Effects of varying doses of ß-nerve growth factor on the timing of ovulation, plasma progesterone concentration and corpus luteum size in female alpacas (Vicugna pacos).

Ovulation in camelids is induced by the seminal plasma protein ovulation-inducing factor (OIF), recently identified as ß-nerve growth factor (ß-NGF). The present study measured the total protein concentration in alpaca seminal plasma using a bicinchoninic acid (BCA) protein quantification assay and found it to be 22.2±2.0mgmL(-1). To measure the effects of varying doses of ß-NGF on the incidence and timing of ovulation, corpus luteum (CL) size and plasma progesterone concentration, 24 female alpacas were synchronised and treated with either: (1) 1mL 0.9% saline (n=5); (2) 4µg buserelin (n=5); (3) 1mg ß-NGF protein (n=5); (4) 0.1mg ß-NGF (n=5); or (5) 0.01mg ß-NGF (n=4). Females were examined by transrectal ultrasonography at 1-2-h intervals between 20 and 45h after treatment or until ovulation occurred, as well as on Day 8 to observe the size of the CL, at which time blood was collected to measure plasma progesterone concentrations. Ovulation was detected in 0/5, 5/5, 5/5, 3/5 and 0/4 female alpacas treated with saline, buserelin, 1, 0.1 and 0.01mg ß-NGF, respectively. Mean ovulation interval (P=0.76), CL diameter (P=0.96) and plasma progesterone concentration (P=0.96) did not differ between treatments. Mean ovulation interval overall was 26.2±1.0h. In conclusion, buserelin and 1mg ß-NGF are equally effective at inducing ovulation in female alpacas, but at doses ≤0.1mg, ß-NGF is not a reliable method for the induction of ovulation.

Proteomic characterization and cross species comparison of mammalian seminal plasma.

Seminal plasma contains a large protein component which has been implicated in the function, transit and survival of spermatozoa within the female reproductive tract. However, the identity of the majority of these proteins remains unknown and a direct comparison between the major domestic mammalian species has yet to be made. As such, the present study characterized and compared the seminal plasma proteomes of cattle, horse, sheep, pig, goat, camel and alpaca. GeLC-MS/MS and shotgun proteomic analysis by 2D-LC-MS/MS identified a total of 302 proteins in the seminal plasma of the chosen mammalian species. Nucleobindin 1 and RSVP14, a member of the BSP (binder of sperm protein) family, were identified in all species. Beta nerve growth factor (bNGF), previously identified as an ovulation inducing factor in alpacas and llamas, was identified in this study in alpaca and camel (induced ovulators), cattle, sheep and horse (spontaneous ovulators) seminal plasma. These findings indicate that while the mammalian species studied have common ancestry as ungulates, their seminal plasma is divergent in protein composition, which may explain variation in reproductive capacity and function. The identification of major specific proteins within seminal plasma facilitates future investigation of the role of each protein in mammalian reproduction. BIOLOGICAL SIGNIFICANCE: This proteomic study is the first study to compare the protein composition of seminal plasma from seven mammalian species including two camelid species. Beta nerve growth factor, previously described as the ovulation inducing factor in camelids is shown to be the major protein in alpaca and camel seminal plasma and also present in small amounts in bull, ram, and horse seminal plasma.

The effect of glycosaminoglycan enzymes and proteases on the viscosity of alpaca seminal plasma and sperm function.

In order to advance the development of cryopreservation and other assisted reproductive technologies in camelids it is necessary to eliminate the viscous component of the seminal plasma without impairing sperm function. It has been postulated that glycosaminoglycans (GAGs) or proteoglycans are responsible for this viscosity. This study investigated the effect of the GAG enzymes hyaluronidase, chondroitinase ABC and keratanase and the proteases papain and proteinase K on seminal plasma viscosity and sperm function in order to aid identification of the cause of seminal plasma viscosity and propose methods for the reduction of viscosity. Sperm motility, DNA integrity, acrosome integrity and viability were assessed during 2h incubation. All enzymes reduced seminal plasma viscosity compared to control (P<0.001) although papain was most effective, completely eliminating viscosity within 30 min of treatment. Sperm motility and DNA integrity was not affected by enzyme treatment. The proportion of viable, acrosome intact sperm was reduced in all enzyme treated samples except those treated with papain (P<0.001). These findings suggest that proteins, not GAGs are the main cause of alpaca seminal plasma viscosity. Papain treatment of alpaca semen may be a suitable technique for reduction of seminal plasma viscosity prior to sperm cryopreservation.

