorbol ester was even higher than that of freshly isolated PMN and was preserved up to 6 h after thawing. The results of this study indicate that cryopreservation of PMN may provide viable and functionally competent neutrophils for therapeutic use in mares susceptible to endometritis.     

How much variation in tree mortality is predicted by soil and topography in Central Amazonia?

Tree mortality in Amazonia has been related to regional variation in soil, topography and climatic disturbances, but the magnitude of the effect of these factors on tree mortality at local and mesoscales remains poorly determined. We investigated tree mortality in 72 1-ha permanent plots spanning 64 km² of tropical moist forest in Reserva Ducke, Manaus, Brazil. Plots were censused three times (2000-2003, 2003-2005, and 2005-2008), resulting in two census intervals. The relationships of soil and topography to tree mortality were dependent on tree size. Small- and medium-sized trees (1 ? dbh < 30 cm) had similar relationships of mortality with soil and topography, while large trees (dbh ? 30 cm) showed different (or no) relationships. The effects of soil and topography on tree mortality also varied temporally. In the second census interval after storms, soil and topography explained about one-fourth of the spatial variation in mortality of small- and medium-sized trees (<30 cm dbh), whereas no effects were detected in the first census interval. In particular, soil fertility was the most important predictor of tree mortality in the study area. Topography alone (altitude and slope) was associated with only 12% of the spatial variation in tree mortality and the magnitude of the effect of soil and topography on tree mortality also increased after storms. In general, plots on more fertile soils, on steep slopes and sandy soils in valleys showed greater tree mortality than those on plateau with well-drained clayey soils. Therefore, disturbance history and tree size should be included when scaling up tree mortality from local to regional scales. As much variation remains unexplained, other landscape features, such as watershed morphology and wind exposure, may be necessary to make more precise predictions on patterns of tree mortality in Central Amazonia.     

Characterization of the LH peak after short and long fixed‐time artificial insemination protocols in sheep raised in the tropics

The aim of this study was to evaluate the peak in luteinizing hormone (LH) and the pregnancy rate of sheep (Texel × Santa Inês) in the tropics using short‐ (6 days) and long‐term (12 days) progesterone protocols followed by artificial insemination (AI) both in and out of the breeding season. Experiment 1 was conducted within (IN) the breeding season (autumn, n = 36), and experiment 2 was conducted outside (OUT) of the breeding season (spring, n = 43). In each experiment, the sheep were divided into two groups (6 or 12 days) according to the duration of treatment with a single‐use progesterone release vaginal device (CIDR^®, Pfizer, São Paulo, SP, Brazil), and blood samples were collected from 10 animals per group every 4 hr to measure the LH and progesterone concentrations. In the spring, the characteristics of the LH peak did not differ between groups; but in the autumn, there were differences between groups at the beginning (G‐6 IN: 36.44 ± 5.46 hr; G‐12 IN: 26.57 ± 4.99 hr) and end of the LH peak (G‐6 IN: 46.22 ± 7.51 hr; G‐12 IN: 34.86 ± 8.86 hr). The results showed alterations in the LH peak during the breeding season only in the sheep undergoing the short‐term protocol.     

Supplementation with sunflower seeds in beef cattle did not impact on oocyte and in vitro embryo production

Supplementation with compounds rich in linoleic acid, including sunflower seed supplementation, promotes increase in conception rates in cows. We aimed to evaluate whether the sunflower seed (linoleic acid source) supplementation in beef donor females alters the plasma concentrations of cholesterol, triglycerides, HDL and LDL, increases the number and quality of oocytes, increases the cleavage rates and determines an improvement in number and quality of in vitro produced blastocysts. Thus, Nelore females were divided into two groups of 15 animals to receive supplementation with or without sunflower seed for 57 days. Females underwent follicular aspiration and the oocytes were subjected to in vitro embryo production. There was no difference (p > .1) between control group and group supplemented with sunflower seed on the number of displayed follicles; number of aspired oocytes; recovery rate; cleavage rate; number of embryos; number of blastocysts; embryos number of grades I and II; plasma concentrations of total cholesterol, triglycerides; HDL and LDL. Therefore, sunflower seed supplementation in oocyte donors did not increase the number and quality of oocytes, cleavage rates and the number and quality of blastocysts produced in vitro.     

Assessment of primers designed for the subspecies-specific discrimination among Babesia canis canis, Babesia canis vogeli and Babesia canis rossi by PCR assay

Canine babesiosis is an infectious disease caused by either Babesia gibsoni or Babesia canis protozoans. The latter is also classified under three different phylogenetic groups, referred to as subspecies B. canis canis, B. canis vogeli and B. canis rossi. The objective of the present study was to validate and standardize a PCR assay to discriminate the organisms at the subspecies level. First, the reference sequences of the 18S rRNA, 5.8S rRNA and 28S rRNA genes, including the internal transcribed spacer 1 (ITS1) and 2 (ITS2) of the most common species and subspecies of the genus Babesia were retrieved from the GenBank database. Subspecies-specific primers (BAB3, BAB4 and BAB5) and one genus-specific primer were designed from the alignment of the sequences. The PCR assays were evaluated in three different combinations of primer pairs in order to assure complete specificity for each reaction. The results of the tests had demonstrated effectiveness of the novel primer pairs BAB1/BAB3, BAB1/BAB4 and BAB1/BAB5 for the amplification of the subspecies-specific target fragments of 746 bp (B. c. canis), 546 bp (B. c. vogeli) and 342 bp (B. c. rossi) by PCR. The original enzymatic amplification assays with novel primers reported in this paper were confirmed to be a reliable tool for the specific discrimination among B. canis subspecies by single-step PCR assays.     

