Determination of Feeder Cell‐Based Cellular Niches Supporting the Colonization and Maintenance of Spermatogonial Stem Cells from Prepubertal Domestic Cat Testes

Recently, isolation and in vitro culture of putative spermatogonial stem cells (SSCs) in the domestic cat have been conducted. However, the cellular niche conditions that facilitate the establishment and long‐term maintenance of feline SSCs (FSSCs) have not been described. Therefore, we investigated the type of feeder cells used to stimulate colony formation and growth of FSSCs among the various factors in the FSSC niche. Spermatogonial stem cells isolated from feline testes were cultured on mitotically inactivated testicular stromal cells (TSCs) derived from cats, dogs and mice, and mouse embryonic fibroblasts (MEFs). The formation and growth of colonies derived from SSCs cultured on each type of feeder cell were identified at passage 0, and the morphology, alkaline phosphatase (AP) activity and expression of SSC‐specific genes in surviving colonies were investigated at passage 4. Among these diverse feeder cells, TSCs from cat showed the greatest colony formation, growth and maintenance of FSSCs, and SSC colonies cultured by passage 4 showed a typical dome‐shaped morphology, AP activity and expression of SSC‐specific genes (NANOG, OCT4, SOX2 and CD9). Accordingly, these results demonstrate that feline TSCs could be used as feeder cells to support the establishment and maintenance of SSCs from domestic cats.     

Prevalence,Serovars and Antimicrobial Susceptibility of Salmonella spp. from Wild and Domestic Green Iguanas (Iguana iguana) in Grenada,West Indies

Cloacal swabs from 62 green iguanas (Iguana iguana), including 47 wild and 15 domestic ones from five parishes of Grenada, were sampled during a 4‐month period of January to April 2013 and examined by enrichment and selective culture for the presence of Salmonella spp. Fifty‐five per cent of the animals were positive, and eight serovars of Salmonella were isolated. The most common serovar was Rubislaw (58.8%), a serovar found recently in many cane toads in Grenada, followed by Oranienburg (14.7%), a serovar that has been causing serious human disease outbreaks in Japan. Serovar IV:48:g,z₅₁:‐ (formerly, S. Marina) highly invasive and known for serious infections in children in the United States, constituted 11.8% of the isolates, all of them being from domestic green iguanas. Salmonella Newport, a serovar recently found in a blue land crab in Grenada, comprised 11.8% of the isolates from the green iguanas. The remaining four less frequent serovars included S. Javiana and S. Glostrup. Antimicrobial susceptibility tests conducted by a disc diffusion method against amoxicillin–clavulanic acid, ampicillin, cefotaxime, ceftazidime, ciprofloxacin, enrofloxacin, gentamicin, nalidixic acid, streptomycin, tetracycline and trimethoprim–sulfamethoxazole showed that drug resistance is minimal, with intermediate susceptibility, mainly to streptomycin, tetracycline and cefotaxime. This is the first report of isolation and antimicrobial susceptibilities of various Salmonella serovars from wild and domestic green iguanas in Grenada, West Indies.     

Australian sperm whales from different whaling stocks belong to the same population

1. Understanding the factors driving population structure in marine mammals is needed to evaluate the impacts of previous exploitation, current anthropogenic threats, conservation status, and success of population recovery efforts.
2. Sperm whales are characterized by a worldwide distribution, low genetic diversity, complex patterns of social and genetic structure that differ significantly within and between ocean basins, and a long history of being commercially whaled. In Australia, sperm whales from the (International Whaling Commission assigned) southern hemisphere ‘Division 5’ stock were very heavily exploited by whaling.
3. The present study assessed the potential effects of whaling on the genetic diversity of sperm whales in Australia and the population genetic structure of these whales within a global context. A combination of historical and contemporary sperm whale samples (n = 157) were analysed across six regions, from south-eastern Australia (‘Division 6’ stock in the Pacific Ocean) to south-western Australia (‘Division 5’ stock in the Indian Ocean).
4. Sperm whales sampled from the ‘Division 5’ and ‘Division 6’ stocks belong to the same population based on nuclear and mitochondrial DNA (mtDNA) analyses. Four novel sperm whale mtDNA haplotypes were identified in animals from Australian waters. Levels of genetic diversity were low in Australian sperm whales but were similar to those previously reported for populations in the Indian and Pacific Oceans.
5. Given the genetic distinctiveness of sperm whales in Australian waters from other regions in the Pacific and Indian Oceans, and the lack of recovery in population numbers, further scientific studies are needed to increase our understanding of population dynamics and the effectiveness of threat management strategies in this species.