ß-Nerve growth factor is a major component of alpaca seminal plasma and induces ovulation in female alpacas.

Ovulation in camelids is induced by an unidentified protein in the seminal plasma of the male termed 'ovulation-inducing factor'. This protein has been reported to be a 14-kDa protein under reducing conditions, which, when purified from seminal plasma, induces ovulation in llamas. The identification of this protein and investigation of its potential to induce ovulation in camelids may aid the development of protocols for the induction of ovulation. In the present study, alpaca seminal plasma proteins were separated using one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the most abundant protein of 14 kDa was identified as ß-nerve growth factor (ß-NGF) by liquid chromatography mass spectrometry. Female alpacas (n = 5 per group) were given intramuscular injections of: (1) 1 mL of 0.9% saline; (2) 4 µg buserelin, a gonadotrophin-releasing hormone agonist; (3) 2 mL alpaca seminal plasma; or (4) 1mg human ß-NGF. Ovulation was detected by transrectal ultrasonography 8 days after treatment and confirmed by plasma progesterone concentrations. Ovulation occurred in 0%, 80%, 80% and 80% of animals treated with saline, buserelin, seminal plasma and ß-NGF, respectively. Treatment type did not affect the diameter of the corpus luteum, but plasma progesterone concentrations were lower in saline-treated animals than in the other treatment groups owing to the lack of a corpus luteum. The present study is the first to identify the ovulation-inducing factor protein in alpacas. ß-NGF successfully induces ovulation in alpacas and this finding may lead to new methods for the induction of ovulation in camelids.

Seminal plasma components in camelids and comparisons with other species.

Camelid semen is characterized by a highly viscous, low-volume ejaculate with a low concentration of spermatozoa that exhibit low progressive motility. The viscous seminal plasma is currently the major impediment to the development of assisted reproductive technologies (ARTs) in camelids. To advance ARTs such as sperm cryopreservation and artificial insemination in camelids, it is necessary to identify the cause of the viscosity and gain an understanding of the role of seminal plasma components on sperm function and fertility. Numerous compounds and proteins have been identified as mediators of sperm function and predictors of fertility in other livestock species, and understanding the importance of specific proteins has progressed the success of ARTs in these species. Current knowledge on the components of camelid seminal plasma is outlined, together with the implications of these components for the development of ARTs in camelids. The cause of semen viscosity, as well as proteins that are present in camelid seminal plasma, is described for the first time. Seminal plasma components are compared with those of other species to hypothesize their role in sperm function and fertility.

The effect of seminal plasma on alpaca sperm function.

In order to advance the development of assisted reproductive technologies in alpacas and other Camelids, the objective of this study was to explore the role of seminal plasma concentration on motility and functional integrity of alpaca sperm. Sixteen male alpacas > 3 y of age were used. In Experiment 1, epididymal sperm were incubated for 0 to 6 h in 0, 10, 25, 50, or 100% seminal plasma and motility was assessed. In Experiment 2, epididymal sperm were incubated in 0, 10, or 100% seminal plasma for 3 h and motility, acrosome integrity and DNA integrity were assessed. In Experiment 3, ejaculated sperm were incubated in 10, 25, 50, or 100% seminal plasma for 0 to 6 h and motility assessed. In Experiment 4, ejaculated sperm were incubated in 10 or 100% seminal plasma for 3 h and motility, acrosome integrity, DNA integrity, and viability were assessed. Epididymal and ejaculated sperm maintained motility longer when incubated in the presence of 10% seminal plasma compared to 0, 25, 50, or 100% seminal plasma (P < 0.001). The mean ± SEM percentage of epididymal sperm with intact acrosomes was less (P < 0.001) in samples incubated in 0% seminal plasma (39.4 ± 3.73) compared to 10% (75.3 ± 1.20) or 100% (77.4 ± 0.90) within 1 h after incubation. However, DNA integrity of ejaculated and epididymal sperm was not significantly affected by seminal plasma concentration. The mean viability of ejaculated sperm was reduced in the presence of 100 (12.7 ± 2.33) compared to 10% (36.2 ± 4.68) seminal plasma (P < 0.001) within 1 h of incubation. We concluded that alpaca semen should be diluted to a final concentration of 10% seminal plasma to prolong motility, preserve acrosome integrity, and maintain viability of sperm.

