Immunolocalization of the Cholesterol Transporters ABCA1 and ABCG1 in Canine Reproductive Tract Tissues and Spermatozoa

The mammalian sperm membrane undergoes cholesterol efflux during maturation and fertilization. Although ATP‐binding cassette (ABC) transporters are known to transport cholesterol through cell membranes in other organs, their presence in canine testis, epididymis and sperm has not been proven to date. Hence, the aim of the present study was to localize the ABC transporters ABCA1 and ABCG1 in canine testicular and epididymidal tissue as well as in spermatozoa membranes. To this end, semen samples from 12 dogs as well as testicles and epididymides of four young and healthy dogs were prepared for immunohistochemistry, respectively. Capacitation and acrosome reaction (AR) were induced in aliquots of the semen samples before immunostaining to assess changes in the expression of ABCA1 and ABCG1. Evaluation by confocal microscopy revealed the presence of both ABCA1 and ABCG1 in canine testicles and of ABCA1 in the epididymides. In spermatozoa, only ABCA1 immunoreactivity was detected, mainly in the region of the acrosome and midpiece. After induction of capacitation, ABCA1 signal persisted in the acrosome but disappeared after AR, indicating a loss of ABCA1 with the loss of the acrosome. We conclude that ABCA1 and ABCG1 are expressed in canine testis, whereas only ABCA1 is expressed in epididymis and spermatozoa membrane, both transporters probably contributing to the regulation of membrane cholesterol content.     

Effects of the supplementation with an high‐polyphenols extra‐virgin olive oil on kinetic sperm features and seminal plasma oxidative status in healthy dogs

The aim of this work was to evaluate the effects of the supplementation of two extra‐virgin olive oils (EVOO) having different polyphenols content, on canine spermatozoa kinetic parameters and seminal plasma oxidative status. The study was conducted on 12 clinically healthy dogs of different breeds (2–7 years, 5–48 kg of body weight) divided into two groups: an experimental group supplemented with EVOO (Coratina cultivar) high in polyphenols (H‐P) and a control group fed EVOO (Cima di Bitonto cultivar) low in polyphenols (L‐P). The oil was daily administered per os (1 ml/3 kg BW) before meal. Semen collection was made twice at 15 days distance (D0₁ and D0₂) and then at 30 (D30), 60 (D60) and 90 (D90) days. Semen concentration and kinetic parameters were measured using computer‐assisted sperm analysis (CASA) system to evaluate: sperm total count, sperm motile (MOT%), progressive motility (PROGR%) and its fractions, straight‐line velocity (VSL, μm/s), curvilinear velocity (VCL, μm/s), average path velocity (VAP, μm/s), amplitude of lateral head displacement (ALH, μm), beat cross frequency (BCF, Hz), straightness (STR%) and linearity (LIN%). On seminal plasma, reactive oxygen species (ROS) and biological antioxidant potential (BAP) were tested. From findings, no differences were found for sperm MOT, VSL, VCL, VAP, ALH, BCF, STR, LIN and BAP. A gradual enhancement of PROGR% was observed in H‐P group (p < .01). The ROS levels were higher in dogs H‐P compared to the other group (p < .05). In conclusion, our results highlight the positive effects of EVOO polyphenols on sperm PROGR% in healthy dogs.     

Dynamic use of coastal areas by bull sharks and the conciliation of conservation and management of negative human–wildlife interactions

1. Knowledge about spatial and temporal variability in the distribution and abundance of predators is necessary to adapt measures to mitigate human–wildlife interactions.
2. Acoustic telemetry and network analyses were used to investigate the spatial ecology of bull sharks, the species responsible for most shark bites in Reunion Island, one of the world's shark bite hotspots.
3. The west coast of the island was not used uniformly by every individual, with size predicting the movements of sharks along the coast.
4. Node-based metrics – closeness, node strength, and cumulated continuous residency times – derived from up to 181 monthly movement networks from 20 individuals, revealed that smaller sharks (<250 cm total length) primarily used the south-west coast while larger individuals spent most of their time in the northern region with regular visits to multiple areas along the coast.
5. This study provides essential knowledge on bull shark behaviour and central areas used at different periods of the year, which correlates well with the dynamics of observed shark bites. Our approach provides a non-invasive alternative to help predicting and anticipating human–shark conflicts and avoid shark culling programmes detrimental to the conservation of large predators such as sharks.

     

Egg quality criteria in Pacific red snapper (Lutjanus peru)

The aim of this study was to evaluate the biochemical parameters used as possible determinants of egg quality in Pacific red snapper (Lutjanus peru). Fertilized eggs of eight spawns were obtained by hormonal induction. Egg quality criteria, including abnormal cleavage (AC), hatching percentage (HR) and survival percentage at first feeding (SR) were recorded. Samples were taken during embryonic development and from yolk‐sac larvae. Proteins, energetic metabolite concentrations and metabolic and digestive enzyme activities were determined using colorimetric methods. Pearson's correlation, and simple and multiple regression models were performed using the biochemical parameters as the independent variables and AC, HR and SR as the dependant variables. Glucose‐6‐phosphatase activity (AC r = 0.87; HR r = ?0.65; SR r = ?0.67) and fructose concentration (AC r = ?0.64; HR r = 0.54; SR r = 0.64) were the only biochemical parameters to be strongly correlated with the three egg quality criteria. The use of multiple regression models increased the regression coefficient of the three quality criteria. Fructose, glucose and glucose‐6‐phosphatase were involved in all multiple regression models. The models proposed in this study may be used to explain egg quality for Pacific red snapper and their use as predictors of egg quality is discussed.     

