Identification of the SP22 Sperm Protein in Santa Inês and Dorper Rams

The sperm membrane protein referred to as SP22 has been identified in different species and, at least in rats, is highly correlated with fertility. The goals of this study were to identify and to quantify the SP22 protein on spermatozoa from adult rams (Dorper and Santa Inês breeds), and to correlate its levels to morphological and kinematics parameters. SP22 on ram sperm was effectively quantified by both enzyme‐linked immunosorbent assay (ELISA) and fluorescein isothiocyanate immunostaining analysis and the two methods were significantly correlated (R² = 0.70). Clustering analysis of motility parameters obtained by computer‐assisted semen analysis system was used to establish that three distinct kinematic subpopulations with different vigour and progressiveness coexistent within ejaculate. While there were significant differences in the distribution of the three subpopulations in the rams, there was no significant correlation between the proportion of each subpopulation in the rams and the SP22 levels. Quantification of SP22 immunostaining intensity was not correlated with any of the sperm parameters. However, SP22 levels obtained by ELISA were negatively correlated with morphological abnormalities and positively correlated with membrane integrity (three variable R² = 0.47). Future breeding studies are now needed to validate that this protein is a biomarker of fertility in this species.     

Detection and Characterization of the Cumulene Carbenes H2C5 and H2C6

Two cumulene carbenes, H2C5 and H2C6, were detected in a supersonic molecular beam by Fourier transform microwave spectroscopy. Their rotational and leading centrifugal distortion constants were determined with high accuracy, such that the entire radio spectrum can now be calculated. Like the known carbenes H2C3 and H2C4, both molecules have singlet electronic ground states and linear carbon-chain backbones. They can be produced in sufficiently high concentrations in the laboratory that their electronic spectra, expected to lie in the visible, should be readily detectable by laser spectroscopy. The microwave spectra of other, more exotic isomers may be detectable as well.     

First identification and characterization of Streptococcus iniae obtained from tilapia (Oreochromis aureus) farmed in Mexico

This is the first study to isolate, identify and characterize Streptococcus iniae as the causative disease agent in two tilapia (Oreochromis aureus) populations. The populations were geographically isolated, of distinct origins, and did not share water sources. Affected fish showed various external (e.g., exophthalmia and cachexia, among others) and internal (e.g., granulomatous septicaemia and interstitial nephritis, among others) signs. All internal organ samples produced pure cultures, two of which (one from each farm, termed S‐1 and S‐2) were subjected to biochemical, PCR and 16S rRNA sequencing (99.5% similarity) analyses, confirming S. iniae identification. The two isolates presented genetic homogeneity regardless of technique (i.e., RAPD, REP‐PCR and ERIC‐PCR analyses). Pathogenic potentials were assessed through intraperitoneal injection challenges in rainbow trout (Oncorhynchus mykiss) and zebrafish (Danio rerio). Rainbow trout mortalities were respectively 40% and 70% at 10⁴ and 10⁶ CFU per fish with the S‐1 isolate, while 100% mortality rates were recorded in zebrafish at 10² and 10⁴ CFU per fish with the S‐2 isolate. The obtained data clearly indicate a relationship between intensified aquaculture activities in Mexico and new disease appearances. Future studies should establish clinical significances for the tilapia industry.     

Comparison of mortality and viral load in rainbow trout (Oncorhynchus mykiss) infected with infectious pancreatic necrosis virus (IPNV) genogroups 1 and 5

Infectious pancreatic necrosis virus (IPNV) is the aetiological agent of a highly contagious disease that affects farmed salmonids. IPNV isolates have been phylogenetically classified into eight genogroups, of which two are present in Chile, genogroups 1 and 5. Here, we compare the mortality rate caused by isolates from both genogroups in rainbow trout (Oncorhynchus mykiss) fry to determine if there is an association between host susceptibility and phylogenetic characterization of IPNV. Fish were challenged by immersion with one of four isolates (two for each genogroup), and mortality curves were assessed after 30 days. Viral load was measured in all mortalities and in live fish sampled at 1, 7 and 20 days post-infection. Although mortality was low throughout the challenge, differences were found between fish infected with different isolates. Both isolates from genogroup 1 caused greater cumulative mortalities than either of the isolates from genogroup 5. When combined, the overall mortality rate of fish challenged with genogroup 1 isolates was significantly higher than those infected with genogroup 5. However, viral load was lower on trout infected with genogroup 1 isolates. These results suggest that rainbow trout are more susceptible to IPNV isolates from genogroup 1 than genogroup 5.     

Effects of thyroxine administration on the growth and survival of pike silverside (Chirostoma estor) juveniles

The pike silverside has high aquaculture potential despite its slow growth. In this study, thyroxine (T₄) concentrations (0.13, 1.3 and 13 nM), a control with no hormonal supplement and a negative control containing 4.5 mM methimazole (MMI) were tested to evaluate the growth of this species. Juveniles (0.2 g) were exposed by immersion to these treatments for 8 h every second day for 120 days, and growth evaluations were performed monthly over the entire trial period. In addition, tissue samples from each treatment group were assayed for triiodothyronine (T₃) and deiodinase type 2 (D2) activity . The survival rates in all T₄ groups were high, and a significant increase in growth was observed (average of 58%). The MMI treatment caused an increase in mortality and a reduction in the final body weight compared with the control. T₄ administration did not affect the tissue levels of T₃, and it decreased muscular D2 activity only after 30 days of exposure. These results demonstrated that low concentrations of T₄ in the culture environment could improve the growth of this species without affecting tissue hormone levels. The technique may have useful applications for early‐stage aquaculture of this and other economically important species.     