Intra-cervical application of Misoprostol at estrus alters the content of cervical hyaluronan and the mRNA expression of follicle stimulating hormone receptor (FSHR), luteinizing hormone receptor (LHR) and cyclooxygenase-2 in the ewe.

The complex anatomy the of ovine cervix limits the success of transcervical artificial insemination in sheep, but Misoprostol (a PGE(1) analogue) relaxes the cervix and facilitates transcervical artificial insemination. However, the mechanism by which Misoprostol causes cervical relaxation is not known. This study examined if intra-cervical Misoprostol altered the hyaluronan content and the mRNA expression of COX-2, LHR, or FSHR in the cervix of the estrus ewe. Estrus was synchronized in cyclic ewes with progestagen pessaries and 48 h after sponge removal ewes were treated intra-cervically with 0 (controls), 200, or 400 microg Misoprostol. Hyaluronan content was determined by ELISA and mRNA expression of LHR, FSHR, and COX-2 was analyzed by in situ hybridization using digoxigenin-11-uridine-5'-triphosphate labeled riboprobes. The hyaluronan content of the cervix was significantly higher in sheep that received 200 (P<0.05) or 400 (P<0.05) microg Misoprostol compared to controls. Moreover, it was significantly (P<0.05) higher in the vaginal region compared to mid and uterine regions. Misoprostol increased (P<0.05) the mRNA expression of LHR and COX-2 but not FSHR. The expression for all three genes was highest in the vaginal region and lowest in uterine region. The luminal epithelium and circular smooth muscle layers had higher (P<0.05) expression for LHR, FSHR, and COX-2 mRNAs, and the sub-epithelial stroma had the lowest (P<0.05). We propose that the intra-cervical application of Misoprostol induces the mRNA expression of LHR, FSHR, and COX-2 through a positive feedback loop. The data suggest that softening of the cervix by Misoprostol is caused by an increase in the hyaluronan content of the cervix.

The effect of estradiol on COX-2, EP2, and EP4 mRNA expression and the extracellular matrix in the cervix of the hypogonadotrophic, ovariectomized ewe.

There is a degree of cervical relaxation in the ewe at estrus that is regulated by changes in prostaglandin synthesis, prostaglandin receptor expression, and changes in the cervical extracellular matrix. It is likely that these are regulated by changes in periovulatory hormones, particularly estradiol. This study determined the effect of estradiol benzoate on the mRNA expression of cyclooxygenase-2 (COX-2) and the prostaglandin E receptors EP(2) and EP(4), the concentration of cervical hyaluronan, and the proportion of smooth muscle and collagen in the cervix of the hypogonadotrophic ovariectomized ewe (Ovis aries). Ovariectomized hypogonadotrophic ewes were given 100 microg estradiol benzoate, and their cervices were collected 0, 24, and 48 h thereafter to determine the expression of cervical COX-2, EP(2), and EP(4) mRNA by in situ hybridization, the concentration of hyaluronan by ELISA, and the proportion of smooth muscle and collagen by Masson's trichrome staining. Estradiol benzoate increased the mRNA expression of COX-2 and EP(4) within 24h after treatment (P<0.05), whereas EP(2) mRNA, hyaluronan, and the ratio of smooth muscle to collagen did not change within 48 h after treatment. The COX-2, EP(2), and EP(4) mRNA expression were greatest in the smooth muscle layers (P<0.05) and least in the luminal epithelium (P<0.05). In conclusion, we inferred that estradiol regulates cervical COX-2 and EP(4) mRNA expression and may regulate cervical relaxation via the synthesis of prostaglandin E(2) and activation of the PGE(2) receptors EP(2) and EP(4).

The mRNA expression of prostaglandin E receptors EP2 and EP4 and the changes in glycosaminoglycans in the sheep cervix during the estrous cycle.