Vitrification with Glutamine Improves Maturation Rate of Vitrified / Warmed Immature Bovine Oocytes

The current study examined the protective effects of l ‐glutamine and cytochalasin B during vitrification of immature bovine oocytes. Oocyte vitrification solution (PBS supplemented with 10% FCS, 25% EG, 25% DMSO and 0.5 m trehalose) was the vitrification control. Treatments were the addition of 7 μg/ml cytochalasin B, 80 mm glutamine or both cytochalasin and glutaminine for 30 s. After warming, oocytes were matured in vitro for 24 h, fixed and stained with Hoechst (33342) for nuclear maturation evaluation. l ‐glutamine improved the vitrified/warmed immature bovine oocytes viability (32.8%), increasing the nuclear maturation rates compared to other treatments and the no treatment vitrified control (17.4%). There was, however, no effect of cytochalasin B on in vitro maturation (14.4%).     

Pregnancy Rates to Fixed Embryo Transfer of Vitrified IVP Bos indicus,Bos taurus or Bos indicus × Bos taurus Embryos

The pregnancy rates obtained after the transfer of cryopreserved in vitro‐produced (IVP) embryos are usually low and/or inconsistent. The objective of this study was to evaluate the pregnancy rates of Holstein, Gyr and Holstein × Gyr cattle after the transfer of vitrified IVP embryos produced with X‐sorted sperm. Seventy‐two Gyr and 703 Holstein females were subjected to ovum pickup (OPU) sessions, followed by in vitro embryo production using semen from sires of the same breeds. Embryos (1636 Holstein, 241 Gyr and 1515 Holstein × Gyr) were exposed to forskolin for 48 h prior to vitrification. The pregnancy rate achieved with Gyr dam and sire was 46.1%, which was similar (p = 0.11) to that of Holstein dam and Gyr sire (40.3%). Crossing Gyr dams with Holstein sires resulted in a pregnancy rate of 38.9% and did not differ (p = 0.58) from the pregnancy rate obtained with the cross between Holstein dams and Gyr sires. The rate obtained with Holstein dam and sire was 32.5%. The average pregnancy rate was 36.6%, and no difference was found in the proportion of female foetuses (88.8%, in average) among breeds (p > 0.05). In conclusion, transfer of cryopreserved X‐sorted embryos represents an interesting choice for dairy cattle. Despite the small differences between pregnancy rates, we highlight the efficiency of this strategy for all of the racial groups studied.     

Effect of Salinity on Growth and Survival in Juvenile Opossum Pipefish,Microphis brachyurus,in Culture Conditions

Natural populations of the opossum pipefish, Microphis brachyurus, are affected in Mexico by anthropogenic alteration of their habitat and unregulated fishing as this species is traded as an aquarium fish without formal records as occurs with other syngnathids around the world. M. brachyurus is an estuarine fish adapted to salinity fluctuations. The aim of this study was to examine the effect on growth, condition, and survival of juvenile pipefish cultured for 4 wk at salinities of 0, 8, and 16 g/L. Pipefish were fed enriched Artemia nauplii maintaining a ration rate of 14% body weight per day (dry weight Artemia: wet weight fish). Pipefish cultured at 16 g/L showed lower survival (40%) than those at 0 and 8 g/L (100 and 96%, respectively), while juveniles cultured in 8 and 16 g/L were heavier (0.393 and 0.388 g, respectively), longer (9.9 cm), and grew faster (1.4) than those in 0 g/L (0.294 g, 9.1 cm, and 0.44, respectively). This study is the first to report that a salinity of 16 g/L compromises M. brachyurus culture potential and a salinity of 0 g/L caused poor growth while the optimal salinity for the species appears to be around 8 g/L.     

Pathological investigations of pigs with porcine multisystemic wasting syndrome (PMWS)

Extract

Pathological changes and immunohistochemical (IHC) examination for porcine circovirus-2 (PCV-2) in nine weaner pigs from a property with PMWS are reported. The pigs were in moderate to poor body condition, and several had diarrhoea and noisy respiration. The outstanding pathological changes related to microscopic lesions in lymphoid tissues, including lymph nodes, palatine tonsils, Peyer's patches, peribronchial lymphoid aggregates and spleen. While there was variability of expression of change between and within cases, the main features of lesions could be summarised as depletion of lymphoid cells, infiltration of histiocytes (often syncytial), and the presence of botryoid intra-cytoplasmic inclusion bodies in macrophages. The changes in lymphoid tissue were typical of those described in PMWS. Also characteristic of this syndrome were histiocyte-dominated cellular inflammatory infiltrates in kidney, liver and lung. There was strong correlation between IHC positivity for PCV-2 and the lesions in lymphoid, pulmonary and renal tissues. Concurrent diseases that are probably promoted by immune suppression may complicate the diagnosis of PMWS. In the present cases, salmonellosis, attaching and effacing Escherichia coli infections, and secondary bacterial bronchopneumonia were identified.