     

Effect of hydrodynamic dispersion on phosphatase reactions in a soil column

Columns of artificially-prepared soil crumbs were perfused with solutions of p-nitrophenyl phosphate and rates of hydrolysis of this substrate as catalyzed by soil phosphatase were measured. Hydrolysis proceeded by a zero-order or by a first-order reaction at high and low concentrations of substrate, respectively. Steady-state concentrations of substrate and products from the columns can be described essentially in terms of plug flow kinetics with but minor corrections for hydrodynamic dispersion. Values of the zero and first-order rate constants corrected for dispersion are 3 and 9 per cent greater, respectively, than those assuming plug flow kinetics. The apparent value of the first-order rate constant is about 70 percent higher than the true value because of electrostatic repulsion between the negatively-charged substrate and the soil particles.     

Monitoring distances travelled by horses using GPS tracking collars

Objective The aims of this work were to (1) develop a low-cost equine movement tracking collar based on readily available components, (2) conduct preliminary studies assessing the effects of both paddock size and internal fence design on the movements of domestic horses, with and without foals at foot, and (3) describe distances moved by mares and their foals. Additional monitoring of free-ranging feral horses was conducted to allow preliminary comparisons with the movement of confined domestic horses. Procedures A lightweight global positioning system (GPS) data logger modified from a personal/vehicle tracker and mounted on a collar was used to monitor the movement of domestic horses in a range of paddock sizes and internal fence designs for 6.5-day periods. Results In the paddocks used (0.8–16 ha), groups of domestic horses exhibited a logarithmic response in mean daily distance travelled as a function of increasing paddock size, tending asymptotically towards approximately 7.5 km/day. The distance moved by newborn foals was similar to their dams, with total distance travelled also dependent on paddock size. Without altering available paddock area, paddock design, with the exception of a spiral design, did not significantly affect mean daily distance travelled. Feral horses (17.9 km/day) travelled substantially greater mean daily distances than domestic horses (7.2 km/day in 16-ha paddock), even when allowing for larger paddock size. Conclusions Horses kept in stables or small yards and paddocks are quite sedentary in comparison with their feral relatives. For a given paddock area, most designs did not significantly affect mean daily distance travelled.     

Effects of Superior Check Desmotomy on Flexor Tendon and Suspensory Ligament Strain in Equine Cadaver Limbs

OBJECTIVE: To measure the effects of transection of the accessory ligament of the superficial digital flexor (SDF) muscle (superior check desmotomy) on flexor tendon and suspensory ligament (SL) strain in vitro. STUDY DESIGN: In vitro experimental biomechanical investigation. ANIMALS USED: Ten equine cadaver forelimbs. METHODS: The effects of superior check desmotomy were determined using equine cadaver forelimbs secured in a servocontrolled hydraulic testing machine. Strain sensors were used to measure strain on the superficial and deep digital flexor tendons and SL, and a goniometer was used to measure joint angles when the limb was loaded at 890 N and 3,115 N before desmotomy, and at 3,115 N after desmotomy. RESULTS: Superior check desmotomy was associated with significantly increased strains on the SDF tendon and SL, and significant alterations in the angles of the metacarpophalangeal and carpal joints. CONCLUSIONS: The superior check ligament has an important role in maintaining joint angles and load distribution in the forelimb. Lengthening of the SDF musculotendinous unit after superior check desmotomy may be associated with increased strain on the SL. CLINICAL RELEVANCE: Transection of the accessory ligament of the SDF muscle may predispose horses to SL desmitis postoperatively.     