Transcervical artificial insemination in sheep is limited by the inability to completely penetrate the cervix with an inseminating pipette. Penetration is partially enhanced at estrus due to a degree of cervical relaxation, which is probably regulated by cervical prostaglandin synthesis and extracellular matrix remodeling. Prostaglandin E(2) acts via prostaglandin E receptors EP(1) to EP(4), and EP(2) and EP(4) stimulate smooth muscle relaxation and glycosaminoglycan synthesis. This study investigated the expression of EP(2) and EP(4) mRNA and glycosaminoglycans in the sheep cervix during the estrous cycle. Sheep cervices were collected prior to, during, and after the luteinizing hormone (LH) surge and during the luteal phase. The mRNA expression of EP(2) and EP(4) was determined by in situ hybridization, glycosaminoglycan composition was assessed by Alcian blue staining, and hyaluronan concentration was investigated by ELISA. The expression of EP(2) mRNA was greatest prior to the LH surge (P=0.02), although EP(2) and EP(4) were expressed throughout the estrous cycle. Hyaluronan was the predominant glycosaminoglycan, and hyaluronan content increased prior to the LH surge (P<0.05). Cervical EP(2) mRNA expression changed throughout the estrous cycle and was greatest prior to the LH surge. We propose that prostaglandin E(2) binds to EP(2) and EP(4) stimulating hyaluronan synthesis, which may cause remodeling of the cervical extracellular matrix, culminating in cervical relaxation.

Activity participation and affect among nursing home residents: support for a behavioural model of depression.

One focus of interventions being developed for depression in nursing homes is increasing pleasant events. The conceptual foundation of such interventions requires establishing links among activity levels, depressive symptoms and positive affect. We observed activity and affect among 28 nursing home residents, five-days-a-week for four weeks, and concurrently collected self-reported affect and depression data. Inclusion of intra-individual analyses using random effects models demonstrated considerable individual variability in the relationship between positive affect and activity, although the results broadly supported the association. Participants differed with regard to what types of events were pleasurable and with regard to the relationships among pleasure, interest and activity. Depression levels were related to some of these individual differences. Activity was related primarily to interest rather than pleasure and higher interest was related to fewer depressive symptoms. Our findings emphasize the importance of individually tailoring pleasant events interventions and suggest that prescriptive group activities are unlikely to prove effective as means for improving positive affect.

Simple, novel physical activity maintains proximal femur bone mineral density, and improves muscle strength and balance in sedentary, postmenopausal Caucasian women.

UNLABELLED: A simple, appealing, physical activity program can be prescribed to reduce the risk of falls in sedentary, postmenopausal, independent-living, Caucasian women. Foot stamping, progressively loaded squats, and in-line dancing positively influence proximal femoral bone mineral density, lower extremity strength, and static and dynamic balance. INTRODUCTION: Foot stamping, squats exercises, and in-line dancing together create a suitable activity program for sedentary, independent-living older women. METHODS: Forty-five postmenopausal women not taking medications for bone health were randomly assigned to one of three groups. All groups attended one line dance class per week. Two groups additionally performed progressively loaded squats five times per week. One group also performed four foot stamps, twice daily, five times per week. Broadband ultrasound attenuation (BUA), proximal femur (PF) and lumbar spine (LS) bone mineral density (BMD), squats number, and balance variables were measured. RESULTS: There were no differences within or between groups in baseline and follow-up BUA, PF or LS BMD; however, a strong stamp compliance effect was apparent for BUA (r = 0.73) and PF BMD (r = 0.79). Number of squats (p < 0.01) and single leg stance time (p < 0.01) increased, while timed up and go time decreased (p < 0.01) for all participants. CONCLUSIONS: Line dancing, particularly in concert with regular squats and foot stamping, is a simple and appealing strategy that may be employed to reduce lower extremity bone loss, and improve lower limb muscle strength and balance, in independent living, otherwise healthy, postmenopausal Caucasian women.

Deep-sea ecology. Developmental arrest in vent worm embryos.

Temperature is a key factor in controlling the distribution of marine organisms and is particularly important at hydrothermal vents, where steep thermal gradients are present over a scale of centimetres. The thermophilic worm Alvinella pompejana, which is found at the vents of the East Pacific Rise (2,500-m depth), has an unusually broad thermotolerance (20-80 degrees C) as an adult, but we show here that the temperature range required by the developing embryo is very different from that tolerated by adults. Our results indicate that early embryos may disperse through cold abyssal water in a state of developmental arrest, completing their development only when they encounter water that is warm enough for their growth and survival.