Coastal geomorphology and oceanographic features shape subtidal benthic communities in management areas of central Chile

1. The spatial variation in community structure among 30 areas for the management and exploitation of benthic resources (AMEBRs) in central Chile was assessed from surveys conducted during the same spring–summer season. One hundred taxa identified in the subtidal surveys were grouped into 10 functional groups.
2. The influence and relative importance of external factors over benthic associations were assessed through nonmetric multidimensional scaling (NMDS) and path analysis. Three main categories of external factors were considered: ‘terrain traits’ (e.g. coastline exposure), ‘surface-ocean conditions’ (e.g. sea surface temperature, SST), and ‘fishery-related variables’ (e.g. cumulative landings).
3. The NMDS revealed a strong association between benthic community structure and both terrain and surface-ocean conditions (42% of variance explained by permutational multivariate analysis of variance), but not with fishery-related variables. Assemblages typical of rocky substrates were associated with larger seasonal fluctuations in SST and surface chlorophyll a. The presence of soft-bottom functional groups was associated with higher levels of surface fluorescence and larger seasonal fluctuations in a satellite-derived proxy for river-plume influence (nLw645).
4. A path analysis model for hard-bottom communities reached a 49% goodness of fit (GOF), with the presence of epifaunal filter-feeders, grazers, predators, and scavengers being significantly correlated with leathery and corticated foliose algae, which in turn depend on the fraction of rocky substrate in the AMEBR; the fraction of rocky substrate is positively correlated with coastline exposure.
5. The model for soft-bottom communities reached a 52% GOF with the presence of infaunal filter-feeders and epifaunal detritivores depending directly on SST and surface chlorophyll a. The fishery-related variables considered in the model had no apparent effect on the structure of either type of subtidal community.
6. These results provide a hierarchy of physical–biological associations determining the functional composition of subtidal benthic communities in management areas along central Chile. This knowledge may improve future assessments of ecosystem response to changes in environmental conditions and management strategies.

     

Morphology of alpaca (Vicugna pacos) embryos in the first third of pregnancy

The breeding of South American camelids is the main economic activity of the high Andean region of South America and it, is potentially, the most profitable resource in of the Puna environmental conditions of the Puna. The duration of the gestation in alpaca is 339.7 ± 12 days. The objective of the present work was to macroscopically and microscopically describe the ontogenic development of the splanchnic cavities of the alpaca and to determine the gestational time in which the post‐cranial ossification centers are observed in the embryos/fetuses of this species, from day 21 to 107 of gestation. The documentation of normal ontogenic development, which is vacant for this period, is of the utmost importance to understand the consequences of the alterations at the different gestational times, as well as for the estimation of the gestational age in the case of abortions. Forty‐seven alpaca specimens of both sexes, at different times of their gestational development, collected during slaughter at local slaughterhouses of the Department of Huancavelica, Peru, were evaluated. Specimens were assigned to seven groups according to their morphological characteristics. The embryogenesis in the alpaca was characterized by a series of changes comparable to those occurring in other mammals with similar gestational periods. Despite these similarities, species differences were found in some organs as stomach, which are observed too in adult individuals.     

Identification of Apoptotic Bodies in Equine Semen

Apoptosis in the testis is required to ensure an efficient spermatogenesis. However, sometimes, defective germ cells that are marked for elimination during this process escape elimination in the testes, giving rise to ejaculates with increased percentages of abnormal and apoptotic spermatozoa and a high percentage of apoptotic bodies. Apoptosis markers in the ejaculate have been associated with low fertility, either in animals or humans. Therefore, the goal of this study was to investigate whether fresh equine semen contains apoptotic bodies [initially named Merocyanine 540 (M540) bodies] and to study the relationship between the quantity of these bodies and cell concentration, the volume of ejaculate, viability and motility. Moreover, we also studied whether the presence apoptotic bodies in fresh semen was related to the resistance of the stallion spermatozoa to being incubated at 37°C or being frozen and thawed. Fresh equine semen was stained with fluorescent dyes such as M540 and Annexin‐V. Active Caspase 3 was studied in fresh semen through Western blotting and immunofluorescence with a specific antibody. Sperm kinematics was assessed in fresh, incubated and thawed samples using computer‐assisted semen analysis, and viability was evaluated with the LIVE/DEAD Sperm Viability Kit. Overall, our results demonstrate for the first time the presence of apoptotic bodies in equine semen. The quantity of apoptotic bodies was highly variable among stallions and was positively correlated with Caspase 3 activity in fresh samples and negatively correlated with the viability and motility of stallion spermatozoa after the cryopreservation process.     

Locomotor effects in sheep of alkaloids identified in Australian Tribulus terrestris

Fresh, mature, ungrazed Tribulus terrestris plant material was subjected to a standard alkaloid extraction procedure. The extract was fractionated by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). Two major alkaloid fractions were demonstrated. These fractions were identified by means of TLC, ultraviolet spectrofluorimetry (UVS) and HPLC, as the beta-carboline indoleamines harmane and norharmane. The extractable alkaloid content was determined to be 44 mg/kg dry matter. Synthetic harmane and norharmane were administered subcutaneously to sheep at a dose rate of 54 mg/kg. Both compounds caused similar nervous effects. The main effect observed was limb paresis, which in some sheep was body side blased. The clinical signs observed in the experimental sheep were consistent with those described for naturally occurring cases of Tribulus terrestris staggers. It was proposed that harmane and norharmane accumulate in tryptamine-associated neurones of the central nervous system, during months of tribulus ingestion, and gradually interact irreversibly with a specific neuronal gene DNA sequence.