Successful Recovery of Motility and Fertility of Cryopreserved Cane Toad (Bufo marinus) Sperm

The recent decline and extinction of amphibian species is a worldwide phenomenon without an identified cause or solution. Assisted reproductive technologies, including sperm cryopreservation, are required to manage endangered amphibian species and preserve their genetic diversity. This study on the Anuran amphibian (Bufo marinus) was undertaken to determine the feasibility of cryopreservation of amphibian sperm. Sperm suspensions for cryopreservation were prepared by macerating testes in cryoprotective additives of 10% (w/v) sucrose or 10% (w/v) sucrose containing either 10, 15, or 20% (v/v) glycerol or 10, 15, or 20% (v/v) dimethyl sulfoxide (Me₂SO). Suspensions were then cooled to −85°C using a controlled rate cooler, stored in LN₂, and thawed in air. The motility and fertilization rate of cryopreserved suspensions and unfrozen control suspensions in Simplified Amphibian Ringer were compared. Sucrose alone had no cryoprotective effect. All other treatments showed varying degrees of recovery of motility and fertilizing capacity. High rates of recovery of motility and fertilizing capacity were observed with 15% Me₂SO (68.9 ± 3.8 and 60.5 ± 4.7%) and 20% glycerol (58.0 ± 5.9 and 81.4 ± 4.3%), respectively. Motility and fertilization rates were similar with Me₂SO but diverged with glycerol as cryoprotectant. The data demonstrate the feasibility of using sperm cryopreservation with amphibian species.     

Comparative analysis of the zeta-crystallin/quinone reductase gene in guinea pig and mouse

zeta-Crystallin is a novel nicotinamide adenine dinucleotide phosphate:quinone reductase, present at enzymatic levels in various tissues of different species, which is highly expressed in the lens of some hystricomorph rodents and camelids. We report here the complementary DNA (cDNA) cloning of zeta-crystallin from liver libraries in guinea pig (Cavia porcellus), where zeta-crystallin is highly expressed in the lens, and in the laboratory mouse (Mus musculus), where expression in the lens occurs only at enzymatic levels. A 5' untranslated sequence different from the one previously reported for the guinea pig lens cDNA was found in these clones. We also report the isolation of genomic clones including the complete guinea pig zeta-crystallin gene and the 5' region of this gene in mouse. These results show the presence of two promoters in the guinea pig zeta-crystallin gene, one responsible for expression at enzymatic levels and the other responsible for the high expression in the lens. The guinea pig lens promoter is not present in the mouse gene. This is the first example in which the recruitment of an enzyme as a lens crystallin can be explained by the acquisition of an alternative lens- specific promoter.      

Predator Presence and Vegetation Density Affect Capture Rates and Detectability of Litoria aurea Tadpoles: Wide-Ranging Implications for a Common Survey Technique

Trapping is a common sampling technique used to estimate fundamental population metrics of animal species such as abundance, survival and distribution. However, capture success for any trapping method can be heavily influenced by individuals’ behavioural plasticity, which in turn affects the accuracy of any population estimates derived from the data. Funnel trapping is one of the most common methods for sampling aquatic vertebrates, although, apart from fish studies, almost nothing is known about the effects of behavioural plasticity on trapping success. We used a full factorial experiment to investigate the effects that two common environmental parameters (predator presence and vegetation density) have on the trapping success of tadpoles. We estimated that the odds of tadpoles being captured in traps was 4.3 times higher when predators were absent compared to present and 2.1 times higher when vegetation density was high compared to low, using odds ratios based on fitted model means. The odds of tadpoles being detected in traps were also 2.9 times higher in predator-free environments. These results indicate that common environmental factors can trigger behavioural plasticity in tadpoles that biases trapping success. We issue a warning to researchers and surveyors that trapping biases may be commonplace when conducting surveys such as these, and urge caution in interpreting data without consideration of important environmental factors present in the study system. Left unconsidered, trapping biases in capture success have the potential to lead to incorrect interpretations of data sets, and misdirection of limited resources for managing species.     

Preliminary evidence for a two‐for‐one deal: Wetland restoration for a threatened frog may benefit a threatened bat

Habitat restoration is an integral feature of wildlife conservation. However, funding and opportunities for habitat restoration are limited, and therefore, it is useful for targeted restoration to provide positive outcomes for non‐target species. Here, we investigate the possibility of habitat creation and management benefitting two threatened wetland specialists: the Green and Golden Bell Frog (Litoria aurea) and the Large‐footed Myotis (Myotis macropus). This study involved two components: (i) assessing co‐occurrence patterns of these species in a wetland complex created for the Green and Golden Bell Frog (n = 9) using counts, and (ii) comparing foraging activity of Large‐footed Myotis in wetlands with low and high aquatic vegetation (n = 6 and 7, respectively) using echolocation metres. Since Large‐footed Myotis possesses a unique foraging behaviour of trawling for aquatic prey, we hypothesised that foraging activity of this species would be higher in wetlands with low aquatic vegetation coverage. Additionally, we provide observations of its potential prey items. We identified one created wetland where both species were found in relatively high numbers, and this wetland had a permanent hydrology, was free of the introduced fish Gambusia (Gambusia holbrooki) and had low aquatic vegetation coverage. We also found that Myotis feeding activity was significantly higher in low aquatic vegetation coverage wetlands (x? = 65.72 ± 27.56 SE) compared to high (x? = 0.33 ± 0.33 SE, P = 0.0000). Although this is a preliminary study, it seems likely that Green and Golden Bell Frog and Large‐footed Myotis would gain mutual benefit from wetlands that are constructed to be permanent, that are Gambusia free, low in aquatic vegetation coverage, and are located in close to suitable roosting habitat for Large‐footed Myotis. We encourage adaptive aquatic vegetation removal for Green and Golden Bell frog as this may have benefits for Large‐footed Myotis. The evidence suggests that the former may be a suitable umbrella species for the latter.     