Olanzapine therapy in treatment-resistant psychotic mood disorders: a long-term follow-up study.

BACKGROUND: Recent studies suggest a role for the atypical antipsychotic olanzapine in the acute treatment of psychotic mood disorders, but long-term data are unavailable. The purpose of this naturalistic study was to determine the long-term effectiveness and tolerability of olanzapine as add-on therapy in psychotic mood disorders. METHOD: Hospital records were reviewed for 125 inpatients at the state psychiatric hospital in Buffalo, N.Y., who received at least 6 weeks of add-on olanzapine treatment for psychotic mood disorders (schizoaffective disorders [bipolar and depressive type], bipolar disorders [I, II, and NOS], and major depressive disorder). A group of schizophrenic patients served as a control group (N = 50). Baseline measures, including age, gender, number of hospitalizations in the 2 years prior to olanzapine treatment, concomitant medications, the Clinical Global Impressions scale (CGI), and the Global Assessment of Functioning-Equivalent (GAF-EQ) and Kennedy Axis V psychological impairment, violence, social skills, and activities of daily living subscale scores, were obtained. Follow-up information was obtained from the patients at least 6 months after initiation of olanzapine or by chart review and discussion with the treating psychiatrist. Patients with a diagnosis of psychotic mood disorders were compared with patients with the non-affective psychotic disorder (schizophrenia) on a variety of outcome measures. RESULTS: Follow-up information was available on 102 patients (82%). Mean follow-up was 15 months; 50 (49%) of the 102 patients remained on olanzapine treatment at follow-up (32 psychotic mood disorder, 18 schizophrenic). The primary reason for discontinuation in both groups was lack of response. Both the psychotic mood disorder and schizophrenic groups had comparable outcomes on the CGI and GAF-EQ. Improvement on the Kennedy Axis V psychological impairment and social skills subscales was seen only in the psychotic mood disorders group (p < .01); both groups showed significant (p < .02) improvement in the violence subscale. Sustained mood-stabilizing effect was evident in only 7/27 (26%) of the psychotic mood disorders patients continuing on add-on olanzapine treatment at follow-up. CONCLUSION: Lack of response was the primary reason for discontinuation of add-on olanzapine in both groups. Mood symptoms predicted a better response to add-on olanzapine in patients with psychotic mood disorders on selective outcome measures. However, only 26% of the patients with psychotic mood disorders sustained a clinically meaningful mood-stabilizing effect with add-on olanzapine treatment at follow-up.

Larval dispersal potential of the tubeworm Riftia pachyptila at deep-sea hydrothermal vents.

Hydrothermal vents are ephemeral because of frequent volcanic and tectonic activities associated with crust formation. Although the larvae of hydrothermal vent fauna can rapidly colonize new vent sites separated by tens to hundreds of kilometres, the mechanisms by which these larvae disperse and recruit are not understood. Here we integrate physiological, developmental and hydrodynamic data to estimate the dispersal potential of larvae of the giant tubeworm Riftia pachyptila. At in situ temperatures and pressures (2 degrees C and 250 atm), we estimate that the metabolic lifespan for a larva of R. pachyptila averages 38 days. In the measured flow regime at a fast-spreading ridge axis (9 degrees 50' N; East Pacific Rise), this lifespan results in potential along-ridge dispersal distances that rarely exceed 100 km. This limited dispersal results not from the physiological performance of the embryos and larvae, but instead from transport limitations imposed by periodic reversals in along-ridge flows and sustained episodes of across-ridge flow. The lifespan presented for these larvae can now be used to predict dispersal under current regimes at other hydrothermal vent sites.

The sagittal band: anatomic and biomechanical study.