Unusual osteochondral lesions of the talus in a horse

A 2-year-old Thoroughbred gelding was evaluated for a grade 3 out of 5 unilateral hind limb lameness. Flexion of the right hock and stifle joints (spavin test) exacerbated the lameness. Response to intra-articular and perineural anaesthesia isolated the source of lameness to the tarsocrural area, despite an absence of tarsocrural joint effusion. Routine radiographic examination of the hock did not reveal any significant abnormalities. Skeletal nuclear scintigraphic evaluation revealed a focal region of increased bone activity in the proximal medial trochlear ridge of the talus. Flexed lateromedial radiographic views identified three discrete semicircular lytic lesions at the proximal articular margin of the medial trochlear ridge of the talus. Conservative management of the lesions was associated with a successful return to racing. The location and appearance of the osteochondral lesions of this report have not been previously reported and may be a manifestation of developmental orthopaedic disease and abnormal endochondral ossification. Nuclear scintigraphy and flexed lateromedial radiographic views facilitated identification of the lesions. This radiographic view is recommended when lameness is isolated to the tarsocrural joint and standard radio-graphic projections fail to identify a cause.     

Influence of premature induction of a luteinizing hormone surge with gonadotropin-releasing hormone on ovulation, luteal function, and fertility in cattle

We tested the hypothesis that luteal function and fertility would be reduced in cattle induced to ovulate prematurely compared with those ovulating spontaneously. Estrus was synchronized in 56 beef cows (24 that were nonlactating and 32 that were nursing calves). At 6.4 +/- 0.1 d after estrus, all follicles > or = 5 mm were aspirated (day of aspiration = d 0) with a 17-gauge needle using the ultrasound-guided transvaginal approach. On d 1.5 and 2, cows were administered 2 luteolytic doses of PGF2alpha. Ovarian structures were monitored by transrectal ultrasonography from d -2 to 12, or ovulation. Emergence of a new follicular wave occurred on d 1.7 +/- 0.1. When the largest follicle of the newly emerged wave was 10 mm in diameter (d 4.8 +/- 0.1), cows were assigned on an alternating basis to receive 100 microg of GnRH (GnRH-10; n = 29) to induce ovulation or, upon detection of spontaneous estrus, to the spontaneous (SPON) treatment (n = 24). Cows were bred by AI at 12 h after GnRH (GnRH-10) or 12 h after the onset of estrus (SPON) as detected using an electronic surveillance system. Blood samples were collected every other day beginning 2 d after ovulation until pregnancy diagnosis 30 d after AI. Ovulation and AI occurred in 29/29 cows in the GnRH-10 and in 24/24 cows in the SPON treatment. Ovulation occurred later (P < 0.05) in the SPON (d 7.7 +/- 0.1) than GnRH-10 (d 6.8 +/- 0.1) treatment. Double ovulations were detected in 47% of cows, resulting in 1.5 +/- 0.1 ovulations per cow. Diameters of the ovulatory and the second ovulatory (in cows with 2 ovulations) follicles were greater (P < 0.05) in the SPON (12.0 +/- 0.3 mm and 10.5 +/- 0.4 mm, respectively) than in the GnRH-10 (10.7 +/- 0.1 mm and 9.2 +/- 0.3 mm) treatment. Cross-sectional areas of luteal tissue and plasma concentrations of progesterone during the midluteal phase were greater (P < 0.05) in the SPON (3.62 +/- 0.2 cm2 and 6.4 +/- 0.3 ng/mL) than in the GnRH-10 (3.0 +/- 0.2 cm2 and 5.4 +/- 0.2 ng/mL) treatment. The conception rate to AI in the SPON (100%) treatment was greater (P < 0.05) than in the GnRH-10 (76%) treatment. The animal model used in this study resulted in unusually high conception rates and double ovulations. In conclusion, premature induction of the LH surge reduced the diameter of ovulatory follicle(s), the luteal function, and the conception rate to AI.