Ovarian activity and plasma sex steroid levels of Distichodus antonii in relation to environmental conditions in the upper basin of the Congo River

Distichodus antonii is an endemic fish species of the Congo River basin in which the stocks of wild populations are threatened by overfishing pressure. Knowledge of its reproductive biology would be useful in consideration of conservation and management options for the species. Therefore, this study investigated changes in ovarian activity and levels of steroid profiles in wild populations in relation to variation in temperature and rainfall. Adult females (n = 101, body weight of 3 183 ± 14.75 g, SE) were captured monthly over one year (2013–2014). Apart from evaluation of oocyte diameters and gonad developmental stages, gonado-, hepato-, lipososomatic indices (GSI, HSI, LSI) and plasma levels of sex steroids (testosterone-T, estradiol-17β-E2) were determined. The results suggested a synchronous development of oocytes with two annual reproductive seasons over the one-year study. Plasma T and E2 levels peaked during spawning periods likely reflecting active oogenesis. The highest values of morphosomatic indices were observed during the longest rainfall period in September, and were associated with high steroidogenic activity evidenced by increased E2 production. In addition, more vitellogenic oocytes (September and October) were observed during the latter season than during the short rainy season (in May).     

AIR: A batch-oriented web program package for construction of supermatrices ready for phylogenomic analyses

Background  

Large multigene sequence alignments have over recent years been increasingly employed for phylogenomic reconstruction of the eukaryote tree of life. Such supermatrices of sequence data are preferred over single gene alignments as they contain vastly more information about ancient sequence characteristics, and are thus more suitable for resolving deeply diverging relationships. However, as alignments are expanded, increasingly numbers of sites with misleading phylogenetic information are also added. Therefore, a major goal in phylogenomic analyses is to maximize the ratio of information to noise; this can be achieved by the reduction of fast evolving sites.     

A comparison between freezing methods for the cryopreservation of stallion spermatozoa

The effects of sperm freezing concentration (40 × 10⁶ mL⁻¹ vs. 400 × 10⁶ mL⁻¹), straw size (0.25 mL vs. 0.5 mL) and freezing method (liquid nitrogen vapour in a Styrofoam^® box vs. programmable freezing machine) were evaluated in a 2 × 2 × 2 factorial experimental design using 3 split ejaculates from each of 4 stallions. Immediately after thawing, the total motility and forward progressive motility of spermatozoa frozen at a concentration of 40 × 10⁶ mL⁻¹ was higher than for spermatozoa frozen at 400 × 10⁶ mL⁻¹. No significant differences were observed in the semen parameters assessed after cryopreservation in either 0.25 or 0.5 mL straws. However, the programmable freezer provided a more consistent and reliable freezing rate than liquid nitrogen vapour. We conclude that an effective protocol for the cryopreservation of stallion spermatozoa at low concentrations would include concentrations of 40 × 10⁶ mL⁻¹ in 0.25 mL straws using a programmable freezer. This freezing protocol would be suitable for emerging sperm technologies such as sex-preselection of stallion spermatozoa as the sorting process yields only low numbers of spermatozoa in a small volume available for either immediate insemination or cryopreservation.     

Invasion status of Florida bass Micropterus floridanus (Lesueur, 1822) in South Africa

Largemouth bass Micropterus salmoides are a popular North American angling species that was introduced into South Africa in 1928. To enhance the largemouth bass fisheries, Florida bass Micropterus floridanus were introduced into KwaZulu Natal, South Africa, in 1980. Knowledge on the status of M. floridanus in South Africa is required, because it lives longer and reaches larger sizes than M. salmoides, which may result in heightened impacts on native biota. Because M. floridanus are morphologically similar, but genetically distinct from M. salmoides, the distribution of this species was assessed by genetically screening 185 Micropterus sp. individuals sampled from 20 localities across South Africa using the mitochondrial ND2 gene. Individuals with mitochondrial DNA matching M. salmoides were recovered from 16 localities, whereas M. floridanus mitochondrial DNA was recovered from 13 localities. At nine localities (45%), the mitochondrial DNA of both species was detected. These results demonstrate M. floridanus dispersal to multiple sites across South Africa.     

Growth factor activity in luminal fluids from the male reproductive tract of the ram, rat, tammar wallaby (Macropus eugenii) and Japanese quail (Coturnix coturnix japonica)

Growth factor activity in luminal fluids from the male reproductive tract was assayed by measuring the stimulation of [3H]thymidine incorporation into BALB/c 3T3 fibroblasts. The potency of fluids from the rete testis of the rat, ram, tammar wallaby and Japanese quail was much the same. However, about 90% of the activity in fluid from the rete testis of the rat and tammar was lost during its passage through the epididymis.