Forty-eight digits from 12 human adult fresh-frozen and formalin-preserved cadaveric hands were used to study the anatomy and biomechanics of the sagittal band (SB) and to investigate the mechanism of its injury. The SB was observed to be part of a complex retinacular system in proximity to the metacarpophalangeal (MCP) joint collateral ligaments and the palmar plate. Dynamic changes in SB fiber orientation were observed with different positions of the MCP and wrist joints. The fibers were perpendicular (0 degrees ) to the extensor tendon in neutral position, distally angulated 25 degrees at 45 degrees of MCP flexion, and 55 degrees with full flexion. Swan-Ganz catheter measurements were obtained deep to the SB in varying positions of the MCP joint. The average pressure generation was greatest (50 mm Hg) during full MCP joint flexion and least (30 mm Hg) during 45 degrees flexion. When MCP joint radial or ulnar deviation was added the average measurement was greatest (57) in neutral MCP position and least (35 mm Hg) in 45 degrees flexion. Serial sectioning of the ulnar SB produced no extensor tendon instability. Partial proximal but not distal sectioning of the radial SB produced tendon subluxation. Complete sectioning of the radial SB produced tendon dislocation. Wrist flexion increased tendon instability after radial SB sectioning. We conclude that (1) extensor tendon instability following SB disruption is most common in the long finger and least common in the small finger; (2) ulnar instability of the extensor tendon is due to partial or complete radial SB disruption, (3) the degree of extensor tendon instability is determined by the extent of SB disruption, (4) proximal rather than distal SB compromise contributes to extensor tendon instability, (5) great forces are inflicted on the SB while the MCP joint is in full extension or less frequently in full flexion, which may be the mechanism of its injury, and (6) wrist flexion contributes to extensor tendon instability after SB disruption and may exacerbate the severity of its injury.

Larval development of the tropical deep-sea echinoid Aspidodiadema jacobyi: phylogenetic implications.

The complete larval development of an echinoid in the family Aspidodiadematidae is described for the first time from in vitro cultures of Aspidodiadema jacobyi, a bathyal species from the Bahamian Slope. Over a period of 5 months, embryos grew from small (98-micron) eggs to very large (3071-micron) and complex planktotrophic echinopluteus larvae. The fully developed larva has five pairs of red-pigmented arms (preoral, anterolateral, postoral, posterodorsal, and posterolateral); fenestrated triangular plates at the bases of fenestrated postoral and posterodorsal arms; a complex dorsal arch; posterodorsal vibratile lobes; a ring of cilia around the region of the preoral and anterolateral arms; and a long, unpaired posterior process containing a fenestrated rod. The presence of a posterior process and posterodorsal arms makes the larva of Aspidodiadema jacobyi much more similar to larvae of irregular urchins in the order Spatangoidea than to other families of the order Diadematoida, to which the family is normally assigned. This unexpected larval form lends support to a recommendation that the Aspidodiadematidae should be either elevated to ordinal status as a sister group of the order Diadematoida, or split off as a sister group of the other families within the order. In either case, if we accept the parsimonious hypothesis that the aboral process and posterodorsal arms were derived only once in the evolutionary history of euechinoids, then the larval data suggest that the Aspidodiadematidae may be very near the node where the irregular and regular euechinoids first diverged.

Methane ice worms: Hesiocaeca methanicola colonizing fossil fuel reserves.

During a research cruise in July 1997 in the Gulf of Mexico we discovered a gas hydrate approximately 1 m thick and over 2 m in diameter which had recently breached the sea floor at a depth of 540 m. The hydrate surface visible from the submarine was considerably greater than that of any other reported hydrate. Two distinct color bands of hydrate were present in the same mound, and the entire exposed surface of the hydrate was infested (2500 individuals/m2) with 2 to 4 cm-long worms, since described as a new species, Hesiocaeca methanicola, in the polychaete family Hesionidae (Desbruyères and Toulmond 1998). H. methanicola tissue stable isotope values are consistent with a chemo-autotrophic food source. No evidence of chemo-autotrophic symbionts was detected, but geochemical data support the presence of abundant free living bacteria on the hydrate. The activities of the polychaetes, grazing on the hydrate bacteria and supplying oxygen to their habitats, appears to contribute to the dissolution of hydrates in surface sediments.

Epidermal lesions and mortality caused by vibriosis in deep-sea Bahamian echinoids: a laboratory study.

When significant mortality of the bathyal spatangoid echinoid Paleopneustes cristatus occurred under laboratory conditions, we investigated the cause and course of the disease by culturing and identifying internal pathogens, then experimentally infecting healthy urchins with isolates of the suspected disease organism. The pathogen was determined to be the gram-negative halophilic bacterium Vibrio alginolyticus. This species was also recovered from frozen post-challenge specimens of P. cristatus and from moribund individuals of Archaeopneustes hystrix, another spatangoid reared under similar in vitro conditions. This is the first experimental study of bacterial disease in any deep-sea invertebrate.