Satellite tagging and cardiac physiology reveal niche expansion in salmon sharks

Shark populations are declining globally, yet the movements and habitats of most species are unknown. We used a satellite tag attached to the dorsal fin to track salmon sharks (Lamna ditropis) for up to 3.2 years. Here we show that salmon sharks have a subarctic-to-subtropical niche, ranging from 2 degrees to 24 degrees C, and they spend winter periods in waters as cold as 2 degrees to 8 degrees C. Functional assays and protein gels reveal that the expression of excitation-contraction coupling proteins is enhanced in salmon shark hearts, which may underlie the shark's ability to maintain heart function at cold temperatures and their niche expansion into subarctic seas.     

Dolphin population specialized in foraging with artisanal fishers requires zero-bycatch management to persist

1. The small population paradigm assumes that populations with low numbers of individuals intrinsically have a high probability of extinction. The small population of Lahille’s bottlenose dolphins Tursiops truncatus gephyreus that specializes in foraging with artisanal fishers in Laguna, southern Brazil, faces human pressures including bycatch in fishing gear. The viability of this population was modelled over 30 and 100 years under different levels of bycatch, including the current scenario of two bycatches every year, two scenarios with higher incidence of bycatches and three management scenarios. The sensitivity of predicted growth rates to fixed-proportion and observed-variation changes in life history parameters was explored.
2. The current scenario predicted a declining population (r = −0.014; λ = 0.986) with a high probability of extinction in the long term (PE = 0.71). A small increase in bycatches would result in a marked increase in the probability of extinction. Management scenarios seem promising, but only the zero-bycatch management scenario would make the difference between a declining and an increasing population.
3. As expected for slow-growing species, population growth rate was most sensitive to proportional changes in adult female and juvenile survival. However, considering observed variation in vital rates, population dynamics were most influenced by variation in reproductive rates.
4. To determine the highest priority for management action, another simulation was made of how additional threat scenarios of recognized human activities (i.e. bycatch influencing adult survival and increased underwater noise or pollution influencing calf survival) would affect population dynamics. Population growth rate was very sensitive to changes in adult bycatch (especially females), as expected, and only subtly sensitive to a reduction in calf survival.
5. The current level of bycatch is unsustainable. Bycatch needs to be eliminated to maximize the probability of long-term persistence of this dolphin population. However, this population’s persistence could be threatened by natural variation in reproductive rates.

     

Colour Doppler Ultrasonography as a Tool to Assess Luteal Function in Santa Inês Ewes

The aim of this study was to evaluate luteal dynamics in the Santa Inês ewes using colour Doppler (CD) ultrasonography. Oestrus was synchronized in nulliparous females (n = 18), and subsequently, they were only teased (n = 6) or teased and mated (n = 12). Blood samples were collected daily for plasma progesterone (P₄) concentrations. Ultrasonographic images of corpora lutea (CL) in CD mode were obtained for further analysis in its largest diameter. The CD mode allowed an early sequential monitoring of CL that was visualized by the first time 0.77 ± 0.62 days after ovulation, with luteal area 29.68 ± 13.21 mm². During the luteogenesis, a progressive increase was observed, followed by a plateau of luteal area, vascularization area and plasma concentrations of P₄ reaching maximum values in D11 (124.0 ± 38.0 mm², 52.78 ± 24.08 mm² and 11.23 ± 4.89 ng/ml, respectively). In the luteolysis, the plasma concentrations of P₄ decreased sharply, whereas luteal and vascularization area gradually. The vascularization area was positively correlated with plasma concentrations of P₄ during the luteogenesis (r = 0.22) and luteolysis (r = 0.48). The luteal dynamics of Santa Inês ewes showed patterns similar to those observed in other sheep breeds studied. The CD ultrasonography has the potential to be used as a tool to assess luteal function in sheep.     

Effect of Marcela Extract (Achyroclines satureiodes) on the Shelf Life of Minced Tilapia (Oreochromis niloticus) Sausages

The effect of marcela extract (ME) on the shelf life of minced tilapia sausages was evaluated in this study. The level of phenolic compounds in the ME was 114.1 ± 2.9 mg GAE/g extract, and the DPPH free radical scavenging activity was 94.95 ± 1.34%. The ME protected the samples from color changes during storage. The addition of 0.5 and 0.75% ME significantly decreased lipid oxidation and microbial growth. The ME did not impact the sausages’ sensory quality, and the addition of 0.75% prevented sensory rejection during storage.